JP2018538249A - Dipeptidyl peptidase-4 and periostin as predictors of clinical response to eosinophil targeted therapeutics in eosinophilic diseases - Google Patents

Abstract

The present disclosure relates to protein and / or gene expression levels of dipeptidyl peptidase-4 (DPP4 / CD26) and periostin (POSTN) as peripheral biomarkers for eosinophilic diseases or disorders such as moderate to severe asthma. Regarding use. The level of DPP4 and / or POSTN above or below a given DPP4 or POSTN threshold level is, for example: To determine patient eligibility for treatment, (ii) specific treatment of eosinophilic disease or disorder with specific IL-5R antagonists should be initiated, discontinued or improved To determine whether (iii) eosinophilic disease or disorder is treatable with a specific IL-5R antagonist, or (iv) specific For predicting the outcome of treatment of an eosinophilic disease or disorder with a typical IL-5R antagonist. [Selection figure] None

Description

Reference to electronically submitted sequence listing Submitted electronically in an ASCII text file (name: DPP4_sequence listing_ST25_ascii.txt, size: 52,739 bytes, and creation date: November 4, 2015) filed with this application. The contents of the sequence listing are incorporated herein by reference in their entirety.
TECHNICAL FIELD The present invention relates to a title.

  Eosinophils are involved in various diseases including allergic diseases, and are considered to play an important role in causing morbidity of allergic diseases such as chronic bronchial asthma and atopic dermatitis. Adv. Immunol. 39, 177 (1986), Immunol. Today, 13, 501 (1992)]. In addition to the above-mentioned diseases, eosinophils are also commonly referred to as hyperacidic syndrome (HES), such as hypereosinocytosis, eosinophilic gastroenteritis, eosinophilic leukemia, eosinophils Involved in granuloma and Kimura disease. Ann. Intern. Med. 97, 78 (1982).

  Bronchial asthma is a common persistent inflammatory disease of the lung characterized by airway hyperresponsiveness, mucus overproduction, fibrosis, and elevated serum IgE levels. Airway hyperresponsiveness (AHR) is an exaggerated constriction of the airway to nonspecific stimuli such as cold air. Both AHR and mucus overproduction are thought to contribute to variable airway obstruction that results in shortness of breath characterized by asthma attacks (exacerbations) and is responsible for the mortality associated with the disease.

  The guidelines of the current British Thoracic Society (BTS) and the Global Initiative for Asthma (GINA) suggest a step-wise approach towards the treatment of asthma (Society, BT). , Thorax, 2003.58 Suppl 1: 1-94; GINA, Global Strategic for Asthma Management and Prevention, 2002, National Institute of Health). Mild to moderate asthma can usually be controlled by the use of inhaled corticosteroids combined with beta stimulants or leukotriene inhibitors. However, due to recorded side effects of corticosteroids, patients tend to not follow the treatment plan, which reduces the effectiveness of the treatment (Milgrom, H. et al. Ann Allergy Asthma Immunol, 2002.88: 429-31; Fish, L. and C. L. Lung, Ann Allergy Asthma Immunol, 2001.86: 24-30; Bender, BGJ Allergy Clin. Immunol, 2002.109: S554-9). Asthma exhibits significant heterogeneity in response to various treatments, thereby necessitating the development of more effective treatments for this disease or the identification of biomarkers that predict response to specific treatments Sex is emphasized.

  The terms "precision medicine", "individualized medicine" or "targeted treatment" are interchangeable to describe efforts to adapt the therapy to a diverse set of molecules and clinical features such as patient subgroups with asthma It is often used for. Current treatment of asthma is governed by inhalation and oral corticosteroids combined with bronchodilators. Although highly effective in most patients, about 10-20% of patients with asthma remain in poor control with current standard treatment (Bousequet et al., Allergy Clin Immunol 126, 926-938 ( 2010)). Recently, significant advances have been made through the discovery of new biomarkers using the “omics” approach in connection with a clinical phenotype called asthma “endotype” with molecular biomarkers (Anderson, Lancet 372). , 1107-1119 (2008); Lotvall et al. J Allergy Clin Immunol 127, 355-360 (2011); Wenzel, Pulm Pharmacol Ther 26, 710-715 (2013)).

  In some cases, these biomarkers are useful predictors of therapeutic response. For example, patients with high levels of periostin as a surrogate marker for interleukin (IL) -13 activity in increased IgE, eosinophilic inflammation or asthma have been identified by immunoglobulin E (Di Domenico et al., Inflamm Allergy Drug Targets). 10, 2-12 (2011)), IL-5 / IL-5Rα (Castro et al., Lancet Respir Med 3,355-366 (2015)), or IL-13 / IL-4Rα (Wenzel, Pulm Pharmacol Ther. 26, 710-715 (2013)) each responds preferentially to monoclonal antibodies targeting. However, the treatment of end-type dynamic changes over time, clinical phenotypes and ultimately new end-types that allow the development of new therapies that fit patients with asthma that remains uncontrolled A better understanding of the association with discovery is required (Holgate et al., Nat Rev Drug Discover 14, 367-368 (2015)).

  The present disclosure is a method for treating eosinophilic disease or disorder in a patient in need thereof, wherein the patient has a predetermined DPP4 or POSTN threshold in one or more samples taken from the patient. Determined or identified as having a dipeptidyl peptidase-4 (DPP4) and / or periostin (POSTN) level that is lower or reduced compared to a level or compared to a DPP4 or POSTN threshold level in one or more control samples A method is provided comprising administering to the patient an eosinophil targeted therapeutic.

  Further provided is a method of treating a patient having an eosinophilic disease or disorder, wherein the patient is compared to a predetermined DPP4 or POSTN threshold level in one or more samples taken from the patient. Or to patients with eosinophil targeted therapeutics if determined or identified as having higher or increased DPP4 and / or POSTN levels compared to DPP4 or POSTN threshold levels in one or more control samples A method comprising discontinuing or not starting administration of

  The present disclosure also provides for the treatment of eosinophilic diseases or disorders in patients in need thereof who have failed or have been unresponsive or intolerant to treatment with a therapeutic agent. A method wherein the patient is more than one DPP4 or POSTN threshold level in one or more samples taken from a patient or more than a DPP4 or POSTN threshold level in one or more control samples. If determined or identified as having low or decreased DPP4 and / or POSTN levels, a method is provided comprising administering to the patient an eosinophil targeted therapeutic.

  Further provided is a method of determining whether a patient having an eosinophilic disease or disorder should be treated with an eosinophil targeted therapeutic agent, wherein the patient is collected from the patient. Determined to have a DPP4 and / or POSTN level in the above samples that is lower or reduced compared to a predetermined DPP4 or POSTN threshold level or compared to a DPP4 or POSTN threshold level in one or more control samples or If identified, a method comprising determining to treat a patient.

  The disclosure also provides a method of selecting a patient diagnosed with an eosinophilic disease or disorder as a candidate for treatment with an eosinophil targeted therapeutic agent, wherein the patient is taken from the patient. Determined or identified as having a DPP4 and / or POSTN level lower or reduced compared to a predetermined DPP4 or POSTN threshold level or compared to a DPP4 or POSTN threshold level in one or more control samples If so, a method is provided comprising selecting a patient for treatment.

In some aspects, the methods disclosed above measure the level of DPP4 and / or POSTN in one or more of the samples obtained from a patient, or measure the level of DPP4 and / or POSTN in a sample Instructing a clinical laboratory or health care provider and / or measuring one or more samples obtained from a patient to the clinical laboratory or health care provider to determine the level of DPP4 and / or POSTN in the sample The method further includes submitting to: In some aspects, the methods disclosed above further comprise determining the level of DPP4 and / or POSTN in one or more samples obtained from the patient. In some aspects, the method disclosed above comprises:
(A) the patient is lower in one or more samples taken from the patient compared to a predetermined DPP4 or POSTN threshold level or compared to a DPP4 or POSTN threshold level in one or more control samples; or If it is determined or identified as having decreased DPP4 and / or POSTN levels, an eosinophil targeted therapeutic is administered to the patient, or (b) the patient is in one or more samples taken from the patient Is determined or identified as having a higher or increased DPP4 and / or POSTN level compared to a predetermined DPP4 or POSTN threshold level or compared to a DPP4 or POSTN threshold level in one or more control samples Discontinue, do not initiate administration of eosinophil-targeted therapeutics to patients, or Further comprising the step of advising the health care provider to not. In some aspects, the patient has elevated blood eosinophils.

  In some aspects, the eosinophil targeted therapeutic agent is a monoclonal antibody or antigen-binding fragment thereof. In some aspects, the monoclonal antibody or antigen-binding fragment thereof specifically binds IL-5 or IL-5R. In some aspects, the monoclonal antibody or antigen-binding fragment thereof that specifically binds IL-5 is mepolizumab, reslizumab, or an antigen-binding fragment thereof.

  In some aspects, the monoclonal antibody or antigen-binding fragment thereof that specifically binds to IL-5R specifically binds to the alpha subunit of IL-5R (SEQ ID NO: 4). In some aspects, the monoclonal antibody or antigen-binding fragment thereof that specifically binds to the α subunit of IL-5R is benralizumab (MED-563) or an antigen-binding fragment thereof. In some aspects, the monoclonal antibody or antigen-binding fragment thereof that specifically binds to the α subunit of IL-5R comprises SEQ ID NO: 12, consists of SEQ ID NO: 12, or consists essentially of SEQ ID NO: 12. A heavy chain variable region (VH) and / or a light chain variable region (VL) consisting of, consisting of SEQ ID NO: 13, or consisting essentially of SEQ ID NO: 13.

  In some aspects, the monoclonal antibody or antigen-binding fragment thereof that specifically binds to the α subunit of IL-5R is one, two, or three complementarity determining regions selected from SEQ ID NOs: 14-16. And / or one, two or three complementarity determining regions selected from SEQ ID NOs: 17-19. In some aspects, the monoclonal antibody or antigen-binding fragment thereof that specifically binds to the α subunit of IL-5R is at least one, two, three, four, selected from SEQ ID NOs: 14-19, Includes 5 or 6 complementarity determining regions.

  In some embodiments, the monoclonal antibody or antigen-binding fragment thereof binds to the same epitope as mepolizumab, reslizuzumab, or venralizumab, or competitively binds mepolizumab, reslizumab, or venralizumab to their respective target epitopes. Inhibit. In some aspects, the eosinophil targeted therapeutic is a fusion protein or complex comprising mepolizumab, reslizumab, or venralizumab, or an antigen-binding fragment thereof. In some aspects, the fusion protein or complex comprises at least one heterologous therapeutic moiety and / or a half-life enhancing moiety.

  In some aspects, the eosinophil targeted therapeutic agent is a polynucleotide. In some aspects, the polynucleotide is DNA or RNA. In some aspects, the polynucleotide is (i) mRNA or a combination thereof, or (ii) an antisense oligonucleotide or a combination thereof. In some aspects, the antisense oligonucleotide or combination thereof is ASM8, PXS1100, or PXS2200. In some aspects, the polynucleotide comprises at least a nucleotide analog.

  In some aspects, the eosinophil targeted therapeutic (eg, benralizumab / MEDI-563) is administered at a fixed dose. In some aspects, the constant dose is 1-1000 mg / dose. In some aspects, the eosinophil targeted therapeutic (eg, benralizumab / MEDI-563) is administered in two or more doses. In some aspects, the eosinophil targeted therapeutic (eg, benralizumab / MEDI-563) is administered weekly, biweekly or monthly. In some aspects, the eosinophil targeted therapeutic (eg, benralizumab / MEDI-563) is administered at about 2 mg to about 100 mg / dose. In some aspects, the eosinophil targeted therapeutic (eg, benralizumab / MEDI-563) is administered at about 20 mg / dose, about 30 mg / dose, or about 100 mg / dose. In some aspects, the eosinophil targeted therapeutic (eg, benralizumab / MEDI-563) is administered once every 4 weeks to once every 12 weeks. In some aspects, the eosinophil targeted therapeutic (eg, benralizumab / MEDI-563) is administered once every 4 weeks. In some aspects, the eosinophil targeted therapeutic (eg, benralizumab / MEDI-563) is administered once every 8 weeks. In some aspects, the eosinophil targeted therapeutic (eg, benralizumab / MEDI-563) is administered once every 4 weeks for 12 weeks and then once every 8 weeks.

  In some aspects, the eosinophil targeted therapeutic (eg, benralizumab / MEDI-563) is administered orally, by inhalation, intravenously, intramuscularly, subcutaneously, or a combination thereof.

  In some aspects, the eosinophilic disease or disorder is a pulmonary disease or disorder. In some aspects, the pulmonary disease or disorder is asthma or COPD. In some embodiments, the asthma is allergic asthma, atopic asthma, corticosteroid naive asthma, chronic asthma, corticosteroid resistant asthma, corticosteroid intractable asthma, asthma resulting from smoking, or corticosteroid Asthma not controlled by. In some aspects, the asthma is mild to moderate asthma (defined as GINA1-3) or severe asthma (defined as GINA4 +).

  In some aspects, the eosinophilic disease or disorder is chronic bronchitis, chronic eosinophilic pneumonia (CEP), nasal polyp, atopic dermatitis (eczema), eosinophilic esophagitis, hyperacidity Syndrome (HES), eosinophilic granulomatosis and polyangiitis (Charg Strauss syndrome), eosinophilic gastritis, eosinophilic enteritis, eosinophilic colitis, allergic rhinoconjunctivitis (hay fever) ), Leukemia, lymphoma, mastocytosis, atheroembolic disease, hyper IgE syndrome, Omen syndrome, thymoma, transplant rejection, adrenal hypofunction, bullous pemphigoid, pemphigus vulgaris, herpes dermatitis, Drug-induced lesions, urticaria, eosinophilic panniculitis, angioedema with eosinophilia, Kimura disease, Schulman syndrome, Wells syndrome, eosinophilic ulcer of the oral mucosa, Eosinophil Pustular folliculitis, recurrent cutaneous necrotic eosinophilic vasculitis, drug / toxin-induced eosinophilic pulmonary disease, Leffler syndrome, allergic bronchopulmonary aspergillosis, eosinophilic granuloma, pleural eosinophilic Encephalopathy, gastroesophageal reflux disease, parasitic infection, fungal infection, Helicobacter pylori infection, inflammatory bowel disease (ulcerative colitis and Crohn's disease), food allergic disorder, protein induction Enteropathies, protein-induced enteritis, allergic colitis, celiac disease, proliferative pemphigus, leiomyomatosis, connective tissue disorder, vasculitis disorder, chronic subdural hematoma, central nervous system infection, ventricular abdominal cavity Short-circuit surgery, congenital heart disease (septal defect or aortic stenosis), eosinophilic urinary disease with renal infection, interstitial nephritis, or eosinophilic cystitis.

  In some aspects, the patient uses one or more additional therapeutic agents for the treatment of an eosinophilic disease or disorder, using an eosinophil targeted therapeutic agent (eg, benralizumab / MEDI-563). Treated either before, during, after, or alternatively against. In some aspects, the one or more additional therapeutic agents further comprise a steroid, a bronchodilator, or both. In some embodiments, the steroid is fluticasone or budesonide. In some aspects, the bronchodilator is salbutamol or salmeterol. In some aspects, the one or more additional therapeutic agents are administered by inhalation, by oral administration, by injection, or a combination thereof. In some aspects, inhalation administration is performed using a metered dose inhaler (MDI) or a dry powder inhaler (DPI). In some aspects, the steroid is administered at a high dose. In some embodiments, the steroid is an inhaled corticosteroid such as beclomethasone or mometasone.

  In some aspects, the one or more samples and / or one or more control samples taken from the patient are whole blood, serum, plasma, saliva, sputum, bronchoalveolar lavage fluid, lung epithelial cells, urine, skin , Nasal fins, or combinations thereof. In some embodiments, the sample taken from the patient is serum. In some aspects, the one or more control samples are (a) one or more samples, (i) a normal healthy person, (ii) a patient with a subset of asthma, (iii) corticosteroid treatment One or more samples obtained from a naïve asthma patient or an asthma patient treated with corticosteroids, (b) a predetermined standard amount of isolated DPP4 or POSTN, or (c) (a) and (b ) In any combination sample.

  In some aspects, the methods disclosed above determine or submit a sample taken from a patient, or (i) a level of a patient's IgE level, (ii) a patient's preference. Eosinophil count, (iii) patient exhaled nitric oxide concentration (FENO), (iv) patient eosinophil / lymphocyte and eosinophil / neutrophil (ELEN) index, (v) patient EOS index, (Vi) further comprising instructing the clinical laboratory to determine the percentage of wall area (WA%) of the subzone airway from the CT scan of the lung, (vii) a combination of two or more thereof. See, for example, International Publication No. 20121588954, which is incorporated herein by reference in its entirety.

  In some aspects, the patient's eosinophil count is ≧ 300 eosinophils / μL. In some aspects, ≧ 300 eosinophils / μL is set as a threshold in patients with mild to moderate asthma. In some aspects, the patient's eosinophil count is ≧ 400 eosinophils / μL. In some aspects, ≧ 400 eosinophils / μL is set as a threshold in patients with severe asthma. In some aspects, the patient's eosinophil count is ≧ 150 eosinophils / μL. In some aspects, ≧ 150 eosinophils / μL is set as a threshold in patients undergoing treatment with, for example, an anti-IL5 antibody (eg, mepolizumab).

  In some aspects, a patient's DPP4 level is measured with a DPP4 detection assay, an immunoassay. In some aspects, a patient's POSTN level is measured with a POSTN detection assay, which is an immunoassay.

  In some aspects, the DPP4 detection immunoassay is (i) the immunoassay described in Example 2, or (ii) the immunoassay disclosed in Table 1, or (ii) a multiplex immunoassay. In some aspects, the multiplex immunoassay is an EMD Millipore MILLIPLEX (TM), Bio-Rad BIO-PLEX (TM), Life Technologies NOVEX MULTIPLEX (TM), Thermo FisherEPI (TM) M Affymetrix (eBioscience) PROCARTA ™, or R & D Systems LUMINEX® assay. In some aspects, the POSTN detection assay is an assay disclosed in WO2015015171A1 or WO2015120185, both of which are hereby incorporated by reference in their entirety.

  In some aspects, the method disclosed above determines, submits for determination, a sample taken from a patient, or IL-22, IL-25, IL-33, TSLP, LCN2, CCL20, sCTLA-3, sCD28, CCL5, CCL11, CCL22, CST1, CCL26, CLCA1, CST2, PRR4, SERPINB2, CEACAM5, iNOS, SERPINB4, CST4, PRB4, TPSD1, TPSG1, MFSD2, CPA3, GPR105, CPA3, GPR105 C2ORF32, TRACH20000196 (TMEM71), DNAJC12, RGS13, SLC18A2, SERPINB10, SH3RF2, FCER1B, RUNX2, PTGS1, ALOX15, or combinations thereof Further comprising the step of instructing the laboratory to determine the expression level or activity of other molecular biomarkers.

In some aspects, the predetermined DPP4 or POSTN threshold level is:
(A) an approximate average DPP4 level or POSTN level measured in serum from multiple patients using an immunoassay;
(B) approximately median DPP4 or POSTN levels measured in sera from multiple patients using immunoassay, and (c) approximately first measured in sera from multiple patients using immunoassay. A second, third, fourth, fifth, sixth, seventh, eighth, or ninth quartile baseline DPP4 level or POSTN level, wherein the patient is (i) normal Healthy volunteers, (ii) patients with mild to moderate asthma, and / or (iii) patients with severe asthma. In some aspects, sera from multiple patients are pooled samples or individual samples.

  In some aspects, the predetermined DPP4 threshold level is at least about 103 ng / mL to at least about 867 ng / mL as measured using an immunoassay. In some aspects, the predetermined DPP4 threshold level is at least about 363 ng / mL as measured using an immunoassay. In some aspects, the predetermined DPP4 threshold level is at least about 376 ng / mL as measured using an immunoassay. In some aspects, the immunoassay is the DPP4 detection assay described in Example 2 or the DPP4 detection assay disclosed in Table 1.

  In some aspects, the predetermined POSTN threshold is at least about 7 ng / mL to at least about 104 ng / mL as measured using an immunoassay. In some aspects, the predetermined POSTN threshold level is at least about 23 ng / mL as measured using an immunoassay. In some aspects, the predetermined POSTN threshold level is at least about 26 ng / mL as measured using an immunoassay. In some aspects, the POSTN immunoassay is the assay disclosed in WO2015015171A1 or WO2015120185, or a commercially available POSTN assay.

  In some aspects, administration of eosinophil targeted therapeutics (a) decreased AER (acute exacerbation rate), (b) increased FEV1 (forced expiratory volume per second), (c) improved ACQ-6 (asthma management questionnaire, 6 item version) value, or (d) results in a combination thereof.

  In some aspects, the decrease in AER is at least about 15%, 20%, 25%, 30%, 35%, 40% compared to an AER baseline value observed in a population of patients treated with placebo. %, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%. In some aspects, the increase in FEV1 is at least about 50, 75, 100, 125, 150, 175, 200, 225, compared to the FEV1 baseline value observed in a population of patients treated with placebo. 250, 275, 300, 325, 350, 400, 450, or 500 mL. In some aspects, the improved ACQ-6 value is at least about -0.3, -baseline from an ACQ-6 baseline value observed in a population of patients treated with placebo,- A change of 0.4, -0.5, -0.6, -0.7, -0.8, -0.9, -1, -1.1, or -1.2.

1 shows the design of Benralizumab Phase 2b study CP220 (NCT01238861) in severely controlled asthma. ACQ-6, Asthma Management Questionnaire-6; AER, annual exacerbation rate (all observed exacerbations up to 52 weeks divided by total duration of person-year follow-up); CBC, complete blood count with differentiation; Fe _NO , exhaled nitric oxide concentration; FEV1, forced expiratory volume for 1 second; ICS, inhaled corticosteroid; ppb, parts per billion; SC, subcutaneous. The ELEN index is calculated according to Khatry et al. Am J Respir Crit Care Med 189: A mathematical algorithm for predicting eosinophils from CBC data as described in A4257 (2014). It is the table | surface which put together the data regarding DPP4 and POSTN in the serum obtained by CP220 test (NCT0123861). A “high DPP4” value (ie, a higher or increased DPP4 level) is a value above the median. The “high POSTN” value is greater than or equal to the median value. A “low DPP4” value (ie, a lower or decreased DPP4 level) is a value less than the median. The “low POSTN” value is less than the median value. The percentages of subgroup high and low DPP4 (A) and high and low POSTN (panel B) with blood eosinophils <or ≧ 300 cells / uL are shown. Lack of correlation between serum DPP4 and serum POSTN levels in subjects participating in CP220 (panel A), serum POSTN correlates with blood eosinophil count (panel B), and serum DPP4 is blood eosinophil It shows that it does not correlate with the number of spheres (panel C). Shows a comparison of the effects of benralizumab on reducing the exacerbation rate in the combined 20 mg + 100 mg dose group versus placebo in the high and low DPP4 subgroup (panel A) and the high and low POSTN subgroup (panel B). ^* = P ≦ 0.05. Numbers above the bar represent n in the placebo / venralizumab treatment group. The percentage number at the bottom of the x-axis represents the proportion of each subgroup. Shows a comparison of the effects of benralizumab on FEV1 in the combined 20 mg + 100 mg dose group versus placebo in the high and low DPP4 subgroup (panel A) and in the high and low POSTN subgroup (panel B). ^* = P ≦ 0.05. Numbers above the bar represent n in the placebo / venralizumab treatment group. The percentage number at the bottom of the x-axis represents the proportion of each subgroup. Shows a comparison of the effects of benralizumab on ACQ-6 in the combined 20 mg + 100 mg group versus placebo in the high and low DPP4 subgroup (panel A) and the high and low POSTN subgroup (panel B). ^* = P ≦ 0.05. Numbers below the bar represent n in the placebo / venralizumab treatment group. The percentage number on the x-axis display represents the proportion of each subgroup.

  Eosinophils are a major effector cell in the pathology of asthma, and more than 50% of patients with severe asthma are associated with persistent eosinophilic inflammation despite treatment with corticosteroids. IL-13, Th2 cytokines produced by mast cells, basophils and eosinophils contribute to the main features of asthma through a number of different mechanisms. Benralizumab (MED-563), a humanized anti-IL-5Ra mAb that binds with high affinity to the alpha chain of IL-5R to block IL-5 function, through antibody-dependent cell-mediated cytotoxicity (ADCC). Induces apoptosis of eosinophils and basophils, thereby depleting eosinophils and basophils.

  In a phase 2b clinical trial (CP220), benralizumab has been shown to be maximally effective in reducing asthma exacerbations in patients with elevated blood eosinophil levels, which is currently in phase III clinical It has been further evaluated in trials. It has been recognized that there may be overlap between severe asthma patients with eosinophilic inflammation and severe asthma patients with IL-13 pathway activation. The inventors have therefore sought to identify biomarkers that may be more selective in identifying patients who may respond to the use of anti-eosinophil therapy, such as benralizumab.

  Serum samples collected in a phase 2b trial of benralizumab (CP220) evaluate the expected usefulness of baseline DPP4 and POSTN levels for reduced exacerbations compared to placebo treatment in subjects treated with benralizumab Was analyzed for.

  Blood eosinophils and POSTN did not correlate with serum DPP4 concentration. In contrast, serum POSTN correlated with blood eosinophil count. Reduced exacerbation rate when patients in the group treated with benralizumab and those in the group treated with placebo are divided by high levels of DPP4 or POSTN (greater than the median) or low levels of DPP4 or POSTN (less than the median) The efficacy of benralizumab against was greatest in subjects with low levels of serum DPP4. This effect at low levels of DPP4 was evident in subjects with both high (≧ 300 cells / microliter) and low (<300 cells / microliter) blood eosinophil levels. The effect with periostin was comparable at high and low levels in subjects with high blood eosinophils, but the effect was highest in subjects with low blood eosinophils in subjects with low levels of periostin. It was.

  The primary observation in this disclosure is that asthmatic patients who also have an IL13-driven disease with elevated blood eosinophils will benefit less from benralizumab as assessed by exceeding median POSTN or DPP4 levels. This decrease in effectiveness is more apparent at high DPP4 than at high POSTN and can be explained by the correlation between POSTN and eosinophils. In addition, as assessed by below median POSTN or DPP4 levels, asthmatic patients with low eosinophils and low IL13-driven disease benefit from treatment with benralizumab. In other words, it is not the eosinophilic disease alone that determines the efficacy of benralizumab, and potentially the anti-IL5 antibody. Rather, it is suggested that IL-13 driven disease or its deficiency is assessed by the presence of low DPP4 and / or POSTN levels that appear to predict the efficacy of benralizumab.

  The present disclosure relates to the use of DPP4 and / or periostin (POSTN) levels as biomarkers for eosinophilic diseases or disorders, such as asthma (and especially moderate and severe asthma). The present disclosure is a method for diagnosing and treating a subject having, for example, an eosinophilic disease or disorder, wherein the level of DPP4 or POSTN in one or more samples taken from a patient is (i) DPP4 Or eosinophils if below the limit of detection of a POSTN detection assay, (ii) below a given DPP4 or POSTN threshold level, or (ii) below a DPP4 or POSTN level in one or more control samples A method is provided comprising administering to a patient a targeted therapeutic agent, eg, an anti-IL-5R antibody such as benralizumab (MED-563).

  In some aspects, above or below a predetermined DPP4 or POSTN threshold level in a sample (eg, serum or sputum) obtained from a patient suffering from an eosinophilic disease or disorder (eg, a lung disease or disorder such as asthma) The presence of DPP4 and / or POSTN levels (protein or RNA expression levels), for example, (ii) to determine whether a patient is eligible or not eligible for treatment with a particular therapeutic agent, (ii) a particular treatment (Iii) to diagnose whether a disease or disorder is treatable or not treatable with a particular therapeutic agent, to determine whether the disease should be initiated, discontinued, or changed (Iv) can be used to predict the outcome of treatment of a disease or disorder using a particular therapeutic agent, etc. In some aspects, the particular therapeutic agent is an eosinophil targeted therapeutic agent, eg, an anti-IL-5R antibody such as benralizumab (MED-563).

In other embodiments, DPP4 above or below a predetermined DPP4 or POSTN threshold level in a sample (eg, serum or sputum) obtained from a patient suffering from an eosinophilic disease or disorder (eg, a lung disease or disorder such as asthma). And / or the presence of POSTN levels, for example,
(I) a patient suffering from an eosinophilic disease or disorder (eg pulmonary disease or disorder such as asthma) uses an eosinophil-targeted therapeutic agent, eg an anti-IL-5R antibody such as benralizumab (medi-563) And / or (ii) to determine whether a particular treatment should be started, discontinued or changed, and / or (Iii) to diagnose whether a disease or disorder is treatable or untreatable with a particular therapeutic agent and / or (iv) to predict the outcome of treatment of a disease or disorder with a particular therapeutic agent In addition,
(I) one or more molecular biomarkers, and / or (ii) one or more clinical biomarkers, and / or (iii) high eosinophil cell count (eg, blood eosinophil count ≧ 300 cells / μL) ), And / or (iv) high Th2 (high Th2 is defined, for example, as IgE> 100 IU / mL and blood eosinophils ≧ 0.14 × 10 ⁹ / L), and / or (v) short-acting Reversibility of FEV1 to β2 agonists, eg ≧ 12%, and / or (vi)% of sub-district airway wall area (WA%), eg about 68% as measured via CT scan of lung, and / or (Vii) It can be used in combination with those combinations.

  In order that the present disclosure may be more readily understood, certain terms are first defined. Additional definitions are set forth throughout the detailed description.

I. Definitions In this specification and the appended claims, the singular forms “a”, “an” and “the” refer to the plural unless the context clearly dictates otherwise. including. The terms “a” (or “an”) and the terms “one or more” and “at least one” may be used interchangeably herein.

  Further, “and / or” as used herein is to be construed as a specific disclosure of each of the two specific features or components, whether or not including the other. Should be. Thus, the term “and / or” as used herein in phrases such as “A and / or B”, “A and B”, “A or B”, “A” (alone), And “B” (alone). Similarly, the term “and / or” when used in a phrase such as “A, B, and / or C” includes the following aspects: A, B, and C; A, B, or C A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone) .

  Whenever an aspect is described herein by the term “comprising”, it is described by the term “consisting of” and / or “consisting essentially of” Other similar aspects are also provided.

  The term “about” is used throughout the specification and claims in the context of numerical values and is well known to those skilled in the art and represents an acceptable accuracy interval. In general, the interval requiring accuracy is ± 15%.

  Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure relates. For example, Concise Dictionary of Biomedicine and Molecular Biology, Juo, Pei-Show, 2nd ed. , 2002, CRC Press; The Dictionary of Cell and Molecular Biology, 3rd ed. , 1999, Academic Press; and Oxford Dictionary Of Biochemistry And Molecular Biology, Revised, 2000, Oxford University Press provides those skilled in the art with a general dictionary of many terms used in this disclosure.

  Units, prefixes, and symbols are represented in a form approved by their System International de Units (SI).

  Numeric ranges are inclusive of the numbers defining the range. Where a range of values is listed, each intervening integer value, and each fraction, between the listed upper and lower limits, are also specifically disclosed along with each subrange between such values. It should be understood that The upper and lower limits of any range can independently be included or excluded from the range and include any limit, none of the limit, or both Each range that includes these limits is also included within the scope of the present invention. When values are explicitly listed, it should be understood that values that are approximately the same amount as the listed values are within the scope of the present invention. Where a combination is disclosed, each subcombination of the elements of the combination is also specifically disclosed and is within the scope of the invention. Conversely, when different elements or groups of elements are disclosed individually, combinations thereof are also disclosed. Where any element of the invention is disclosed as having a plurality of alternatives, examples of the invention in which each alternative is excluded alone or in any combination with other alternatives are also disclosed herein . Multiple elements of the invention may have such exclusions, and all combinations of elements having such exclusions are disclosed herein.

  Amino acids are referred to herein by either their commonly known three letter symbols or by the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission. Unless otherwise indicated, amino acid sequences are written left to right in the amino to carboxy direction.

  Nucleotides are referenced by their generally accepted single letter code. Unless otherwise indicated, nucleic acids are written left to right in the 5 'to 3' direction. Nucleotides are referred to herein by their one-letter symbols recommended by their commonly known IUPAC-IUB Biochemical Nomenclature Committee. Thus, A represents adenine, C represents cytosine, G represents guanine, T represents thymine, and U represents uracil.

  The terms “polynucleotide”, “oligonucleotide”, “nucleic acid”, “nucleic acid molecule”, and “gene” refer to polymers of nucleotides of any length, and ribonucleotides, deoxyribonucleotides, analogs thereof, or mixtures thereof. Are used interchangeably herein to refer to.

  The phrase “DNA sequence” refers to a contiguous nucleic acid sequence. The sequence can be either single stranded or double stranded, either DNA or RNA, but double stranded DNA sequences are preferred. The sequence can be a full-length genomic sequence from 6-20 nucleotides in length to thousands or hundreds of thousands of base pairs.

  The terms “polypeptide”, “peptide”, and “protein” are used interchangeably herein to refer to polymers of amino acids of any length. The polymer can be linear or branched, it can contain modified amino acids, and it can be interrupted by non-amino acids. The term is also modified naturally or by intervention (eg, disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation or modification, eg, conjugation with a labeling component). Includes amino acid polymers. Furthermore, within its definition, for example, a polypeptide containing one or more analogs of an amino acid (including non-natural amino acids such as homocysteine, ornithine, p-acetylphenylalanine, D-amino acid, and creatine), As well as other modifications known in the art.

  An “isolated” polypeptide, antibody, polynucleotide, vector, cell, or composition is a polypeptide, polynucleotide, or composition in a form not found in nature. Isolated polypeptides, polynucleotides, or compositions include those that have been purified to the extent that they are no longer found in nature. In some aspects, the isolated polypeptide, polynucleotide, or composition is substantially pure.

  The term “amino acid substitution” refers to the replacement of an amino acid residue present in a parent sequence with another amino acid residue. Amino acids can be substituted within a parent sequence, for example, through chemical peptide synthesis or through recombinant methods known in the art. Amino acid substitutions also occur in natural variants.

  Thus, reference to “substitution at position X” or “substitution at position X” refers to substitution of the amino acid present at position X with another amino acid residue. In some aspects, the substitution pattern can be described according to schema AXY, where A is a single letter code corresponding to the naturally occurring amino acid at position X and Y is a substitution of an amino acid residue. In other aspects, the substitution pattern may be described according to schema XY, where Y is a single letter code corresponding to an amino acid residue that substitutes a naturally occurring amino acid at position X.

  A “conservative amino acid substitution” is one in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Basic side chains (eg, lysine, arginine, histidine), acidic side chains (eg, aspartic acid, glutamic acid), uncharged polar side chains (eg, glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), non- Polar side chains (eg alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), β-branched side chains (eg threonine, valine, isoleucine) and aromatic side chains (eg tyrosine, phenylalanine, tryptophan, A family of amino acid residues with similar side chains, including histidine, is defined in the art. Thus, if an amino acid in a polypeptide is replaced with another amino acid from the same side chain family, the substitution is considered conservative. In another aspect, a string of amino acids can be conservatively replaced with a composition of structurally similar strings and / or side chain family members that are out of order.

  Non-conservative substitutions are: (ii) when a residue with a positively charged side chain (eg, Arg, His or Lys) is replaced with or by a negatively charged residue (eg, Glu or Asp) When a hydrophilic residue (eg Ser or Thr) is substituted with or by a hydrophobic residue (eg Ala, Leu, Ile, Phe or Val), (iii) cysteine or proline is any other residue When substituted with or by a group, or (iv) a residue having a large hydrophobic or aromatic side chain (eg Val, His, Ile or Trp) having a smaller side chain ( For example, Ala, Ser) or those having no side chain (for example, Gly) or substituted by it.

  Other substitutions can be readily identified by those skilled in the art. For example, for the amino acid alanine, the substitution can be obtained from any one of D-alanine, glycine, β-alanine, L-cysteine and D-cysteine. For lysine, the substitution can be any one of D-lysine, arginine, D-arginine, homo-arginine, methionine, D-methionine, ornithine, or D-ornithine. In general, substitutions within functionally important regions that can be envisaged to induce changes in the properties of an isolated polypeptide include: (i) polar residues such as serine or threonine are hydrophobic residues, For example, when substituted with (or by) leucine, isoleucine, phenylalanine, or alanine, (ii) when a cysteine residue is substituted (or by) any other residue, (iii) positive charge When a residue having a side chain, such as lysine, arginine or histidine is substituted (or by) a residue having a negatively charged side chain, such as glutamic acid or aspartic acid, or (iv) a large side chain Residues that have, for example, phenylalanine are substituted (or thereby) with those that do not have such side chains, such as glycine. It is the case. The possibility that one of the aforementioned non-conservative substitutions may alter the functional properties of the protein also correlates with the position of the substitution relative to a functionally important region of the protein, and thus some non-conservative substitutions May have little or no effect on biological properties.

  The term “percent sequence identity” between two polypeptide or polynucleotide sequences is a sequence over a comparison window that allows for additions or deletions (ie, gaps) that must be introduced for optimal alignment of the two sequences. Refers to the number of identical matching positions shared by. A matched position is any position where the same nucleotide or amino acid is present in both the target and reference sequences. Gaps present in the target sequence are not counted because they are neither nucleotides nor amino acids. Similarly, gaps present in the reference sequence are not counted because the nucleotides or amino acids of the target sequence are counted, not the nucleotides or amino acids from the reference sequence.

  The percentage of sequence identity is determined by determining the number of positions where the same amino acid residue or nucleobase is present in both sequences to obtain the number of matching positions, and the number of matching positions is the number of positions in the comparison window. Calculated by dividing by the total number and multiplying the result by 100 to obtain the percentage of sequence identity. Comparison of sequences between two sequences and determination of percent sequence identity can be accomplished using readily available software, both online and downloadable. Suitable software programs are available from various sources and for alignment of both proteins and nucleotide sequences. One suitable program for determining percent sequence identity is a program available from the National Government for Biotechnology Information BLAST website (blast.ncbi.nlm.nih.gov). This is a part bl2seq of BLAST package software. bl2seq performs a comparison between two sequences using either the BLASTN or BLASTP algorithm. BLASTN is used to compare nucleic acid sequences, while BLASTP is used to compare amino acid sequences. Other suitable programs are for example Needle, Stretcher, Water, or Matcher, part of the EMBOSS package software of the bioinformatics program, and also the European Bioinformatics Institute (EBI), www. ebi. ac. Available from uk / Tools / psa.

  Different regions within a single polynucleotide or polypeptide target sequence that align with a polynucleotide or polypeptide reference sequence may each have their own percent sequence identity. It is observed that the percent sequence identity value is rounded to the nearest decimal place. For example, 80.11, 80.12, 80.13, and 80.14 are rounded to 80.1, while 80.15, 80.16, 80.17, 80.18, and 80.19 are Rounded to 80.2. It is also accepted that the length value is always an integer.

  In certain aspects, the percent identity “X” of the first amino acid sequence to the second sequence amino acid is 100 × (Y / Z), wherein Y is (alignment by visual inspection or a specific sequence alignment program As) and the number of amino acid residues scored as identical matches in the alignment of the first and second sequences (and Z is the total number of residues in the second sequence). If the length of the first sequence is longer than the second sequence, the percent identity of the first sequence to the second sequence will be higher than the percent identity of the second sequence to the first sequence. .

  One skilled in the art will appreciate that the generation of sequence alignments for percent sequence identity calculations is not limited to binary sequence-sequence comparisons performed exclusively with primary sequence data. The sequence alignment can be derived from multiple sequence alignments. One suitable program for generating multiple sequence alignments is ClustalW2, www. christal. org. Another suitable program is MUSCLE, www. drive5. com / mouse /. ClustalW2 and MUSCLE are alternatively available from, for example, EBI.

  As used herein, the term “antibody” (or fragment, variant, or derivative thereof) refers to an antibody that can bind to an antigen in the context of a typical antibody produced by a B cell. It refers to at least a minimal portion, such as at least the variable domain (VH) of the heavy chain and the variable domain (VL) of the light chain. The basic antibody structure in vertebrate systems is relatively well understood. For example, Harlow et al. , Antibodies: A Laboratory Manual (Cold Spring Harbor Laboratory Press, 2nd ed. 1988).

  Antibodies or antigen-binding fragments, variants, or derivatives thereof include but are not limited to polyclonal, monoclonal, human, humanized, or chimeric antibodies, single chain antibodies, epitope binding fragments such as Fab, Fab ′ and F ( ab ′) 2, Fd, Fvs, single chain Fvs (scFv), single chain antibodies, disulfide-bonded Fvs (sdFv), fragments containing either VL or VH domains, fragments produced by Fab expression libraries . ScFv molecules are known in the art and are described, for example, in US Pat. No. 5,892,019. The immunoglobulins or antibody molecules encompassed by this disclosure can be any type of immunoglobulin molecule (eg, IgG, IgE, IgM, IgD, IgA, and IgY), class (eg, IgG1, IgG2, IgG3, IgG4, IgA1). And IgA2) or a subclass.

  “Specifically binds” generally means that an antibody or fragment, variant, or derivative thereof binds to an epitope via its antigen binding domain, and that binding is complementary between the antigen binding domain and the epitope. Means that it is partly accompanied. According to this definition, an antibody is said to “specifically bind” to an epitope when it binds to that epitope more easily than if it binds to a random unrelated epitope via its antigen-binding domain. .

  If the antibody or fragment, variant, or derivative thereof preferentially binds to its epitope to the extent that it blocks binding of the reference antibody or antigen-binding fragment to the epitope to some extent, the reference antibody or antigen-binding fragment is It is said to competitively inhibit binding to a given epitope. Competitive inhibition can be measured by any method known in the art, such as a competitive ELISA assay. A binding molecule has at least 90%, at least 85%, at least 80%, at least 75%, at least 70%, at least 65%, at least 60%, at least 55% binding of a reference antibody or antigen-binding fragment to a given epitope. Or at least 50% competitive inhibition.

  An antibody or antigen-binding fragment, variant, or derivative thereof disclosed herein can be described or identified in terms of an epitope or portion of an antigen, such as a target polysaccharide to which they recognize or specifically bind. . For example, the portion of the α subunit of human IL-5R that specifically interacts with the antigen binding domain of an anti-IL-5R antibody, such as benralizumab (MEDI-563), is an “epitope”.

  The term “epitope”, as used herein, is an antigenic protein determinant capable of binding to an antibody disclosed herein (eg, an anti-IL-5R antibody) or fragment, variant, or derivative thereof. Point to. In some aspects, the term epitope refers to a protein determinant (eg, amino acid sequence) of a subunit of an IL-5R protein, such as the α subunit of IL-5R. In some aspects, the term epitope refers to a protein determinant (eg, amino acid sequence) of IL-5. In some aspects, binding of the disclosed antibodies to either IL-5R or IL-5 blocks the interaction of IL-5R with IL-5.

  Epitopes usually consist of chemically active surface molecule groups, such as amino acids or sugar side chains, and usually have specific three-dimensional structural characteristics and specific charge characteristics. The part of the antibody or binding molecule that recognizes the epitope is called a paratope. The epitopes of protein antigens are divided into two categories, conformational epitopes and linear epitopes, based on their structure and interaction with paratopes. A conformational epitope consists of discontinuous sections of the amino acid sequence of the antigen. These epitopes interact with paratopes based on the 3-D surface characteristics and shape or tertiary structure of the antigen. In contrast, linear epitopes interact with paratopes based on their primary structure. A linear epitope is formed by a contiguous sequence of amino acids from an antigen.

  The term “antibody binding site” refers to a region in an antigen (eg, the amino acid sequence of IL-5 or IL-5R) that contains continuous or discontinuous sites (ie, epitopes) to which a complementary antibody specifically binds. Thus, the antibody binding site may exceed the epitope and determine properties such as binding affinity and / or stability or affect properties such as antigenic enzyme activity or dimerization in the antigen. Additional regions may be included. Thus, even if two antibodies bind to the same epitope within the antigen, such antibody will bind to a well-defined antibody binding site if the antibody molecule establishes a clear intermolecular contact with amino acids outside the epitope. It is done.

  An antibody or fragment, variant, or derivative thereof blocks the binding of a reference antibody or antigen-binding fragment to a given epitope, which in part blocks the binding of the reference antibody or antigen-binding fragment to the epitope. When preferentially binding to a degree, it is said to inhibit competitively. Competitive inhibition can be measured by any method known in the art, such as a competitive ELISA assay. A binding molecule has at least 90%, at least 85%, at least 80%, at least 75%, at least 70%, at least 65%, at least 60%, at least 55% binding of a reference antibody or antigen-binding fragment to a given epitope. , At least 50%, at least 45%, at least 40%, at least 35%, at least 30%, at least 25%, or at least 20%.

As used herein, the term “eosinophilic disease or disorder” refers to any disease or disorder characterized by elevated levels of eosinophils in the blood, tissue, or organ. Normal levels in the blood are on the order of 250 eosinophils / mm ³ . The term eosinophilic disease or disorder also includes eosinophilic inflammation. Blood levels above about 300 / mm ³ are considered elevated. Examples of eosinophilic diseases and disorders include pulmonary diseases and disorders such as bronchial asthma, chronic bronchitis (in COPD), or chronic eosinophilic pneumonia (CEP). Furthermore, the term eosinophilic disease or disorder includes nasal polyps, atopic dermatitis, eosinophilic esophagitis, hyperacidic syndrome (HES), eosinophilic granulomatosis and polyangiitis (EGPA). , Formerly Churg-Strauss syndrome), eosinophilic gastritis, eosinophilic enteritis, eosinophilic colitis and other diseases and conditions. Normal and abnormal (eg, elevated) eosinophil levels in eosinophilic diseases or disorders disclosed herein are known in the art.

  In some aspects, the eosinophilic disease or disorder can be a pulmonary disease or disorder. As used herein, the term “pulmonary disease or disorder” refers to any pathology affecting the lung or respiratory system that is characterized at least in part by elevated levels of eosinophils. Non-limiting examples include asthma, idiopathic pulmonary fibrosis (IPF), chronic obstructive pulmonary disease (COPD), allergic rhinitis, or chronic sinusitis.

  The term “asthma” refers to a disease that exhibits reversible airway obstruction and / or bronchial hyperresponsiveness when associated or not associated with underlying inflammation. As an example of asthma, see, for example, Expert Panel Report 3: Guidelines for the Diagnosis and Management of Asthma, National Asthma Education and Prevention Program (specifications in EP) ) As mentioned in, allergic asthma, atopic asthma, corticosteroid naive asthma, chronic asthma, corticosteroid resistant asthma, corticosteroid refractory asthma, asthma due to smoking, not controlled by corticosteroids Asthma and other asthma.

The term “COPD” as used herein refers to chronic obstructive pulmonary disease. The term “COPD” includes two main conditions, emphysema and chronic obstructive bronchitis. Thus, in the broadest sense, the term COPD, as used herein, refers to chronic bronchitis and emphysema as its subconditions in addition to COPD itself. The Global Initiative for Chronic Obstructive Lung Disease (GOLD) for chronic obstructive pulmonary disease classifies COPD into four different stages. GOLD classification in COPD stage 0: Risk of COPD. Patients may have normal spirometry, although they may present with chronic cough and sputum symptoms. Stage I: Mild COPD. Characterized by FEV ₁ ≧ 80%, FEV ₁ / FVC <70%. Patients may or may not have an increased incidence of chronic cough and sputum. Stage II: moderate COPD. Characterized by airflow deterioration (30% ≧ FEV ₁ > 80%). Patients with stage II disease are symptomatic, seek medical attention, and often have shortness of breath during exertion. Stage II has two subcategories, IIA and IIB. IIA patients have 50% to 80% FEV ₁ , and stage IIB patients have 30% to 50% FEV ₁ . Patients with FEV ₁ less than 50% are particularly prone to acute exacerbations of the disease. Stage III: Severe COPD. Characterized by less than 30% FEV ₁ . Patients are also included in Stage III if they have respiratory failure or right heart failure. In these patients, the quality of life is significantly impaired. Acute exacerbations in this patient population often require hospitalization, which is often life threatening.

  The term “idiopathic pulmonary fibrosis” (IPF) refers to a disease characterized by progressive scarring of the lung, or fibrosis. It is a specific type of interstitial lung disease in which the alveoli gradually replace fibrotic tissue. In the case of IPF, due to progressive scarring, thin and soft tissue usually thickens and stiffens, making it more difficult for the lungs to expand and preventing oxygen from entering the bloodstream easily. For example, Am. J. et al. Respir. Crit. Care Med. 2000.161: 646-664.

  The term eosinophilic disease or disorder also encompasses the diseases and conditions disclosed below that are characterized by the presence of high eosinophil levels. Normal and abnormal (eg elevated) eosinophil levels in eosinophilic diseases or disorders disclosed below are also known in the art or measured using methods known in the art Can be done.

High eosinophil levels are observed in the following diseases or disorders:
(I) Allergic disorders: Allergic disorders are classically characterized by the presence of eosinophils. Allergic rhinoconjunctivitis (hay fever) has elevated levels of eosinophils in the nasal mucosa. Asthma shows an increase in the number of eosinophils in the lung after exacerbation,
(Ii) Drug reaction: Any drug / medicine has the potential to cause a reaction. Some of these reactions are allergic in nature, and eosinophils may increase in the blood or in tissues where drugs are concentrated,
(Iii) Infectious diseases: parasitic infections (helminthiasis-worms), fungal infections and some other types of infections are accompanied by an increase in eosinophil counts,
(Iv) Hematological disorders: Hematological disorders with elevated levels of eosinophils include hyperacidic syndrome, leukemia, lymphoma, tumor, mastocytosis, and atheroembolic disease,
(V) Immune disorders and reactions: Hyper-IgE syndrome, Omen's syndrome, thymoma, and transplant rejection are only some types of conditions with increased eosinophil counts,
(Vi) Endocrine disorders: Adrenal hypofunction is associated with an increase in the level of eosinophils in the blood.

Eosinophils have also been found to be increased or pathologically present in the following diseases or disorders:
(I) Skin and subcutaneous disorders: atopic dermatitis (eczema), bullous pemphigoid, pemphigus vulgaris, herpes dermatitis, drug-induced lesions, hives, eosinophilic panniculitis, eosinic acid Angioedema with hypercytosis, Kimura disease, Schulman syndrome, Wells syndrome, eosinophilic ulcer of the oral mucosa, eosinophilic pustular folliculitis, and recurrent cutaneous necrotizing eosinophilic vasculitis,
(Ii) Lung disease: drug / toxin-induced eosinophilic lung disease, Leffler syndrome, allergic bronchopulmonary aspergillosis, eosinophilic pneumonia, Churg-Strauss syndrome, eosinophilic granuloma, and pleural acid Polycytosis,
(Iii) Gastrointestinal disease: gastroesophageal reflux disease, parasitic infection, fungal infection, Helicobacter pylori infection, inflammatory bowel disease (ulcerative colitis and Crohn's disease), food allergic disorder, protein Induced enteropathy and protein-induced enterocolitis, allergic colitis, celiac disease, proliferative pemphigus (MR) and primary eosinophilic esophagitis, gastroenteritis, and colitis. Rare tumor (leiomyomatosis), connective tissue disorder, and vasculitis disorder,
(Iv) neuropathy: organized chronic subdural hematoma membrane, central nervous system infection, ventricular shunt, and drug-induced adverse reactions,
(V) Heart condition: Heart disorders have been reported secondary to systemic disorders such as hyperacidic syndrome or Churg-Strauss syndrome. Certain congenital heart diseases (septal defects, aortic stenosis) are accompanied by elevated levels of eosinophils in the blood,
(Vi) Kidney disease: eosinophilic urinary disease (urinary eosinophil) with infection, interstitial nephritis, eosinophilic cystitis.

  As used herein, the term “exacerbation” refers to an exacerbation of the symptoms of an eosinophilic disease or disorder as compared to the patient's baseline condition. In certain aspects, an “asthma exacerbation” is an event in the natural course of a disease characterized by changes in the patient's baseline lung function, dyspnea, cough, and / or sputum beyond normal daily fluctuations And may warrant medication changes in patients with acute onset and basal asthma. In certain embodiments, the exacerbation of asthma may be a rapid increase in symptoms of shortness of breath and / or wheezing and / or an increase in sputum production.

  As used herein, the term “treat”, “treatment” or “treatment of” (eg, in the phrase “treat a patient with an eosinophilic disease or disorder”) (i ) Reducing the likelihood of an eosinophilic disease or disorder (eg, a pulmonary disease or disorder such as asthma), (ii) reducing the incidence of an eosinophilic disease or disorder, and (iii) eosinophils. Reducing the severity of the sexually transmitted disease or disorder, preferably to the extent that the subject no longer suffers from discomfort and / or altered function (eg asthma in the case of asthma compared to an untreated patient) Relative reduction in exacerbation), or (iv) a combination thereof.

  For example, treatment cures the ability of the treatment, when administered to a subject, to prevent the occurrence of an eosinophilic disease or disorder, and / or the symptoms, signs, or causes of an eosinophilic disease or disorder. Or the ability to alleviate. Treatment also refers to alleviation or reduction of at least one clinical symptom, and / or inhibition or delay in the progression of the condition, and / or prevention or delay of the onset of the disease or condition. Thus, the terms “treat”, “treating” or “treatment of” (or grammatically equivalent terms) refer to both prophylactic and therapeutic treatment plans. Point to.

  The present disclosure provides methods and systems that provide a therapeutic effect in the treatment of eosinophilic diseases or disorders (eg, lung diseases or disorders such as asthma). The therapeutic effect is not necessarily a cure for a particular eosinophilic disease or disorder, but most typically (i) alleviation of eosinophilic disease or disorder or increased survival, (ii) eosinophilic Elimination of disease or disorder, reduction of symptoms associated with eosinophilic disease or disorder, (iii) prevention or alleviation of secondary disease, (iv) disorder resulting from occurrence of primary eosinophilic disease or disorder Or a condition, (v) prevention of an eosinophilic disease or disorder, or (v) a result comprising a combination thereof.

  The term “subject” or “patient” as used herein refers to any subject whose diagnosis, prognosis, or treatment of an eosinophilic disease or disorder (eg, a pulmonary disease or disorder such as asthma) is desired. Refers to a subject, particularly a mammalian subject. As used herein, the term “subject” or “patient” includes any human or non-human animal. The term “non-human animal” includes all vertebrates, eg, mammals and non-mammals, eg, non-human primates, sheep, dogs, cats, horses, cows, bears, chickens, amphibians, reptiles, and the like. As used herein, phrases such as “a patient with an eosinophilic disease or disorder” refer to treatment, imaging or other for that eosinophilic disease or disorder (eg, a pulmonary disease or disorder such as asthma). Includes subjects who will benefit from diagnostic methods and / or the implementation of prophylactic treatment, eg, mammalian subjects. The term “normal healthy volunteer” corresponds to “healthy subject” or “healthy volunteer”. One skilled in the art will recognize that the term “healthy volunteers” is generally described in the United States Pharmacopoeia, p. 2645 (U.S. Pharmaceutical Covention, Inc., 28th ed., 2005).

  In some aspects of the present disclosure, the subject is a naïve subject. A naive subject is a subject who has not been treated, eg, treated. In some embodiments, the naïve subject has not been treated with a therapeutic agent prior to being diagnosed as having an eosinophilic disease or disorder (eg, a pulmonary disease or disorder such as asthma).

  In another aspect, a subject has the ability to modulate a therapeutic and / or therapeutic agent (eg, an inflammatory response associated with an eosinophilic disease or disorder prior to being diagnosed as having an eosinophilic disease or disorder. Taking one or more treatments).

  In some aspects, the subject has received at least one therapeutically effective amount of an oral or inhaled corticosteroid. In some aspects, inhalation administration is performed using a metered dose inhaler (MDI) or a dry powder inhaler (DPI). In some aspects, the steroid is administered at a high dose. The term high dose may refer to at least 500 μg ICS (eg, fluticasone) as DPI or at least 440 μg ICS as MDI when applied to inhaled corticosteroids (ICS), for example, in a total daily dose. In some aspects, the high total daily dose of ICS, as DPI, is at least about 300 μg, at least about 350 μg, at least about 400 μg, at least about 450 μg, at least about 500 μg, at least about 550 μg, at least about 600 μg, at least about 650 μg, At least about 700 μg, at least about 750 μg, at least about 800 μg, at least about 850 μg, at least about 900 μg, at least about 950 μg, or at least 1000 μg of ICS (eg, fluticasone). In some aspects, the high total daily dose of ICS is at least about 300 μg, at least about 350 μg, at least about 400 μg, at least about 450 μg, at least about 500 μg, at least about 550 μg, at least about 600 μg, at least about 650 μg, as MPI. At least about 700 μg, at least about 750 μg, at least about 800 μg, at least about 850 μg, at least about 900 μg, at least about 950 μg, or at least 1000 μg of ICS (eg, fluticasone).

  The term “high dose” means when applied to inhaled corticosteroids (ICS) (eg fluticasone) in combination therapy (eg with bronchodilators such as salmeterol), for example, two inhalations at a dose twice a day. It may refer to about 230 μg fluticasone and about 21 μg salmeterol as MDI, or about 500 μg fluticasone and about 50 μg salmeterol as DPI in a single dose. The concentrations of corticosteroids that are considered high doses alone and in combination with other therapeutic agents are well known in the art.

  In some embodiments, the subject has received multiple therapeutically effective doses of oral or inhaled corticosteroids. In some aspects, the subject is a chronic oral corticosteroid (OCS) user.

  In certain embodiments, the subject is receiving a long-acting β2 adrenergic agent, such as salmeterol xinafoate. In some aspects, the subject is receiving a synthetic glucocorticoid, such as fluticasone propionate. In certain embodiments, the subject has received a combination of salmeterol xinafoate and fluticasone propionate (ADVAIR®). In certain embodiments, the subject is receiving a β2 adrenergic bronchodilator, such as albuterol sulfate.

  The term “treatment” as used herein refers to any means for healing, alleviation, or prevention of eosinophilic diseases or disorders (eg, pulmonary diseases or disorders such as asthma), eg, therapeutic agents ( For example, eosinophil targeted therapeutics), instrumentation, symptomatic therapy, and surgical or rehabilitation procedures. In this context, the term treatment encompasses any protocol, method and / or therapeutic or diagnostic agent that can be used in the prevention, management, treatment, and / or amelioration of an eosinophilic disease or disorder. .

  In some aspects, a patient is considered positive for DPP4 or POSTN when a measurement for a biomarker exceeds a predetermined threshold level. A patient is considered to have a high or elevated DPP4 level (high DPP4) when the measured DPP4 level in the sample exceeds a predetermined threshold level or exceeds the level of DPP4 in one or more control samples. Conversely, a patient has a low or reduced DPP4 level (low DPP4) when the measured DPP4 level in the sample is below a predetermined threshold level or below the level of DPP4 in one or more control samples Conceivable.

  Similarly, a patient has a high or elevated POSTN level (high POSTN) when the measured POSTN level in the sample exceeds a predetermined threshold level or exceeds the level of POSTN in one or more control samples. Conceivable. A patient is considered to have a low or reduced POSTN level (low POSTN) when the measured POSTN level in the sample is below a predetermined threshold level or below the level of POSTN in one or more control samples.

  One skilled in the art will appreciate that it is possible to combine quantitative and qualitative means. For example, the presence of DPP4 and / or POSTN in a sample may be determined using a qualitative assay that simply detects the presence or absence of a biomarker that exceeds a certain quantification limit in the assay, whereas other biomarkers The level may be measured based on a quantitative assay that indicates that the level of the biomarker is above or below a certain predetermined threshold or within a certain range.

  In some aspects, the DPP4 level (eg, protein or nucleic acid level) of the subject in the sample is below a predetermined DPP4 threshold level, or the DPP4 level is reduced compared to the DPP4 level in one or more control samples. In some cases, the subject may be administered at least one therapeutically effective amount of the eosinophil targeted therapeutic agent. In other embodiments, the POSTN level (eg, protein or nucleic acid level) of the subject in the sample is below a predetermined POSTN threshold level, or the POSTN level is reduced compared to the POSTN level in one or more control samples. The subject can be administered at least one therapeutically effective amount of the eosinophil targeted therapeutic agent.

  The term “therapeutic agent” as used herein is also administered to a subject having an eosinophilic disease or disorder (eg, a pulmonary disease or disorder such as asthma) to produce the desired normal beneficial effect. Refers to any therapeutically active substance. The term therapeutic agent includes, for example, without limitation, classic low molecular weight therapeutic agents commonly referred to as small molecule drugs, antibodies or active fragments thereof, peptides, lipids, protein drugs, protein complex drugs, And biologics including enzymes, oligonucleotides, ribozymes, genetic material, prions, viruses, bacteria, and eukaryotic cells.

  The therapeutic agent can also be a prodrug that metabolizes to the desired therapeutically active agent when administered to a subject. In some aspects, the therapeutic agent is a prophylactic agent. Further, the therapeutic agent can be formulated as a drug. The therapeutic agent can also be or include an agent activated by a radioisotope, or some other form of energy such as light or ultrasound energy, or other circulating molecules that can be administered systemically. obtain.

  In one aspect, the therapeutic agent is a small molecule agent. In a specific embodiment, the therapeutic agent is a corticosteroid. In another aspect, the therapeutic agent can be a leukotriene modifier such as montelukast, zafirlukast or zileuton. In a further aspect, the therapeutic agent can be methylxanthine (eg, theophylline) or chromone (eg, sodium cromoglycate and nedocromil). In another aspect, the therapeutic agent can be a long acting β2 stimulant such as salmeterol, formoterol, or indacaterol. In a further aspect, the therapeutic agent can be methotrexate or cyclosporine.

  In certain embodiments, the therapeutic agent can be an agent used to prevent, treat, manage, or ameliorate an eosinophilic disease or disorder, such as a pulmonary disease or disorder such as asthma. . Non-limiting examples of therapeutic agents for asthma include anticholinergic agents (eg, ipratropium bromide and oxitropium bromide), β2 antagonists (eg, albuterol (PROVENTIL® or VENTOLIN®)), vitorterol (TOMALATE (R)), fenoterol, formoterol, isoetarine, metaproterenol, pibuterol (MAXAIR (R)), salbutamol, salbutamol terbutaline, and salmeterol, terbutaline (BRETHAIRE (R))), corticosteroids (e.g. , Prednisone, beclomethasone dipropionate (VANCERIL® or BECLOVEENT®), triamcinolone acetonide (AZMA) ORF (registered trademark)), flunisolide (AEROBID (registered trademark)), mometasone (NASONEX (registered trademark), ASMANTEX (registered trademark)), and fluticasone propionate (FLOVENT (registered trademark))), leukotriene antagonists (for example Montelukast, zafirlukast, and zileuton), theophylline (THEO-DUR®, UNIDUR® tablets, and SLO-BID® Gyrocaps), and salmeterol (SEREVENT®), cromolyn, and nedocromil (INTAL® and TILADE®)), IgE antagonists, IL-4 antagonists (including antibodies), IL-5 antagonists (including antibodies), PDE4 inhibitors, NF-κB inhibitors , IL 13 antagonists (including antibodies), CpG, CD23 antagonists, selectin antagonists (eg, TBC1269), mast cell protease inhibitors (eg, tryptase kinase inhibitors (eg, GW-45, GW-58, and genistein) , Phosphatidylinostide-3 ′ (PI3) -kinase inhibitors (eg, calfostin C), and other kinase inhibitors (eg, staurosporine), C2a receptor antagonists (including antibodies), and supportive breathing Therapies include auxiliary mechanical ventilation.

  An eosinophil targeted therapeutic agent, as used herein, can be any “therapeutic agent” as defined herein, either directly or indirectly, (i) Can inhibit, reduce or neutralize the activity of eosinophils in a patient, or (ii) inhibit and reduce eosinophil levels in a patient systemically or within a specific tissue or organ Or (iii) can reduce the half-life of eosinophils in a patient, or (iv) can prevent exacerbation of symptoms associated with elevated levels of eosinophils Or (v) a combination thereof.

  In some aspects of the present disclosure, the eosinophil targeted therapeutic agent is (i) an antibody that targets, for example, IL-5R or IL-5 or an antigen-binding fragment thereof, (ii) the therapeutic agent disclosed on Any of (eg, corticosteroids), or (iii) any combination thereof.

  A “therapeutically effective” amount, as used herein, is an amount of a therapeutic agent that provides some improvement or benefit to a subject having an eosinophilic disease or disorder (eg, a pulmonary disease or disorder such as asthma). . Thus, a “therapeutically effective” amount is an amount that provides some relief, alleviation, and / or reduction in at least one clinical symptom of an eosinophilic disease or disorder (eg, a pulmonary disease or disorder such as asthma).

  Clinical symptoms associated with eosinophilic diseases or disorders (eg, pulmonary diseases or disorders such as asthma) can be treated by the methods and systems of the present disclosure well known to those skilled in the art. Furthermore, those skilled in the art will appreciate that the therapeutic effect need not be complete or radical as long as some benefit is provided to the subject. In some aspects, the term “therapeutically effective” refers to a therapeutic agent capable of altering a biomarker level, eg, DPP4 level and / or POSTN level, and / or a patient's eosinophil count in a patient in need thereof. Refers to the amount.

  As used herein, a “sufficient amount” or “sufficient amount to obtain” special results in a patient having an eosinophilic disease or disorder (eg, a pulmonary disease or disorder such as asthma) is desired Effect, optionally the amount of a therapeutic agent (eg, an anti-IL-5R such as benralizumab) that is effective to produce a therapeutic effect (ie, by a therapeutically effective amount). In some aspects, such a special result is a decrease in the patient's eosinophil count.

  The term “sample” as used herein includes any bodily fluid or tissue obtained from a subject, such as whole blood, serum, sputum, saliva, or epithelial tissue (eg, lung tissue). The sample can be any body fluid or tissue, such as whole blood, serum, saliva, urine, nasal discharge, sputum, bronchoalveolar lavage fluid, lung tissue, peripheral blood mononuclear cells, total white blood cells, lymph node cells, spleen cells, tonsil cells Including skin, nasal mucosa, lung epithelial cells and the like. In some specific embodiments, the sample is blood or a fraction thereof. The sample can be obtained by any means known in the art.

  In some aspects, the sample may be obtained by taking biological samples from multiple subjects and pooling them or pooling an aliquot of each subject's biological sample. it can. Pooled samples can be treated as samples from a single subject. The term sample also includes all experimentally separated fractions described above. For example, a blood sample can be fractionated into serum or into fractions with special types of cells. In some aspects, the sample can be a combination of samples from an individual, eg, a combination of tissue and fluid samples.

  As used herein, the term “control” when used to characterize a subject refers to a healthy subject or a patient who has been diagnosed with a particular disease other than an eosinophilic disease or disorder. The term “control sample” refers to one or more biological samples obtained from a healthy subject or a patient diagnosed with a disease other than an eosinophilic disease or disorder.

  To apply the methods and systems of the present disclosure, a patient-derived sample can be obtained before or after the treatment is applied to treat an eosinophilic disease or disorder (eg, a pulmonary disease or disorder such as asthma). . In some cases, a series of samples can be obtained from a patient after treatment has begun or after treatment has been completed. The sample can be requested, for example, by a health care provider (eg, a physician) or a health care benefits provider, obtained by the same or different health care provider (eg, nurse, hospital) or clinical laboratory and / or Or after processing, the results can be sent to the original health care provider or yet another health care provider, health care provider or patient.

  Similarly, quantification of the expression levels of the biomarkers disclosed herein, eg DPP4 and / or POSTN, alone or in combination with at least one additional biomarker, eg, measurement of a patient's eosinophil count A comparison and / or ratio between expression levels of a biomarker gene or protein, an assessment of the absence or presence of a biomarker, a measurement of a biomarker level relative to a specific threshold, a therapeutic decision, or a combination thereof is 1 It may be performed by one or more health care providers, health care benefits providers, and / or clinical laboratories.

  As used herein, the term “medical provider” refers to an individual or facility that interacts and administers directly to a living subject, eg, a human patient. Non-limiting examples of health care providers include doctors, nurses, technicians, therapists, pharmacists, counselors, alternative medical practitioners, medical facilities, clinics, hospitals, emergency rooms, clinics, emergency medical centers, alternative medical clinics / facilities, And general and / or professional treatment, evaluation, maintenance, including but not limited to general medicine, professional medicine, surgery, and / or any other type of treatment, evaluation, maintenance, treatment, medication and / or advice, Any other entity that provides treatment, medication, and / or advice relating to all or any part of the patient's health.

  As used herein, the term “clinical laboratory” refers to a facility for the examination or processing of material from a living subject, eg, a human. Non-limiting examples of treatment include, for example, from a human body intended to provide information about the diagnosis, prevention, or treatment of any disease or disorder in a living subject, eg, a human, or assessment of its health Biological examination of material, biochemical examination, serological examination, chemical examination, immunohematological examination, hematology examination, biophysical examination, cytological examination, pathological examination, genetic Or other tests. These tests may also be taken on samples obtained or otherwise obtained, or the presence or absence of various substances in a living subject, such as the human body, or a sample obtained from a living subject, such as the human body. Procedures for preparing, determining, measuring or otherwise describing.

  As used herein, the term “medical benefit provider” refers to an individual entity, organization, or group that provides patients with one or more medical benefits, benefit plans, health insurance, and / or medical expense accounts. The use of the program, in whole or in part, includes providing, presenting, soliciting, paying, or otherwise participating in providing.

  In some aspects, a health care provider manages or directs another health care provider to administer a treatment that treats an eosinophilic disease or disorder (eg, a pulmonary disease or disorder such as asthma). be able to. Healthcare providers obtain samples, process samples, submit samples, receive samples, transfer samples, analyze or measure samples, quantify samples, analyze / measure / quantify samples One or more samples that receive results obtained from analysis / measurement / quantification of the sample, provide results obtained, compare / score the results obtained from analysis / measurement / quantification of one or more samples Providing a comparison / score from one or more samples, administering a treatment (eg, administration of an eosinophil-targeted therapeutic agent such as benralizumab / MEDI-563) Start, end treatment, continue treatment, temporarily suspend treatment, increase the amount of therapeutic agent administered, reduce the amount of therapeutic agent administered, yes Continue to administer the amount of therapeutic agent Increase the frequency of administration of a therapeutic agent, reduce the frequency of administration of a therapeutic agent, maintain the same frequency of administration for a therapeutic agent, replace one treatment or therapeutic agent with at least another treatment or therapeutic agent, Acts such as combining one treatment or therapeutic agent with at least another treatment or additional therapeutic agent can be performed or directed to another health care provider or patient to perform.

  In some aspects, the health care provider provides, for example, sample collection, sample processing, sample submission, sample receipt, sample transfer, sample analysis or measurement, sample quantification, sample analysis / measurement. / Providing results obtained from quantification, Sample analysis / measurement / Transfer of results obtained from quantification, Comparison / scoring of results obtained from analysis / measurement / quantification of one or more samples, one or more Comparison / score transfer from a sample, treatment or treatment administration, initiation of treatment or treatment administration, interruption of treatment or treatment administration, continuation of treatment or treatment administration, treatment or treatment administration Temporarily interrupted, increased amount of therapeutic agent administered, decreased amount of therapeutic agent administered, continued administration of a certain amount of therapeutic agent, increased frequency of therapeutic agent administration, frequency of therapeutic agent administration Decrease in Approve or deny maintaining frequency, replacing one treatment or treatment with at least another treatment or treatment, or combining one treatment or treatment with at least another treatment or additional treatment it can.

  In addition, health care providers may, for example, authorize or deny treatment prescriptions, authorize or deny treatment coverage, authorize or deny reimbursement of treatment costs, determine or deny eligibility for treatment, etc. Can do.

  In some aspects, a clinical laboratory, for example, takes or obtains a sample, processes a sample, submits a sample, receives a sample, transfers a sample, analyzes or measures a sample, quantifies a sample Receive results obtained from sample analysis / measurement / quantification, provide results obtained from sample analysis / measurement / quantification, compare results obtained from analysis / measurement / quantification of one or more samples / Scoring, providing a comparison / score from one or more samples, obtaining a comparison / score from one or more samples, or taking other related actions.

  As used herein, the term “computed tomography” or “CT” refers to an imaging method that uses a tomographic image (virtual “slice”) of a specific region of a scanned organ, tissue or object. Digital geometry processing is used to create a three-dimensional (3D) image of the interior of a subject or organ from a series of two-dimensional (2D) radiographs taken around a single axis of rotation.

  As used herein, the term “computed tomography scan” or “CT scan” includes but is not limited to x-ray, multi-detector computed tomography (MDCT), high resolution computed tomography (HRCT), Positron emission tomography (PET), positron emission tomography / computed tomography (PET-CT), single photon emission computed tomography (SPECT), magnetic resonance imaging (MRI), computer X-ray body axis tomography (CAT) Scan), computer-assisted tomography, xenon ventilated computed tomography, and hyperpolarized gas MRI lung ventilation imaging (hyperpolarized f gas lung MRI ventilation im refers to the generation of tomographic images obtained using any suitable method.

II. Low DPP4 and POSTN as biomarkers for eosinophilic disease
In general, the methods disclosed herein are:
(A) 1, 2, 3, or more changes in the level of DPP4 and / or POSTN alone or in patients with eosinophilic diseases or disorders (eg lung diseases or disorders such as asthma) Detecting in combination with detecting changes in biomarker levels, such as blood eosinophil count,
(B) predicting an increased clinical response to treatment with an eosinophil-targeted therapeutic agent, eg, an anti-IL-5R antibody such as benralizumab (MED-563), based on the detected DPP4 and / or POSTN levels;
(C) an eosinophil targeted therapeutic if DPP4 and / or POSTN levels, alone or in combination with other biomarkers, indicate that the patient would benefit from treatment with an eosinophil targeted therapeutic To the patient. If DPP4 and / or POSTN levels, alone or in combination with other biomarkers, indicate that the patient does not benefit from treatment with an eosinophil targeted therapeutic, treatment is interrupted, eg, Can be discontinued and changed (eg, increased dose or frequency of administration)
based on.

  In other words, specific levels of DPP4 and / or POSTN alone or in combination with other molecules or clinical biomarkers (eg, patient eosinophil count) correlate with the clinical efficacy of the therapeutic agent, Useful for predicting clinical outcome in a particular population of patients suffering from an acidophilic disease or disorder (eg, a pulmonary disease or disorder such as asthma).

  The term “DPP4” as used herein refers to the dipeptidyl peptidase IV protein (EC 3.4.14.5; Uniprot: P27487) encoded by the DPP4 gene (cDNA sequence corresponds to SEQ ID NO: 3). Point to. DPP4 is also known as DPP-IV, adenosine deaminase complex protein 2, or CD26 (surface antigen classification 26). DPP4 relates to attractin, FAP, DPP8 and DPP9. DPP4 is a highly conserved multifunctional type II transmembrane glycoprotein that exists both in the circulation (plasma) and on the surface of several cell types including epithelial, endothelial and lymphoid cells. Thus, DPP4 exists in both a membrane bound form (SEQ ID NO: 1) and a soluble form (SEQ ID NO: 2).

  DPP4 is part of the serine protease family involved in T cell costimulation, chemokine biology, type II diabetes, and tumor biology (Zhong et al., Atherosclerosis 2013; 226: 305-314). The role of DPP4 in inflammatory respiratory disease-like asthma was found to be an increase in DPP4 transcripts (and other Th2 signature genes) in the nasal mucosal epithelium of children with dust mite allergic rhinitis with uncontrolled asthma, Giovanni -Chami (Giovannini-Cami et al., European Republic Journal. 2012 May; 39 (5): 1197-205). The term DPP4 also includes fragments, variants (eg, K1R, V7I, S437I, T557I, D663E variants known in the art), and derivatives thereof (eg, glycosylated or deglycosylated protein forms of DPP4 protein, Or other chemically modified forms of proteins).

  In some aspects, in addition to or in addition to measuring the level of DPP4, the methods disclosed herein determine a sample taken from a patient, submit for determination, or periostin Instructing a clinical laboratory to determine expression levels or activity may be included. The use of periostin as a biomarker in pulmonary diseases such as asthma is described, for example, in Jia, et al. , J Allergy Clin. Immunol 2012 130: 647-654; Takayama, et al. , J Allergy Clin Immunol 2006 118: 98-104; and PCT Publication No. WO2012 / 083132, each of which is incorporated herein by reference in its entirety.

  The term “POSTN” as used herein refers to the osteoblast specific factor protein periostin (Uniprot: Q15063; SEQ ID NO: 20) encoded by the POSTN gene. POSTN is also known as osteoblast specific factor 2 (OSF-2). POSTN functions as a ligand for α-V / β-3 and α-V / β-5 integrins to support epithelial cell adhesion and migration. POSTN is a gla domain vitamin K-dependent factor. The term POSTN also includes fragments, variants (eg, isoforms produced by alternative splicing), and derivatives thereof (eg, glycosylated or deglycosylated protein forms of proteins, or otherwise chemically modified forms of proteins). including.

  Seven POSTN isoforms produced by alternative splicing: also known as OSF-2OS, 836 amino acids long isoform 1 (Uniprot: Q15063-1); also known as OSF-2p1, 779 amino acids long Is isoform 2 (Uniprot: Q15063-2); is isoform 3 (Uniprot: Q15063-3) is 781 amino acids long; is isoform 4 (Uniprot: Q15063-4) is 809 amino acids long; Isoform 5 (Uniprot: Q15063-5); isoform 6 (Uniprot: Q15063-6) which is 749 amino acids long; and isoform 7 (Uniprot: Q1506) which is 721 amino acids long; -7) it is known in the art. Known POSTN variants include those having any of the following sequence differences related to the reference isoform 1 sequence: I290F, D421V, T339I, or V814M.

  In some aspects, the DPP4 and POSTN biomarkers disclosed herein can be combined with other biomarkers for eosinophilic disease or disorder, such as the patient's eosinophil count, and then inflammatory One or more molecular biomarkers in the pathway or clinical biomarkers known in the art can be substituted or combined.

  The term “biomarker” as used herein refers to a factor that is a characteristic indicator of a biological process, biological event, and / or pathological condition, such as an eosinophil targeted therapeutic agent, Refers to predictors of clinical response to treatment with anti-IL-5R antibodies such as, for example, benralizumab (MED-563). As used herein, the term biomarker encompasses both clinical markers and molecular biomarkers (biological markers). Thus, in the context of the present disclosure, the term “biomarker” refers to a “biological biomarker” or “molecular biomarker”, such as, for example, the DPP4 and POSTN biomarkers disclosed herein, alone or Included in combination with molecular biomarkers associated with the IL-13 pathway, molecular biomarkers associated with certain eosinophilic diseases or disorders (eg, pulmonary diseases or disorders such as asthma), and combinations thereof. The biological markers disclosed herein also include genes (DNA and / or RNA) that encode those proteins, and metabolites.

  As disclosed above, the term “biomarker” can also predict response to a biotherapeutic drug, “clinical biomarker” also referred to as “clinical condition marker”, eg, gender, age, concomitant drug Smoking status, body mass index (BMI) and the like. For example, U.S. Patent Application Publication No. 20150065530, U.S. Patent Application Publication No. 20140141990, U.S. Patent Application Publication No. 20130005596, U.S. Patent Application Publication No. 20090233304, U.S. Patent Application Publication No. 201401199709. U.S. Patent Application Publication No. 20130303398 and United States Patent Application Publication No. 201110212104, which are hereby incorporated by reference in their entirety.

  The DPP4 molecule biomarkers disclosed herein are also at least about 70%, 71%, relative to the wild-type sequence in either its membrane bound form (SEQ ID NO: 1) or soluble form (SEQ ID NO: 2), 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88% 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity protein or fragment thereof, and membrane binding of DPP4 or At least about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, for each wild-type nucleic acid sequence encoding a soluble form, 81%, 82%, 83%, 84%, 85%, 86%, 8 %, Including 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, a nucleic acid having 98% or 99% sequence identity.

  The POSTN molecular biomarkers disclosed herein are also at least about 70%, 71%, 72%, 73%, relative to the wild type sequence POST (SEQ ID NO: 20) or POSTN isoforms known in the art. 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90% , 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% of sequence identity or a wild-type nucleic acid sequence of SEQ ID NO: 20 or the art At least about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82% relative to POSTN isoforms known in the art , 83%, 8 %, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity A nucleic acid having

  The DPP4 and POSTN molecular biomarkers disclosed herein also include fragments, variants, and derivatives thereof. As used herein, a “variant” biomarker includes at least one amino acid sequence modification relative to the amino acid sequence of the corresponding wild-type polypeptide. The amino acid sequence modification may be, for example, one or more amino acid substitutions, deletions or insertions, preferably conservative substitutions. Variant biomarkers can have any combination of amino acid substitutions, deletions or insertions. In one aspect, the variant polypeptide of the biomarker has at least 60, 70, 80, 85, 90, 95, 97, 98, 99, 99.5 or the amino acid sequence of the wild-type polypeptide to which the amino acid sequence corresponds. There may be some integer number of amino acid modifications to share 100% identity.

  In some aspects of the disclosure, the methods disclosed herein use DPP4 exclusively as a biomarker, POSTTN exclusively as a biomarker, and a combination of DPP4 and POSTN, Or, optionally, by incorporating additional clinical and / or molecular biomarkers such as the patient's eosinophil count, it can be applied to eosinophilic diseases or disorders (eg lung diseases or disorders such as asthma).

  In some aspects, the methods disclosed herein include additional biomarkers such as blood eosinophil cell count, patient IgE level levels, pre- or post-bronchodilator FEV1 reversibility, pulmonary Using sub-area airway wall area percentage (WA%) from CT scan data, or combinations thereof may be included. To predict a therapeutic response (eg, improvement in airway resistance and / or FEV1),% wall area (WA%) as measured using a CT scan of lungs with subzone airways (WA%) can be used.

  For example, the presence of eosinophils above normal baseline levels in the blood can be used as a biomarker in combination with DPP4 levels and / or POSTN levels. Severe asthma patients have frequent exacerbations and hospitalizations, accounting for the majority of disease costs and most of their mortality (Gaga et al., 2009). Inflammation, an important feature in severe asthma, exhibits a different phenotype that can be characterized by varying degrees of eosinophil and neutrophil infiltration persistence (Balzar et al., 2002). The presence of eosinophils in asthma is well documented via airway biopsy. The clinical significance of eosinophils in asthma is demonstrated by the findings of frequent exacerbations of asthma in patients with sputum eosinophil counts> 3%. Furthermore, clinical trials designed to adapt inhaled anti-inflammatory therapies to maintain the eosinophil count at <3% have resulted in reduced asthma exacerbations (Green et al., 2002). Symptomatic asthma patients with sputum eosinophilia refractory to standard treatment also improve after monoclonal antibody treatment (mepolizumab) that depletes airway eosinophils (Nair et al., 2009; Haldar et al., 2009).

  To date, the only accurate and reliable method for identifying patients with eosinophilic asthma has been limited to obtaining sputum samples derived from patients (Molfino, 2012). The hemorrhoid guidance procedure is a cumbersome and complex process that requires skilled technicians and equipment not readily available in practice. Even with these shortcomings, induced sputum continues to be a criterion for assessing the cellular inflammatory processes that occur in asthma (Lieberman, 2007). Panelists convened from the National Institutes of Health and federal agencies to raise biomarkers to assess disease progression and response to treatment classify patients as eosinophilic asthma patients 2% eosinophils in sputum are recommended as a cut-off (Szefler et al., 2012).

A cut-off approach based on absolute values, eg, 痰 EOS% cut-off, is used as a criterion for prediction and classification in pulmonary diseases and disorders such as eosinophilic asthma. In one aspect, the EOS% cutoff refers to classifying a patient as eosinophils with eosinophilicity of 2% or greater. The 1%, 2%, 2.5%, and 3% sputum eosinophil EOS% cut-off points are reported as distinguishing between eosinophilic and non-eosinophil patients Yes. For example, Green et al. , 2002 and Jayaram et al. , 2006. Belda et al. 2000 showed a mean + 2 standard deviation for 痰 EOS% in healthy subjects was 2.2%. To date, attempts to predict and classify eosinophilic asthma have examined the correlation between individual scales (such as blood eosinophil count and FE _NO ) and 痰 EOS%.

  The cut-off disclosed above with respect to eosinophil count, ie, 1%, 2%, 2.2%, 2.5%, or 3% percentage of sputum eosinophils or% EOS Can be used as a threshold value in the method disclosed in.

The term “wall area” as used herein refers to the cross-sectional area of the bronchial wall (eg, areas in the upper lobe and sub-area bronchi). The wall area percentage (WA%) is calculated as follows: 100 ^* wall area / (wall area + lumen area). Tools for measuring wall area and wall area percentage are well known in the art. For example, Gupta et al. J Allergy Clin Immunol. 133 (3): 729-738 (2014); Gupta et al. , Thorax. 65 (9): 775-81 (2010). In some aspects, the airway dimensions are measured from lung computed tomography (CT) imaging data. Such imaging data may be processed using commercially available software such as, for example, VIDA Apollo (eg, Volumetric Information Display and Analysis (VIDA) Pulmonary Workstation, VIDA Diagnostics, Coralville, Iowa).

  In other aspects, the methods disclosed herein are also known as CTLA-3 (soluble CTLA-3; cytotoxic T lymphocyte-associated serine esterase 3, granzyme A, or granzyme 1; Uniprot: P12544). SCD28 (soluble CD28; also known as surface antigen classification 28 or Tp44; Uniprot: P10747), CCL5 (chemokine CC motif ligand 5; also known as RANTES; Uniprot: P13501), CCL11 (CC motif chemokine 11; Also known as eosinophil chemotactic protein or eotaxin-1; Uniprot: P51671), CCL22 (C-C motif chemokine 22; Uniprot: O00626), or combinations thereof. Such may include the use of additional molecular biomarkers in combination with DPP4 and / or POSTN level. These biomarkers have been used in IL-13 mediated lung disease, for example, see Lun et al. , J .; Clin. Immunol. 2007 27: 430-437.

  In some aspects, the methods disclosed herein may also include CCL26, FZD5, DOK1, CST2, ZNF436, C20orf100, NAGS, CST1, CDH13, HRH1, TMEM132B, NTRK1, SLCO2A1, IgE, FETUB, KRT31IKRT38, C6or1 , ATP5J, TUBAL3, JAM2, NOVA2, NOS2A, HS3ST4, GRM8, IL1R2, CTDSPL, CEP72, LOC199800, LYPD1, DISP1, NKX1-2, C4orf38, LOXL4, PRKD1, CPR1H, PAM124B, GPR44C , ALOX15, AADAC, ACTA1, ODC1, DKFZp434F142, ACHE CSF3, LOC100132552, C12orf27, ZNF331, GK5, DUSP1IDUSP4, LRWD1, PGLYRP4, GUSBL2, CLGN, NR1I2, EST, LRRC37B, SAA4, SLC12A3, TMEM45A, FLJ37464, MJ4464 Use additional molecular biomarkers in combination with DPP4 and / or POSTN levels, such as the expression level or activity of PTP4A1, PCYT2, RHOV, PROS1, C11orf63, TCTN1, PIP5K1B, OSBPL6, NSUM7, GJB7, IRS2, or combinations thereof Can include. These genes are described in Choi et al. J. et al. Immunol. 186 (3): 1861-9 (2011) and WO2000091490, both of which are incorporated herein by reference in their entirety, are part of a Th-2 signature.

  In some aspects, the methods disclosed herein also include cystatin-SN (CST1); chemokine (CC motif) ligand 26 (CCL26); calcium-activated chloride channel regulator (1CLCA1); cystatin-SA. (CST2); proline rich protein 4 (PRR4); also known as plasminogen activator inhibitor 2 (placental PAI), HsT1201, PAI, PAI-2, PAI2 or PLANH2 (SERPINB2); carcinoembryonic antigen-related cell adhesion molecule 5 ( CEACAM5), also known as surface antigen class 66 (CD66e); inducible NOS (iNOS, NOS2), serpin peptidase inhibitor, clade B (ovalbumin), member 4 (SERPINB4, LEUPIN, PI11, S CA-2, SCCA1, SCCA2), cystatin S (CST4), basic salivary proline rich protein 4 (PRB4), tryptase delta 11 (TPSD1), tryptase gamma 1 (TPSG1), 2A protein having a major facilitator superfamily domain ( MFSD2), carboxypeptidase A3 (CPA3), G protein-coupled receptor 105 (GPR105), cadherin 26 (CDH26), gelsolin (GSN), protein 11 interacting with cannabinoid receptors (C2ORF32), transmembrane protein 711 (TRACH20000196) , TMEM71), DnaJ (Hsp40) homolog, subfamily C, member 121 (DNAJC12), RGS13 (G protein signaling) 3 regulators), solute carrier family 18 member 2 (SLC18A2), serpin peptidase inhibitor, clade B (obalbumin), member 10 (SERPINB10), SH3 ring finger 2 protein (SH3RF2), high affinity immunoglobulin epsilon receptor sub DPP4 and / or, such as the expression level or activity of unit beta (FCER1B), runt-related transcription factor 2 (RUNX2), prostaglandin endoperoxide synthase 1 (PTGS1), arachidonic acid 15-lipoxygenase (ALOX15), and combinations thereof It may include using additional molecular biomarkers in combination with POSTN levels.

  In some aspects, determining that a patient's DPP4 level is high (higher or increased) results in a measured level of DPP4 of about ≧ about as measured by a DDP4 detection immunoassay including the immunoassay disclosed in Example 2. It is required to be 376 ng / mL. In some aspects, determining that a patient's DPP4 level is low (lower or lower) results in a measured level of DPP4 of <about about 30% as measured by a DDP4 detection immunoassay, including the immunoassay disclosed in Example 2. It is required to be 376 ng / mL.

  In some aspects, determining that a patient's DPP4 level is high (higher or increased) has a measured level of DPP4 of about ≧ about as measured by a DDP4 detection immunoassay including the immunoassay disclosed in Example 2. It is required to be 363 ng / mL. In some aspects, determining that a patient's DPP4 level is low (lower or lower) results in a measured level of DPP4 of about < It is required to be 363 ng / mL.

  In some aspects, determining that a patient's POSTN level is high (higher or increased) is a measurement of POSTN as measured by a POSTN detection immunoassay, including the POSTN immunoassay disclosed in WO2015120185. The level is required to be ≧ about 25.8 ng / mL. In some aspects, determining that a patient's POSTN level is low (lower or reduced) is a measurement of DPP4 as measured by a POSTN detection immunoassay, including the POSTN immunoassay disclosed in WO2015120185. The level is required to be <about 25.8 ng / mL.

  In some aspects, determining that a patient's POSTN level is high (higher or increased) is a measurement of POSTN as measured by a POSTN detection immunoassay, including the POSTN immunoassay disclosed in WO2015120185. The level is required to be ≧ about 23.5 ng / mL. In some aspects, determining that a patient's POSTN level is low (lower or reduced) is measured by POSTN as measured by a POSTN detection immunoassay, including the POSTN immunoassay disclosed in WO2015120185. The level is required to be <about 23.5 ng / mL.

  In some aspects, determining that a patient's blood eosinophil count is high (higher or increased) requires a measured level of blood eosinophils to be ≧ 150 cells / μL. In some aspects, determining that a patient's blood eosinophil count is low (lower or lower) requires a measured level of blood eosinophils to be <150 cells / μL.

  In some aspects, determining that a patient's blood eosinophil count is high (higher or increased) requires a measured level of blood eosinophils to be ≧ 300 cells / μL. In some aspects, determining that a patient's blood eosinophil count is low (lower or lower) requires a measured level of blood eosinophils to be <300 cells / μL.

  In some aspects, determining that a patient's blood eosinophil count is high (higher or increased) requires a measured level of blood eosinophils to be ≧ 400 cells / μL. In some aspects, determining that a patient's blood eosinophil count is low (lower or lower) requires a measured level of blood eosinophils to be <400 cells / μL.

  In some aspects, determining that a patient's blood eosinophil count is high (higher or increased) has a measured level of blood eosinophils of at least about 150 cells / μL, 160 cells / μL, 170 Cells / μL, 180 cells / μL, 190 cells / μL, 200 cells / μL, 210 cells / μL, 220 cells / μL, 230 cells / μL, 240 cells / μL, 250 cells / μL, 260 cells / μL, 270 Cells / μL, 280 cells / μL, 290 cells / μL, 300 cells / μL, 310 cells / μL, 320 cells / μL, 330 cells / μL, 340 cells / μL, 350 cells / μL, 360 cells / μL, 370 It is required to be cells / μL, 380 cells / μL, 390 cells / μL, or 400 cells / μL.

  In some aspects, determining that a patient's blood eosinophil count is low (lower or lower), the measured level of blood eosinophils is at least less than 150 cells / μL, 150 cells / μL, 160 Cells / μL, 170 cells / μL, 180 cells / μL, 190 cells / μL, 200 cells / μL, 210 cells / μL, 220 cells / μL, 230 cells / μL, 240 cells / μL, 250 cells / μL, 260 Cells / μL, 270 cells / μL, 280 cells / μL, 290 cells / μL, 300 cells / μL, 310 cells / μL, 320 cells / μL, 330 cells / μL, 340 cells / μL, 350 cells / μL, 360 It is required to be cells / μL, 370 cells / μL, 380 cells / μL, 390 cells / μL, or 400 cells / μL.

III. Measurement of DPP4, POSTN, eosinophil count, and other biomarkers Levels of DPP4, POSTN, and other biomarkers disclosed herein (the level of their expressed proteins, or their respective nucleic acid levels, Any mRNA level, for example) can be detected and quantified by any of several methods well known to those skilled in the art. These methods include analytical, biochemical methods such as electrophoresis, capillary electrophoresis, high performance liquid chromatography (HPLC), thin layer chromatography (TLC), superdiffusion chromatography, mass spectrometry, or fluid or gel precipitation. Reaction, immunodiffusion (single or double), immunohistochemistry, affinity chromatography, immunoelectrophoresis, radioimmunoassay (RIA), enzyme-linked immunosorbent assay (ELISA), chemiluminescent immunoassay (CLIA), immunofluorescence assay Including various immunological methods such as Western blotting.

  In some aspects, the methods used to detect and / or quantify DPP4, POSTN, or other molecular biomarkers disclosed herein are encoded by a gene or gene segment in a sample Measuring the level, concentration, or amount of RNA, eg, mRNA. The level of RNA, eg, mRNA, is any technique known in the art, including but not limited to Northern blotting or quantitative PCR (qPCR), including methods such as reverse transcription qPCR, real-time qPCR, and endpoint qPCR. May be measured. Alternatively, “tag-based” techniques such as continuous analysis of gene expression (SAGE) and RNA-Seq may be performed to provide a relative measure of cell concentration of different mRNAs.

  In some aspects, DPP4, POSTN, and other molecular biomarkers disclosed herein can be detected and / or quantified in electrophoretic polypeptide separation (eg, one-dimensional or two-dimensional electrophoresis). . Means for detecting polypeptides using electrophoretic techniques are well known to those skilled in the art (in general, R. Scopes (1982) Polypeptide Purification, Springer-Verlag, NY; Deutscher, (1990) Methods in Enzymology Vol. 182: Guide to Polypeptide Purification, Academic Press, Inc., NY).

  In some aspects, Western blot (immunoblot) analysis is used to detect and quantify the presence or absence of DPP4, POSTN, or any other molecular biomarker disclosed herein in a sample. It is done. This technique generally involves separating a sample polypeptide by gel electrophoresis based on molecular weight and transferring the separated polypeptide to a suitable solid support (such as a nitrocellulose filter, nylon filter, or derivatized nylon filter). And incubating the sample with an antibody that specifically binds to the analyte. The antibody that specifically binds to the analyte may be directly labeled, or alternatively using a labeled antibody that specifically binds to the domain of the primary antibody (eg, a labeled sheep anti-mouse antibody). May be detected.

  In some aspects, DPP4, POSTN, and other molecular biomarkers disclosed herein can be detected and / or quantified in a biological sample using an immunoassay. For a general review of immunoassays, see Methods in Cell Biology Volume 37: Antibodies in Cell Biology, Asai, ed. Academic Press, Inc. New York (1993); Basic and Clinical Immunology 7th Edition, States & Terr, eds. See also (1991). For example, US Patent Application Publication No. 2007 / 0212723A1, Shang et al. , Circulation Research 101: 1146-1154 (2007); and International Patent Application Publication Nos. WO 2012/094651 and WO 2010/129964.

  In some aspects, the immunoassay can use one or more antibodies or antigen-binding fragments thereof that recognize molecular biomarkers (eg, human DPP4 or human POSTN). In some aspects, the immunoassay comprises a sandwich immunoassay, such as an enzyme linked immunosorbent assay (ELISA) or a sandwich electrochemiluminescence (ECL) assay, in which a primary antibody to a molecular biomarker (eg, DPP4 or POSTN) or its Antigen-binding fragments are used as “capture” antibodies. The capture antibody is attached to a solid support, the antigen from the sample or standard is capable of binding to the capture antibody, and then a secondary label or antigen binding thereof to the same biomarker including a detectable label, ie a “detection” antibody. Fragments are added. The detection antibody can be detected by either enzymatic reaction, ECL reaction, radioactivity, or any other detection method known in the art.

  In some aspects, the immunoassay comprises the following steps: first, the capture antibody or fragment thereof is bound to a solid support, such as a multi-well plate or other assay device known to those skilled in the art. To be. The capture antibody is allowed to attach for a period of time, eg, overnight, and then unbound antibody is removed. The plate can then be washed to remove any unbound capture antibody. The plate can then be treated with a blocking solution, allowing non-specific proteins to bind to any unbound regions of the solid support.

  Typical blocking solutions contain irrelevant proteins such as nonfat dry milk or serum albumin. The plate can then be washed again to remove any unbound blocking solution. A sample suspected of containing a molecular biomarker (eg, DPP4 or POSTN) is then added to the plate. Samples are typically serially diluted in duplicate or triplicate. Also included are controls, such as standard amounts of biomarkers (eg, pure recombinant DPP4 or pure recombinant POSTN) or suitable fragments thereof and various negative controls. The antigen is allowed to bind to the capture antibody for a period of room temperature, for example 1 hour. After incubation, the plate can then be washed to remove any unbound antigen.

  A detection antibody is then added. The detection antibody is typically an anti-antibody that specifically binds to an epitope of a molecular biomarker (eg, DPP4 or POSTN) epitope that is different from the epitope of interest to which the capture antibody binds. The detection antibody can be labeled or unlabeled. If the detection antibody is unlabeled, an additional step of adding a labeled secondary antibody will be required, as is well known by those skilled in the art.

  The detection antibody can be directly labeled with an enzyme, such as horseradish peroxidase or alkaline phosphatase, or can be labeled with a tag that allows binding of the enzyme. For example, the detection antibody can be coupled to biotin and the enzyme can be attached in subsequent steps by allowing binding of enzyme-bound streptavidin to a biotin tag. Alternatively, the detection antibody can be coupled to a chemiluminescent, fluorescent, or ECL tag. An example of the latter is ruthenium chelate. After incubation, the plate can then be washed to remove any unbound detection antibody. Detection of the detection antibody can be accomplished by methods that vary based on the type of detection antibody used.

  When the detection antibody is tagged with biotin, enzyme-conjugated streptavidin is added, unbound streptavidin is washed away, and the substrate is added, thereby producing a colorimetric reaction that can be read, for example, with a spectrophotometer. can get. When the detection antibody is bound to the ruthenium chelate, the plate receives an electric current and the light emission is measured.

  Immunoassays for detecting molecular biomarkers (eg, DPP4 or POSTN) can be either competitive or non-competitive. Non-competitive immunoassays are assays in which the amount of analyte captured is directly measured. In a competitive assay, the amount of analyte in the sample measures the amount of labeled analyte added (exogenous) that is displaced (or competitively excluded) from the capture agent by the analyte present in the sample. Indirectly measured. In one competitive assay, a known amount of, eg, a labeled molecular biomarker (eg, DPP4 or POSTN) is added to the sample, and then the sample is contacted with a capture agent. The amount of labeled molecular biomarker (eg, DPP4 or POSTN) bound to the antibody is inversely proportional to the concentration of molecular biomarker (eg, DPP4 or POSTN) present in the sample.

  In some aspects, the method directly measures the level of DPP4, POSTN, or other biomarker disclosed herein in a patient sample, where the absolute level is, for example, purified Calculated by plotting the immunoassay results on a standard curve using full length DPP4 or POSTN, or DPP4 or POSTN fragment (or full length biomarker or fragment thereof disclosed herein). The detected signal from the detection antibody can then be quantified based on various standards and controls included on the plate. By plotting the results on a calibration curve, the absolute level of DPP4, POTN, or any other biomarker disclosed herein is, for example, pg or ng biomarker / mL, or It can be calculated in pg or ng biomarker / mg total protein.

  Detection assays in DPP4, POSTN, or other molecular biomarkers disclosed herein can be scored (positive or negative or as an amount of analyte) according to standard methods well known to those skilled in the art. The particular method of scoring will depend on the assay format and the choice of label. For example, a Western blot assay can be scored by visualizing the colored product produced by the enzyme label. A clearly visible colored band or spot at the correct molecular weight is scored as a positive result, while the absence of a clearly visible spot or band is scored as negative. The intensity of the band or spot can provide a quantitative measure of analyte concentration.

  In some aspects, the measured expression level of a molecular biomarker (eg, DPP4 or POSTN) represents an average expression level or a mean expression level based on two or more measurements of the expression level. In some aspects, the measured expression level is the average or mean of several measurements of the expression level of the same sample. In some aspects, the measured expression level is an average or mean of several measurements of expression levels of different samples containing the same component from the same subject. In some aspects, the level of expression measured is quantile normalized as is done in RNA Seq technology using techniques well known by those skilled in the art.

  The term “DPP4 detection assay” as used herein refers to both quantitative and qualitative assays capable of detecting the presence or absence of DPP4 in a biological sample. The term DPP4 detection assay encompassed an immunoassay such as an ELISA.

  The term “POSTN detection assay” as used herein refers to both quantitative and qualitative assays capable of detecting the presence or absence of POSTN in a biological sample. The term POSTN detection assay included an immunoassay such as an ELISA. POSTN detection assays are well known in the art. In some aspects, the POSTN detection assay is an immunoassay disclosed in WO20150151711A1 or WO2015120185, or a commercially available POSTN assay.

  In some aspects, the DPP4 detection assay or POSTN detection assay is a multiplex immunoassay capable of detecting the expression level of DPP4 and / or POSTN in a sample, eg, Bio-Rad BIO-PLEX ™, EMD Millipore MILLIPLEX (trademark), Life Technologies NOVEX MULTIPLEX (trademark), Thermo Fisher Scientific LUMINEX (trademark), PerkinElmer ALPHAREX (trademark), PerkinElmer ALPHAREX (trademark)

  In some aspects, the DPP4 detection assay is an assay included in Table 1 or a variation thereof.

  When the limit of detection (LOD) of an assay is used as a predetermined DPP4 or POSTN threshold level, the predetermined DPP4 or POSTN threshold level becomes the specific LOD in such an assay.

  A given DPP4 or POSTN when the level of DPP4 or POSTN expression in a control subject or subjects (or subpopulations thereof) suffering from a particular eosinophilic disease or condition is used as the threshold level for a given DPP4 or POSTN biomarker The threshold level is the level of molecular biomarker reported for each population (eg, healthy controls, patients with moderate asthma, or patients with severe asthma) in the manufacturer's instructions for the assay.

  The term “level” when applied to a biomarker disclosed herein, eg, DPP4, POSTN, or a biomarker set disclosed herein, is a biomarker in one or more biological samples or Refers to one or more measurements obtained using any analytical method to detect the presence / absence or expression / absence of biomarker set expression (protein expression or gene expression) The presence, absence, absolute amount or concentration, relative amount or concentration, titer, expression level, ratio of measured levels, etc. of one or more biomarkers in a biological sample Show.

  The exact nature of the “value” or “level” depends on the particular assay (eg, immunoassay, mass spectrometry, in vivo) used to detect DPP4, POSTN, or another biomarker disclosed herein. Depending on the specific design and components of molecular imaging, gene expression profiling, aptamer based assays, etc.). See, for example, US Patent Application Publication No. 2010/00221752.

  As used herein with respect to DPP4, POSTN, and other biomarkers disclosed herein, the terms “elevated”, “increased” or “higher” apply when applied to a biomarker level. , Refers to the expression level or range of a biomarker measured in a control sample (“normal level”), or a level in a biological sample (eg, serum or sputum) that is above a particular threshold disclosed herein. These thresholds are, for example, about 363 ng / mL or about 376 ng / mL for DPP4 serum protein as measured using the DPP4 immunoassay described in Example 2 or a DPP4 detection assay including one of the DPP4 assays disclosed in Table 1. including. These thresholds can also be measured, for example, about 23.5 ng / mL for POSTN serum proteins as measured using the POSTN immunoassay described in WO2015015185 or any POSTN assay known in the art. Or about 25.8 ng / mL.

  As used herein with respect to DPP4, POSTN, and other biomarkers disclosed herein, the terms “reduced”, “reduced” or “lower” apply when applied to a biomarker level. , Refers to the expression level or range of a biomarker measured in a control sample (“normal level”), or a level in a biological sample (eg, serum or sputum) that is below a particular threshold disclosed herein. These thresholds are, for example, about 363 ng / mL or about 376 ng / mL for DPP4 serum protein as measured using a DPP4 immunoassay described in Example 2 or a DPP4 detection assay including one of the DPP4 immunoassays disclosed in Table 1. including. These thresholds can also be measured, for example, about 23.5 ng / mL for POSTN serum proteins as measured using the POSTN immunoassay described in WO2015015185 or any POSTN assay known in the art. Or about 25.8 ng / mL.

  Normal levels or ranges in DPP4, POSTN, and other biomarkers disclosed herein can be defined according to standard conventions. Thus, the level measured in a particular biological sample can be compared to the level or range of levels measured in a similar normal sample. In this context, a normal sample or a control sample is a sample obtained from an individual having undetectable symptoms of, for example, an eosinophilic disease or disorder (eg, a pulmonary disease or disorder such as asthma). Thus, for example, a level of DPP4 or POSTN is when the level of DPP4 or POSTN is present in a test sample in a normal sample, in a control sample, or at a level or range lower than a particular threshold level disclosed herein. , Low level, lower level, reduced level, reduced level, or grammatical variants thereof.

  Conversely, for example, the level of DPP4 or POSTN is present in a level or range in which the level of DPP4 or POSTN is higher in the test sample than in a normal sample, in a control sample, or a particular threshold level disclosed herein. Sometimes it is said to be a high level, a higher level, an increased level, an elevated level, or a grammatical variant thereof. These thresholds are for example about 363 ng / mL or about 376 ng / mL for serum DPP4 protein as measured using a DDP4 immunoassay described in Example 2 or a DDP4 immunoassay comprising one of the DPP4 detection assays disclosed in Table 2. including. These thresholds can also be measured, for example, about 23.5 ng / mL for POSTN serum proteins as measured using the POSTN immunoassay described in WO2015015185 or any POSTN assay known in the art. Or about 25.8 ng / mL.

  In some aspects, the level of a molecular biomarker disclosed herein (eg, DPP4 or POSTN) is at least about 5% above a normal or control sample, or a particular threshold level disclosed herein. 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90 %, 95%, 100%, 105%, 110%, 115%, 120%, 125%, 130%, 135%, 140%, 145%, 150%, 155%, 160%, 165%, 170%, 175%, 180%, 185%, 190%, 195%, 200%, 205%, 210%, 215%, 220%, 225%, 230%, 235%, 240%, 245%, 250%, 255% 260%, 2 5%, 270%, 275%, 280%, 285%, 290%, 295%, 300%, 305%, 310%, 315%, 320%, 325%, 330%, 335%, 340%, 345% 350%, 355%, 360%, 365%, 370%, 375%, 380%, 385%, 390%, 395%, 400%, 405%, 410%, 415%, 420%, 425%, 420 %, 435%, 440%, 445%, 450%, 455%, 460%, 465%, 470%, 475%, 480%, 485%, 490%, 495%, or 500% higher? Or considered expensive.

  In some aspects, the level of a molecular biomarker disclosed herein (eg, DPP4 or POSTN) is at least about 5% above a normal or control sample, or a particular threshold level disclosed herein. 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90 %, 95%, 100%, 105%, 110%, 115%, 120%, 125%, 130%, 135%, 140%, 145%, 150%, 155%, 160%, 165%, 170%, 175%, 180%, 185%, 190%, 195%, 200%, 205%, 210%, 215%, 220%, 225%, 230%, 235%, 240%, 245%, 250%, 255% 260%, 2 5%, 270%, 275%, 280%, 285%, 290%, 295%, 300%, 305%, 310%, 315%, 320%, 325%, 330%, 335%, 340%, 345% 350%, 355%, 360%, 365%, 370%, 375%, 380%, 385%, 390%, 395%, 400%, 405%, 410%, 415%, 420%, 425%, 420 %, 435%, 440%, 445%, 450%, 455%, 460%, 465%, 470%, 475%, 480%, 485%, 490%, 495%, or 500% lower or lower? Or considered low.

  As used herein, the term “threshold level” (or alternatively “threshold” or “predetermined threshold level” herein) refers to DPP4, POSTN, or Refers to the level of any other biomarker disclosed in. In some aspects, the threshold level is a protein or nucleic acid expression level expressed as an average of the level of protein or nucleic acid expression level from a sample taken from a control population of healthy (non-disease) subjects. Also good.

  In some aspects, the threshold level may be a different time in the same subject, such as a level before the assay, such as a level measured before the subject develops a disease or before the start of treatment. In general, samples are normalized by a common factor. For example, body fluid samples are normalized by body fluid volume, and cell-containing samples are normalized by protein content or cell number. In another aspect, the threshold level is also the same in the corresponding control sample or control group of subjects who do not respond to treatment with an anti-IL-5R antibody such as, for example, an eosinophil targeted therapeutic, eg, benralizumab (MED-563). It may refer to the level of biomarker expression.

  In some aspects, the expression level of DPP4, POSTN, or any other biomarker disclosed herein is compared to a threshold level (or alternatively “predetermined threshold level” herein). Is done. Thus, as used herein, the term “threshold level” or “predetermined threshold level” is a cutoff or threshold when the measured expression level of a protein or nucleic acid is compared.

  Based on a comparison with a known control sample, the “threshold level” in DPP4 or any other biomarker disclosed herein can be determined and is a test sample that falls above or below the threshold level of the biomarker Indicates that the patient from whom the sample is derived may or may not benefit from treatment with an eosinophil-targeted therapeutic agent, eg, an anti-IL-5R antibody such as benralizumab (MED-563).

  For example, if the DPP4 or POSTN level exceeds its threshold level in the sample, the patient does not benefit from treatment with an eosinophil-targeted therapeutic agent, for example, an anti-IL-5R antibody such as benralizumab (MEDIA-563) It is shown that there is. Conversely, for samples where DPP4 or POSTN levels are below that threshold level, the patient may benefit from treatment with an eosinophil-targeted therapeutic agent, such as an anti-IL-5R antibody such as benralizumab (MED-563). It is shown that there is.

  In some aspects, the threshold level (eg, protein expression level or gene expression level) in DPP4, POSTN, or any other biomarker disclosed herein is predetermined and the sample type (eg, serum , Lung tissue, sputum), the type of eosinophilic disease or disorder (eg, asthma, IPF, COPD) and optionally the assay used.

  Serum samples from a population of mild to moderate asthma patients quantified using the immunoassay described in Example 2, one of the DPP4 detection immunoassays disclosed in Table 1, or the multiplex immunoassay disclosed above Lower than 376 ng / mL indicates that patients may have an increased clinical response to anti-IL-5R antibodies such as eosinophil targeted therapeutics such as benralizumab (MED-563) Show. Thus, in some aspects of the methods disclosed herein, the predetermined DPP4 threshold level is about 376 ng / mL of expressed DPP4 protein in serum as measured using the immunoassay described in Example 2. .

  Derived from a population of mild to moderate asthma patients quantified using the immunoassay described in WO20150120185, or any POSTN detection known in the art, or the multiplex immunoassay disclosed above A POSTN level from a serum sample below 25.8 ng / mL indicates that the patient has an increased clinical response to an anti-IL-5R antibody such as eosinophil targeted therapeutics such as benralizumab (MEDI-563). Show that you can. Thus, in some aspects of the methods disclosed herein, the predetermined POSTN threshold level is about 25. 5 for the expressed POSTN protein in serum as measured using the immunoassay described in WO2015015185. 8 ng / mL.

  In some other aspects, the predetermined DPP4 threshold level is determined by eosinophilic disease or disorder (eg, asthma), for example as measured according to either the DPP4 immunoassay described in Example 2 or the DPP4 immunoassay disclosed in Table 1. Based on median biomarker levels in serum measured from multiple patients with lung disease or disorder).

  Thus, in some aspects, the predetermined threshold level of DPP4 is determined by measuring an eosinophilic disease or disorder (eg, Serum median DPP4 values measured from multiple patients with pulmonary diseases or disorders such as asthma).

  In some aspects, the predetermined DPP4 threshold level is about 376 ng / mL +/− 25 pg / mL of expressed DPP4 protein in serum as measured using the DPP4 immunoassay described in Example 2.

  In some aspects, the DPP4 threshold level can be from about 100 to about 870 ng / mL. Accordingly, the DPP4 threshold level is approximately 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550, 560, 570, 580, 590, 600, 610, 620, 630, 640, 650, 660, 670, 680, 690, 700, 710, 720, 730, 740, 750, 760, 770, 780, 790, 800, 810, 820, 830, 840, 850, 860, or 870n / May be mL.

  In some aspects, a “low level DPP4” is defined as a value below one of these threshold levels, while a “high level DPP4” is defined as a value greater than or equal to the same threshold level (ie, , If the threshold level is 250 ng / mL, the low level DPP4 will be less than 250 ng / mL and the high level DPP4 will be greater than 250 ng / mL).

  In some aspects, the predetermined DPP4 threshold level is a first, second, third, fourth, second, expression of DPP4 in the serum of a control patient as measured using the DPP4 immunoassay described in Example 2. Corresponds to the fifth, sixth, seventh, eighth, ninth or tenth decile DPP4 baseline level. In some aspects, the predetermined DPP4 threshold level is the first, second, third of expression of DPP4 in the serum of mild to moderate asthma patients as measured using the DPP4 immunoassay described in Example 2. , 4th, 5th, 6th, 7th, 8th, 9th or 10th decile corresponding to the DPP4 baseline level. In some aspects, the predetermined DPP4 threshold level is the first, second, third, fourth, fourth, fourth, fourth, fourth, and fourth levels of DPP4 expression in the serum of patients with severe asthma as measured using the DPP4 immunoassay described in Example 2. Corresponds to the fifth, sixth, seventh, eighth, ninth or tenth decile DPP4 baseline level.

  In some aspects, a “lower or decreased DPP4 level” is defined as a value below one of these threshold levels, while a “higher or increased DPP4 level” is a value greater than or equal to the same threshold level. Is defined.

  In some other aspects, the predetermined POSTN threshold level is an eosinophilic disease or disorder as measured according to the POSTN immunoassay described in, for example, WO2015015185, or any POSTN detection known in the art. Based on median biomarker levels in serum measured from multiple patients (eg, pulmonary diseases or disorders such as asthma).

  Thus, in some aspects, the predetermined threshold level of POSTN is determined by eosinophilic disease or as measured using the POSTN immunoassay described in WO2015015185, or any POSTN detection known in the art. Serum median POSTN value in serum measured from multiple patients with a disorder (eg, pulmonary disease or disorder such as asthma).

  In some aspects, the predetermined POSTN threshold level is expressed in the POSTN protein expressed in serum as measured using the POSTN immunoassay described in WO2015015185, or any POSTN detection known in the art. About 26 ng / mL +/− 250 pg / mL.

  In some aspects, the POSTN threshold level is about 7-105 ng / mL. Accordingly, the POSTN threshold level is approximately 7, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100 or 105 ng / Can be mL.

  In some aspects, “low level POSTN” is defined as a value below one of these threshold levels, while “high level POSTN” is defined as a value greater than or equal to the same threshold level (ie, , If the threshold level is 250 ng / mL, the low level POSTN will be less than 250 ng / mL and the high level POSTN will be greater than 250 ng / mL).

  In some aspects, the predetermined POSTN threshold level is a level of POSTN in the serum of a control patient, as measured using the POSTN immunoassay described in WO20151201585, or any POSTN detection known in the art. Corresponds to the POSTN baseline level of the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth or tenth quartile of expression. In some aspects, the predetermined POSTN threshold level is the first, second, and second expression of POSTN in the serum of mild to moderate asthma patients as measured using the POSTN immunoassay described in WO20150151855. , 3rd, 4th, 5th, 6th, 7th, 8th, 9th or 10th percentile corresponding to the POSTN baseline level. In some aspects, the predetermined POSTN threshold level is a first, second, third, third, or third expression of POSTN expression in the sera of patients with severe asthma as measured using the POSTN immunoassay described in WO2015015185. Corresponds to the fourth, fifth, sixth, seventh, eighth, ninth, or tenth percentile POSTN baseline level.

  In some aspects, a “lower or decreased POSTN level” is defined as a value below one of these threshold levels, while a “higher or increased POSTN level” is a value greater than or equal to the same threshold level. Is defined.

  In some aspects, the predetermined DPP4 or POSTN threshold level corresponds to, for example, the assay used for DPP4, the LOD of the immunoassay described in Example 2, or the LOD of any of the DPP4 assays in Table 2. In some aspects, the DPP4 or POSTN detection assay may be adapted for use as a LOD-based method by diluting a sample such that a new DPP4 or POSTN threshold level corresponds to the LOD. For example, if the initial predetermined DPP4 threshold level in an undiluted sample is 350 ng / mL and the LPP of the DPP4 immunoassay is 50 ng / mL, the patient sample is 1: 7 using a suitable buffer. Can be diluted to (ratio between DPP4 threshold and LOD). After such dilution, only samples having a DPP4 level above a predetermined DPP4 threshold level in an undiluted sample will have a DPP4 level above the LOD of the assay.

  In some aspects, the threshold level is determined by the manufacturer of a commercial immunoassay used to detect the presence or absence of DPP4, POSTN, or other biomarkers that can be combined with DPP4 and / or POSTN in a sample. As reported in the instructions for use, the average or mean expression level measured from a sample obtained from a healthy volunteer.

  In some aspects, the expression level of a molecular biomarker disclosed herein (eg, DPP4 or POSTN) as measured in a sample is above or below a threshold level or threshold at each biomarker. In these embodiments where the expression level of each biomarker disclosed herein (eg, DPP4 or POSTN) measured in the sample is above or below a threshold level, the expression level is determined from the sample source Patients (eg, mild to moderate asthma patients or severe asthma patients) would benefit from treatment with an eosinophil-targeted therapeutic agent, eg, an anti-IL-5R antibody such as benralizumab (MED-563) or It may indicate that it may not be possible. The degree to which the measured expression level is above or below the threshold level or threshold may range to any degree.

  In exemplary aspects, the measured expression level of a biomarker disclosed herein (eg, DPP4 or POSTN) is at least or approximately 10% higher or lower than the threshold level, eg, at least above the threshold level. Or about 15% higher or lower, at least or about 20% higher or lower than the threshold level, at least or about 25% higher or lower than the threshold level, and at least or about 30% higher or lower than the threshold level , At least or about 35% higher or lower than the threshold level, at least or about 40% higher or lower than the threshold level, at least or about 45% higher or lower than the threshold level, at least or about lower than the threshold level 50% higher Or at least or about 55% higher or lower than the threshold level, at least or about 60% higher or lower than the threshold level, at least or about 65% higher or lower than the threshold level, and higher than the threshold level At least or approximately 70% higher or lower, at least or approximately 75% higher or lower than the threshold level, at least or approximately 80% higher or lower than the threshold level, and at least or approximately 85% higher than the threshold level or Low, at least or about 90% higher or lower than the threshold level, and at least or about 95% higher or lower than the threshold level.

  In an exemplary aspect, the measured expression level of a biomarker disclosed herein (eg, DPP4 or POSTN) is at least 2-fold higher or lower than the threshold level and at least 3-fold higher than the threshold level Or low, at least 4 times higher or lower than the threshold level, at least 5 times higher or lower than the threshold level, at least 6 times higher or lower than the threshold level, and at least 7 times higher or lower than the threshold level , At least 8 times higher or lower than the threshold level, at least 9 times higher or lower than the threshold level, or at least 10 times higher or lower than the threshold level.

  As discussed above, the level of a biomarker disclosed herein (eg, DPP4 or POSTN) can be measured using methods known in the art. Those skilled in the art, in addition to the assays disclosed above, can use a large number of methods available in the art that allow those skilled in the art to measure threshold levels as described throughout this section, for example, It will be appreciated that there are DPP4 detection methods and immunoassays described in Example 2 and Table 1.

  In some aspects, the predetermined threshold level of a biomarker disclosed herein (eg, DPP4 or POSTN) is a population of subjects (eg, a plurality of normal healthy volunteers, or an eosinophilic disease or disorder, Defined in terms of specific percentile values within a plurality of patients (eg, patients with pulmonary diseases or disorders such as asthma). In some aspects, the plurality of patients having a pulmonary disease or disorder is a plurality of patients having mild to moderate asthma, or a plurality of patients having severe asthma.

  In some aspects, the predetermined threshold level in DPP4 or POSTN is a DPP4 or POSTN protein in a plurality of normal healthy volunteers or a plurality of patients having a pulmonary disease or disorder such as eosinophilic disease or disorder, eg, asthma 10th, 15th, 20th, 25th, 30th, 35th, 40th, 45th, 50th, 50th, 55th, 60th, 65th, 70th, 75th, This corresponds to the 80th, 85th, or 90th percentile. In some aspects, the plurality of patients having a pulmonary disease or disorder is a plurality of patients having mild to moderate asthma, or a plurality of patients having severe asthma.

  The threshold level (eg, protein expression level or gene expression level) of a biomarker disclosed herein (eg, DPP4 or POSTN) is determined by the nature of the assay, eg, capture and detection antibodies used, source, purity, and It may vary based on the composition of the standard substance. In one aspect, any threshold level for determining whether a patient can benefit from treatment with an eosinophil-targeted therapeutic agent, eg, an anti-IL-5R antibody such as benralizumab (MEDI-563). Instead, the patient's DPP4 and / or POSTN levels can be compared to one or more control levels. According to this embodiment, a test sample (eg, a sample from a patient suffering from a pulmonary disease or disorder such as asthma) is taken from one or more control samples, eg, a sample taken from a normal healthy person, the same patient. An earlier sample, a sample taken from a patient with a subset of the patient's disease (eg, asthma or COPD), a predetermined standard amount of an isolated biomarker protein or gene or fragment thereof, or a combination thereof The

  The results can be expressed as a ratio to the control sample to determine a percent increase or decrease in the patient's biomarker level (eg, protein expression level or gene expression level) relative to the control biomarker level. The control sample is a patient sample, for example whole blood if the patient sample is whole blood, serum if the patient sample is serum, plasma if the patient sample is plasma, and if the patient sample is saliva Saliva, urine if patient sample is urine, sputum if patient sample is sputum, bronchoalveolar lavage fluid if patient sample is bronchoalveolar lavage fluid, patient sample is lung tissue In some cases, it can be a matched pair with one or more of the lung tissues. Once determined, the biomarker expression level can be recorded in the patient's medical record.

  In some aspects, as disclosed above, DPP4 and / or POSTN levels can be combined with other biomarkers such as eosinophil levels. Thus, in some aspects, high levels of at least about 150 cells / μL, at least about 300 cells / μL, or at least about 400 cells / μL combined with DPP4 and / or POSTN levels below the threshold levels disclosed above Eosinophils (eg peripheral blood eosinophils) are eosinophilic targeted therapeutics in patients with eosinophilic diseases or disorders (eg pulmonary diseases or disorders such as asthma) such as benralizumab (MED-563) Is predictive for positive clinical response to anti-IL-5R antibodies such as

  In some aspects, a blood analyte for use as a biomarker, eg, eosinophil level, can be determined from a complete blood count (CBC) with differentiation. The term “CBC with differentiation” as used herein refers to complete blood count (CBC) with white blood cell (WBC) differentiation. The term “leukocyte” includes, for example, neutrophils, lymphocytes, monocytes, eosinophils, and basophils. The terms “eosinophil” and “eosinophils” can be abbreviated as “EOS”. The term “white blood cell count” as used herein refers to the number of white blood cells from any sample, eg, complete blood count (CBC) with white blood cell (WBC) differentiation (CBC with differentiation). Obtaining CBC with differentiation can be accomplished using any suitable technique available in the art, eg, by automated hematology analyzer or blood coulter counter (eg, flow cytometry), or by hand-manipulating cells. It can be achieved by counting at work (eg using a microscope). CBC with differentiation is one of the most widely ordered clinical laboratory tests worldwide.

  In some aspects, the sample may be analyzed using an automated hematology analyzer, such as SIEMENS ADVIA 120; ABBOTT CELL DYN 3500; BECKMAN COULTER LH750; SYSMEX X SERIES; In some aspects, in order to maximize the accuracy of the disclosed method, the readout information from the automated hematology analyzer may have an absolute eosinophil count of at least two orders of magnitude (eg, 150, 220, 340 cells). / ΜL) and at least 3 orders of magnitude for lymphocytes and neutrophils (eg, 1,530, 2,340, 3,410 cells / μL). In some aspects, before analyzing the blood sample, the tube is several times, eg, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more than 10 times, or manufacturer's instructions It needs to be reversed according to the book. In some aspects, the sample can be manually analyzed.

  In some embodiments, eosinophil levels can be measured by flow cytometry. In some aspects, the disclosed method comprises WBC, eosinophil count, neutrophil count, lymphocyte count, eosinophil precursor count, basophil precursor count, eotaxin-2 level, and their Any combination or ratio may be included. In a further aspect, a method or system of the present disclosure may include a log of blood eosinophil / WBC ratio, blood eosinophil / blood lymphocyte ratio, and blood eosinophil / blood neutrophil ratio.

The methods or systems of the present disclosure can be any one or any of the following non-limiting examples of physiological biomarkers: post-albuterol ΔFEV1, tiotropium bromide ΔFEV1, FEV1, FEV / FVC, ΔAM / PM PEF variation, and FE _NO May be included. These biomarkers are known in the art and can be measured according to standard medical protocols. The disclosed method or system also provides biomarkers of patient symptoms such as, but not limited to, ACQ score, AQLQ score, Berlin questionnaire (examination of sleep apnea), Borg score (evaluation of dyspnea), previous Any one or any combination of sinus surgery, atopy history, intubation history, aspirin sensitivity history, history of corticosteroid burst for the past 3 or 12 months, or history of ER visits for the past 3 years Can be included. A method or system of the present disclosure may also include any one or any combination of the following parameters: gender, age, weight, race, height, or body mass index (BMI).

  In some aspects, a method or system of the present disclosure may include a value corresponding to the average of several measurements from multiple samples collected at different time intervals. Thus, in some aspects, multiple samples are collected at different intervals, eg, about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, or about 7 days. obtain. In some aspects, the plurality of samples is about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about 10 weeks. , About 11 weeks, or about 12 weeks apart. In some embodiments, the plurality of samples is about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months , About 9 months, about 10 months, about 11 months, or about 12 months. In some aspects, multiple samples may be collected at intervals greater than 12 months. Optionally, more than two samples, eg, 3 samples, 4 samples, 5 samples, 6 samples, 7 samples, 8 samples, 9 samples, 10 samples, or more are averaged. In some aspects, samples are collected at regular intervals. In other embodiments, samples are not collected at regular intervals. Optionally, the sample is collected in response to an event, such as an exacerbation of symptoms.

In some aspects, administration of an eosinophil targeted therapeutic agent according to the methods disclosed herein, eg, an anti-IL-5R antibody such as benralizumab (MEDI-563) comprises:
(A) decrease in AER (acute exacerbation rate),
(B) Increase in FEV1 (forced expiratory volume per second),
(C) Results of improved ACQ-6 (Asthma Management Questionnaire, 6 item version),
(D) As a result of improved AQLQ (Asthma QOL Questionnaire),
(E) decreased cytokine secretion by type 2 innate lymphocytes (ILC2),
(F) reduced ILC2 levels;
(G) resulting in reduced levels of eosinophils and / or basophils, or (h) resulting in combinations thereof.

  In some aspects, administration of an eosinophil-targeted therapeutic agent, eg, anti-IL-5R antibody such as benralizumab (MEDI-563), according to the methods disclosed herein is associated with health known in the art. Bring improvements in other methods of detecting quality of life.

IV. Eosinophil targeted therapeutics in combination with other clinical or molecular biomarkers disclosed herein (eg, blood eosinophil levels) and / or with other clinical or molecular biomarkers known in the art The combined DPP4 and / or POSTN can e.g. eosinophilic diseases or disorders using anti-IL-5R antibodies such as eosinophilic targeted therapeutics, e.g. benralizumab (MEDI-563) or lung diseases such as asthma or For patients suffering from (disability), it can be used to determine whether to treat, select for treatment, monitor treatment, or initiate, improve or end treatment.

The term “eosinophil targeted therapeutic agent” as used herein refers to a therapeutic agent capable of reducing the number of eosinophils in a patient in need thereof. In some aspects, eosinophil targeted therapeutics can also drastically reduce basophil count. In certain embodiments, the eosinophil targeted therapeutic is capable of reducing eosinophil counts and / or basophil counts in patients in need thereof (i) a monoclonal antibody, or (ii) an antigen thereof A binding fragment, or (iii) a monoclonal antibody comprising one or more additional therapeutic moieties, or a compound comprising antigen binding thereof, or (iv) a combination thereof.

  In other embodiments, the eosinophil targeted therapeutic agent can be a small molecule. In some aspects, the eosinophil targeted therapeutic agent is a combination of eosinophil targeted therapeutic agents (eg, at least one antibody or fragment thereof and at least one small molecule, or at least antibody or fragment thereof and At least one nucleic acid, or a combination therapy comprising at least one nucleic acid and at least one small molecule).

  In another aspect, the therapeutic agent is a biological agent. In certain embodiments, a biological agent is any substance made by an organism or product thereof, a substance made using recombinant DNA technology, nucleotides made by any means, nucleotide analogs, oligonucleotides, oligonucleotide analogs Body, peptide, or peptide analog. In a specific embodiment, the biological agent is an antibody or antibody fragment, antibody mimetic, soluble receptor polypeptide, soluble receptor fusion polypeptide, interleukin, interleukin fusion polypeptide, antisense molecule, siRNA or miRNA. obtain.

  In some aspects, an eosinophil targeted therapeutic of the present disclosure (eg, an antibody that specifically binds to IL-5R or IL-5) is specific for a reference molecule (eg, IL-5R or IL-5). Competitively binding to a given target site to the extent that it partially blocks the binding of the reference molecule to the target site. Inhibit. Competitive inhibition can be determined by any method known in the art, such as a competitive ELISA assay.

  Eosinophil targeted therapeutics of the present disclosure (eg, antibodies that specifically bind to IL-5R or IL-5) have at least about 90%, at least about 85% binding of a reference molecule to a given epitope. , At least about 80%, at least about 75%, at least about 70%, at least about 65%, at least about 60%, at least about 55%, or at least 50%.

In some aspects, the eosinophil targeted therapeutic agent is
(I) is capable of inhibiting or reducing the level of IL-5 or IL-5R;
(Ii) is capable of inhibiting or reducing expression of molecules that interact with IL-5 or IL-5R;
(Iii) is capable of increasing the expression of molecules that interact with IL-5 or IL-5R,
(Iv) is capable of inhibiting or reducing the expression of molecules upregulated by IL-5 or IL-5R;
(V) has the ability to inhibit or inhibit the expression of molecules that interact with molecules up-regulated by IL-5 or IL-5R, or (vi) is a therapeutic agent that combines them.

  In some aspects, the eosinophil targeted therapeutic is a compound included in Table 2.

  Eosinophil targeted therapeutics, including the antibodies disclosed above, are disclosed on intact immunoglobulins as well as antigen binding fragments, variants or derivatives thereof, antibodies or fragments thereof that bind to the same epitope. Or antibodies or fragments thereof that competitively inhibit the binding of the antibodies disclosed above to their respective targets. Consider antibodies (or fragments thereof) that are identical or similar in amino acid sequence to the antibodies disclosed in Table 2, particularly within the variable region or within its CDRs (but mutations within the constant region are also contemplated) Is done. For example, in one aspect, the eosinophil targeted therapeutic agent is about 70%, about 75% relative to the polypeptide sequence of the antibodies disclosed in Table 2 (ie, antibody heavy and / or light chain). A polypeptide having an amino acid sequence that is about 80%, about 85%, about 90%, about 92%, about 95%, about 98%, about 99% or 100% identical.

In some aspects, an eosinophil targeted therapeutic agent is disclosed above, comprising at least one, two, three, four, or six complementarity determining regions of the antibodies disclosed above. An isolated antigen binding protein that targets any of the antigens. In various aspects, the eosinophil targeted therapeutic disclosed herein comprises an antigen-binding fragment of an antibody, eg, any fragment of an antagonist antibody referred to herein, eg, IL-5. (SEQ ID NO: 9) or an antibody that specifically binds to IL-5R, such as the alpha subunit of IL-5R (SEQ ID NOs: 4-8, corresponding to the five isoforms of subunits made by alternative splicing) including. Such fragments include, but are not limited to, Fab, Fab ′, Fab′-SH, Fv, scFv, F (ab ′) ₂ , Nanobodies, and Diabodies.

  In some aspects, the eosinophil targeted therapeutic agent is a therapeutic agent capable of specifically binding to IL-5R, eg, an anti-IL-5R antibody such as benralizumab or an antigen-binding fragment thereof, or benralizumab or an antigen thereof. Anti-IL-5R molecule containing binding fragment. In some aspects, the eosinophil targeted therapeutic agent is a therapeutic agent capable of specifically binding to IL-5, for example, an anti-IL-5 antibody such as reslizumab, mepolizumab, or an antigen-binding fragment thereof, or reslizumab , Mepolizumab, or an antigen-binding fragment thereof, an anti-IL-5 molecule. In some aspects, the anti-IL-5R molecule or anti-IL-5 molecule is an antibody-drug complex (ADC).

  In one aspect, the eosinophil targeted therapeutic can be an anti-cytokine antibody or an anti-cytokine receptor antibody that binds to a cytokine other than IL-5 or IL-5R and its receptor. In one aspect, the eosinophil targeted therapeutic can be an anti-IL-5 or anti-IL5R antibody capable of blocking IL-5-mediated signal transduction through IL-5 receptor.

  In one aspect, the anti-IL5R antibody specifically binds to the α subunit of IL-5R (SEQ ID NO: 4). Non-limiting examples of anti-human IL-5 antibodies include reslizumab and mepolizumab. Non-limiting examples of anti-human IL-5 receptor α antibodies of the present disclosure include US Pat. No. 7,179,464, US Pat. No. 6,538,111, US Pat. No. 6,018,032. And US Patent Application Publication No. 2004 / 0136996A1, US Patent Application Publication No. 2005 / 0226867A1, or International Publication No. WO 2008/143878. In one aspect, the eosinophil-targeted therapeutic agent can be an antibody specific for IL-5, for example, reslizumab, mepolizumab, and any combination thereof.

  Unless bound by a particular theory, eosinophil targeted therapeutics used according to the methods and systems described herein are anti-IL-5R antibodies capable of mediating eosinophil depletion in vivo. obtain. In one aspect, depletion in vivo can be mediated by ADCC, CDC or antibody-mediated phagocytosis. In a specific aspect, the therapeutic agent can be an anti-IL-5R antibody having ADCC activity.

  In another specific aspect, the therapeutic agent can be an anti-IL-5R antibody with enhanced ADCC activity. A non-limiting example of an anti-IL-5R antibody with enhanced ADCC activity is also referred to herein as “medi-563” as described in International Publication No. WO 2008/143878. ) Benralizumab. Benralizumab is a disulfide, dimer (230-230 ″: 233-233 ″)-bis with humanized mouse monoclonal MEDI-523γ1 heavy chain (224-214 ′)-humanized mouse monoclonal MEDI-523κ light chain. It is an immunoglobulin G1 antibody containing a disulfide. See U.S. Pat. No. 8,501,176, U.S. Patent Application Publication No. 20150044204, and International Publication No. WO2015502504, all of which are hereby incorporated by reference in their entirety. .

  In another specific embodiment, the therapeutic agent can be an anti-IL-5R antibody that binds to the same epitope as venralizumab or competes with venralizumab for binding to IL-5R. Benralizumab epitopes are described in International Publication No. WO 2008/143878, the disclosure of which is hereby incorporated by reference in its entirety. In one aspect, the anti-IL-5R antibody or antigen-binding fragment comprises SEQ ID NO: 12, consists of SEQ ID NO: 12, or comprises a heavy chain variable region (VH) consisting essentially of SEQ ID NO: 12. In one aspect, the anti-IL-5R antibody or antigen-binding fragment comprises a light chain variable region (VL) comprising SEQ ID NO: 13, consisting of SEQ ID NO: 13, or consisting essentially of SEQ ID NO: 13. In some aspects, the anti-IL-5R antibody or antigen-binding fragment comprises at least one, two, three, four, five or six complementarity determining regions selected from SEQ ID NOs: 14-19. .

  In some aspects, the eosinophil targeted therapeutic agent is a fusion protein or complex comprising one of the monoclonal antibodies in Table 2 or an antigen binding fragment thereof. In some aspects, the eosinophil targeted therapeutic agent is a fusion protein or complex comprising one of the monoclonal antibodies or antigen-binding fragments thereof according to any one of claims 12-18. In some aspects, the fusion protein or complex comprises at least one heterologous therapeutic moiety and / or a half-life enhancing moiety. In some aspects, the term eosinophil targeted therapeutic also includes, for example, an antagonist of IL-5 or IL-5R, such as an aptamer, peptide, mRNA, iRNA, shRNA, or small molecule inhibitor. To do.

  In some aspects, the eosinophil targeted therapeutic is a polynucleotide, such as DNA or RNA. In some aspects, the polynucleotide is (i) mRNA or a combination thereof, or (ii) an antisense oligonucleotide or a combination thereof. In some aspects, the polynucleotide comprises at least a nucleotide analog. In some aspects, the antisense oligonucleotide or combination thereof is an oligonucleotide that targets a gene encoding the β subunit of IL-5R, eg, ASM8, PXS1100, or PXS2200. See, for example, International Publication No. WO 20066042202 (incorporated herein by reference in its entirety). ASM8 is a combination of two oligonucleotides (TOP004 and TOP005). TOP004 is a 19-mer specific for β subunit mRNA common to IL-3, IL-5, and GM-CSF receptors.

  In one aspect, the eosinophil targeted therapeutic agent can be an antibody specific for IL-13 / IL-4α. In specific embodiments, the therapeutic agent can be Aerovant ™ (Aerovance), GSK-679586 (GSK), IMA-026 (Wyeth), or MILR1444A (Genentech).

  In a further aspect, the eosinophil targeted therapeutic is an antibody specific for the IL-2 receptor. In a specific embodiment, the eosinophil targeting agent can be daclizumab (ZENAPAX®). Daclizumab is a therapeutic humanized monoclonal antibody to the α subunit of the IL-2 receptor of T cells. In another aspect, the eosinophil targeted therapeutic agent can be an anti-IgE antibody. In a specific embodiment, the eosinophil targeted therapeutic can be omalizumab (XOLAIR®). Omalizumab is a recombinant DNA-derived humanized IgG1k monoclonal antibody that selectively binds to human immunoglobulin E (IgE). Omalizumab is approved by the FDA to treat patients with moderate to severe allergic asthma. Some studies have shown that it reduces airway eosinophil count, but it has not been specifically approved for the treatment of eosinophilic asthma. In another aspect, the eosinophil targeted therapeutic agent can be a recombinantly produced cytokine. In a specific embodiment, the eosinophil targeted therapeutic agent can be interferon alpha. Non-limiting examples of interferon α therapeutics include PEGASYS® (peginterferon α-2a) and ALBUFFERON® / ZALBIN® (Albuinterferon α-2b). The eosinophil targeted therapeutics of the present disclosure are also not limited to IL-13, IL-17A, IL-17F, IL-25, IL-33, TNF-α, IL-1β, IL-6, Or it can be used in combination with one or more antagonists (eg, antibodies) of other cytokines associated with inflammation, including TGF-β.

  In certain embodiments, the half-life of antibodies used in accordance with the disclosed methods and systems can be extended by methods known in the art. Such an extension can then reduce the amount and / or frequency of administration of the antibody composition. Antibodies with improved in vivo half-life and methods for preparing them are described in US Pat. No. 6,277,375; and WO 98/23289 and WO 97/3461. No. pamphlet.

V. DPP4 and POSTN Levels in Diagnosis and Treatment of Eosinophilic Disease DPP4 and POSTN are variously expressed in subjects with eosinophilic diseases or disorders, eg pulmonary diseases or disorders such as asthma. DPP4 and POSTN are present at different levels in samples from patients with mild to moderate asthma relative to healthy controls, or patients with severe asthma relative to healthy controls.

  For example, differences in DPP4 and POSTN levels observed in healthy individuals, patients with mild to moderate asthma, and patients with severe asthma may indicate that the patient has a particular treatment, such as eosinophil targeted therapeutics such as benralizumab ( It can be applied to predict clinical outcome when treated with anti-IL-5R antibodies such as MEDI-563). Thus, if a subject's DPP4 and / or POSTN levels exceed a certain threshold, the subject will receive a specific treatment, eg, an eosinophil-targeted therapeutic agent, eg, an anti-IL-5R antibody such as benralizumab (MEDI-563). Candidates for treatment with the treatment used.

  In some aspects, treatment with a particular treatment is simply the measurement that DPP4 and / or POSTN is expressed below a predetermined threshold level (eg, using a diluted sample) or below a detectable level. For example, it would be sufficient to identify a subject as a candidate for treatment with an anti-IL-5R antibody such as an eosinophil targeted therapeutic agent, eg, benralizumab (MED-563).

  In some aspects of the methods disclosed herein, the patient is moderate asthma or a mild to moderate asthmatic patient (GINA1-3). In another aspect of the methods disclosed herein, the patient is a severe asthmatic patient (GINA4 +). In other embodiments, the patient suffers from COPD.

  In some aspects of the methods disclosed herein, the DPP4 level or POSTN level can be used alone. In other aspects, DPP4 and / or POSTN levels can be combined with other molecules or clinical biomarkers such as blood eosinophil counts. In other aspects, additional biomarkers, such as other biomarkers indicative of inflammation or clinical biomarkers (eg, age, smoking status, body mass index, etc.) can be combined with DPP4 and / or POSTN.

  This finding can be found, for example, in new ways to determine treatment (eg, by selecting patients as specific treatment candidates), eosinophilic diseases or disorders (eg, mild to moderate asthma, or severe asthma). Pulmonary diseases or disorders), methods for monitoring the effectiveness of therapeutic agents (eg, benralizumab / MEDI-563) for treating eosinophilic diseases or disorders, or formulations, dosing schedules, or administrations It can be applied to devise a method for adjusting the pathway.

  The methods disclosed herein are based at least in part on the expression level of DPP4 and / or POSTN in a subject, alone or in combination with one or more additional biomarkers (eg, eosinophil count). Prescribing, initiating and / or altering prevention and / or treatment, eg, for eosinophilic diseases or disorders (eg, pulmonary diseases or disorders such as mild to moderate asthma or severe asthma).

  The present disclosure relates to patients with eosinophilic diseases or disorders (eg pulmonary diseases or disorders such as mild to moderate asthma or severe asthma) such as eosinophilic targeted therapeutics such as benralizumab (MEDIA-563). A method for determining whether to be treated with a treatment regimen comprising administration of an anti-IL-5R antibody, comprising: (a) DPP4 and / or POSTN levels, optionally blood eosinophils in a sample taken from a patient Measuring a level of an additional biomarker such as a sphere or instructing a clinical laboratory to measure, and (b) instructing a health care provider to treat or treat a patient, or One or more biomarkers each compared to a predetermined one or more threshold levels of each biomarker or in one or more control samples Eosinophil target if determined to have higher or lower DPP4 and / or POSTN levels in the sample, optionally levels of additional biomarkers such as blood eosinophils, compared to the bell Instruct the health care provider to discontinue treatment with a treatment plan that includes the administration of a chemotherapeutic agent, not initiate treatment, refuse treatment, or discontinue treatment, do not initiate treatment, or refuse treatment And providing a method.

  In one aspect, the disclosure provides for patients having an eosinophilic disease or disorder (eg, a pulmonary disease or disorder such as mild to moderate asthma or severe asthma) and an eosinophil targeted therapeutic agent such as benralizumab ( A method for determining whether to be treated with a treatment regimen comprising administration of an anti-IL-5R antibody, such as MEDI-563), (a) DPP4 and / or POSTN levels, optionally taken from a patient Measuring or instructing a clinical laboratory to measure the level of an additional biomarker, such as blood eosinophils, in the sample; Lower or decreased DPP4 and / or in a sample compared to a threshold level or compared to one or more biomarker levels in one or more control samples Or treatment comprising administration of an eosinophil targeted therapeutic if it is determined to have a higher or increased level of at least one optional additional biomarker such as POSTN level, eosinophil count, etc. Treating a patient with a plan or instructing a health care provider to treat the patient.

  In one aspect, the disclosure provides for patients having an eosinophilic disease or disorder (eg, a pulmonary disease or disorder such as mild to moderate asthma or severe asthma) and an eosinophil targeted therapeutic agent such as benralizumab ( A method for determining whether to be treated with a treatment regimen comprising administration of an anti-IL-5R antibody, such as MEDI-563), (a) DPP4 and / or POSTN levels, optionally taken from a patient Measuring or instructing a clinical laboratory to measure the level of an additional biomarker, such as blood eosinophils, in the sample; and (b) the patient has one or more of the predetermined biomarkers Higher or increased DPP4 and / or in a sample compared to a threshold level or compared to one or more biomarker levels in one or more control samples Or if it is determined to have a lower or reduced level of POSTN levels, or at least one optional additional biomarker such as blood eosinophils, Instructing the health care provider to discontinue treatment, do not initiate treatment, refuse treatment, or discontinue, do not initiate or refuse treatment on a treatment plan that includes administration And a method comprising:

  Further provided is that patients diagnosed with an eosinophilic disease or disorder (eg, a pulmonary disease or disorder such as mild to moderate asthma or severe asthma) can be treated with an eosinophil targeted therapeutic agent such as benralizumab A method of selecting as a candidate for treatment with an anti-IL-5R antibody such as (MEDIA-563), comprising: (a) DPP4 and / or POSTN levels in a sample taken from a patient, optionally blood eosinophils Measuring a level of an additional biomarker, such as a sphere, or instructing a clinical laboratory to measure, and (b) comparing the patient to one or more predetermined threshold levels or one Lower or decreased DPP4 and / or POSTN levels in the sample, blood compared to one or more biomarker levels in the control sample Medical treatment to treat or treat a patient with an eosinophil targeted therapeutic if it is determined to have a higher or increased level of at least one optional additional biomarker such as eosinophils And instructing the provider.

  Further provided is that patients diagnosed with an eosinophilic disease or disorder (eg, a pulmonary disease or disorder such as mild to moderate asthma or severe asthma) can be treated with an eosinophil targeted therapeutic agent such as benralizumab. A method of selecting as a candidate for treatment with an anti-IL-5R antibody such as (MEDIA-563), comprising: (a) DPP4 and / or POSTN levels in a sample taken from a patient, optionally blood Measuring or instructing a clinical laboratory to measure the level of an additional biomarker, such as an acid sphere, and (b) the patient is compared to a predetermined threshold level or levels, or 1 Higher or increased DPP4 and / or POSTN levels in the sample compared to one or more biomarker levels in one or more control samples; Discontinue treatment of the patient with an eosinophil targeted therapeutic to the patient if determined to have a lower or reduced level of at least one optional additional biomarker such as blood eosinophils Or instructing the health care provider to start treatment, not start treatment, refuse or stop treatment, do not start or refuse treatment.

  In some aspects, the disclosed methods may require ordering and / or performing one or more additional assays. For example, if the level of DPP4 and / or POSTN (eg, protein expression level or gene expression level) is measured within the normal range (ie does not increase), the DPP4 or POSTN detection assay excludes false negative results May be repeated and / or one or more additional DPP4 or POSTN detection assays may be performed to monitor the condition of the subject.

  Conversely, if an increase in DPP4 and / or POSTN levels (eg, protein expression level or gene expression level) is measured, it may be desirable to repeat the DPP4 or POSTN detection assay to exclude false positive results. is there.

  In some aspects, the predetermined DPP4 threshold level is at least about 363 ng / mL (as mild to moderate asthmatic patients, for example, as measured in serum using a DPP4 detection assay including the immunoassay described in Example 2. Median), or at least about 376 ng / mL (mean value in patients with mild to moderate asthma). Therefore, DPP4 expression levels above those values are considered high or elevated, and DPP4 expression levels below those values are considered low or reduced.

  In some aspects, the predetermined POSTN threshold level is at least about 23.5 ng / mL (from mild to low) as measured in serum using, for example, a POSTN detection assay, including the immunoassay described in WO2015015185. Median in patients with moderate asthma), or at least about 25.8 ng / mL (mean value in patients with mild to moderate asthma). Thus, POSTN expression levels above those values are considered high or elevated, and POSTN expression levels below those values are considered low or reduced.

  In some aspects, the predetermined blood eosinophil level is 150 cells / μL. In some aspects, the predetermined blood eosinophil level is 300 cells / μL. In another aspect, the blood eosinophil threshold level is 400 cells / μL. Thus, blood eosinophil levels above those values are considered high or elevated, and blood eosinophil levels below those values are considered low or reduced.

  In some aspects, DPP4 and / or POSTN levels above or below a predetermined threshold level in patients with eosinophilic diseases or disorders (eg, pulmonary diseases or disorders such as mild to moderate asthma or severe asthma) The presence of can be used in combination with one or more clinical or molecular biomarkers specific for such diseases.

For example, in patients with asthma, measuring DPP4 and / or POSTN levels can be combined with measuring other biomarker levels (eg, blood eosinophils). Thus, in one aspect, the level of DPP4 and / or POSTN is, for example,
(I) Whether a patient suffering from a pulmonary disease (eg asthma or COPD) is eligible or not eligible for a specific treatment with an eosinophil targeted therapeutic agent, eg an anti-IL-5R antibody such as benralizumab (MED-563) To determine whether or not to respond to a particular treatment,
(Ii) To determine whether a particular treatment (eg, with an eosinophil targeted therapeutic) should be started, stopped, or improved,
(Iii) to diagnose whether a disease (eg, asthma or COPD) is treatable or untreatable with a particular therapeutic agent, or (iv) the outcome of treatment of a disease (eg, asthma or COPD) with a particular therapeutic agent To predict or predict,
For example, it can be combined with blood eosinophils in any of the methods disclosed herein.

  Those skilled in the art will appreciate that DPP4 and / or POSTN levels (eg, protein expression levels or gene expression levels) include, but are not limited to, positive selectors, including treatment, diagnosis, and monitoring methods. The DPP4 and / or POSTN level (eg protein expression level or gene expression level) in a sample taken from a patient is below a predetermined DPP4 or POSTN threshold level, Alternatively, a specific effect is exerted when reduced compared to DPP4 or POSTN levels in one or more control samples (eg, eosinophilic targeted therapy in patients with eosinophilic diseases or disorders such as asthma). Understand that it is treated with drugs) Wax.

  One skilled in the art will recognize that DPP4 and / or POSTN levels (eg, protein expression levels or gene expression levels) as negative selectors, including but not limited to therapeutic, diagnostic, and monitoring methods herein. Can be used in accordance with the disclosed methods, ie, DPP4 and / or POSTN levels (eg, protein expression levels or gene expression levels) in a sample taken from a patient are above a predetermined DPP4 or POSTN threshold level, or A specific effect is exerted when increased compared to DPP4 or POSTN levels in one or more control samples (eg, treating patients with pulmonary diseases such as asthma with eosinophil targeted therapeutics) You will understand further.

  In one aspect, the present disclosure relates to whether a patient will benefit from treatment with an eosinophil targeted therapeutic agent, eg, an anti-IL-5R antibody such as benralizumab (MED-563). , Methods, assays, and kits for facilitating decisions by health care providers, or clinical laboratories. The methods, assays and kits provided herein also provide medical care regarding whether the patient will benefit from treatment with any other eosinophil targeted therapeutic known to those of skill in the art. Facilitate decisions by health care providers, health care providers, or clinical laboratories.

  In one aspect, the methods disclosed herein include making a diagnosis that may be a differential diagnosis based at least in part on the level of DPP4 and / or POSTN of the patient. In some aspects, the methods disclosed herein include informing a subject of a DPP4 and / or POSTN detection assay result and / or a diagnosis based at least in part on DPP4 and / or POSTN levels. . The patient may be notified verbally, in writing and / or electronically.

  This diagnosis can also be recorded in the patient's medical record. For example, in various aspects, a diagnosis of a pulmonary disease (eg, asthma) that can be treated with a specific eosinophil-targeted therapeutic agent, eg, an anti-IL-5R antibody such as benralizumab (MEDI-563), is recorded in a medical record. .

  The term “medical record” or “patient medical record” typically refers to one of a patient's medical history and complaints, physician physical findings, results of diagnostic tests and procedures, and pharmacotherapy and treatment procedures for a patient. Refers to patient examination and / or treatment reports, including the above. Medical records are typically created by one or more physicians and / or physician assistants, and also various diseases or injuries and / or inoculations and / or allergies that require medical attention, and / or Or written, transcribed, or otherwise recorded records and / or history of treatment, and / or prognosis, and / or often health information related to parents, siblings, and / or occupations is there. The record can be reviewed by a physician in diagnosing the condition.

  The medical record may be in paper form and / or maintained on a computer readable medium. Medical records may be maintained by laboratories, clinics, hospitals, health maintenance organizations, insurance companies, and / or personal medical records websites. In some aspects, a diagnosis based on at least partially measured DPP4 levels and / or POSTN levels may be performed using medical alert articles (medicals) such as cards, worn articles, and / or radio frequency identification (RFID) tags. recorded on or in the alert article). As used herein, the term “wearing article” refers to any article that can be worn on a subject's body, including but not limited to a tag, bracelet, necklace, armband, or headband.

  As used herein, the term “diagnosis” means detecting a disease or determining the stage or degree of a disease. Usually, the diagnosis of a disease is based on an assessment of one or more factors and / or symptoms indicative of the disease. That is, a diagnosis can be made based on the presence, absence or amount of a factor that indicates the presence or absence of a disease or disorder. Each factor or symptom considered to be indicative for the diagnosis of a particular disease need not be exclusively related to the particular disease, for example, a differential diagnosis that can be inferred from the diagnostic factor or symptom may be made. Similarly, a factor or symptom indicative of a particular disease may be present in an individual who does not have the particular disease.

  The term “diagnosis” also includes determining the therapeutic effect of drug therapy or predicting the pattern of response to drug therapy. The diagnostic method may be used independently or in combination with other diagnostic and / or stage classification methods known in the medical art for a particular disease.

  As used herein, the term “differential diagnosis” refers to a determination based on an analysis of clinical data which of two or more diseases with similar symptoms are likely to be associated with a subject's symptoms. The term indicates whether a patient is sensitive to treatment with an eosinophil-targeted therapeutic agent, such as an anti-IL-5R antibody such as benralizumab (MED-563), and the measured DPP4 level in a patient sample. Is also used to refer to determining whether or not is above a predetermined threshold level or whether it is elevated relative to the level in one or more control samples.

  The term “prognosis”, as used herein, refers to a probable course of disease or disease and prediction of outcome. Prognosis is usually determined by assessing a disease factor or symptom that indicates a favorable or unfavorable course or outcome of the disease. The phrase “determining prognosis” as used herein refers to a process by which one of ordinary skill in the art can predict the course or outcome of a condition in a patient. The term “prognosis” does not refer to the ability to predict the course or outcome of a condition with 100% accuracy. Instead, those skilled in the art will recognize that the term “prognosis” increases the probability that a particular course or outcome will occur, i.e., in a patient whose course or outcome exhibits a given condition, compared to an individual who does not exhibit that condition. It will be understood that when done, it refers to an increased likelihood of occurring.

  The terms “preferred prognosis” and “positive prognosis” or “unfavorable prognosis” and “negative prognosis” as used herein are for the prediction of a probable course of a condition or disease and / or possible outcome. Is a relative term. A favorable or positive prognosis predicts a better outcome than an unfavorable or negative prognosis in the condition. In a general sense, a “preferred prognosis” is a relatively better outcome than a number of other possible prognoses that may be associated with a particular condition, while an unfavorable prognosis is a number of cases that may be associated with a particular condition A relatively poor outcome is expected than other possible prognoses. Typical examples of preferred or positive prognosis include better than average remission rate, lower tendency to metastasis, longer life than expected life expectancy, differentiation of benign processes from cancer processes, and the like.

  The disclosure includes methods of treating eosinophilic diseases or disorders in a subject (eg, pulmonary diseases or disorders such as mild to moderate asthma or severe asthma) based on changes in the expression of DPP4 and / or POSTN. The present disclosure is a method of treating a patient having an eosinophilic disease or disorder (eg, a pulmonary disease or disorder such as mild to moderate asthma or severe asthma), wherein the patient has a predetermined DPP4 or POSTN threshold level. Determined to have lower or decreased DPP4 and / or POSTN levels in one or more samples taken from a patient compared to DPP4 or POSTN levels in one or more control samples In some cases, a method is provided comprising the step of administering to a patient an eosinophil-targeted therapeutic agent, eg, an anti-IL-5R antibody such as benralizumab (MEDI-563). In some aspects, the sample is obtained from a patient and submitted for measurement of the level of DPP4 and / or POSTN in the sample.

  The disclosure also provides a method of treating a patient having an eosinophilic disease or disorder (eg, a pulmonary disease or disorder such as mild to moderate asthma or severe asthma), wherein (a) a sample is taken from the patient Submitting for measurement of DPP4 and / or POSTN levels in a sample, and (b) the patient compares DPP4 or POSTN with a predetermined DPP4 or POSTN threshold level or in one or more control samples If the sample has a lower or decreased level of DPP4 and / or POSTN in the collected sample, an anti-IL-5R antibody, such as an eosinophil-targeted therapeutic agent such as benralizumab (MEDI-563) Administering to a patient.

  Further provided is a method of treating a patient having an eosinophilic disease or disorder (eg, a pulmonary disease or disorder such as mild to moderate asthma or severe asthma), which is collected from (a) the patient. Submitting a sample for determination of DPP4 and / or POSTN levels in the sample; and (b) DPP4 in a patient compared to a predetermined DPP4 or POSTN threshold level or in one or more control samples Or patients with anti-IL-5R antibodies, such as eosinophil-targeted therapeutics, eg, benralizumab (medi-563), if they have higher or increased levels of DPP4 and / or POSTN in the sample compared to the level of POSTN Discontinuing or not starting administration to a method.

  The disclosure also provides a method of treating a patient having an eosinophilic disease or disorder (eg, a pulmonary disease or disorder such as mild to moderate asthma or severe asthma), the sample obtained from the patient (a) Measuring the level of DPP4 and / or POSTN in the sample; and (b) the level of DPP4 and / or POSTN in the sample is higher or increased or lower than the predetermined DPP4 or POSTN threshold level or Determining whether to decrease; (c) the patient is lower or decreased in the sample compared to a predetermined DPP4 or POSTN threshold level or compared to a DPP4 or POSTN level in one or more control samples Eosinophil targeting if determined to have DPP4 and / or POSTN levels A therapeutic, eg, anti-IL-5R antibody such as benralizumab (MEDI-563) is administered to the patient, or the patient compares DPP4 or in one or more control samples or compared to a predetermined DPP4 or POSTN threshold level. If determined to have higher or increased DPP4 and / or POSTN levels in the sample compared to the POSTN level, the health care provider should be discontinued or refused to administer the eosinophil targeted therapeutic to the patient. A method is provided including a step of advising.

  Further provided is a method of treating a patient having an eosinophilic disease or disorder (eg, a pulmonary disease or disorder such as mild to moderate asthma or severe asthma), which is collected from (a) the patient. Submitting a sample for measurement of the level of DPP4 and / or POSTN in a sample obtained from the patient; and (b) the patient is compared to a predetermined DPP4 or POSTN threshold level, or one or more If it is determined to have a lower or decreased DPP4 and / or POSNT level in the sample compared to the DPP4 or POSTN level in the control sample, eosinophilic targeted therapeutics such as benralizumab (MEDI-563) Anti-IL-5R antibody such as) or the patient has a predetermined DPP4 or POSTN threshold level Eosinophil targeted therapy if determined to have higher or increased DPP4 and / or POSTN levels in the sample compared to or compared to DPP4 or POSTN levels in one or more control samples Discontinuing, not starting, or refusing administration of the drug to the patient.

  In some aspects, the patient's DPP4 and / or POSTN levels are as described herein using an antibody or antigen-binding fragment thereof that recognizes human DPP4, POSTN, or an antigen-binding fragment, variant or derivative thereof. Measured by a simple immunoassay. In some aspects, the sample is obtained from a patient and submitted to a clinical laboratory, for example, for measurement of DPP4 and / or POSTN levels in the sample.

  In some aspects of the above therapeutic methods, the patient's DPP4 and / or POSTN level (eg, DNA or RNA level) is one or more capable of specifically measuring the expression level of the DPP4 gene and / or the POSTN gene. In an assay using oligonucleotide probes.

  In some aspects, a molecular biomarker (eg, DPP4 or POSTN) detection assay (eg, an immunoassay) (eg, a DPP4 immunoassay described in Example 2 assembled as a “point of care” diagnostic kit, eg, Performed on a sample obtained from a patient by a medical professional treating the patient (using an immunoassay as described herein, including the POSTN assay described in US2015120185).

  In some aspects, the sample is obtained from a patient (eg, for example, either the DPP4 immunoassay described in Example 2 or the immunoassay disclosed in Table 1 or the POSTN assay described in WO2015015185). (For example, to a clinical laboratory for measurement of DPP4 and / or POSTN levels in a sample according to medical professional instructions).

  In some aspects, the clinical laboratory performing the assay may allow the patient to use DPP4 and / or the patient's DPP4 and / or from a treatment with an eosinophil-targeted therapeutic agent, eg, an anti-IL-5R antibody such as benralizumab (MEDI-563). Or, the health care provider will be advised on whether a benefit can be gained based on whether the POSTN level is above a predetermined DPP4 or POSTN threshold or increased relative to one or more control samples.

  In some aspects, the disclosure provides a method of treating a patient with an eosinophilic disease or disorder (eg, a pulmonary disease or disorder such as mild to moderate asthma or severe asthma) over a long period of time, from the patient Measure the level of DPP4 and / or POSTN (eg, protein expression level or gene expression level) in the first sample collected, or the first sample collected from the patient is DPP4 and / or in the sample Or submitting for measurement of POSTN level and the patient's DPP4 and / or POSTN level in the first sample is below a predetermined DPP4 or POSTN threshold level, or in one or more control samples Eosinophil targeted therapeutics such as benralizumab (ME) when decreased compared to DPP4 or POSTN levels Administering an anti-IL-5R antibody such as I-563) to a patient, wherein the patient's DPP4 and / or POSTN levels are disclosed, for example, in the DPP4 immunoassay described in Example 2 or Table 1, for example. A DPP4 immunoassay or an immunoassay comprising the POSTN assay described in WO2015015185. The test may be performed by a health care provider or clinical laboratory as described above.

  In some aspects, the results of an immunoassay as provided herein are used to determine whether a patient's insurance is for treatment with an eosinophil-targeted therapeutic. Can be submitted to.

  In some aspects, the disclosure provides a method of treating a patient having an eosinophilic disease or disorder (eg, a pulmonary disease or disorder such as mild to moderate asthma or severe asthma), such as in a clinical laboratory And DPP4 and / or POSTN levels (eg protein expression level or gene expression level) and eosinophilic acid in a first sample obtained from a patient having an eosinophilic disease or disorder, eg, a sample provided by a health care provider Measuring the sphere level and the patient's DPP4 and / or POSTN level in the first sample is below a predetermined DPP4 or POSTN threshold level or compared to the DPP4 or POSTN level in one or more control samples Measuring DPP4 and / or POSTN levels in patients An anti-eosinophil targeted therapeutic, such as benralizumab (MED-563), if it falls below a given DPP4 or POSTN threshold level or decreases compared to the DPP4 or POSTN level in one or more control samples. Advising a healthcare provider to administer an IL-5R antibody to a patient, wherein the DPP4 and / or POSTN level of the patient in the first sample is, for example, as described in Example 2 Or a DPP4 immunoassay disclosed in Table 1 or an immunoassay comprising a POSTN assay as described in WO2015015185.

  In some aspects, the sample is obtained from a patient, eg, using an immunoassay, alone or at least in combination with the level of another biomarker, such as blood eosinophils, in the sample using DPP4 and / or POSTN levels. Or a combination thereof (eg, protein expression level or gene expression level) is submitted to a clinical laboratory, for example.

  In some aspects, the clinical laboratory performing the assay is such that the patient has a DPP4 and / or POSTN level (eg, protein expression level or gene expression level) of the patient below a predetermined DPP4 or POSTN threshold, or one Based on whether it is low compared to the above control samples, the health care provider will be advised as to whether or not it can benefit from treatment with eosinophil targeted therapeutics.

  In some aspects, the method of treatment discussed herein (eg, for pulmonary diseases or disorders such as asthma) comprises an eosinophil targeted therapeutic agent and an eosinophil targeted therapy per serum volume. Administering a specific amount of the drug to the subject in an amount and / or at sufficient intervals to be achieved and / or maintained using, for example, an assay as described herein.

  For example, an antibody (eg, targeting IL-5 or IL-5R) or antigen-binding fragment thereof has at least about 15 ng / ml, 20 ng / ml, 25 ng / ml, 30 ng / ml, 35 ng / ml in serum, 40 ng / ml, 45 ng / ml, 50 ng / ml, 55 ng / ml, 60 ng / ml, 65 ng / ml, 70 ng / ml, 75 ng / ml, 80 ng / ml, 85 ng / ml, 90 ng / ml, 95 ng / ml, 100 ng / ml, 120 ng / ml, 140 ng / ml, 160 ng / ml, 180 ng / ml, 200 ng / ml, 220 ng / ml, 240 ng / ml, 260 ng / ml, 280 ng / ml, 300 ng / ml, 320 ng / ml, 340 ng / ml, 360 ng / ml, 380 ng / ml, 400 ng / ml, 20 ng / ml, 440 ng / ml, 460 ng / ml, 480 ng / ml, 500 ng / ml, 520 ng / ml, 540 ng / ml, 560 ng / ml, 580 ng / ml, 600 ng / ml, 620 ng / ml, 640 ng / ml, 660 ng / ml, 680 ng / ml, 700 ng / ml, 720 ng / ml, 740 ng / ml, 760 ng / ml, 780 ng / ml, 800 ng / ml, 820 ng / ml, 840 ng / ml, 860 ng / ml, 880 ng / ml, 900 ng / ml, It can be administered to achieve 920 ng / ml, 940 ng / ml, 960 ng / ml, 980 ng / ml, or 1000 ng / ml.

  In further embodiments, the antibody or antigen-binding fragment thereof (eg, targeting IL-5 or IL-5R) achieves a concentration of antibody in the serum of about 12.5 ng / ml to about 1000 ng / ml. Can be administered. One skilled in the art will recognize that when the amount administered here applies to a full-length antibody or immunoglobulin molecule and an antigen-binding fragment thereof is used, the absolute amount is administered in a manner that can be calculated based on the molecular weight of the fragment. You will understand the difference.

  In some aspects, the methods of treatment discussed herein, for example, for eosinophilic diseases or disorders (eg, pulmonary diseases or disorders such as mild to moderate asthma or severe asthma) include eosinophil targeting Anti-IL-5R antibodies, such as benralizumab (MED-563), every 15-54 mg every 0.5-1.5 months, every 55-149 mg every 1.5-4.5 months, Administering to the subject in amounts and intervals of 150-299 mg every 4-8 months, or 300-1100 mg every 4-12 months.

  In some embodiments, the amount and interval is 15 to 21 mg every 0.5 to 1.0 months, 55 to 70 mg every 1.5 to 3.0 months, 150 to 260 mg every 4 to 6 months Or every 4-8 months at 300-700 mg. In some aspects, the amounts and intervals are 21 mg monthly, 70 mg every 3 months, 210 mg every 6 months, or 700 mg every 6 months. In some aspects, the amounts and intervals are 210 mg every 3 months or 700 mg every 3 months. In some aspects, the amounts and intervals are 210 mg every month or 700 mg every month.

  In some aspects, the eosinophil targeted therapeutic (eg, benralizumab / MEDI-563) is administered at about 2 mg to about 100 mg / dose. In some aspects, the eosinophil targeted therapeutic (eg, benralizumab / MEDI-563) is administered at about 20 mg / dose, about 30 mg / dose, or about 100 mg / dose. In some aspects, the eosinophil targeted therapeutic (eg, benralizumab / MEDI-563) is administered once every 4 weeks to once every 12 weeks. In some aspects, the eosinophil targeted therapeutic (eg, benralizumab / MEDI-563) is administered once every 4 weeks. In some aspects, the eosinophil targeted therapeutic (eg, benralizumab / MEDI-563) is administered once every 8 weeks. In some aspects, the eosinophil targeted therapeutic (eg, benralizumab / MEDI-563) is administered once every 4 weeks for 12 weeks and then once every 8 weeks.

  In some aspects of the method, an eosinophil targeted therapeutic, eg, an anti-IL-5R antibody such as benralizumab (MED-563) is administered intravenously (IV). In some aspects of the method, an eosinophil targeted therapeutic, eg, an anti-IL-5R antibody such as benralizumab (MED-563) is administered subcutaneously (SC).

  In some aspects, an eosinophil targeted therapeutic, eg, an anti-IL-5R antibody such as benralizumab (MED-563) is administered in one or more fixed doses. In some aspects, the dose is weekly, every 2 weeks, every 3 weeks, every 4 weeks, every 5 weeks, every 6 weeks, every 7 weeks, every 8 weeks, every 9 weeks, every 10 weeks, or every 12 weeks Every dose. In some aspects, the dosage is about 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, 350 mg, 360 mg, 370 mg, 380 mg, 390 mg, 400 mg, 410 mg, 420 mg, 430 mg, 440 mg, 450 mg, 460 mg, 470 mg, 480 mg, 4 0 mg, 500 mg, 510 mg, 520 mg, 530 mg, 540 mg, 550 mg, 560 mg, 570 mg, 580 mg, 590 mg, 600 mg, 610 mg, 620 mg, 630 mg, 640 mg, 650 mg, 660 mg, 670 mg, 680 mg, 690 mg, 700 mg, 710 mg, 720 mg, 730 mg, 740 mg, 750 mg, 760 mg, 770 mg, 780 mg, 790 mg, 800 mg, 810 mg, 820 mg, 830 mg, 840 mg, 850 mg, 860 mg, 870 mg, 880 mg, 890 mg, 900 mg, 910 mg, 920 mg, 930 mg, 940 mg, 950 mg, 960 mg, 980 mg, 980 mg Contains 990 mg or 1000 mg. In some aspects, the dose is greater than 1000 mg.

  In some aspects 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49 or 50 doses are administered.

  The formulation, dosing schedule, and route of administration of anti-IL-5R antibodies, such as eosinophil targeted therapeutic agents, eg, benralizumab (MED-563), for optimal therapeutic response according to the methods disclosed herein. It can be adjusted to provide an effective amount. For administration of an eosinophil targeted therapeutic agent, the therapeutic agent may be administered through any suitable means, composition and route known in the art. With respect to the dosing regimen, a single bolus can be administered, several divided doses can be administered over time, or the dose can be proportionally reduced or increased as indicated in the emergency of the therapeutic situation.

  An eosinophil targeted therapeutic, eg, an anti-IL-5R antibody such as benralizumab (MED-563) is administered by any suitable technique, such as, but not limited to, parenterally, topically, or by inhalation. May be. When injected, the pharmaceutical composition includes, for example, intra-articular, intravenous, intramuscular, intralesional, intraperitoneal or dermal routes (intradermal, transdermal or sub-dermal, and subcutaneous). ) Via bolus injection or continuous infusion. In some aspects, the pharmaceutical composition is administered by the intravenous route. In some aspects, the pharmaceutical composition is administered by the subcutaneous route.

  In further embodiments, the composition is administered by oral, buccal, rectal, intratracheal, gastric, or intracranial route. Localized administration is contemplated, for example, at the site of disease or injury, for example by enema or suppositories for conditions involving the gastrointestinal tract. Further considered is transdermal delivery and sustained release from implants. Delivery by inhalation includes, for example, nasal or oral inhalation, use of a nebulizer, inhalation of an antagonist in aerosol form, and the like. Other options include eye drops; oral formulations including pills, syrups, lozenges or chewing gum; and topical formulations such as lotions, gels, sprays, and ointments, prefilled syringes and autoinjectors .

  Advantageously, the eosinophil-targeted therapeutic agent, eg, an anti-IL-5R antibody such as benralizumab (MEDI-563), is one or more additional components such as a physiologically acceptable carrier, excipient or diluent. Can be administered in the form of a composition comprising Optionally, the composition further comprises one or more physiologically active agents as a combination therapy.

  The pharmaceutical composition comprises an eosinophil-targeted therapeutic agent, eg, an anti-IL-5R antibody such as benralizumab (MEDI-563), a buffer, an antioxidant such as ascorbic acid, a low molecular weight polypeptide (less than 10 amino acids). And so on), one or more substances selected from the group consisting of proteins, amino acids, carbohydrates such as glucose, sucrose or dextrin, chelating agents such as EDTA, glutathione, stabilizers and excipients.

  Neutral buffered saline or saline mixed with the same kind of serum albumin is an example of a suitable diluent. In accordance with appropriate industry standards, preservatives such as benzyl alcohol may also be added. The composition may be formulated as a lyophilizate using appropriate excipient solutions (eg, sucrose) as diluents.

  In some aspects, the eosinophil targeted therapeutic agent, eg, an anti-IL-5R antibody such as benralizumab (MED-563) is about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175 , 180, 185, 190, 195 or 200 mg / ml.

  Exemplary formulations useful in the present invention include sucrose at a suitable concentration such as glutamate, citrate or acetate buffer, 1-20% (w / v) at a suitable pH of 4.5-5.2. , Excipients such as glycine, proline, glycerol, and / or sorbitol, and polysorbate (polysorbate 20 or 80) or poloxamer (poloxamer 1888) at an appropriate concentration of 0.001% to 0.1% (w / v) ) Containing a surfactant such as a nonionic surfactant. Such formulations are disclosed in US Pat. No. 6,171,586 and WIPO published applications WO20120002766 and WO201108120. In some aspects, the formulation comprises sodium acetate, sucrose and polysorbate 20.

In some aspects, administration of the patient to an anti-IL-5R antibody, such as an eosinophil targeted therapeutic agent, eg, benralizumab (MED-563), comprises:
(A) decrease in AER (acute exacerbation rate),
(B) Increase in FEV1 (forced expiratory volume per second),
(C) Results of improved ACQ-6 (Asthma Management Questionnaire, 6 item version),
(D) As a result of improved AQLQ (Asthma QOL Questionnaire),
(E) decreased cytokine secretion by type 2 innate lymphocytes (ILC2),
(F) reduced ILC2 levels;
(G) resulting in a decrease in eosinophil count and / or basophil count, or (h) a combination thereof.

  In some aspects, a decrease in AER following administration of an eosinophil-targeted therapeutic agent, for example, an anti-IL-5R antibody such as benralizumab (MEDI-563), is associated with an AER observed in a population of patients treated with placebo. Compared to at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, or at least 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least About 100%.

In some aspects, an increase in FEV ₁ following administration of an anti-IL-5R antibody, such as an eosinophil targeted therapeutic, eg, benralizumab (MED-563), is observed in a population of patients treated with placebo. ₁ at least about 3%, at least about 5%, at least about 7%, at least about 9%, at least about 11%, at least about 13%, at least about 15%, at least about 17%, at least about 19%, At least about 21%, at least about 23%, at least about 25%, at least about 27%, at least about 29%, at least about 31%, at least about 33%, or at least about 35%.

In some aspects, an increase in FEV ₁ following administration of an anti-IL-5R antibody, such as an eosinophil targeted therapeutic, eg, benralizumab (MED-563), is observed in a population of patients treated with placebo. ₁ at least about 25 mL, at least about 50 mL, at least about 75 mL, at least about 100 mL, at least about 125 mL, at least about 150 mL, at least about 175 mL, at least about 200 mL, at least about 225 mL, at least about 250 mL, at least about 275 mL, at least About 300 mL, at least about 325 mL, or at least about 350 mL.

  In some aspects, the change in ACQ-6 following administration of an anti-IL-5R antibody such as an eosinophil targeted therapeutic, eg, benralizumab (MEDID-563), is observed in a population of patients treated with placebo Compared to the average ACQ-6, about -0.2, -0.3, -0.4, -0.5, -0.6, -0.7, -0.8, -0.9,- 1, -1.1, or -1.2.

  In some aspects, the decrease in cytokines secreted by type 2 innate lymphocytes (ILC2) following administration of an anti-IL-5R antibody, such as an eosinophil targeted therapeutic, eg, benralizumab (MED-563) is At least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30 compared to the levels of cytokines secreted by ILC2 found in the population of patients treated with %, At least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80 %, At least about 85%, at least about 90%, at least about 95%.

  In some aspects, a decrease in eosinophils after administration of an eosinophil targeted therapeutic, eg, an anti-IL-5R antibody such as benralizumab (MEDI-563), is observed in a population of patients treated with placebo. At least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least About 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least About 95%.

  In some aspects, a decrease in basophils following administration of an anti-IL-5R antibody, such as an eosinophil targeted therapeutic, eg, benralizumab (MED-563), is observed in a population of patients treated with placebo. At least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least About 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least About 95%.

VII. Kits for Detection and Quantification of DPP4 or POSTN The present disclosure also provides kits for detecting DPP4 and / or POSTN levels, for example through immunoassay methods. Such kits include containers each having one or more of various reagents (eg, concentrated forms) utilized in the method, including, for example, one or more anti-DPP4 antibodies (ie, antibodies capable of detecting DPP4). obtain. One or more anti-DPP4 or anti-POSTN antibodies, eg, capture antibodies capable of detecting DPP4 or POSTN, can be provided already attached to a solid support. One or more antibodies, eg, detection antibodies, can be provided that is already conjugated to a detectable label, eg, biotin or ruthenium chelate.

  The kit also provides reagents, buffers, and / or reagents and instruments to support the performance of the assays provided herein for coupling the detectable label to the antibody (and the label itself). obtain. In certain embodiments, a labeled secondary antibody that binds to the detection antibody can be provided. Kits provided in accordance with the present disclosure may further include suitable containers, plates, and any other reagents or materials necessary to perform the assays provided herein.

  In some aspects, the kit comprises one or more nucleic acid probes (eg, oligonucleotides comprising natural and / or chemically modified nucleotide units) capable of hybridizing a partial sequence of a DPP4 or POSTN gene sequence under high stringency conditions. )including. In some aspects, one or more nucleic acid probes (eg, oligonucleotides comprising natural and / or chemically modified nucleotide units) capable of hybridizing a subsequence of a DPP4 or POSTN gene sequence under high stringency conditions are: Coupled to a microarray chip.

  A kit provided in accordance with the present disclosure may also include a brochure or instructions for use describing the method. In DPP4 or POSTN detection immunoassays (eg, single or multiplex assays to detect DPP4 or POSTN), particularly sandwich immunoassays, eg, ELISA or ECL assays, sandwich immunoassays are performed on a solid support. Bound primary “capture” antibodies (eg, antibodies that specifically bind to DPP4 or POSTN) or antigen-binding fragments thereof, and secondary anti-DPP4 “detection” antibodies or antigen-binding fragments thereof. Immunoassays can be performed by the methods provided herein or by methods well known and understood by those skilled in the art.

  In one aspect, the immunoassay method involves binding a capture antibody or fragment thereof to a solid support and applying a test or control sample, where DPP4 or POSTN (if present in the sample) is the capture antibody or fragment thereof. Applying a detection antibody or fragment thereof that can bind to DPP4 or POSTN already bound to a capture antibody or fragment thereof; and a detection antibody or fragment thereof bound to DPP4 or POSTN. Measuring the amount of. In certain aspects, the assay may further comprise a washing step, a blocking step and an incubation step.

  The test kit may include instructions for performing one or more DPP4 and / or POSTN detection assays, eg, an immunoassay or nucleic acid detection assay to detect the presence / absence or level of DPP4 or POSTN . The instructions included in the kit can be attached to the packaging material or included as a package insert. The instruction manual is typically a handwritten material or a printed material, but is not limited thereto. Any medium capable of storing such instructions and communicating them to the end user is contemplated. Such media include, but are not limited to, electronic storage media (eg, magnetic disks, tapes, cartridges, chips), optical media (eg, CD-ROM), and the like. As used herein, the term “instruction for use” may include the address of an Internet site that provides instructions for use.

VIII. Companion Diagnostic System The method disclosed herein can be provided as a companion diagnostic available, for example, via a web server, to inform the clinician or patient about promising treatment options. The methods disclosed herein include the step of collecting or otherwise obtaining a biological sample, and DPP4 and / or POSTN levels (eg, full-length or soluble DPP4 protein expression levels, POSTN expression levels, or DPP4 or POSTN Analysis to detect and measure the level of gene expression in) alone or in combination with other biomarkers (eg, gene signatures such as Th-2 signature or gene panel used to derive patient eosinophil counts) Carrying out the method.

  As molecular biomarkers that can be combined with DPP4 and / or POSTN, IL-33, IL-25, TSLP, IL-22, CCL20, LCN2, sCTLA-3, sCD28, CCL5, CCL11, CCL22, CCL26, FZD5, DOK1, CST2, ZNF436, C20orf100, NAGS, CST1, CDH13, HRH1, TMEM132B, NTRK1, SLCO2A1, IgE, FETUB, KRT31IKRT34, C6orf138, ATP5J, TUBAL3, JAM2, RVA3C, NOVA2C, NOVA2C LYPD1, DISP1, NKX1-2, C4orf38, LOXL4, PRKD1, PAM124B GPR44, HIGD1B, CLCA1, SEPT11, CYYR1, CD36, ALOX15, AADAC, ACTA1, ODC1, DKFZp434F142, ACHE, CSF3, LOC100132552, C12orf27, ZNF331, GK5, DUSP1IDUSP4, LW1, LW4 SAA4, SLC12A3, TMEM45A, FLJ37464, MUC5B, CXCL6, GLRB, DKFp686K01114, FOLR1, TSPAN6, AKR1C1, KIAA0232, PTP4A1, PCYT2, RHOV, PROS1, T11N63P Or combinations thereof. Lun et al. , J .; Clin. Immunol. 27: 430-437 (2007), Choi et al, J. MoI. Immunol. 186 (3): 1861-9 (2011), and WO2009124090A1, which are hereby incorporated by reference in their entirety.

  Standard names, aliases, etc., of proteins and genes named by identifiers used throughout this application (eg, PIP5K1B) can be identified, for example, via Genecards (www.genecards.org) or Uniprot (www.uniprot.org).

  Molecular biomarkers that can be combined with DPP4 and / or POSTN include, for example, IL-33, IL-25, TSLP, IL-22, CCL20, LCN2, CST1, CCL26, CLCAl, CST2, PRR4, SERPINB2, CEACAM5, iNOS, SERPINB4, CST4, PRB4, TPSD1, TPSG1, MFSD2, CPA3, GPR105, CDH26, GSN, C2ORF32, TRACH20000196 (TMEM71), DNAJC12, RGS13, SLC18A2, SERPINB10, SH3RF2, AFR1 Can be mentioned.

  A measured DPP4 and / or POSTN level-derived DPP4 and / or POSTN level (eg, protein expression level or gene expression level) or normalized score alone (eg, for therapeutic, diagnostic, prognostic, or monitoring purposes) Alternatively, it can be used in combination with levels or normalized scores from other biomarkers (eg, gene signatures such as the Th-2 signature or a panel of genes used to induce patient eosinophil counts).

These scores can also be used to obtain a diagnostic score, for example: (i) the patient's IgE level, (ii) the patient's eosinophil or basophil count, (iii) the patient's exhaled nitric oxide concentration ( FE _NO ), (iv) patient eosinophil / lymphocyte and eosinophil / neutrophil (ELEN) index, (v) patient EOS index, (vi) patient IgE level, (vii) bronchodilator. FVC measurements or reversibility before or after administration of FEV1, (viii) percentage of subarea airway wall area (WA%) from lung CT scan, or (ix) a score corresponding to a combination of two or more thereof Can be combined.

  In this approach, the diagnostic score is the sum of all marker calculations compared to a preset DPP4 or POSTN threshold that is an indicator of the presence or absence of a disease (or a specific disease state, eg, moderate or severe asthma) It may be a single number determined from Or the diagnostic score may be a series of bars, each representing a biomarker value, and the pattern of response may be compared to a preset pattern for the determination of the presence or absence of disease.

  Because at least some aspects of the methods described herein are complicated in the calculations involved, of DPP4 and / or POSTN levels (eg, levels in serum) as biomarkers disclosed herein Methods involving use can be performed using a computer. In some aspects, a computer system includes a processor, an input device, an output device, a storage device, a computer readable storage media reader, a communication system, a processing acceleration (eg, a DSP or a dedicated processor), and a memory. Including hardware elements electrically coupled via A computer readable storage media reader is a computer readable storage medium, ie remote, local, fixed and / or removable so as to temporarily and / or more permanently contain computer readable information Can be further coupled to combinations that collectively represent storage devices + storage media, memory, etc., and may include storage devices, memory, and / or any other such accessible system resources.

  A single architecture may be used when equipped with one or more servers that can be further designed according to the currently desired protocols, protocol variations, extensions, and the like. However, it will be apparent to those skilled in the art that the embodiments may be fully utilized depending on the requirements of more specific applications. Even customized hardware can be used and / or certain elements can be implemented in hardware, software or both. In addition, connections to other computer devices (not shown) such as network input / output devices are available, and one or more other connections to wired, wireless, modem, and / or other computer devices. However, it should be understood that it can be used.

  In one aspect, the system further includes one or more devices to provide input data to one or more processors. The system further includes a memory for storing a dataset of ranked data elements. In another aspect, the apparatus for providing input data includes a detector for detecting the characteristics of the data element, such as a fluorescence plate reader, a mass spectrometer, or a gene chip reader.

  Further, the system may include a database management system. The user's request or query can be formatted in an appropriate language understood by the database management system that processes the query to extract relevant information from the training set database. The system may be connectable to a network to which a network server and one or more clients are connected. The network may be a local area network (LAN) or a wide area network (WAN) as is known in the art. Preferably, the server includes the hardware necessary to execute a computer program product (eg, software) for accessing data in the database to process user requests. The system can communicate with an input device for providing data regarding the data element (eg, an expression value) to the system. In one aspect, the input device may include a gene expression profiling system including, for example, a mass spectrometer, gene chip, or array reader.

  Some aspects described herein may be implemented to include a computer program product. A computer program product may include a computer readable medium having computer readable program code embodied in the medium for causing an application program to run on a computer having a database. As used herein, a “computer program product” is included on a physical medium of any nature (eg, secretary, electronic, magnetic, optical or otherwise) and is a computer or other automated data processing An ordered set of instructions in the form of natural or programming language statements that can be used in the system. Such programming language statements, when executed by a computer or data processing system, direct the computer or data processing system to function according to the particular content of the statement.

  Computer program products include, but are not limited to, programs in source and object code and / or test or data libraries embedded in a computer readable medium. Further, computer program products that allow a computer system or data processing device to function in a preselected manner include, but are not limited to, original source code, assembly code, object code, machine language, their encryption or It may be provided in a number of formats including a compressed version and any and all equivalents.

  In one aspect, for performing the therapeutic, diagnostic, prognostic, or monitoring methods disclosed herein, for example, the level of DPP4 and / or POSTN in one or more samples taken from a patient is predetermined An eosinophil-targeted therapeutic agent (eg, an IL- such as benralizumab) if it is lower or decreased compared to the DPP4 or POSTN threshold level of or a DPP4 or POSTN level in one or more control samples. A computer program product is provided for determining whether an antibody that specifically binds to 5R, or an antigen-binding fragment thereof) should be administered to a patient in need thereof.

A computer program product includes a computer readable medium that embodies program code that is executable by a processor of a computer device or system, the program code including:
(A) Code for retrieving data belonging to a biological sample from a subject, where the data is DPP4 and / or POSTN level data (or data typically derived from DPP4 and / or POSTN level values) Alone or in conjunction with values corresponding to other biomarkers in a biological sample (eg, a gene signature, eg, a gene panel used to induce a Th-2 signature). These values may also include, for example, (i) the level of the patient's IgE level, (ii) the patient's eosinophil count or basophil count, (iii) the patient's exhaled nitric oxide concentration (FE _NO ), (iv ) Patient eosinophil / lymphocyte and eosinophil / neutrophil (ELEN) index, (v) Patient EOS index, (vi) Peripheral airway wall area percentage from lung CT scan data (WA%) ), (Vii) the patient's IgE level, (viii) FEV1, before or after administration of bronchodilator, FVC measurements or reversibility, or (ix) a value corresponding to a combination of two or more thereof .
(B) Code that executes a classification method that indicates whether, for example, an eosinophil-targeted therapeutic should be administered to a patient in need thereof.

  While various aspects are described as methods or apparatuses, it is to be understood that the aspects are executable through code coupled to a computer, eg, code resident on or accessible by a computer. is there. For example, software and databases can be utilized when performing many of the methods discussed above. Thus, in addition to those achieved by hardware, these aspects can be achieved through the use of a product comprising a computer usable medium having computer readable program code embodied therein. It is also noted that this enables the functionality disclosed herein. Accordingly, it is desirable that aspects also be considered protected by this patent in their program code means as well.

  Further, some aspects store code in a virtually any type of computer readable storage device including, but not limited to, RAM, ROM, magnetic media, optical media, or magneto-optical media. obtain. Even more generally, some aspects may be implemented in software, or hardware, or any combination thereof, such as, but not limited to, software running on a general purpose processor, microcode, PLA, or ASIC.

  It is also envisioned that some aspects may be achieved as a computer signal embodied in a carrier wave and a signal propagating in a transmission medium (eg, electricity and light). Thus, the various types of information discussed above can be formatted in some structure, eg, a data structure, transmitted as an electrical signal in a transmission medium, or stored on a computer readable medium.

Example 1
Phase 2b dose range study to assess the efficacy and safety of benralizumab (MED-563) in adults with uncontrolled asthma The Ph2b study (CP220) was conducted in a population of moderate to severe uncontrolled asthma patients. It was carried out using benralizumab (MED-563) (FIG. 1). Two data sets: (i) Data set using samples taken at baseline (pre-dose on week 1 day 1) measured for POSTN and DPP4 levels and (ii) -1 week measured for POSTN and DPP4 levels Post-search analysis was performed using data sets using samples taken at (screening) and at 40 weeks. DPP4 was measured using the R & D Systems DPP4 detection kit. POSTN was measured using the periostin detection method developed as a companion diagnostic for traloquinumab (see WO 2015120185 (incorporated herein by reference in its entirety)).

  POSTN is a downstream activation marker for IL-13 pathway activation and is under development as a promising biomarker to identify patients who can respond to anti-IL-13 treatment. It is also used as a promising marker for identifying patients with evidence of increased Th2 pathway activation. DPP4 is also a downstream activation marker for IL-13 pathway activation.

  The patient population consisted of uncontrolled asthma patients for medium or high doses of inhaled corticosteroids and long acting beta agonists. The study included 609 randomized subjects. 324 were included in the EOS + group (PBO = 80; benralizumab 2 mg = 81; 20 mg = 81 and 100 mg = 82). 285 were included in the EOS-group (PBO = 143; benralizumab 100 mg = 142). The study evaluates benralizumab 3 dose versus placebo in subjects with high blood eosinophil levels (as determined by ELEN index) and 1 dose in subjects with low blood eosinophil levels (as determined by ELEN index). There was a double-blind placebo control in patients with moderate to severe asthma evaluating placebo.

  In order to understand the patient population that can respond to treatment with benralizumab, the CP220 patient population can be transferred to other known patient subgroups and other to obtain a better understanding of biomarkers or other promising responder groups. It was proposed to segment by clinical features. For samples measured in data set A (ie, when POSTN and DPP4 were measured in samples collected on day 1 of week 1), the following search objectives were: (i) Blood eosinophil levels and base To assess the relationship between serum POSTN levels and serum DPP4 levels in the line (exploratory outcome variables were blood eosinophils, serum POSTN, and serum DPP4); (ii) for exacerbations, lung function and symptoms Assessing the efficacy of benralizumab by baseline POSTN (exploratory outcome variables were serum periostin, annual exacerbation rate, FEV1, and ACQ); (iii) efficacy of benralizumab on exacerbations, lung function and symptoms Assessed by baseline DPP4 (exploratory outcome variables are serum DPP4, annual exacerbation rate, FE (Iv) Evaluating the efficacy of benralizumab on exacerbations, lung function and symptoms by baseline DPP4 along with baseline eosinophil levels (the exploratory outcome variable is blood eosinophils) , Serum periostin, annual exacerbation rate, FEV1, and ACQ); and (v) Evaluating the efficacy of benralizumab on exacerbations, lung function and symptoms by baseline POSTN along with baseline eosinophil levels (exploration The outcome variables were planned for blood eosinophils, serum DPP4, annual exacerbation rate, FEV1, and ACQ).

  For samples measured in Data Set B (ie, when POSTN and DPP4 were measured in samples collected at -1 and 40 weeks), the following objectives were: (i) Serum within the mITT population Assessing the effect of treatment with benralizumab on periostin levels by baseline blood eosinophils and baseline POSTN levels (exploratory outcome variables were blood eosinophils and serum POSTN); and (ii) mITT population Planned to evaluate the effect of treatment with benralizumab on serum DPP4 levels in vivo by baseline blood eosinophils and baseline DPP4 levels (exploratory outcome variables were blood eosinophils and serum DPP4) .

  To assess the therapeutic effect of benralizumab on efficacy endpoints, the following analyzes were performed: (i) any dose response effects were evaluated, placebo (combined EOS + and EOS−), 2 mg, 20 mg and 100 mg (combined) Evaluation by EOS + and EOS−); and (ii) Evaluation by placebo (combined EOS + and EOS−), pooled group 20 mg and 100 mg dose groups (EOS + and EOS−).

  In an optional subgroup analysis for a specific biomarker cutoff, a high eosinophil population is defined as blood eosinophils ≧ 300 cells / μL and a low eosinophil population is blood eosinophils <300 cells / μL. It is defined as High POSTN and high DPP4 are defined as subjects with a level ≧ median, and low POSTN and low DPP4 are defined as subjects with a level <median.

The current and predicted output for Dataset A is as follows:
(A) Descriptive statistics table of median, mean, standard deviation and range of periostin and DPP4 within the mITT population to assess POSTN and DPP4 levels within the study population and within other clinical subgroups of interest (Fig. 2).
(B) Median, mean, standard deviation and range of POSTN and DPP4 in reversible and irreversible and ICS medium and high ICS subpopulations and in blood eosinophils ≧ 300 and <300 cells / μL subpopulations A table of descriptive statistics will be created.
(C) Demographic and clinical characteristics of subjects with high POSTN, low POSTN, high DPP4 and low DPP4 to show relevance to specific clinical features or treatment when DPP4 or POSTN is divided into subgroups A feature table will be created. Demographic and clinical features for use in the tables are age, gender, BMI, FEV1 (L), FEV1 (% prediction), reversibility, ACQ, ICS dose, OCS dose, (from the asthma history questionnaire) Asthma exacerbations over the past 12 months, asthma symptom diary card score, serum IgE, and blood eosinophil count.
(D) A plot of baseline serum POSTN vs. baseline DPP4 correlation will be generated to show the relationship between POSTN and DPP4 within the baseline study population.
(E) A plot of baseline serum POSTN vs. baseline blood eosinophils will be generated to show the relationship between POSTN and blood eosinophils within the baseline study population.
(F) To show the relationship between DPP4 and blood eosinophils within the baseline test population, a plot of baseline serum DPP4 versus baseline blood eosinophils will be generated.
(G) High POSTN and low POSTN subjects with mITT, pulmonary function reversible subjects, pulmonary function irreversible subjects, high ICS subjects and ICS medium subjects to assess overlap between different subgroups within the study population Drawing of overlapping blood eosinophils> 300, <300 cells / μL.
(H) High DPP4 and low DPP4 subjects by mITT, pulmonary function reversible subjects, pulmonary function irreversible subjects, high ICS subjects and ICS medium subjects to assess overlap between different subgroups within the study population Drawing of overlapping blood eosinophils> 300, <300 cells / μL.
(I) (i) rate of exacerbation by baseline serum POSTN by treatment group, (ii) change in FEV1 by baseline serum POSTN by treatment group, (iii) change in ACQ by baseline serum POSTN by treatment group, (iv) Rate of exacerbation by baseline serum DPP4 by treatment group, (v) Change in FEV1 by baseline serum DPP4 by treatment group, (vi) Change in ACQ by baseline serum DPP4 by treatment group, (vii) Baseline serum by treatment group POSTN vs. baseline blood eosinophil exacerbation rate, (viii) Baseline serum DPP4 vs. baseline blood eosinophil exacerbation rate by treatment group, (ix) Baseline serum POSTN vs. baseline blood eosinophil by treatment group FE by (X) change in ACQ by baseline serum POSTN vs. baseline blood eosinophils by treatment group, (xi) change in FEV1 by baseline serum DPP4 vs. baseline blood eosinophils by treatment group, and ( xii) A plot will be generated showing the effect of benralizumab on baseline serum DPP4 by treatment group vs. change in ACQ by baseline blood eosinophils.
(J) To assess the effect of benralizumab on efficacy endpoints within a particular subpopulation, use high or low baseline serum POSTN for exacerbation rates, changes in FEV1 from baseline and changes in ACQ from baseline. A plot of the effects of benralizumab in subjects with was made for the following subgroups:
(I) high POSTN and low POSTN (mITT only),
(Ii) high and low POSTN in subjects with reversible lung function;
(Iii) Each of the above groups in the high POSTN + eosinophil ≧ 300 and <300 cells / μL subgroup and the low POSTN + eosinophil ≧ 300 and <300 cells / μL subgroup.
(K) To assess the effects of benralizumab on efficacy endpoints within a particular subpopulation, use high or low baseline serum POSTN for exacerbation rates, changes in FEV1 from baseline and changes in ACQ from baseline A plot of the effects of benralizumab in the subject will be generated for the following subgroups:
(I) high and low POSTN in subjects with irreversible lung function,
(Ii) high and low POSTN in subjects using high dose ICS,
(Iii) high POSTN and low POSTN in subjects using medium dose ICS,
(Iv) high and low POSTN in subjects not using chronic OCS;
(V) high and low POSTN in subjects who are not using chronic OCS and are ≧ 12% reversible at baseline,
(Vi) Each of the above groups in the high POSTN + eosinophil ≧ 300 and <300 cells / μL subgroup and the low POSTN + eosinophil ≧ 300 and <300 cells / μL subgroup.
(L) To assess the effects of benralizumab on efficacy endpoints within a particular subpopulation, use high or low baseline serum DPP4 for exacerbation rates, changes in FEV1 from baseline and changes in ACQ from baseline. A plot of the effects of benralizumab in subjects with was made for the following subgroups:
(I) high DPP4 and low DPP4 (mITT only),
(Ii) high and low DPP4 in subjects with reversible lung function,
(Iii) Each of the above at high DPP4 + eosinophils ≧ 300 and <300 cells / μL and low DPP4 + eosinophils ≧ 300 and <300 cells / μL.

To assess the effects of benralizumab on efficacy endpoints within a particular subpopulation in subjects with high or low baseline serum DPP4 to exacerbation rate, changes in FEV1 from baseline and changes in ACQ from baseline A plot of the effects of benralizumab will be generated for the following subgroups:
(I) high and low DPP4 in subjects with irreversible lung function,
(Ii) high and low DPP4 in subjects using high dose ICS,
(Iii) high and low DPP4 in subjects using medium dose ICS,
(Iv) high and low DPP4 in subjects not using chronic OCS,
(V) high and low DPP4 in subjects who are not using chronic OCS and are ≧ 12% reversible at baseline,
(Vi) Each of the above at high DPP4 + eosinophils ≧ 300 and <300 cells / μL and low DPP4 + eosinophils ≧ 300 and <300 cells / μL.

The predicted output in data set B is as follows.
(A) Descriptive statistics showing POSTN levels within the study population at baseline and after treatment with benralizumab and within other clinical subgroups of interest are within the mITT population, at -1 and 40 weeks The median, average, standard deviation and range of POSTN will be presented in a descriptive statistics table.
(B) Descriptive statistics showing DPP4 levels in the study population at baseline and after treatment with benralizumab and within other clinical subgroups of interest are within the mITT population, at -1 and 40 weeks. The DPP4 median, mean, standard deviation and range will be presented in a descriptive statistics table.
(C) Changes in baseline POSTTN versus baseline blood eosinophil levels by treatment group (ie after benralizumab treatment and placebo) to show the effect of benralizumab on serum POSTN levels by raising baseline blood eosinophil levels Will create a plot.
(D) Changes in baseline DPP4 versus baseline blood eosinophil levels by treatment group (ie after benralizumab treatment and placebo) to show the effect of benralizumab on serum DPP4 levels by increasing baseline blood eosinophil levels Will create a plot.
(E) To show the effect of treatment with benralizumab on serum POSTN levels by test population and subgroup, the following populations were: (i) mITT, (ii) blood eosinophils ≧ 300 and <300 cells / μL, (iii) ) FEV1 reversible and irreversible subjects, (iv) A plot of the effect of benralizumab on POSTN levels from weeks -1 to 40 in high ICS and ICS medium will be generated.
(F) To demonstrate the effect of treatment with benralizumab on serum DPP4 levels by test population and subgroup, the following populations were: (i) mITT, (ii) blood eosinophils ≧ 300 and <300 cells / μL, (iii) A plot of the effects of benralizumab on DPP4 levels from week -1 to week 40 in (iv) high ICS and ICS medium and FIV1 reversible and irreversible subjects will be generated.

Example 2
DPP4 ELISA Immunoassay The DPP4 detection assay disclosed herein is a quantitative ELISA based immunoassay. A mouse monoclonal antibody specific for human DPP4 was pre-coated on a microplate (R & D Systems, catalog number DC260). After first adding 100 μL of assay diluent to the wells of the microplate, 50 μL of standard, control and 1:50 diluted serum samples were added. The plate was incubated at room temperature for 2 hours ± 15 minutes to allow DPP4 to bind to the capture antibody on the plate.

  To remove unbound material, the plate was then washed 4 times with 1 × wash buffer (MedImmune) using a plate washer. The plates were then incubated with 200 μL of detection antibody (polyclonal anti-DPP4 antibody-HRP complex, R & D Systems, catalog number DC260) for 2 hours ± 15 minutes at room temperature. The plates were then washed and incubated with 200 μl of chromogenic HRP substrate, hydrogen peroxide / tetramethylbenzidine (R & D Systems, catalog number DC260) for 30 minutes ± 5 minutes at room temperature protected from light.

  The enzyme reaction was stopped by adding 50 μL of stop solution (2N sulfuric acid). To measure the optical density at 450 nm, readings were taken on a SpectraMaxPlus 384 microplate spectrophotometer (Molecular Devices) within 30 minutes after stopping the reaction plate. The intensity of the color produced was directly proportional to the amount of DPP4 bound. The DPP4 concentration in the sample and control was interpolated from a calibration curve of recombinant human DPP4 run on each plate. A secondary model was used for curve fitting. The derived DPP4 concentration was adjusted for initial sample dilution.

  The quantifiable range was 16 ng / mL to 1000 ng / mL DPP4 in 100% serum. The assay was reproducible, accurate, and specific for DPP4. Similarity between the recombinant standard and endogenous serum DPP4 has been demonstrated. The assay has acceptable matrix interference, 3 freeze-thaw cycle stability and 1.5 year long-term stability at -80 ° C.

Example 3
Serum biomarkers with high blood eosinophil baseline or low IL-13 pathway activation predict reduction in exacerbation rate with benralizumab in patients with moderate to severe asthma The humanized monoclonal antibody benralizumab is eosinophilic inflammation Selectively depletes eosinophils and basophils through enhanced antibody-dependent cytotoxicity, reduced exacerbations in the Ph2b test (NCT01238861, see FIG. 1) in patients with moderate to severe asthma with (Castro et al., Lancet Respir Med. 2: 878-90 (2014)). Dipeptidyl peptidase (DPP4) and POSTN (POSTN) are upregulated by IL-13 and are promising biomarkers of response to anti-IL-13 treatment (Brightling et al., Lancet Respir Med. 3: 692-701). (2015)). We explored the ability of baseline levels of serum DPP4 and POSTN to predict a reduction in exacerbation rates with benralizumab in the Ph2 test.

  Serum DPP4 was measured using the immunoassay of Example 2, and POSTN was measured using an immunoassay manufactured by Abbott Diagnostics. Asthmatic subjects have high or low DPP4 or POSTN (defined as serum concentrations ≧ or <median, respectively) or high or low eosinophils (baseline blood eosinophils ≧ or <300 cells / μL, respectively) Divided into subgroups). Groups were further divided to combine high and low DPP4 or high and low POSTN with high and low eosinophils (FIG. 3).

  The effect of benralizumab (20 mg and 100 mg subcutaneous treatment arm combination) was evaluated relative to placebo for exacerbation rate in each subgroup.

  Median serum DPP4 and POSTN concentrations ranged from 363.5 ng / mL (range from 103.3 ng / mL to 867.5 ng / mL) and 23.6 ng / mL (range from 7.5 ng / mL to 104.1 ng / mL, respectively) )Met. Please refer to FIG.

  Serum DPP4 did not correlate with blood eosinophils (correlation coefficient = 0.05, P = 0.22) or serum POSTN (correlation coefficient = 0.03, P = 0.48). Serum POSTN correlated with blood eosinophils (correlation coefficient = 0.33, P <0.001) (FIG. 4).

  High eosinophils, low DPP4 and low POSTN significantly predicted a reduction in exacerbation rates with benralizumab treatment. In the high subgroup of eosinophils, the decrease in exacerbation rate was independent of POSTN levels. In the low subgroup of eosinophils, low POSTN showed a numerically greater exacerbation rate reduction than high POSTN. Low DPP-4 predicted a decrease in exacerbation rate in high eosinophils and identified a numerically greater decrease in exacerbation rate in the low eosinophil subgroup (FIGS. 5, 6 and 7). See also Table 4.

  Reduction in exacerbation rate (ERR) was performed with a Poisson regression model with the logarithm of the follow-up period as the offset period and the baseline ICS status as the covariate. Overdispersion correction was given by Pearson's chi-square statistic divided by degrees of freedom and applied to standard error and likelihood ratio statistics to be adjusted accordingly.

  Conclusion: High baseline blood eosinophil concentrations predicted the efficacy of benralizumab and the effect was independent of POSTN in this subgroup. Baseline DPP-4 concentrations below the median indicating low IL-13 pathway activation predict the efficacy of venralizumab and identify patients with low blood eosinophil responsiveness to benralizumab. DPP4, a biomarker of IL-13 pathway activation, identified a patient subset that would benefit from benralizumab depleting anti-IL-5R / eosinophils (low DPP4) and IL-13 targeted traloquinumab (high DPP4) obtain.

  It should be understood that the detailed description section, rather than the summary and summary section, is intended to be used for interpreting the scope of the claims. The Summary and Summary sections may illustrate one or more, but not all, exemplary embodiments of the invention, as discussed by the inventors, and therefore the invention and the appended claims. It is never intended to limit.

  The present invention has been described above using functional components and exemplifies the implementation of its specific functions and relationships. The boundaries of these functional components are arbitrarily defined herein for convenience of explanation. Another boundary may be defined as long as its specific functions and relationships are properly implemented.

  The foregoing description of specific embodiments will fully demonstrate the general nature of the invention, so that others can apply the knowledge of those skilled in the art without undue experimentation, Various applications, such as specific embodiments, can be easily modified and / or adapted without departing from the general concept of the invention. Accordingly, such adaptations and modifications are intended to be included within the meaning and scope of equivalents of the disclosed embodiments based on the teachings and guidance presented herein. The expressions or terms in this specification are for purposes of explanation and are not intended to be limiting, so the terms or expressions herein are to be interpreted by those skilled in the art in view of the teachings and guidance. It should be understood that it should.

  The breadth and scope of the present invention should not be limited by any of the above-described exemplary embodiments, but should be defined only in accordance with the following claims and their equivalents.

  All publications, patents, patent applications, and / or other documents cited in this application are incorporated by reference for each purpose as if each individual publication, patent, patent application, and / or other document was included. Are incorporated by reference in their entirety for all purposes to the same extent as individually indicated.

Claims (62)

  A method of treating an eosinophilic disease or disorder in a patient, wherein the patient is compared to a predetermined DPP4 threshold level or in one or more controls in one or more samples taken from the patient. Administering to the patient an eosinophil-targeted therapeutic if determined or identified as having a dipeptidyl peptidase-4 (DPP4) level that is lower or reduced compared to a DPP4 threshold level in the sample; Method.   A method of treating a patient having an eosinophilic disease or disorder, wherein the patient is compared to a predetermined DPP4 threshold level in one or more samples taken from the patient, or one or more Discontinuing or not starting administration of the eosinophil targeted therapeutic to said patient if it is determined or identified as having a higher or increased DPP4 level compared to the DPP4 threshold level in the control sample ,Method.   A method of treating an eosinophilic disease or disorder in a patient who has not been successful, unresponsive or intolerant to treatment with a therapeutic agent, wherein the patient is collected from the patient If it is determined or identified to have a DPP4 level that is lower or reduced compared to a predetermined DPP4 threshold level or compared to a DPP4 threshold level in one or more control samples Administering an eosinophil-targeted therapeutic to the patient.   A method for determining whether a patient having an eosinophilic disease or disorder should be treated with an eosinophil targeted therapeutic agent, wherein the patient is in one or more samples taken from the patient. If it is determined or identified to have a lower or decreased DPP4 level compared to a predetermined DPP4 threshold level or compared to a DPP4 threshold level in one or more control samples A method comprising the steps of:   A method of selecting a patient diagnosed with an eosinophilic disease or disorder as a candidate for treatment with an eosinophil-targeted therapeutic agent, wherein the patient is in one or more samples taken from the patient. If the patient is determined or identified as having a DPP4 level lower or reduced compared to a predetermined DPP4 threshold level or compared to a DPP4 threshold level in one or more control samples A method comprising the step of selecting.   Measuring the level of the DPP4 in one or more of the samples obtained from the patient, or instructing a clinical laboratory or health care provider to measure the level of the DPP4 in the sample, and / or 6. The method of any one of claims 1-5, further comprising submitting the one or more samples obtained from the patient to a clinical laboratory or health care provider to measure the level of the DPP4 in the sample. The method according to item.   7. The method of any one of claims 1-6, further comprising determining the level of the DPP4 in the one or more samples obtained from the patient. (A) the patient is lower or reduced in one or more samples taken from the patient compared to a predetermined DPP4 threshold level or compared to a DPP4 threshold level in one or more control samples When determined or identified as having DPP4 levels, an eosinophil targeted therapeutic is administered to the patient, or (b) the patient is pre-determined in one or more samples taken from the patient. Said patient with an eosinophil-targeted therapeutic if it is determined or identified as having a higher or increased DPP4 level compared to a DPP4 threshold level of or a DPP4 threshold level in one or more control samples 8. The method according to any one of claims 1 to 7, further comprising the step of advising a healthcare provider to discontinue, not initiate or refuse the administration to .   9. The method according to any one of claims 1 to 8, wherein the eosinophil targeted therapeutic agent is a monoclonal antibody or an antigen-binding fragment thereof.   10. The method of claim 9, wherein the monoclonal antibody or antigen-binding fragment thereof specifically binds to IL-5 or IL-5R.   11. The method according to claim 10, wherein the monoclonal antibody or antigen-binding fragment thereof that specifically binds to IL-5 is mepolizumab, reslizumab, or an antigen-binding fragment thereof.   11. The method of claim 10, wherein the monoclonal antibody or antigen-binding fragment thereof that specifically binds to IL-5R specifically binds to the α subunit of IL-5R (SEQ ID NO: 4).   13. The method of claim 12, wherein the monoclonal antibody or antigen-binding fragment thereof that specifically binds to the α subunit of IL-5R is benralizumab or an antigen-binding fragment thereof.   The monoclonal antibody or antigen-binding fragment thereof that specifically binds to the α subunit of IL-5R comprises a heavy chain variable region (VH) comprising or consisting of SEQ ID NO: 12, and / or SEQ ID NO: 13 125. The method of claim 124, comprising a light chain variable region (VL) comprising or consisting of SEQ ID NO: 13.   One, two, or three complementarity determining regions selected from SEQ ID NOs: 14-16, and / or wherein the monoclonal antibody or antigen-binding fragment thereof specifically binds to the α subunit of IL-5R, and / or 13. The method of claim 12, comprising one, two or three complementarity determining regions selected from SEQ ID NOs: 17-19.   The monoclonal antibody or antigen-binding fragment thereof that specifically binds to the α subunit of IL-5R is at least one, two, three, four, five, or selected from SEQ ID NOs: 14-19 13. A method according to claim 12, comprising 6 complementarity determining regions.   19. The monoclonal antibody or antigen-binding fragment thereof binds to the same epitope as one of the monoclonal antibodies or antigen-binding fragment thereof according to any one of claims 12-18, or any one of claims 12-18. 10. The method of claim 9, wherein the binding of one of the monoclonal antibodies according to one or antigen binding fragments thereof is competitively inhibited or both.   The eosinophil-targeted therapeutic agent is a fusion protein comprising one of the monoclonal antibodies according to any one of claims 12 to 19 or an antigen-binding fragment thereof. The method described in 1.   18. The eosinophil targeted therapeutic agent is a complex comprising one of the monoclonal antibodies according to any one of claims 12 to 19 or an antigen-binding fragment thereof. The method described in 1.   20. The method of claim 19, wherein the fusion protein or complex comprises at least one heterologous therapeutic moiety and / or a half-life enhancing moiety.   9. The method according to any one of claims 1 to 8, wherein the eosinophil targeted therapeutic agent is a polynucleotide.   The method of claim 21, wherein the polynucleotide is DNA or RNA.   23. The method of claim 21, wherein the polynucleotide is (i) mRNA or a combination thereof, or (ii) an antisense oligonucleotide or a combination thereof.   24. The method of claim 23, wherein the antisense oligonucleotide or combination thereof is ASM8, PXS1100, or PXS2200.   25. The method of any one of claims 21 to 24, wherein the polynucleotide comprises at least a nucleotide analog.   26. The method of any one of claims 1-25, wherein the eosinophil targeted therapeutic is administered at a constant dose.   27. The method of claim 26, wherein the constant dose is 1-1000 mg / dose.   28. The method of any one of claims 1-27, wherein the eosinophil targeted therapeutic is administered in two or more doses.   29. The method of any one of claims 1-28, wherein the eosinophil targeted therapeutic is administered weekly, biweekly, bimonthly, and every 3 months.   30. The eosinophil targeted therapeutic agent is administered orally, by inhalation, intravenously, intramuscularly, subcutaneously, or a combination thereof. Method.   31. A method according to any one of claims 1 to 30, wherein the eosinophilic disease or disorder is a pulmonary disease or disorder.   32. The method of claim 31, wherein the pulmonary disease or disorder is asthma.   The asthma is allergic asthma, atopic asthma, corticosteroid naive asthma, chronic asthma, corticosteroid resistant asthma, corticosteroid refractory asthma, asthma caused by smoking, or asthma not controlled by corticosteroid 35. The method of claim 32, wherein:   35. The method of claim 32, wherein the asthma is mild to moderate asthma or severe asthma.   The eosinophilic disease or disorder is chronic bronchitis, chronic eosinophilic pneumonia (CEP), nasal polyp, atopic dermatitis (eczema), eosinophilic esophagitis, hyperacidic syndrome (HES), Eosinophilic granulomatosis and polyangiitis (Charg Strauss syndrome), eosinophilic gastritis, eosinophilic enteritis, eosinophilic colitis, allergic rhinoconjunctivitis (hay fever), leukemia, lymphoma Mastocytosis, atheroembolic disease, hyper IgE syndrome, omen syndrome, thymoma, transplant rejection, hypoadrenalism, bullous pemphigoid, pemphigus vulgaris, herpes dermatitis, drug-induced lesions, Urticaria, eosinophilic panniculitis, angioedema with eosinophilia, Kimura disease, Schulman syndrome, Wells syndrome, eosinophilic ulcer of the oral mucosa, eosinophilic pustular folliculitis, relapse Skin necrotic eosinophilic vasculitis, drug / Element-induced eosinophilic lung disease, Lefler syndrome, allergic bronchopulmonary aspergillosis, eosinophilic granuloma, pleural eosinophilia, gastroesophageal reflux disease, parasitic infection, fungal infection, Helicobacter Helicobacter pylori infection, inflammatory bowel disease (ulcerative colitis and Crohn's disease), food allergic disorders, protein-induced enteropathy, protein-induced enterocolitis, allergic colitis, celiac disease, proliferative Pemphigus, leiomyomatosis, connective tissue disorder, vasculitis disorder, chronic subdural hematoma, central nervous system infection, ventricular abdominal shunt, congenital heart disease (septal defect or aortic stenosis), renal infection 31. The method according to any one of claims 1 to 30, which is eosinophilic urinary disease, interstitial nephritis, or eosinophilic cystitis.   The patient may use one or more additional therapeutic agents for the treatment of the eosinophilic disease or disorder before, during, after, or alternatively, administration of an eosinophil targeted therapeutic agent 36. The method of any one of claims 1-35, being treated with any of them.   40. The method of claim 36, wherein the one or more additional therapeutic agents further comprise a steroid, a bronchodilator, or both.   38. The method of claim 37, wherein the steroid is fluticasone, budesonide, mometasone, or beclomethasone.   38. The method of claim 37, wherein the bronchodilator is salbutamol or salmeterol.   40. The method of claim 36, wherein the one or more additional therapeutic agents are administered by inhalation, by oral administration, by injection, or a combination thereof.   41. The method of claim 40, wherein inhalation administration is performed using a metered dose inhaler (MDI) or a dry powder inhaler (DPI).   38. The method of claim 37, wherein the steroid is administered at a high dose.   The one or more samples taken from the patient and / or the one or more control samples are whole blood, serum, plasma, saliva, sputum, bronchoalveolar lavage fluid, lung epithelial cells, urine, skin, nasal passages, 43. A method according to any one of claims 1-42, comprising one or more of or a combination thereof.   44. The method of claim 43, wherein the sample taken from the patient is serum. The one or more control samples are
(A) one or more samples,
(I) a normal healthy person,
(Ii) a patient having a subset of asthma,
(Iii) one or more samples obtained from a naïve asthma patient or a corticosteroid treated asthma patient for corticosteroid treatment;
45. The method of any one of claims 1-44, wherein (b) a predetermined standard amount of isolated DPP4, or (c) any combination sample in (a) and (b). Determining a sample taken from the patient, submitting for determination, or (i) a level of the patient's IgE level,
(Ii) the patient's eosinophil count and / or basophil count,
(Iii) the patient's exhaled nitric oxide concentration (FENO),
(Iv) the patient's eosinophil / lymphocyte and eosinophil / neutrophil (ELEN) index;
(V) the patient's EOS index,
(Vi) Percentage wall area percentage (WA%) of the subregion airway from a CT scan of the lung of the patient,
46. The method of any one of claims 1-45, further comprising: (vii) instructing a clinical laboratory to determine a combination of two or more thereof.   47. The method of claim 46, wherein the patient's eosinophil count is ≧ 150 eosinophils / μL, ≧ 300 eosinophils / μL, or ≧ 400 eosinophils / μL.   49. The method of claim 48, wherein the patient has an eosinophil count of 150-400 eosinophils / [mu] L.   49. The method of any one of claims 1-48, wherein the patient's DPP4 level is measured with a DPP4 detection assay which is an immunoassay. The immunoassay comprises
52. The method of claim 51, wherein the method is (i) the immunoassay described in Example 2, or (ii) the immunoassay disclosed in Table 1, or (ii) a multiplex immunoassay.   The multiplex immunoassay is EMD Millipore MILLIPLEX (trademark), Bio-Rad BIO-PLEX (trademark), Life Technologies NOVEX MULTICELPEX (trademark), Thermo Fisher Scientific PLU (trademark). 51. The method of claim 50, which is a (trademark) or R & D Systems LUMINEX (R) assay.   Sample collected from said patient, submitted for determination, or periostin, IL-22, IL-25, IL-33, TSLP, LCN2, CCL20, sCTLA-3, sCD28, CCL5, CCL11 , CCL22, CST1, CCL26, CLCA1, CST2, PRR4, SERPIN2, CEACAM5, iNOS, SERPINB4, CST4, PRB4, TPSD1, TPSG1, MFSD2, CPA3, GPR105, CDH26, GSN, C2ORF32, TRACH200019G13 Expression level or activity of SLC18A2, SERPINB10, SH3RF2, FCER1B, RUNX2, PTGS1, ALOX15, or a combination thereof. Further comprising the step of instructing the laboratory to constant A method according to any one of claims 1 to 51. The predetermined DPP4 threshold level is
(A) an approximate average DPP4 level measured in sera from multiple patients using an immunoassay;
(B) approximately median DPP4 levels measured in sera from multiple patients using an immunoassay, and (c) approximately first, second measured in sera from multiple patients using an immunoassay. , Third, fourth, fifth, sixth, seventh, eighth, or ninth quartile baseline DPP4 level, wherein the patient is
(I) normal healthy volunteers,
53. The method according to any one of claims 1 to 52, wherein the method is (ii) a patient with mild to moderate asthma and / or (iii) a patient with severe asthma.   54. The method of claim 53, wherein the serum from a plurality of patients is a pooled sample or individual samples.   55. The method of any one of claims 1 to 54, wherein the predetermined DPP4 threshold level is at least about 103 ng / mL to at least about 867 ng / mL as measured using an immunoassay.   55. The method of any one of claims 1 to 54, wherein the predetermined DPP4 threshold level is at least about 363 ng / mL as measured using an immunoassay.   55. The method of any one of claims 1 to 54, wherein the predetermined DPP4 threshold level is at least about 376 ng / mL as measured using an immunoassay.   58. The method according to any one of claims 55 to 57, wherein the immunoassay is the DPP4 detection assay described in Example 2 or the DPP4 detection assay disclosed in Table 1. Administration of said eosinophil targeted therapeutic agent comprises:
(A) decrease in AER (acute exacerbation rate),
(B) Increase in FEV1 (forced expiratory volume per second),
59. The method of any one of claims 1 to 58, which results in (c) an improved ACQ-6 (asthma management questionnaire, 6 item version) value, or (d) a combination thereof.   Said decrease in AER is at least about 15%, 20%, 25%, 30%, 35%, 40%, 45%, compared to an AER baseline value observed in a population of patients treated with placebo, 60. The method of claim 59, wherein the method is 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%.   The increase in FEV1 is at least about 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300 compared to FEV1 baseline values observed in a population of patients treated with placebo. 60. The method of claim 59, wherein the method is 325, 350, 375, 400, 425, 450, 475, or 500 mL.   The improved ACQ-6 value is at least about −0.3, −0.4, −− from baseline compared to an ACQ-6 baseline value observed in a population of patients treated with placebo. 60. The method of claim 59, wherein the method is a change of 0.5, -0.6, -0.7, -0.8, -0.9, -1, -1.1, or -1.2.