Anti-CD6 antibodies for the treatment of severe asthma

Abstract

The present invention provides compositions and methods for using the anti-CD6 antibody itolizumab or its antigen-binding portion to treat, improve and prevent asthma, and especially steroid-resistant or refractory severe asthma, wherein the anti-CD6 antibody or its antigen-binding portion is used alone or in combination with other agents that can be used to treat asthma.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to U.S. Provisional Application No. 62 / 636,092, filed on February 27, 2018, the contents of which are hereby incorporated by reference in their entirety.

[0003] Sequence Listing Statement

[0004] The sequence listing associated with this application is provided in text format in lieu of a paper copy and is hereby incorporated by reference into the specification. The name of the text file containing the sequence listing is EQIL_006_01WO_ST25.txt. The text file is 5KB, was created on February 27, 2019, and will be submitted electronically via EFS-Web. Background Art Technical Field

[0005] The present invention relates to compositions and methods for treating severe asthma. Asthma is a disease in which: (i) bronchial constriction; (ii) excessive mucus production; and (iii) inflammation and swelling of the airways occur, causing widespread and variable airflow obstruction, making it difficult for asthmatic patients to breathe. Asthma is a chronic disease primarily characterized by persistent airway inflammation. However, asthma is further characterized by acute episodes of additional airway constriction caused by hyperresponsive airway smooth muscle contraction.

[0006] In asthma, chronic inflammatory processes in the airways play a central role in increasing resistance to airflow within the lungs.

[0007] The chronic nature of asthma can also lead to airway wall remodeling (i.e., structural changes such as thickening or edema), which can further affect the function of the airway wall and influence airway hyperresponsiveness. Other physiological changes associated with asthma include excess mucus production and, if asthma is severe, mucus plugging, and constant epithelial exfoliation and repair. Epithelial exfoliation exposes underlying tissues to substances that would not normally come into contact with them, further enhancing the cycle of cellular damage and inflammatory response.

[0008] In susceptible individuals, asthma symptoms include recurrent episodes of shortness of breath (dyspnea), wheezing, chest tightness and coughing. Currently, asthma is managed through a combination of irritant avoidance and pharmacology, depending largely on the subtype of asthma the patient has.

[0009] Asthma is a heterogeneous disease. Based on the severity of asthma, the National Asthma Education and Prevention Program (NAEPP) of the United States classifies it into four major clinical categories, including: (1) intermittent asthma; (2) mild persistent asthma; (3) moderate persistent asthma; and (4) severe persistent asthma. On the other hand, the 2016 Global Initiative for Asthma (GINA) classifies asthma severity as mild, moderate, or severe, with severity assessed based on the level of treatment required to control symptoms and exacerbations.

[0010] For example, NAEPP has the following clinical features:

[0011] Intermittent asthma is characterized by:

[0012] Symptoms of cough, wheezing, chest tightness, or dyspnea occur less than twice a week; outbreaks are brief but may vary in intensity; nighttime symptoms occur less than twice a month; no symptoms between outbreaks; lung function test FEV1 is 80% of normal or higher; peak flow varies less than 20% from morning to morning or from morning to afternoon each day (worldwide webpage: / / emedicine.medscape.com / article / 296301-guidelines#g2).

[0013] Mild persistent asthma is characterized by:

[0014] Symptoms of cough, wheezing, chest tightness, or dyspnea 3-6 times per week; outbreaks may affect activity level; symptoms at night

[0015] 3-4 times a month; lung function test FEV 1 is above 80% of normal or higher; peak flow changes are less than

[0016] 20%-30% (Id.).

[0017] Moderate persistent asthma is characterized by:

[0018] Symptoms of cough, wheezing, chest tightness, or dyspnea occur daily; flare-ups may affect activity level; symptoms occur more frequently at night

[0019] 5 or more times a month; FEV1 on lung function test is above 60% of normal but below 80% of normal;

[0020] Peak flow rate variation greater than 30% (Id.).

[0021] Severe persistent asthma is characterized by:

[0022] Persistent cough, wheezing, chest tightness, or dyspnea; frequent nocturnal symptoms; lung function test FEV 1

[0023] 60% or less of normal; peak flow variation greater than 30% (Id.).

[0024] In addition, there are four different phenotypes of airway inflammation, depending on the type of inflammatory cells, or lack thereof, in the asthmatic airways: eosinophilic inflammation, neutrophilic inflammation, mixed inflammation, and agranulocytic inflammation. As discussed further below, neutrophilic inflammation, mixed inflammation, and agranulocytic inflammation are forms of non-eosinophilic asthma.

[0025] Eosinophilic inflammation, commonly known as eosinophilic asthma or allergic asthma, is considered to be a product of Th2-mediated inflammatory response (commonly known as type 2 high), accompanied by the production of IL-4, IL-5 and IL-13, resulting in an increase in lung eosinophils. Eosinophils secrete additional cytokines that enhance potential inflammation and worsen asthma symptoms. Clinically, in contrast to the non-eosinophilic form, the differentiation of eosinophilic asthma is mainly based on the level of circulating eosinophils found in blood samples, which is used as a substitute for understanding the level of eosinophils that may be present in the lungs. Typically, if the patient's blood eosinophil count is ≤300 cells / μl, the patient is considered to have a "low" blood eosinophil count. Such patients suffer from non-eosinophilic asthma. In contrast, patients with blood eosinophil counts>300 cells / μl are considered to suffer from eosinophilic asthma.

[0026] Neutrophilic inflammation, commonly known as neutrophilic asthma or non-allergic asthma, is considered to be a product of a Th1 / Th17-mediated inflammatory response (commonly known as type 2 low), accompanied by the production of IFN-γ, IL-6, IL-17, and IL-8, leading to an increase in lung neutrophils. Neutrophils secrete additional cytokines that enhance underlying inflammation and worsen asthma symptoms. Typically, patients with neutrophilic asthma show blood neutrophil counts > 4600 cells / μl, for example (Vedel-Krogh, S. et al., Clin Chem. 2017Apr; 63(4): 823-832; Schleich, F. et al., BMC Pulm Med. 2013; 13: 11).

[0027] Mixed inflammation is a phenotype in which a patient can simultaneously display eosinophilic inflammation and / or neutrophilic inflammation, although the levels of either granulocyte may be low.

[0028] Agranulocytic inflammation is a phenotype in which patients exhibit lung inflammation despite normal levels of eosinophils and neutrophils. Agranulocytic inflammation is also a form of non-allergic asthma and is thought to be the product of a Th1 / Th17-mediated inflammatory response (often referred to as type 2 low), with the production of IFN-γ, IL-6, and IL-17. Of note, patients with agranulocytic asthma typically do not have detectable blood eosinophil counts.

[0029] The heterogeneity of asthma is Figure 7 Figure 1 shows a graph of the relationship between inflammation type (e.g., high Th2 or low Th2) and the current understanding of various asthma phenotypes. Although the phenotypes overlap, Th2-mediated responses are shown on the left side of the figure, and these responses are associated with allergic eosinophilic responses that respond to corticosteroid treatment. In contrast, the non-allergic, non-eosinophilic responses on the right side of the figure show low Th2 responses, have high Th1 responses and / or Th1 / Th17 responses, are neutrophilic or agranulocytic, and do not demonstrate responsiveness to corticosteroid treatment.

[0030] Current treatment / control options for asthma range from irritant avoidance to pharmacological and even surgical interventions.

[0031] Irritation avoidance is achieved by systematically identifying and minimizing exposure to each type of stimulus. However, avoiding all potential irritants may not be practical and is not always helpful.

[0032] Asthma is managed pharmacologically by: (1) long-term control through the use of anti-inflammatory drugs and long-acting bronchodilators; and (2) short-term control of acute exacerbations through the use of short-acting bronchodilators (e.g., beta agonists). Both approaches require repeated and regular use of prescription medications.

[0033] In clinical settings, asthma patients are most often tested for allergic asthma or other forms of asthma by analyzing their blood eosinophil counts to determine the best course of action for disease control. As further discussed herein, for patients with low or no eosinophil counts in the blood, steroid therapy is contraindicated. Although there is a spectrum of the exact boundaries for distinguishing low blood eosinophil counts from normal blood eosinophil counts or high blood eosinophil counts in the art, as used herein, "low" blood eosinophil counts mean that the blood eosinophil count of the object is ≤300 cells / μl. Such patients with low (or no) detectable blood eosinophils represent a high-risk population of severe asthma patients who are in urgent need of effective therapeutic agents.

[0034] Current treatments for asthma include: long-acting beta agonists (LABA), short-acting beta agonists (SABA), such as albuterol (ProAir HFA, Proventil HFA, Ventolin HFA), Metaproterenol, Levalbuterol (Xopenex HFA), and Pirbuterol (Maxair), which are administered through a quick-acting (rescue) inhaler that turns the drug into a mist that can be inhaled deeply into the patient's lungs or through a nebulizer. Corticosteroids are also used to treat asthma. These drugs, which are usually inhaled or taken in pill form, help reduce lung inflammation and control asthma symptoms. Corticosteroids can also be given intravenously, usually to patients who are vomiting or experiencing respiratory failure. Ipratropium (atrovent) is also sometimes used as a bronchodilator to treat severe asthma attacks, especially when albuterol is not fully effective. Table 1 provides a list of various pharmacological agents that are currently approved by the FDA for the treatment of severe asthma or are currently or have been in development for the treatment of asthma.

[0035] Table 1: Agents approved or in development for the treatment of severe asthma

[0036]

[0037] (As of January 2018)

[0038] Finally, if the asthma attack becomes life-threatening, intubation and mechanical ventilation with oxygen to help the person breathe may be needed while other medications are tried to control the asthma.

[0039] However, currently available treatments have drawbacks.

[0040] Difficulties associated with patient compliance with medication control and difficulty avoiding stimuli that trigger asthma are common barriers to successful asthma control. In addition, many of these therapies focus on alleviating the symptoms of asthma but do not address the underlying cause of the disease. Therefore, current control techniques are neither completely successful nor free of side effects.

[0041] High doses of corticosteroid anti-inflammatory drugs can have serious side effects that need to be carefully managed. In addition, some patients are resistant to or refractory to steroid therapy. These patients constitute another category of asthma patients, called severe asthma (SA) patients, who have received great clinical attention due to the severity of their disease and lack of response to traditional asthma treatments. In fact, although only 5% to 10% of asthma patients suffer from steroid-resistant or refractory disease, in the United States, Europe, and Australia, care for these patients accounts for 50%-80% of all asthma-related healthcare costs because these patients are often hospitalized and require emergency care (Hansbro et al., Immunological Reviews. 2017; 278: 41–62).

[0042] Severe asthma is most commonly associated with a nonallergic Th1 / Th17 phenotype, such as neutrophilic asthma or agranulocytic asthma, or with a mixed phenotype of Th1 / Th17-neutrophilic asthma / Th2-eosinophilic asthma, in which Th17 cells and neutrophils respond poorly to corticosteroids and sometimes fail to respond at all ( Figure 7 ).

[0043] Th2-mediated allergic eosinophilic asthma usually responds to corticosteroid therapy ( Figure 7 ).

[0044] Therefore, there is a need for new asthma treatments that selectively treat the underlying pathology of the disease. There is also a need for new asthma therapies that are effective in treating steroid-resistant or refractory forms of asthma (i.e., SA). The present invention provides compositions and methods for treating, preventing, and alleviating severe asthma, and in specific embodiments, severe asthma characterized by low blood eosinophil counts or no blood eosinophil counts. Summary of the invention

[0045] The present invention particularly relates to the treatment, prevention or alleviation of asthma, comprising administering an anti-CD6 antibody to a subject. In a specific embodiment, the anti-CD6 antibody is EQ001. In some embodiments, the asthma is severe asthma. In some embodiments, the asthma is characterized by low eosinophils or no eosinophils. In some embodiments, the asthma is neutrophilic asthma. In some embodiments, the asthma is agranulocytic asthma. In some embodiments, the asthma is mixed inflammatory asthma. In certain embodiments, the asthma is non-allergic asthma. In certain embodiments, the asthma is non-eosinophilic asthma.

[0046] In some embodiments, the present invention provides a method for inhibiting T cell-mediated pulmonary inflammation in a subject suffering from asthma, comprising: administering an anti-CD6 antibody or an antigen-binding fragment thereof to the subject, wherein the anti-CD6 antibody or the antigen-binding fragment thereof comprises: heavy chain and light chain variable regions, the heavy chain and light chain variable regions comprising the amino acid sequences shown in SEQ ID NOs: 1 and 2.

[0047] In some embodiments, the present invention provides a method of inhibiting T cell-mediated pulmonary inflammation in a subject suffering from asthma, comprising: administering an anti-CD6 antibody or antigen-binding fragment thereof to the subject.

[0048] In some embodiments, the present invention provides a method for preventing or reducing the migration of T cells into and through lung tissue in response to an asthma-inducing antigen, comprising: administering an anti-CD6 antibody or antigen-binding fragment thereof to a subject.

[0049] In some embodiments, the present invention provides a method of regulating or reducing the symptoms or severity of asthma, comprising: administering an anti-CD6 antibody or antigen-binding fragment thereof to a subject.

[0050] In some embodiments, the present invention provides a method for regulating or reducing the symptoms or severity of asthma, comprising: contacting T cells with an anti-CD6 antibody or an antigen-binding fragment thereof.

[0051] In some embodiments, the asthma is severe asthma. In some embodiments, the asthma is characterized by low blood eosinophils or absence of blood eosinophils. In some embodiments, the asthma is refractory to steroid therapy. In some embodiments, the asthma is neutrophilic asthma. In some embodiments, the asthma is mixed inflammatory asthma. In some embodiments, the asthma is agranulocytic.

[0052] In some embodiments, the anti-CD6 antibody or antigen-binding fragment thereof binds to a CD6 protein on the surface of a T cell. In some embodiments, the T cell is a Th1 T cell, a Th17 T cell, or a Th1 and Th17 T cell.

[0053] In some embodiments, the anti-CD6 antibody or its antigen-binding fragment is EQ001 or an antigen-binding fragment of EQ001. In some embodiments, the anti-CD6 antibody or its antigen-binding fragment binds to domain 1 or domain 3 on CD6. In some embodiments, the anti-CD6 antibody or its antigen-binding fragment binds to domain 3 on CD6. In some embodiments, the binding of the anti-CD6 antibody or its antigen-binding fragment to the CD6 protein on the surface of T cells regulates the activity and / or migration of T cells. In some embodiments, the anti-CD6 antibody or its antigen-binding fragment is a humanized antibody. In some embodiments, the anti-CD6 antibody or its antigen-binding fragment is selected from the group consisting of: UMCD6 mAb, Itolizumab (EQ001), the anti-CD6 antibodies described in Table 2, and the anti-CD6 antibodies disclosed herein. In some embodiments, the anti-CD6 monoclonal antibody is: an antibody produced by the secretory hybridoma IOR-T1A deposited in ECACC with the deposit number ECACC 96112640; an antibody having the same sequence as the antibody produced by the secretory hybridoma; or an antibody having the same CDR sequence as the antibody produced by the secretory hybridoma.

[0054] In some specific embodiments, any of the methods disclosed herein comprises administering EQ001. In some specific embodiments, any of the methods disclosed herein comprises administering an antigen binding fragment EQ001. In some embodiments, the antigen binding fragment is selected from the group consisting of: Fv, Fab, CDR1, CDR2, CDR3, a combination of CDRs, a variable region, a heavy chain, and a light chain.

[0055] In some embodiments, the anti-CD6 antibody or antigen-binding fragment thereof comprises one or more CDR sequences selected from SEQ ID NOs: 5-10. In some embodiments, the anti-CD6 antibody or antigen-binding fragment thereof comprises heavy and light chain variable regions, which comprise the amino acid sequences shown in SEQ ID NOs: 1 and 2. In some embodiments, SEQ ID NOs: 1 and 2 are encoded by SEQ ID NOs: 3 and 4, respectively. In some embodiments, the anti-CD6 antibody or antigen-binding fragment thereof comprises a VH sequence that is at least 80% identical to the amino acid sequence shown in SEQ ID NO: 1. In some embodiments, the anti-CD6 antibody or antigen-binding fragment thereof comprises a VK sequence that is at least 80% identical to the amino acid sequence shown in SEQ ID NO: 2. In some embodiments, the anti-CD6 antibody or antigen-binding fragment thereof comprises: a VH sequence that is at least 80% identical to the amino acid sequence shown in SEQ ID NO: 1 and a VK sequence that is at least 80% identical to the amino acid sequence shown in SEQ ID NO: 2.

[0056] In some embodiments, any of the methods of the present invention further comprises administering one or more additional agents capable of treating, preventing or alleviating one or more asthma-related symptoms. In some embodiments, additional agents include agents capable of modulating the immune system. In some embodiments, additional agents include agents that are immunosuppressants. In some embodiments, additional agents include long-acting beta agonists, short-acting beta agonists, or a combination thereof. In some embodiments, additional agents include salbutamol. In some embodiments, salbutamol is administered in a dosage form selected from the following: aerosol powder; solution; capsule; and powder suspension. In some embodiments, additional agents include corticosteroids. In some embodiments, corticosteroids are administered as inhalation formulations. In some embodiments, corticosteroids are administered in a dosage form selected from the following: tablets, sustained-release capsules; extended-release tablets; extended-release capsules; syrups; solutions; elixirs; suspensions; sustained-release tablets; liquids; and disintegrating tablets. In some embodiments, additional agents include ipratropium. In some embodiments, ipratropium is administered in a spray dosage form.

[0057] In some embodiments, any of the methods of the invention further comprises administering intubation, mechanical ventilation, and / or oxygen therapy.

[0058] In some embodiments, in any of the methods of the present invention, the anti-CD6 antibody or antigen-binding fragment thereof is administered as a pharmaceutical composition comprising one or more pharmaceutically acceptable salts, excipients or vehicles. In some embodiments, the composition comprises one or more agents selected from the group consisting of: carriers, excipients, diluents, antioxidants, preservatives, colorants, flavorings and diluents, emulsifiers, suspending agents, solvents, fillers, bulking agents, buffers, delivery vehicles, tonicity agents, cosolvents, wetting agents, complexing agents, buffers, antimicrobial agents and / or surfactants.

[0059] In some embodiments, the present invention provides a method for inhibiting T cell-mediated pulmonary inflammation in a subject suffering from asthma, comprising: administering an anti-CD6 antibody or an antigen-binding fragment thereof to the subject, wherein the anti-CD6 antibody or the antigen-binding fragment thereof comprises: heavy chain and light chain variable regions, the heavy chain and light chain variable regions comprising the amino acid sequences shown in SEQ ID NOs: 1 and 2, and wherein the asthma is characterized by low blood eosinophils or no blood eosinophils. In some embodiments, the asthma is resistant to steroid therapy or is refractory to steroid therapy. In some embodiments, the asthma is neutrophilic asthma. In some embodiments, the asthma is mixed inflammatory asthma. In some embodiments, the asthma is agranulocytic asthma. In some embodiments, the T cells are selected from: (i) Th1 T cells; (ii) Th17 T cells; or (iii) Th1 and Th17 T cells. In some embodiments, the subject has a blood eosinophil count of ≤300 cells / μl. In some embodiments, the subject suffers from non-allergic asthma. In some embodiments, the anti-CD6 antibody is EQ001.

[0060] In some embodiments, the present invention provides a method of inhibiting T cell-mediated lung inflammation in a subject having asthma, comprising: administering an anti-CD6 antibody or an antigen-binding fragment thereof to the subject, wherein the asthma is characterized by low or no blood eosinophils.

[0061] In some embodiments, the present invention provides a method for preventing or reducing the migration of T cells into and through lung tissue in response to asthma-inducing antigens, wherein the asthma is characterized by low or no blood eosinophils, the method comprising: administering an anti-CD6 antibody or an antigen-binding fragment thereof to a subject.

[0062] In some embodiments, the present invention provides a method of regulating or reducing the symptoms or severity of asthma, comprising: administering an anti-CD6 antibody or antigen-binding fragment thereof to a subject when the asthma is characterized by low or no blood eosinophils.

[0063] In some embodiments, the present invention provides a method of regulating or reducing the symptoms or severity of asthma, comprising: contacting T cells with an anti-CD6 antibody or an antigen-binding fragment thereof, wherein the asthma is characterized by low or no blood eosinophils.

[0064] In some embodiments, the asthma is resistant to steroid therapy or is refractory to steroid therapy. In some embodiments, the asthma is neutrophilic asthma. In some embodiments, the asthma is mixed inflammatory asthma. In some embodiments, the asthma is agranulocytic. In some embodiments, the T cells are selected from: (i) Th1 T cells; (ii) Th17 T cells; or (iii) Th1 and Th17 T cells. In some embodiments, the subject has a blood eosinophil count of ≤300 cells / μl. In some embodiments, the subject has non-allergic asthma. In some embodiments, the asthma is severe asthma. In some embodiments, the asthma is severe asthma. In some embodiments, the anti-CD6 antibody or antigen-binding fragment thereof is EQ001 or an antigen-binding fragment thereof. In some embodiments, the anti-CD6 antibody is EQ001. In some embodiments, the anti-CD6 antibody or antigen-binding fragment thereof binds to domain 1 or domain 3 on CD6. In some embodiments, the anti-CD6 antibody or antigen-binding fragment thereof binds to domain 3 on CD6. In some embodiments, the anti-CD6 antibody or antigen-binding fragment thereof is selected from the group consisting of: UMCD6 mAb, Itolizumab (EQ001), the anti-CD6 antibodies described in Table 2, and the anti-CD6 antibodies disclosed herein. In some embodiments, the anti-CD6 monoclonal antibody is: an antibody produced by the secretory hybridoma IOR-T1A deposited at ECACC with the deposit number ECACC 96112640; an antibody having the same sequence as the antibody produced by the secretory hybridoma; or an antibody having the same CDR sequence as the antibody produced by the secretory hybridoma. In some embodiments, the anti-CD6 antibody or antigen-binding fragment thereof comprises one or more CDR sequences selected from SEQ ID NOs: 5-10. In some embodiments, the anti-CD6 antibody or antigen-binding fragment thereof comprises heavy and light chain variable regions, which comprise the amino acid sequences shown in SEQ ID NOs: 1 and 2. In some embodiments, SEQ ID NOs: 1 and 2 are encoded by SEQ ID NOs: 3 and 4, respectively. In some embodiments, the anti-CD6 antibody or antigen-binding fragment thereof comprises a VH sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 1. In some embodiments, the anti-CD6 antibody or antigen-binding fragment thereof comprises a VK sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 2. In some embodiments, the anti-CD6 antibody or antigen-binding fragment thereof comprises: a VH sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 1 and a VK sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 2.In some embodiments, the antigen binding fragment is selected from: Fv, Fab, CDR1, CDR2, CDR3, a combination of CDRs, a variable region, a heavy chain, and a light chain. In some embodiments, the anti-CD6 antibody or its antigen binding fragment binds to the CD6 protein on the surface of a T cell. In some embodiments, the binding of the anti-CD6 antibody or its antigen binding fragment to the CD6 protein on the surface of a T cell regulates the activity and / or migration of the T cell.

[0065] In some embodiments, the method further comprises administering one or more additional agents capable of treating, preventing or alleviating one or more asthma-related symptoms. In some embodiments, additional agents include agents capable of regulating the immune system. In some embodiments, additional agents include agents that are immunosuppressants. In some embodiments, additional agents include long-acting beta agonists, short-acting beta agonists, or a combination thereof. In some embodiments, additional agents include salbutamol. In some embodiments, salbutamol is administered in a dosage form selected from the following: aerosol powder; solution; capsule; and powder suspension. In some embodiments, additional agents include corticosteroids. In some embodiments, corticosteroids are administered as inhalation preparations. In some embodiments, additional agents include ipratropium. In some embodiments, ipratropium is administered in a spray formulation. In some embodiments, the method further comprises administering intubation, mechanical ventilation, and / or oxygen therapy. In some embodiments, anti-CD6 antibodies or antigen-binding fragments thereof are administered as pharmaceutical compositions comprising one or more pharmaceutically acceptable salts, excipients, or vehicles. In some embodiments, the composition comprises one or more agents selected from the group consisting of: carriers, excipients, diluents, antioxidants, preservatives, colorants, flavorings and diluents, emulsifiers, suspending agents, solvents, fillers, bulking agents, buffers, delivery vehicles, tonicity agents, co-solvents, wetting agents, complexing agents, buffers, antimicrobial agents and / or surfactants.

[0066] Sequence Description

[0067] SEQ ID NO: 1: amino acid sequence of EQ001 VH sequence;

[0068] SEQ ID NO:2: amino acid sequence of EQ001 VK sequence;

[0069] SEQ ID NO:3: nucleic acid (DNA) sequence of EQ001 VH sequence;

[0070] SEQ ID NO:4: nucleic acid (DNA) sequence of EQ001 VK sequence;

[0071] SEQ ID NO:5: amino acid sequence of EQ001 VH CDR1;

[0072] SEQ ID NO:6: amino acid sequence of EQ001 VH CDR2;

[0073] SEQ ID NO:7: amino acid sequence of EQ001 VH CDR3;

[0074] SEQ ID NO:8: amino acid sequence of EQ001 VK CDR1;

[0075] SEQ ID NO:9: amino acid sequence of EQ001 VK CDR2;

[0076] SEQ ID NO: 10: Amino acid sequence of EQ001 VK CDR3. BRIEF DESCRIPTION OF THE DRAWINGS

[0077] Figure 1. Sequence of the EQ001 anti-CD6 antibody. Figure 1A : Nucleotide sequences of the variable heavy chain (VH) and light chain (VK) of EQ001 derived from plasmid and genomic DNA. Figure 1B : The amino acid sequences of VH and VK of EQ001;

[0078] Figure 2 .CD6 in the lungs of patients with severe asthma + Left column: ALCAM is expressed in the lamina propria of fatal asthmatic lungs (lower left) but not in the lamina propria of normal lungs (upper left). Center column: CD6 + Cells increase. Right column: CD6 + Cells co-localize with the lamina propria expressing ALCAM in asthmatic lungs (bottom right);

[0079] Figure 3 De novo bioinformatics-based analysis compared the expression of CD4, CD6, and various Th17 markers in controls, moderate asthmatics, and severe asthmatics using publicly available RNASeq datasets generated from cells collected by bronchial lavage. Figure 3 A: Significant differences in CD4 expression levels, with severe asthma patients expressing significantly higher levels of CD4. Figure 3 B: Significant differences in CD6 expression levels, with severe asthma patients expressing significantly higher levels of CD6. Figure 3 C to 3G: Significant differences in the expression levels of Th17 markers, with severe asthma patients expressing significantly higher levels of CD6. Figure 3 C: CCR6 expression; Figure 3 D: CCR4 expression; Figure 3 E: KLRB1 expression; Figure 3 F: IL-17A expression; Figure 3 G: IL-17F expression;

[0080] Figure 4 .CD6 was highest in a group of severe asthma patients. Figure 4 A: Unsupervised cluster analysis based on the expression of >1000 genes (not related to asthma severity) showed that asthma patients were divided into 4 major clusters. Figure 4 B: Each cluster exhibited a different average CD6 expression, with the highest CD6 expression occurring in cluster 3 consisting only of severe asthma patients, indicating that this subset of severe asthma patients had a high index;

[0081] Figure 5 .CD6 blockade in a mouse model of allergic asthma reduces Th2 cytokine levels in bronchiolar lavage fluid. Figure 5 A: The table shows the treatment groups and dosing regimen. Figure 5 B: Description of dosage regimen. Figure 5 C: CD6 blockade with mCD6D1 anti-CD6 antibody during challenge resulted in reduced levels of Th2 cytokines IL-4, IL-5, and IL-13 in bronchiolar lavage fluid (BALF) and modest reductions of these cytokines in pneumocytes (data not shown);

[0082] Figure 6 CD6 blockade in the classic OVA-vaccinated Th2 model inhibits Th2-driven IgE production. Figure 6 A: The table shows the treatment groups and dosing regimen. Figure 6 B: Description of dosage regimen. Figure 6 C: Prophylactic CD6 blockade with mCD6D1 anti-CD6 antibody inhibited the production of OVA-specific IgE, demonstrating the influence of the CD6 pathway on Th2 responses;

[0083] Figure 7 The examples show asthma phenotypes associated with inflammatory type (Th2 high or Th2 low) and other variables. CS = corticosteroids; GM-CSF = granulocyte-macrophage colony-stimulating factor. DETAILED DESCRIPTION

[0084] Unless otherwise indicated, the practice of the present invention will employ conventional molecular biology methods and recombinant DNA techniques within the skill of the art, many of which are described below for illustrative purposes. Such techniques are fully explained in the literature. See, for example, Sambrook, et al., Molecular Cloning: A Laboratory Manual (3 rd Edition, 2000); DNA Cloning: A Practical Approach, vol. I & II (D. Glover, ed.); Oligonucleotide Synthesis (N. Gait, ed., 1984); Oligonucleotide Synthesis: Methods and Applications (P. Herdewijn, ed., 2004); Nucleic Acid Hybridization (B.Hames&S.Higgins, eds., 1985); Nucleic Acid Hybridization: Modern Applications (Buzdin and Lukyanov, eds., 2009); Transcription and Translation (B.Hames&S.Higgins, eds., 1984); Animal Cell Culture(R.Freshney,ed.,1986);Freshney,RI(2005)Culture of Animal Cells,a Manual of Basic Technique,5 th Ed.Hoboken NJ, John Wiley&Sons; B.Perbal, A Practical Guide to Molecular Cloning(3 rd Edition2010); Farrell, R., RNA Methodologies: A Laboratory Guide for Isolation and Characterization (3 rdEdition 2005). Poly(ethylene glycol), Chemistry and Biological Applications, ACS, Washington, 1997; Veronese, F., and JM Harris, Eds., Peptide and protein PEGylation, Advanced Drug Delivery Reviews, 54 (4) 453-609 (2002); Zalipsky, S., et al., "Use of functionalized Poly(Ethylene Glycols) for modification of polypeptides in Polyethylene Glycol Chemistry: Biotechnology and Biomedical Applications". The publications discussed above in this application are provided solely for their disclosure prior to the filing date of the present application. Nothing herein should be construed as an admission that the invention is not entitled to antedate such disclosure by virtue of prior invention.

[0085] definition

[0086] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the present invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, preferred methods and materials are described herein. For purposes of the present invention, the following terms are defined as follows.

[0087] As used herein, the articles "a" and "an" refer to one or more than one (ie, at least one) of the grammatical object of the article. For example, "an element" means one element or more than one element.

[0088] Unless otherwise stated, the term "and / or" is used in the present invention to mean "and" or "or".

[0089] The term "such as" as used herein means "for example," and will be understood to imply the inclusion of stated steps or elements, or groups of steps or elements, but not the exclusion of any other steps or elements, or groups of steps or elements.

[0090] "About" means an amount, level, value, number, frequency, percentage, dimension, size, amount, weight or length that differs by at most 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2% or 1% relative to a reference amount, level, value, number, frequency, percentage, dimension, size, amount, weight or length.

[0091] As used herein, the term "administering" refers to transferring, delivering, introducing or transporting a substance, such as a compound (e.g., a drug compound) or other agent (e.g., an antigen) to a subject in any mode. Modes of administration include oral administration, topical contact, intravenous, intraperitoneal, intramuscular, intranasal, or subcutaneous administration. Administration "in combination" with another substance, such as one or more therapeutic agents, includes simultaneous (concurrent) and sequential administration in any order.

[0092] As used herein, the term "binding partner" refers to a substance (e.g., a molecule, particularly a polymer molecule) that can bind to a nucleic acid molecule (e.g., DNA or RNA, including mRNA molecules), and a peptide, protein, sugar, polysaccharide, or lipid by an interaction (usually by non-covalent bonding) sufficient to form a complex between the agent and the nucleic acid molecule, peptide, protein, or sugar, polysaccharide, or lipid. In some embodiments, the binding partner is an immunoglobulin or a protein-like binding molecule having an immunoglobulin-like function as defined below. In some embodiments, the binding partner is an aptamer. In some embodiments, the binding partner is specific for a specific target. In some embodiments, the binding partner includes multiple binding sites, each of which is specific for a specific target. As an illustrative example, the binding partner can be a protein-like agent having an immunoglobulin-like function with two binding sites. For example, the binding partner can be an antigen-binding fragment of an antibody. For example, the binding partner can be a bispecific diabody, such as a bispecific single-chain diabody.

[0093] As used herein, the term "carrier" encompasses vehicles, excipients, and diluents that participate in carrying or transporting a pharmaceutical formulation from one organ or part of a subject's body to another organ or part of a subject's body, and means a material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material.

[0094] As used herein, the term "chimeric antibody" refers to an immunoglobulin polypeptide or domain antibody comprising sequences from more than one species. In a chimeric antibody, a heavy chain or a light chain may comprise a variable region sequence from one species (e.g., human) and a constant region sequence from another species (e.g., mouse). For example, a "chimeric antibody" can be an immunoglobulin having a variable region derived from an animal antibody (e.g., a rat or mouse antibody) fused to another molecule (e.g., a constant domain derived from a human antibody). The term "chimeric antibody" is intended to encompass antibodies in which: (i) the heavy chain is chimeric, but the light chain comprises a V region and a C region from only one species; (ii) the light chain is chimeric, but the heavy chain comprises a V region and a C region from only one species; and (iii) both the heavy chain and the light chain are chimeric.

[0095] When used in conjunction with a compound, an "effective amount" is the amount of a compound (e.g., an anti-CD6 antibody (e.g., EQ001)) required to elicit a desired response. In some embodiments, the desired response is, for example, a biological response in a subject. In some embodiments, a compound (e.g., an anti-CD6 antibody) can be administered to a subject in an effective amount to produce a biological response in the subject. In some embodiments, an effective amount is a "therapeutically effective amount."

[0096] The terms "therapeutically effective amount" and "therapeutic dose" are used interchangeably herein and refer to an amount of a compound, e.g., an anti-CD6 antibody (e.g., EQ001), which, upon administration to a subject, is effective for treating a disease or condition in the subject as described herein.

[0097] The term "prophylactically effective amount" is used herein to refer to an amount of a compound, such as an anti-CD6 antibody (eg, EQ001), which, upon administration to a subject, is effective for preventing or delaying the onset of a disease or disorder in a subject as described herein.

[0098] In this regard, "humanized antibody" as used herein is an immunoglobulin polypeptide or domain antibody containing the structural elements of a human antibody and the antigen binding site of a non-human antibody. "Humanized antibody" contains the minimum number of residues from a non-human antibody, and the humanized antibody is derived from a non-human antibody. For example, a humanized antibody may contain only the CDR region of a non-human antibody, or only those residues that constitute the hypervariable region of a non-human antibody. A humanized antibody may also contain certain residues outside the variable region of a non-human polypeptide, such as residues necessary to simulate the structure of a non-human antibody or to minimize spatial interference. Typically, a humanized antibody comprises a human framework, at least one CDR from a non-human antibody, and any constant region present is substantially identical to a human immunoglobulin constant region, i.e., at least about 85%-90% identical, such as at least 95% identical. Therefore, in some cases, all parts of a humanized immunoglobulin (possibly excluding CDR) are substantially identical to the corresponding parts of one or more natural human immunoglobulin sequences. In addition, a humanized antibody may comprise residues that do not correspond to a human antibody or a non-human antibody.

[0099] As used herein, the term "antibody fragment" refers to any form of antibody other than the full-length form. Antibody fragments herein include antibodies that are smaller components present in full-length antibodies and antibodies that have been engineered. Antibody fragments include, but are not limited to: Fv, Fc, Fab, and (Fab')2, single-chain Fv (scFv), double antibodies, three antibodies, four antibodies, bifunctional hybrid antibodies, CDR1, CDR2, CDR3, CDR combinations, variable regions, framework regions, constant regions, heavy chains, light chains, alternative scaffold non-antibody molecules, and bispecific antibodies. Unless otherwise specifically stated, records and claims using the term one or more "antibodies" may specifically include one or more "antibody fragments".

[0100] The term "asthma" has its ordinary scientific meaning and includes intermittent asthma, mild persistent asthma, moderate persistent asthma, and severe persistent asthma.

[0101] The term "severe asthma" is used herein to describe a separate category of asthma in which symptoms of the disease are poorly controlled with steroids. Severe asthma includes asthma that is steroid-resistant and / or refractory to corticosteroids (CS). In some embodiments, severe asthma (SA) is mainly driven by a response mediated by a neutrophilic Th1 / Th17 T cell. In some embodiments, SA is mainly driven by a neutrophilic Th1 / Th17 T cell-mediated response. In some embodiments, SA is also resistant to one or more other asthma therapeutic agents. For example, in some embodiments, SA is also resistant to one or more of SABA and / or LABA. In some embodiments, severe asthma may be characterized by a response mediated by a neutrophilic Th1 / Th17 T cell, a neutrophilic Th1 / Th17 T cell-mediated response, or a combined neutrophilic Th1 / Th17 and eosinophilic Th2 T cell-mediated response. In certain embodiments, severe asthma may include a neutrophilic T cell response but not an eosinophilic T cell response. In certain embodiments, severe asthma may include an agranulocytic T cell response but not an eosinophilic T cell response.

[0102] The term "steroid-resistant asthma" is used herein to describe asthma for which steroid therapy (e.g., corticosteroids) has limited efficacy. Thus, treatment of steroid-resistant asthma with steroids (e.g., corticosteroids) will produce minimal detectable therapeutic benefit. In some cases, such treatment produces substantially no therapeutic benefit.

[0103] The term "steroid-refractory asthma" is used herein to describe asthma that is refractory to steroid therapy (eg, corticosteroids). Thus, treatment of steroid-refractory asthma with steroids (eg, corticosteroids) will not produce a detectable therapeutic benefit.

[0104] The term "LABA" means a long-acting beta agonist. LABAs are known in the art and include, but are not limited to: formoterol fumarate; salmeterol xinafoate.

[0105] The term "SABA" means a short-acting beta agonist. SABAs are known in the art and include, but are not limited to, albuterol (e.g., albuterol sulfate, albuterol sulfate HFA, albuterol sulfate inhalation solution, albuterol sulfate nebulizer solution, and levalbuterol hydrochloride), metaproterenol, pyrbutaline (e.g., pirbuterol acetate); isoproterenol hydrochloride; isoproterenol hydrochloride; and terbutaline sulfate.

[0106] The term "VH" is used herein to refer to the variable heavy chain of an antibody.

[0107] The term "VK" is used herein to refer to the variable light chain of an antibody.

[0108] The term "antigen-binding fragment" with respect to an antibody refers to any antibody fragment that retains binding affinity for the antigen bound by the parent full-length antibody, and antigen-binding fragments include, but are not limited to, Fv, Fab, (Fab')2, scFv, diabodies, triabodies, tetrabodies, bifunctional hybrid antibodies, CDR1, CDR2, CDR3, CDR combinations, variable regions, heavy chains, light chains, and bispecific antibodies.

[0109] Throughout the specification, unless the context requires otherwise, the words "comprise", "include" and "contain" and will be understood to imply the inclusion of the steps or elements, or steps or groups of elements, rather than excluding any other steps or elements, or steps or groups of elements. "Consisting of" means including and limited to whatever follows the phrase "consisting of". Therefore, the phrase "consisting of" means that the listed elements are necessary or mandatory, and no other elements may be present. "Substantially consisting of" means including any element listed after the phrase, and is limited to other elements that do not interfere with or promote the activity or action specified for the listed elements in the present invention. Therefore, the phrase "substantially consisting of" means that the listed elements are necessary or mandatory, but other elements are optional and may or may not exist, depending on whether they substantially affect the activity or action of the listed elements.

[0110] The term "modulation" includes "increase", "enhancement" or "stimulation" as well as "decrease" or "reduction" compared to a control, usually by a statistically significant or physiologically significant amount. The amount of "increase", "stimulation" or "enhancement" is usually a "statistically significant" amount, and may include an increase of 1.1, 1.2, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30 times or more (e.g., 500 times, 1000 times) (including all integers and decimal points between and above 1.5, 1.6, 1.7, 1.8, etc.) to the amount produced by no composition (e.g., in the absence of any anti-CD6 antibody of the invention) or a control composition, sample or test subject. A "reduced" or "lowered" amount is generally a "statistically significant" amount and can include a 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% decrease (including all integers in between) in the amount produced by the no composition (absence of agent or compound) or the control composition.

[0111] The terms "polypeptide" and "protein" are used interchangeably herein to refer to polymers of amino acid residues, as well as variants and synthetic analogs thereof. Thus, these terms apply to amino acid polymers in which one or more amino acid residues is a synthetic non-naturally occurring amino acid (e.g., a chemical analog of a corresponding naturally occurring amino acid), as well as naturally occurring amino acid polymers.

[0112] As used herein, "subject" or "patient" includes any animal that exhibits symptoms or is at risk of exhibiting symptoms that can be treated or diagnosed with an anti-CD6 antibody or antigen-binding fragment thereof. Suitable subjects (patients) preferably include human patients. Suitable subjects also include experimental animals (e.g., mice, rats, rabbits, or guinea pigs), farm animals (e.g., pigs, horses, cattle), and domestic animals or pets (e.g., cats or dogs). Non-human primates (e.g., monkeys, chimpanzees, baboons, or rhesus monkeys) are also included.

[0113] "Substantially" or "essentially" means nearly completely or entirely, for example, 95% or more of some given amount.

[0114] As used herein, "treating" or "treating" includes any desired effect on the symptoms or pathology of a disease or condition, and may include even minimal changes or improvements in one or more measurable markers of the disease or condition being treated. "Treatment" or "treating" does not necessarily mean complete eradication or cure of the disease or condition or its associated symptoms. A subject receiving such treatment is any subject in need of such treatment. Exemplary markers of clinical improvement will be apparent to those skilled in the art.

[0115] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those of ordinary skill in the field to which the present invention belongs. Although any method, composition, reagent, cell similar or equivalent to those described herein can be used in the practice or test of the present invention, preferred methods and materials are described herein. All publications and references cited in this specification, including but not limited to patents and patent applications, are incorporated herein by reference in their entirety, just as each individual publication or reference is explicitly and individually indicated as being incorporated herein by reference in its entirety. Any patent application for which the present application claims priority is also incorporated herein by reference in its entirety in the above-mentioned manner for publications and references.

[0116] summary

[0117] The present invention relates to the treatment, prevention or alleviation of severe asthma, comprising administering an anti-CD6 antibody to a subject.

[0118] CD6 is an important cell surface protein expressed primarily by a subset of human T cells and B cells, as well as some B cell chronic lymphocytic leukemias and neurons [Aruffo et al., J. Exp. Med. 1991, 174: 949; Kantoun et al., J. Immunol. 1981, 127: 987; Mayer et al., J. Neuroimmunol. 1990. 29: 193]. CD6 is a member of a large protein family characterized by having at least one domain homologous to the cysteine-rich scavenger receptor domain (SRCR) of type I macrophages [Matsumoto, et al., J. Exp. Med. 1991, 173: 55 and Resnick et al., Trends Biochem. Sci. 1994, 19: 5]. Other members of this family include CD5 [Jones et al., Nature. 1986, 323:346]; cyclophilin C [Friedman et al. 1993, PNAS 90:6815]; complement factor I, which binds activated complement proteins C3b and C4b [Goldberger, et al., J. Biol. Chem. 1987, 262:10065]; bovine WC-1, which is expressed by τ / δ T cells [Wijingaard et al., J. Immunol. 1992, 149:3273] and M130 [Law et al., Eur J. Immunol. 1993, 23:2320] and is a marker for macrophage activation.

[0119] The extracellular domain of the mature CD6 protein consists of three SRCR domains (hereinafter referred to as D1, D2 and D3). D3 corresponds to the membrane-proximal SRCR domain, followed by a short stem region of 33 amino acids. These extracellular domains are anchored to the cell membrane via short transmembrane domains, followed by cytoplasmic domains of variable length [Aruffo et al., J. Exp. Med. 1991, 174: 949].

[0120] Studies using CD6-immunoglobulin fusion proteins comprising a selected extracellular domain of CD6 fused to a human IgG1 constant domain (CD6-Rgs) led to the identification and cloning of the CD6 ligand termed "activated leukocyte adhesion molecule" (ALCAM), also known as CD166 [Patel, et al., J. Exp. Med. 1995. 181: 1563-1568; Bowen et al., J. Exp. Med 1995, 181: 2213-2220].

[0121] ALCAM is a 100kD-105kD type I transmembrane glycoprotein that is a member of the immunoglobulin superfamily and contains five extracellular immunoglobulin domains (two NH2-terminal membrane distal variable-(V) type (V1, V2 or D1, D2) Ig folds and three membrane proximal constant-(C2) type [C1, C2, C3] Ig folds), a transmembrane region, and a short cytoplasmic tail region. The N-terminal domain (D1) is only involved in ligand binding, while the membrane proximal domains (C2, C3 or D4, D5) are required for pro-homoantigen interactions.

[0122] ALCAM binds to domain 3 of CD6, which corresponds to the membrane-proximal SRCR domain [Whitney, et. al., J. Biol. Chem 1995, 270: 18187-18190].

[0123] CD6 in T cells (including CD4 + CD6 is expressed on the surface of T cells and may play a role in T cell activation, differentiation, survival, and migration. However, CD6 + Role of T cells in the pathogenesis of severe non-allergic asthma.

[0124] Here we report the surprising finding that patients with steroid-refractory, fatal severe asthma express significantly higher levels of the protein CD6 and its ligand ALCAM ( Figure 2 ). Furthermore, bioinformatics-based de novo sequencing analysis of publicly available RNASeq datasets confirmed this result by demonstrating that severe asthmatics expressed significantly higher levels of CD6 mRNA in bronchial lavage fluid (BALF) compared with moderate asthmatics and non-asthmatics ( Figure 3 B). In addition, bioinformatics-based de novo sequencing analysis of these datasets also showed that severe asthmatics expressed significantly higher Th17 markers and cytokines CCR6, CCR4, KLRB1, IL-17A, and IL-17F in bronchial lavage fluid (BALF) compared with moderate asthmatics and non-asthmatics ( Figure 3 C-3G). Considering the disease in these patients with severe asthma (referring to non-Th2 disease, as Th2-mediated asthma usually responds well to steroid therapy, see Figure 7 ), as well as the high expression of these Th17 markers and cytokines, we concluded that this elevated CD6 expression was primarily attributable to Th17 T cell infiltration of the lung tissue.

[0125] Interestingly, a recent report suggests that eosinophilic Th2-driven allergic asthma may be mediated in part by ALCAM, as mice knocked out of ALCAM and treated with anti-ALCAM antibodies showed reduced Th2 cytokine levels in response to allergen exposure (Kim, et al., Am J Respir Crit Care Med. 2018 Apr 15; 197(8): 994-1008). However, these findings were limited to allergic eosinophilic Th2-mediated responses in allergic asthma models (focusing on Th2 cytokines, including IL-4, IL-5, IL-13, and IgE); in contrast, our results above focused on non-allergic severe asthma cases that exhibited low or no eosinophils. In addition, our results suggest that high levels of Th1 and Th17 T cells (expressing significantly elevated levels of Th17 markers (CCR6, CCR4, KLRB1, IL-17A, and IL-17F)) may be the pathological basis of severe steroid-refractory asthma. Interestingly, our data also differ from the findings reported by Kim et al., who found low levels of resident ALCAM protein in the lung due to ALCAM shedding (via the activity of metalloproteinases of the ADAM family); in contrast, we observed high levels of ALCAM protein in the lungs of patients with non-allergic severe asthma ( Figure 2 ). Thus, based on the report by Kim et al., our data further illustrate the complex differences between eosinophilic and noneosinophilic asthma, which may underlie the often steroid-responsive nature of the former and the often steroid-refractory or resistant nature of the latter.

[0126] Of note, we also report here that CD6 inhibition effectively reduced Th2 cytokines in bronchiolar lavage fluid in a Th2-mediated murine model of allergic asthma (the same model used by Kim et al., supra). Figure 5 C) and inhibited Th2-driven IgE production ( Figure 6), suggesting that eosinophilic Th2 asthma could also be treated with anti-CD6 antibodies. Thus, in contrast to the divergent responses to steroid therapy that is almost the hallmark distinction between eosinophilic and noneosinophilic asthma, our data suggest that CD6 inhibition may provide a one-stop mechanism for simultaneously inhibiting Th2-mediated and Th1 / Th17-mediated asthma. To our knowledge, this is the first report suggesting the use of CD6 antibodies to treat severe asthma characterized by a low or no eosinophilic component (i.e., Th1 / Th17-mediated asthma, rather than Th2-mediated allergic asthma).

[0127] Therefore, certain aspects of the present invention provide methods and compositions directed to inhibiting T cell-mediated pulmonary inflammation (including inhibiting or blocking CD6-signaling pathways) in subjects with asthma. In a specific embodiment, the method and composition are directed to inhibiting T cell-mediated pulmonary inflammation in subjects with asthma, and the asthma is characterized by low blood eosinophils or no blood eosinophils. Methods for determining blood eosinophil counts are well known in the art and can be used according to the present invention (see, for example, Kostikas, et al., Curr Drug Targets. 2018 Dec; 19 (16): 1882–1896, the contents of which are incorporated herein by reference in their entirety).

[0128] As is clear to those of ordinary skill in the art, the presence of high or low eosinophils is the so-called gold standard for distinguishing allergic asthma from other forms of non-allergic asthma. Eosinophils are usually less than 5% of peripheral blood leukocytes, but their production is greatly increased in response to type 2 helper T cells (Th2)-mediated inflammation (such as inflammation present in allergic asthma), and many clinical studies have used measurements of blood eosinophil counts as a surrogate for lung eosinophil levels. See, for example, Kostikas, et al., Curr Drug Targets. 2018 Dec; 19 (16): 1882–1896, the contents of which are incorporated herein by reference in their entirety. As used herein, reference to an asthma subject with "low" eosinophils means that the subject's blood eosinophil count is ≤ 300 cells / μl. Reference to an asthma subject with "no" eosinophils means that the subject does not have detectable blood eosinophils / μl.

[0129] Certain aspects of the invention provide methods and compositions directed to inhibiting T cell-mediated pulmonary inflammation in subjects with asthma, comprising: administering an anti-CD6 antibody (e.g., EQ001) or an antigen-binding fragment thereof to the subject. In some embodiments, the asthma may be severe asthma. In specific embodiments, the asthma may be characterized by low eosinophils or absence of eosinophils. In specific embodiments, the asthma may be neutrophilic asthma. In specific embodiments, the asthma may be agranulocytic asthma. In specific embodiments, the asthma may be mixed inflammatory asthma.

[0130] Certain aspects of the invention provide methods and compositions directed to treating, preventing or mitigating the migration of T cells into and through lung tissue in response to asthma-inducing antigens, comprising: administering an anti-CD6 antibody (e.g., EQ001) or an antigen-binding fragment thereof to a subject. In some embodiments, the asthma may be severe asthma. In specific embodiments, the asthma may be characterized by low eosinophils or absence of eosinophils. In specific embodiments, the asthma may be neutrophilic asthma. In specific embodiments, the asthma may be agranulocytic asthma. In specific embodiments, the asthma may be mixed inflammatory asthma.

[0131] Certain aspects of the invention provide methods and compositions directed to regulating or alleviating the symptoms or severity of asthma, comprising: administering an anti-CD6 antibody (e.g., EQ001) or an antigen-binding fragment thereof to a subject. In some embodiments, the asthma may be severe asthma. In specific embodiments, the asthma may be characterized by low eosinophils or absence of eosinophils. In specific embodiments, the asthma may be neutrophilic asthma. In specific embodiments, the asthma may be agranulocytic asthma. In specific embodiments, the asthma may be mixed inflammatory asthma.

[0132] Certain aspects of the invention provide methods and compositions directed to regulating or alleviating the symptoms or severity of asthma, comprising: contacting T cells with an anti-CD6 antibody (e.g., EQ001) or an antigen-binding fragment thereof. In some embodiments, the asthma may be severe asthma. In specific embodiments, the asthma may be characterized by low eosinophils or absence of eosinophils. In specific embodiments, the asthma may be neutrophilic asthma. In specific embodiments, the asthma may be agranulocytic asthma. In specific embodiments, the asthma may be mixed inflammatory asthma.

[0133] Certain aspects of the invention provide methods and compositions directed to inhibiting T cell-mediated pulmonary inflammation in subjects with asthma, comprising: administering to the subject a binding partner that specifically binds to CD6 on T cells and prevents or inhibits activation of CD6 signals. The binding partner can be an anti-CD6 antibody (e.g., EQ001) or an antigen binding fragment thereof. In some embodiments, asthma can be severe asthma. In specific embodiments, asthma can be characterized by low eosinophils or absence of eosinophils. In specific embodiments, asthma can be neutrophilic asthma. In specific embodiments, asthma can be agranulocytic asthma. In specific embodiments, asthma can be mixed inflammatory asthma.

[0134] Certain aspects of the present invention provide methods and compositions directed to treating, preventing or slowing the migration of T cells into and through lung tissue in response to asthma-inducing antigens, comprising: administering to a subject a binding partner that specifically binds to CD6 on T cells and prevents or inhibits the activation of CD6 signals. The binding partner can be an anti-CD6 antibody (e.g., EQ001) or an antigen-binding fragment thereof. In some embodiments, asthma can be severe asthma. In specific embodiments, asthma can be characterized by low eosinophils or absence of eosinophils. In specific embodiments, asthma can be neutrophilic asthma. In specific embodiments, asthma can be agranulocytic asthma. In specific embodiments, asthma can be mixed inflammatory asthma.

[0135] Certain aspects of the present invention provide methods and compositions directed to regulating or alleviating the symptoms or severity of asthma, comprising: administering to a subject a binding partner that specifically binds to CD6 on T cells and prevents or inhibits activation of CD6 signals. The binding partner can be an anti-CD6 antibody (e.g., EQ001) or an antigen binding fragment thereof. In some embodiments, asthma can be severe asthma. In specific embodiments, asthma can be characterized by low eosinophils or absence of eosinophils. In specific embodiments, asthma can be neutrophilic asthma. In specific embodiments, asthma can be agranulocytic asthma. In specific embodiments, asthma can be mixed inflammatory asthma.

[0136] Certain aspects of the present invention provide methods and compositions directed to regulating or alleviating the symptoms or severity of asthma, comprising: contacting T cells with a binding partner that specifically binds to CD6 on T cells and prevents or inhibits the activation of CD6 signals. The binding partner can be an anti-CD6 antibody (e.g., EQ001) or an antigen-binding fragment thereof. In some embodiments, asthma can be severe asthma. In specific embodiments, asthma can be characterized by low eosinophils or no eosinophils. In specific embodiments, asthma can be neutrophilic asthma. In specific embodiments, asthma can be agranulocytic asthma. In specific embodiments, asthma can be mixed inflammatory asthma.

[0137] Certain aspects of the invention provide methods and compositions directed to treating, preventing or mitigating the migration of T cells into and through lung tissue in response to asthma-inducing antigens, including: inhibiting or blocking the CD6 pathway. In some embodiments, asthma can be severe asthma. In specific embodiments, asthma can be characterized by low eosinophils or no eosinophils. In specific embodiments, asthma can be neutrophilic asthma. In specific embodiments, asthma can be agranulocytic asthma. In specific embodiments, asthma can be mixed inflammatory asthma.

[0138] Certain aspects of the invention provide methods and compositions directed to regulating or alleviating the symptoms or severity of asthma, including: inhibiting or blocking the CD6 pathway. In some embodiments, asthma can be severe asthma. In specific embodiments, asthma can be characterized by low eosinophils or no eosinophils. In specific embodiments, asthma can be neutrophilic asthma. In specific embodiments, asthma can be agranulocytic asthma. In specific embodiments, asthma can be mixed inflammatory asthma.

[0139] Certain aspects of the invention provide methods and compositions directed to regulating or alleviating the symptoms or severity of asthma, including: inhibiting or blocking the CD6 pathway. In some embodiments, asthma can be severe asthma. In specific embodiments, asthma can be characterized by low eosinophils or no eosinophils. In specific embodiments, asthma can be neutrophilic asthma. In specific embodiments, asthma can be agranulocytic asthma. In specific embodiments, asthma can be mixed inflammatory asthma.

[0140] In some embodiments, the method of treating severe asthma comprises modulating the activation, proliferation, differentiation, survival and / or migration of one or more CD6 expressing cells by contacting the cells with an anti-CD6 antibody (e.g., EQ001). For example, the CD6 expressing cells contacted with the anti-CD6 antibody (e.g., EQ001) can be T cells expressing CD6. Thus, in subjects with severe asthma, this property of such T cells can be modulated by using anti-CD6 antibodies. In some embodiments, the T cells so modulated are CD4 + T cells. In some embodiments, the T cells so regulated are T helper 1 (Th1) T cells. In some embodiments, the T cells so regulated are T helper 17 (Th17) T lymphocytes (T cells). In some embodiments, anti-CD6 antibodies (e.g., EQ001) regulate the activation, differentiation, survival and / or migration of Th1 T cells and / or Th17 T cells and another cell expressing CD6. For example, anti-CD6 antibodies (e.g., EQ001) can regulate the activation, differentiation, survival and / or migration of a combination of cells selected from the following: (i) Th17 T cells and Th2 T cells; (ii) Th17 T cells and Th1 T cells; (iii) Th1 T cells and Th2 T cells; and (iv) Th17 T cells, Th1 T cells and Th2 T cells. In some embodiments, asthma can be severe asthma. In specific embodiments, asthma can be characterized by low eosinophils or no eosinophils. In specific embodiments, asthma can be neutrophilic asthma. In specific embodiments, asthma can be agranulocytic asthma. In certain embodiments, the asthma may be mixed inflammatory asthma.

[0141] In some embodiments, the asthma is severe asthma including a neutrophilic phenotype, or a mixed phenotype of eosinophilic and neutrophilic granulocytes. In some embodiments, the asthma is severe asthma including a neutrophilic phenotype, agranulocytic phenotype, or a mixed phenotype of eosinophilic and neutrophilic granulocytes. In some embodiments, the asthma is neutrophilic severe asthma characterized by involvement of CD6 + In airway inflammation, the CD6 + The T cells are Th17 T cells. In some embodiments, the asthma is neutrophilic severe asthma characterized by the involvement of CD6 + In airway inflammation, the CD6 + The T cells are Th1 T cells. In certain specific embodiments, the asthma is neutrophilic severe asthma characterized by the involvement of Th1 CD6 + T cells and Th17 CD6 +In some embodiments, asthma is severe agranulocytic asthma characterized by involvement of CD6 + In airway inflammation, the CD6 + The T cells are Th17 T cells. In some embodiments, the asthma is severe agranulocytic asthma characterized by the involvement of CD6 + In airway inflammation, the CD6 + The T cells are Th1 T cells. In certain specific embodiments, the asthma is severe agranulocytic asthma characterized by the involvement of Th1 CD6 + T cells and Th17 CD6 + In some embodiments, severe asthma is mixed neutrophilic and eosinophilic asthma characterized by the involvement of CD6 T cells, which are Th1 T cells. + T cells, CD6 for Th17 T cells + T cells, and CD6 T cells, which are Th2 T cells + T cells in airway inflammation.

[0142] In a specific embodiment, the invention provides a method comprising administering an anti-CD6 antibody (e.g., EQ001) to a patient with neutrophilic severe asthma characterized by involvement of CD6 T cells that are Th17 T cells. + T cells in airway inflammation.

[0143] In a specific embodiment, the invention provides a method comprising administering an anti-CD6 antibody (e.g., EQ001) to a patient with neutrophilic severe asthma characterized by involvement of CD6 T cells that are Th1 T cells. + T cells in airway inflammation.

[0144] In a specific embodiment, the invention provides a method comprising administering an anti-CD6 antibody (e.g., EQ001) to a patient with neutrophilic severe asthma characterized by involvement of CD6 T cells that are Th17 T cells. + CD6 T cells and Th1 T cells + T cells in airway inflammation.

[0145] In certain embodiments, the invention relates to the use of an anti-CD6 antibody disclosed herein (e.g., EQ001) to treat severe asthma comprising a low eosinophilic T cell response. In certain embodiments, the invention relates to the use of an anti-CD6 antibody disclosed herein (e.g., EQ001) to treat severe asthma comprising an absence of an eosinophilic T cell response.

[0146] In certain embodiments, the present invention relates to the use of an anti-CD6 antibody disclosed herein (e.g., EQ001) to treat severe asthma comprising a neutral T cell response but no eosinophilic T cell response. In certain embodiments, the present invention relates to the use of an anti-CD6 antibody disclosed herein (e.g., EQ001) to treat severe asthma comprising a neutral T cell response but substantially no eosinophilic T cell response. In certain embodiments, the present invention relates to the use of an anti-CD6 antibody disclosed herein (e.g., EQ001) to treat severe asthma comprising a neutral T cell response and a low eosinophilic T cell response. In certain embodiments, the present invention relates to the use of an anti-CD6 antibody disclosed herein (e.g., EQ001) to treat severe asthma comprising a neutral T cell response and a low eosinophilic T cell response. In certain embodiments, the present invention relates to the use of an anti-CD6 antibody disclosed herein (e.g., EQ001) to treat a + In certain embodiments, the invention relates to the use of an anti-CD6 antibody disclosed herein (e.g., EQ001) to treat a disease comprising essentially Th1 and Th17 CD4 + In certain embodiments, the present invention relates to treating asthma (e.g., severe asthma) comprising a T cell response mediated by Th1 and / or Th17 CD4 + Effector cell-mediated T cell responses, the Th1 and / or Th17 CD4 + Effector cells are Th2 CD4 + At least 2x of effector cells or Th2 CD4 + 3x, 4x, 5x, 10x, 15x, 20x, 30x, 40x, 50x, 100x, 1,000x, 10,000x for effector cells.

[0147] The binding partner that specifically binds to CD6 on T cells and prevents or inhibits activation of CD6 signaling can be an anti-CD6 antibody or an antigen-binding portion thereof.

[0148] Anti-CD6 antibodies can be any antibodies that bind to CD6 and block CD6-mediated downstream signals in T cells. For example, blocking studies using anti-CD6 monoclonal antibodies (monoclonal antibodies, mAbs) have shown that CD6 plays an important role in T cell development by regulating the adhesion of T cells to thymic epithelial (TE) cells (Patel et al., J. Exp. Med. (1995) 181: 1563-1568). Other studies have shown that CD6 can play an important role as an auxiliary molecule in T cell activation. For example, some anti-CD6 mAbs directly promote T cell mitosis (Gangemi et al., J. Immunol. (1989) 143:2439; Bott et al., Int. Immunol. (1993) 7:783), while other anti-CD6 mAbs can stimulate T cell proliferation together with anti-CD3, anti-CD2 or PMA (Gangemi et al., J. Immunol. (1989) 143:2439; Morimoto et al., J. Immunol. (1988) 140:2165-2170; Osorio et al., Cell. Immunol. (1994) 154:23). Additional evidence for a role for CD6 in T cell activation comes from studies showing that CD6 is hyperphosphorylated on Ser and Thr residues (Swack et al., Mol. Immunol. (1989) 26:1037-1049; Swack et al., J. Biol. Chem. (1991) 266:7137; Cardenas et al., J. Immunol., 145:1450-1455 (1990)), and phosphorylated on Tyr residues (Wee et al., J. Exp. Med. (1993) 177:219-223). These studies and others implicate CD6 as an important regulator of immature and mature T cell function in vivo, affecting both T cell activation and signal transduction (De Wit, J., et al., Blood (2011) 118:6107-6114), and any antibodies that can prevent these effects are suitable for use in the present invention.

[0149] U.S. Patent No. 6,372,215 discloses antibodies and other binding agents that specifically bind to SRCR domain 3 (D3) of human CD6 (hCD6) or human CD6 stalk domain (CD6S) and inhibit activated leukocyte adhesion molecule (ALCAM) binding to CD6.

[0150] Earlier publications and patents disclosed the sequence of a mouse anti-CD6 (IOR-T1) monoclonal antibody and the amino acid modifications made to humanize IOR-T1 to T1h (humanized IOR-T1). U.S. Pat. No. 5,712,120 and its equivalent EP0699755 disclose specific methods for humanizing mouse monoclonal antibodies and the sequences of IOR-T1 and T1h. U.S. Pat. No. 6,572,857 and its equivalent EP 0807125 disclose the sequences of IOR-T1 and T1h (humanized IOR-T1). PCT / IN2008 / 00562 and the related U.S. Pat. No. 8,524,233 entitled "A Monoclonal Antibody and a Method Thereof" disclose the production of anti-CD6 antibodies in NS0 cells, the anti-CD6 antibodies having the heavy and light chain sequences provided herein as SEQ ID NOs: 1 and 2. The antibody is also referred to as T1h in the patent, although its sequence is different from the original T1h. The INN name of the antibody is itolizumab. Itolizumab is produced in mouse-derived NS0 cell lines and Chinese Hamster Ovary (CHO) cells, and when produced in CHO cells, its trade name EQ001 is cited herein; when produced in NS0 cells, its trade name ALZUMAb is cited herein. EQ001 (i.e. itolizumab produced in CHO cells) is also referred to as "Bmab-600" in the art. In various embodiments herein, regardless of its production method, we refer to the antibody itself by its INN name itolizumab. Therefore, as used herein, the term itolizumab includes ALZUMAb and EQ001, each of which has the same sequence as itolizumab. The amino acid sequences of the variable heavy (VH) and variable light (VK) chains of Itolizumab (and EQ001 / ALZUMAb) are provided herein as SEQ ID NOs: 1 and 2, respectively. The nucleotide (DNA) sequences of the VH and VK chains of Itolizumab (and EQ001 / ALZUMAb) are provided herein as SEQ ID NOs: 3 and 4, respectively. The amino acid sequences of the VHCDR1-3 chains of Itolizumab (and EQ001 / ALZUMAb) are provided herein as SEQ ID NOs: 5-7, respectively. The amino acid sequences of the VKCDR1-3 chains of Itolizumab (and EQ001 / ALZUMAb) are provided herein as SEQ ID NOs: 8-10, respectively.

[0151] Therefore, the anti-CD6 antibody may be an anti-CD6 monoclonal antibody comprising heavy chain and light chain variable regions comprising the amino acid sequences shown in SEQ ID NO:1 and SEQ ID NO:2.

[0152] The anti-CD6 antibody can be an anti-CD6 monoclonal antibody comprising heavy chain and light chain variable regions, wherein the heavy chain and light chain variable regions comprise: the nucleotide sequence shown in SEQ ID NO: 3 or its complementary sequence; and (b) a nucleic acid molecule comprising the nucleotide sequence shown in SEQ ID NO: 4 or its complementary sequence.

[0153] The anti-CD6 antibody may be an anti-CD6 monoclonal antibody comprising heavy and light chain variable regions comprising amino acid sequences at least 80% homologous to the amino acid sequences shown in SEQ ID NO:1 and SEQ ID NO:2.

[0154] The anti-CD6 antibody can be an anti-CD6 monoclonal antibody that specifically binds to CD6 and comprises at least about 65% amino acid sequence identity or homology, at least about 70% amino acid sequence identity or homology, at least about 75% amino acid sequence identity or homology, at least about 80% amino acid sequence identity or homology, at least about 80% amino acid sequence identity or homology, at least about 85% amino acid sequence identity or homology, at least about 90% amino acid sequence identity or homology, at least about 95% amino acid sequence identity or homology, at least about 98% amino acid sequence identity or homology, or at least about 99% amino acid sequence identity or homology in the portion corresponding to the amino acid residues represented by SEQ ID NOs: 1 and 2.

[0155] The anti-CD6 antibody may comprise one or more CDRs selected from the following: EQ001 heavy chain CDR1: GFKFSRYAMS (SEQ ID NO: 5); EQ001 heavy chain CDR2: TISSGGSYIYYPDSVKG (SEQ ID NO: 6); EQ001 heavy chain CDR3: RDYDLDYFDS (SEQ ID NO: 7); EQ001 light chain CDR1: KASRDIRSYLT (SEQ ID NO: 8); EQ001 light chain CDR2: YATSLAD (SEQ ID NO: 9); EQ001 light chain CDR3: LQHGESP (SEQ ID NO: 10); and combinations thereof.

[0156] In a specific embodiment, the anti-CD6 antibody comprises each of the EQ001 CDRs provided in SEQ ID NOs: 5-10. In a specific embodiment, the anti-CD6 antibody is a humanized antibody comprising each of the EQ001 CDRs provided in SEQ ID NOs: 5-10. In a specific embodiment, the anti-CD6 antibody is a humanized IgG antibody comprising each of the EQ001 CDRs provided in SEQ ID NOs: 5-10. In a specific embodiment, the anti-CD6 antibody is a humanized IgG1 antibody comprising each of the EQ001 CDRs provided in SEQ ID NOs: 5-10. In a specific embodiment, the anti-CD6 antibody is a humanized antibody produced in CHO cells, wherein the humanized antibody comprises each of the EQ001 CDRs provided in SEQ ID NOs: 5-10.

[0157] The anti-CD6 antibody may be selected from UMCD6 mAb (Li et al., PNAS March 7, 2017, vol. 114, no. 10, 2687-2692, incorporated herein by reference in its entirety) and any one of the antibodies listed in Table 2:

[0158] Table 2: Anti-CD6 Antibodies

[0159]

[0160] The anti-CD6 antibody may be T1h disclosed in US Pat. No. 8,524,233 (incorporated herein by reference in its entirety).

[0161] The anti-CD6 antibody may be itolizumab. The anti-CD6 antibody may be ALZUMAb. The anti-CD6 antibody may be EQ001.

[0162] The anti-CD6 antibody may be an antibody produced by the secretory hybridoma IOR-T1A deposited in ECACC with the deposit number ECACC 96112640.

[0163] Anti-CD6 antibodies can bind to CD6 on the surface of T cells. Anti-CD6 antibodies can bind to domain 1, domain 2, or domain 3 of CD6 on the surface of T cells. In some aspects, anti-CD6 antibodies bind to domain 1 or domain 3 on CD6. In a specific embodiment, anti-CD6 antibodies bind to domain 3 on CD6. The binding of anti-CD6 antibodies to CD6 on the surface of T cells can regulate the activity of T cells. In some aspects, the binding of anti-CD6 antibodies to CD6 on the surface of T cells regulates the activity and / or migration of T cells. In specific aspects, the binding of anti-CD6 antibodies to CD6 on the surface of T cells regulates the migration of T cells to and through lung tissue.

[0164] An anti-CD6 antibody (eg, EQ001) can be delivered to a subject as an anti-CD6 pharmaceutical composition.

[0165] Pharmaceutical compositions suitable for delivering anti-CD6 antibodies and methods for preparing the same will be apparent to those skilled in the art. Such compositions and methods for preparing the same can be found, for example, in Remington's Pharmaceutical Sciences, 19th Edition (Mack Publishing Company, 1995), the contents of which are incorporated herein by reference in their entirety. Pharmaceutical compositions containing anti-CD6 antibodies are also known in the art. For example, anti-CD6 antibodies can be pharmaceutical compositions disclosed in U.S. Patent Application No. 12 / 525,449 (US20100047242) (incorporated herein in its entirety by reference).

[0166] The pharmaceutical compositions of the present invention may comprise an active pharmaceutical agent (eg, an anti-CD6 antibody, such as EQ001) and one or more pharmaceutically acceptable carriers, excipients, diluents, surfactants, and / or vehicles.

[0167] The pharmaceutical composition may comprise an anti-CD6 antibody (or an antigen-binding fragment thereof) and one or more agents selected from the group consisting of: carriers, excipients, diluents, antioxidants, preservatives, colorants, flavorings and diluents, emulsifiers, suspending agents, solvents, fillers, bulking agents, buffers, delivery vehicles, tonicity agents, cosolvents, wetting agents, complexing agents, buffers, antimicrobial agents and / or surfactants. These agents are known in the art (e.g., see Remington's Pharmaceutical Sciences, 18th edition, Mack Publishing Co., Easton, PA (1990), incorporated herein by reference in its entirety).

[0168] The present invention also includes combination therapy, which includes: administering to a patient an anti-CD6 antibody (e.g., EQ001) or an antigen-binding portion thereof in combination with a second active agent; or a device or procedure capable of treating, preventing or alleviating one or more asthma-related symptoms. In this context, "combined administration" means: (1) part of the same unit dosage form; (2) administered separately, but as part of the same treatment program or regimen, usually but not necessarily on the same day.

[0169] As previously described, EQ001 (or another anti-CD6 antibody) can be administered alone as a monotherapy in some aspects, or as a combination therapy in some aspects. In some aspects, any of the EQ001 (or another anti-CD6 antibody) described herein for administration to a patient according to the methods disclosed herein (e.g., EQ001) can be administered in combination with one or more other therapeutic agents as a combination therapy. For example, EQ001 (or another anti-CD6 antibody) can be administered to a patient as a combination therapy with another agent for the treatment of an inflammatory or autoimmune disease. The combination therapy can include administering EQ001 (or another anti-CD6 antibody) and an agent selected from, for example, but not limited to, a steroid or an immunosuppressant. The steroid can be a corticosteroid. The corticosteroid can be prednisone.

[0170] EQ001 (or another anti-CD6 antibody) can be administered before, after, or simultaneously with one or more such anti-inflammatory or autoimmune disease agents. In some embodiments, such combinations can provide significant advantages, including additive or synergistic activity in therapy.

[0171] In various embodiments, the compositions and methods disclosed herein, e.g., methods for treating asthma, involve administering to a subject an effective amount of EQ001 (or another anti-CD6 antibody) or a composition (e.g., a pharmaceutical composition) comprising EQ001 (or another anti-CD6 antibody).

[0172] EQ001 (or another anti-CD6 antibody) can be administered as a pharmaceutical composition. The CD6-ALCAM pathway inhibitor can be administered before, after, and / or simultaneously with one or more other therapeutic agents. If administered simultaneously with one or more other therapeutic agents, such administration can be simultaneous (e.g., in the form of a single composition), or can be by two or more separate compositions, optionally by the same or different modes of administration (e.g., non-systemic, systemic, oral, intravenous, etc.).

[0173] Administration of EQ001 (or another anti-CD6 antibody) and / or other therapeutic agents can be accomplished by any mode of administration for therapeutic agents. These modes include systemic or non-systemic administration, such as oral, nasal, parenteral, transdermal, subcutaneous, vaginal, buccal, rectal or topical modes of administration.

[0174] For administration in the methods of use described herein, EQ001 (or another anti-CD6 antibody) can be mixed with a non-toxic pharmaceutically acceptable carrier substance (e.g., saline or phosphate-buffered saline) prior to administration and will be administered using any medically appropriate method, such as parenteral administration (e.g., injection), such as by intravenous or intra-arterial injection.

[0175] The formulation of EQ001 (or another anti-CD6 antibody) used according to the present invention can be prepared by mixing the antibody having the desired purity with an optional pharmaceutically acceptable carrier, excipient or stabilizer in the form of a lyophilized formulation or an aqueous solution. Acceptable carriers, excipients, or stabilizers are non-toxic to recipients at the doses and concentrations employed, and include: buffers, such as phosphates, citrates, and other organic acids; antioxidants, including ascorbic acid and methionine; preservatives, such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl alcohol or benzyl alcohol; alkyl parabens, such as methyl paraben or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol and m-cresol; low molecular weight (less than about 1 0 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers, such as polyvinylpyrrolidone; amino acids, such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates, including glucose, mannose, or dextrins; chelating agents, such as EDTA; sugars, such as sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions, such as sodium; metal complexes (such as zinc protein complexes); and / or nonionic surfactants, such as TWEEN TM 、PLURONICS TM , or polyethylene glycol (PEG).

[0176] EQ001 (or another anti-CD6 antibody) can also be prepared by, for example, coacervation techniques or by interfacial polymerization and entrapment in microcapsules, such as hydroxymethylcellulose or gelatin microcapsules and poly(methyl methacrylate) microcapsules in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles and nanocapsules) or macroemulsions. Such techniques are well known in the art.

[0177] Sustained release formulations can be prepared. Suitable examples of sustained release formulations include semipermeable matrices of solid hydrophobic polymers containing EQ001 (or another anti-CD6 antibody), which are in the form of shaped articles, such as films or microcapsules. Examples of sustained release matrices include polyesters, hydrogels, copolymers of L-glutamic acid, non-degradable ethylene-vinyl acetate, and degradable lactic acid-glycolic acid copolymers.

[0178] EQ001 (or another anti-CD6 antibody) can be administered to a subject according to known methods, such as by bolus intravenous administration or continuous infusion over a period of time by intramuscular, intraperitoneal, intraspinal, subcutaneous, intraarticular, intrasynovial, intrathecal or oral routes. In some cases, intravenous or subcutaneous administration of EQ001 (or another anti-CD6 antibody) is preferred.

[0179] Depending on the intended mode of administration, the disclosed compounds or pharmaceutical compositions may be in solid, semisolid or liquid dosage forms, e.g., injections, tablets, suppositories, pills, delayed release capsules, elixirs, tinctures, emulsions, syrups, powders, liquids, suspensions, etc., sometimes in unit doses, and consistent with conventional pharmaceutical practice. Similarly, the disclosed compounds or pharmaceutical compositions may also be administered intravenously (bolus and infusion), intraperitoneally, subcutaneously or intramuscularly, and all in forms familiar to those skilled in the art of pharmacy. Pharmaceutical compositions suitable for delivery of EQ001 (or another anti-CD6 antibody) (alone or, for example, in combination with another therapeutic agent according to the present invention) and methods for their preparation are apparent to those skilled in the art. Such compositions and methods for their preparation may be found, for example, in Remington's Pharmaceutical Sciences, 19th edition (Mark Publishing Company), the contents of which are incorporated herein in their entirety.

[0180] The dosage regimen using EQ001 (or another anti-CD6 antibody) is selected based on a variety of factors, including: the type, species, age, weight, sex and physical condition of the patient; the severity of the disease to be treated; the route of administration; the patient's renal or liver function; and the specific disclosed compound used. A physician or veterinarian of ordinary skill in the art can readily determine and prescribe the effective amount of the drug required to prevent, counter or arrest the progression of the disease.

[0181] An exemplary non-limiting range of a therapeutically effective amount of EQ001 (or another anti-CD6 antibody) used in the present invention is about 0.01 mg / kg-100 mg / kg per subject's body weight, such as about 0.01 mg / kg-50 mg / kg, such as about 0.01 mg / kg-25 mg / kg. A medical technician with ordinary skills in the art can easily determine and prescribe the effective amount of the desired pharmaceutical composition. For example, a physician can start with a dose of EQ001 (or another anti-CD6 antibody) at a level lower than the required level to obtain the desired therapeutic effect, and gradually increase the dose until the desired effect is achieved.

[0182] In one embodiment, EQ001 (or another anti-CD6 antibody) is infused at a weekly dose of 1 mg to 500 mg per kg of subject body weight (e.g., 20 mg / kg to 200 mg / kg). This administration can be repeated, for example, 1-8 times, such as 3-5 times. In an alternative approach, administration can be performed by continuous infusion over a period of 2 hours to 24 hours, such as 2 hours to 12 hours.

[0183] In one embodiment, EQ001 (or another anti-CD6 antibody) is administered up to 7 times, e.g., 4-6 times, at a weekly dose of 0 mg to 200 mg. Administration can be performed by continuous infusion over a period of 2 to 24 hours, e.g., 2 to 12 hours. This regimen can be repeated one or more times as needed, e.g., after 6 months or 2 months.

[0184] In some aspects of these combination therapies, the second active agent is one or more agents capable of modulating the immune system. In some aspects of these combination therapies, the second active agent is one or more immunosuppressants. In some aspects of these combination therapies, the second active agent is one or more beta agonists.

[0185] In some aspects of these combination therapies, the second active agent is one or more short-acting beta agonists. In some aspects of these combination therapies, the second active agent is albuterol. In some aspects, albuterol is administered in a dosage form selected from: aerosol powder; solution; capsule; and powder suspension. In some aspects of these combination therapies, the second active agent is a steroid, such as a corticosteroid. In some aspects, the corticosteroid is administered in a dosage form selected from: tablets, sustained-release capsules; extended-release tablets; extended-release capsules; syrups; solutions; elixirs; suspensions; sustained-release tablets; liquids; and disintegrating tablets.

[0186] In some aspects of these combination therapies, the second active agent is ipratropium. In some aspects, ipratropium is administered in a spray formulation.

[0187] In some aspects of these combination therapies, the second active agent includes one or more agents selected from the group consisting of beclomethasone dipropionate, budesonide, flunisolide, fluticasone propionate, mometasone furoate, triamcinolone acetonide, dexamethasone, hydrocortisone, methylprednisone, prednisolone, prednisone, formoterol fumarate, salmeterol xinafoate, albuterol sulfate, isoetharine hydrochloride, isoproterenol hydrochloride, and sedative. hydrochloride; levalbuterol hydrochloride; pirbuterol acetate; terbutaline sulfate; ipratropium bromide; montelukast sodium; zafirlukast; zileuton; oxytriphylline; theophylline; cromolyn sodium; nedocromil sodium; omalizumab; combination medicines containing fluticasone and salmeterol; combination medicines containing xinafoate (inhaled steroid plus long-acting beta-2 agonist); combination medicines containing budesonide and formoterol fumarate; fumarate (an inhaled steroid plus a long-acting beta-2 agonist); a combination medicine containing fluticasone propionate and salmeterol xinafoate (an inhaled steroid plus a long-acting beta-2 agonist);A combination comprising budesonide and formoterol fumarate (an inhaled steroid plus a long-acting beta-2 agonist); a combination comprising ipratropium bromide and albuterol sulfate; or a combination thereof. ;

[0188] In some aspects, the invention also includes a combination therapy comprising administering an anti-CD6 antibody (e.g., EQ001) or an antigen binding portion thereof to a patient in combination with a procedure selected from intubation, mechanical ventilation, oxygen therapy, and combinations thereof. Such a procedure may also be administered to a subject in combination with any one or more of the aforementioned additional agents useful in the combination therapy described herein.

[0189] In certain aspects, the methods disclosed herein, which can include administering to a subject an anti-CD6 antibody (eg, EQ001), or an antigen-binding portion thereof, can provide treatment for one or more asthma-associated symptoms.

[0190] The optimal dosage and dosage regimen to be administered can be readily determined by those skilled in the art and will vary with the pharmacodynamic characteristics of the specific agent, its time and mode of administration, the strength of the preparation, and the development of the disease state (including the nature of the disease and the extent of the symptoms). In addition, factors related to the specific patient being treated (including the patient's sex, age, weight, diet, physical activity, and concomitant diseases) will result in the need to adjust the dosage and / or regimen.

[0191] All U.S. patents, U.S. patent application publications, U.S. patent applications, PCT patent applications, PCT patent application publications, foreign applications, foreign patent applications, and non-patent publications cited in this specification or listed in any application data sheet are incorporated herein by reference in their entirety. From the foregoing, it will be appreciated that although specific embodiments of the invention have been described herein for illustrative purposes, various modifications may be made without departing from the spirit and scope of the invention.

[0192] Example

[0193] Example 1

[0194] CD6 expression is significantly elevated in human fatal asthma patients

[0195] Summary of results: Lung tissues from patients with fatal asthma showed CD6 + The cells were highly infiltrated into the lamina propria, and ALCAM was overexpressed in this area.

[0196] Fresh lungs were obtained from patients with fatal asthma or non-asthmatic subjects. Tissue samples were fixed and subsequently stained for CD6 ALCAM expression by immunofluorescence. ALCAM expression was elevated in the lamina propria of patients with fatal asthma and absent in the lamina propria of non-asthmatic controls. In these lungs, ALCAM staining was localized to areas of high levels of infiltrating CD6 + In cells ( Figure 2 ). This suggests that severe asthma is associated with CD6 + T cell infiltration, and ALCAM expression may be involved in CD6 T cell infiltration during severe / fatal asthma + Cell migration / infiltration into the lung. In contrast, previous studies have shown that ALCAM is reduced in the lung tissue of animals exposed to allergens in an allergic asthma model due to increased metalloproteinase-mediated ALCAM shedding.

[0197] Example 2

[0198] CD6 expression is significantly elevated in human fatal asthma patients

[0199] Summary of results: De novo sequencing analysis of publicly available RNASeq datasets confirmed that severe asthmatics expressed significantly higher levels of CD6 compared with moderate asthmatics and non-asthmatics.

[0200] RNA data were obtained from cell pellets collected by bronchiolar lavage as part of two longitudinal prospective clinical studies:

[0201] BOBCAT study (Arron et al., Eur Respir J. 2014 Feb; 43(2): 627-9)

[0202] MAST study (Simpson et al., Nat Immunol. 2014 Dec; 15(12): 1162-70)

[0203] The transcriptomes of these samples were determined by RNASeq, and a dataset containing transcription read counts plotted against all individual genes for each sample was obtained in a public database (Sun et al., Sci Signal. 2015 Dec 1; 8(405)).

[0204] We used bioinformatics to mine the literature to compare CD6 expression in controls, moderate asthma, and severe asthma patients, and our analysis showed significant differences in CD6 expression levels, with severe asthma patients expressing significantly higher levels of CD6 ( Figure 3B). Similarly, although a less pronounced increase in CD4 expression was also observed, severe asthmatics expressed significantly higher levels of CD4 than moderate and healthy subjects ( Figure 3 A). Concomitant increase in CD6 + Therefore, these data suggest that CD6 + Increased T cells.

[0205] Comparative analysis of the expression of Th17 markers and cytokines in controls, moderate asthmatics, and severe asthmatics revealed that CCR6 ( Figure 3 C), CCR4( Figure 3 D) KLRB1( Figure 3 E), IL-17A ( Figure 3 F) and IL-17F( Figure 3 G) showed significant differences in the expression levels of Th17 cells, indicating an increased presence of Th17 cells in patients with severe asthma and a correlation between CD6 and Th17 cells.

[0206] Using the same dataset, objective cluster analysis of all asthma patients (regardless of asthma type) revealed a subgroup of severe asthma patients with high CD6 and similar gene expression profiles, suggesting that a unique patient profile may be associated with high CD6 expression ( Figure 4 A and Figure 4 B).

[0207] The authors are not aware of any prior analysis that examined CD6 expression in this dataset or any prior reports that CD6 expression is increased in severe asthma. These data suggest that CD6 is highly expressed in the lungs of patients with severe asthma that is poorly treated with steroids. Therefore, these data appear as the first evidence supporting the use of CD6 inhibition to treat severe asthma (e.g., asthma caused by Th1 / Th17 T cells).

[0208] Of note, allergic asthma also includes a T cell component, although Th2-mediated responses are well responsive to steroid therapy. Therefore, we sought to determine whether CD6 might be a viable target for therapies that are effective against severe and allergic asthma, as this would have the potential to: (1) minimize patient compliance issues due to adverse side effects of long-term steroid use; (2) provide a convenient single-agent therapy for all forms of asthma; and (3) provide a therapy that patients could continue to use even if their asthma endotype changes (e.g., Th2 to Th2 / Th17 or Th1 / Th17).

[0209] Example 3

[0210] CD6 inhibition is a viable treatment for allergic and severe asthma

[0211] Summary of results: In a murine model of allergic asthma, CD6 blockade was demonstrated to be a potent immunomodulator of Th2-mediated disease.

[0212] Figure 5 A and Figure 5 B shows the experimental groups and experimental schemes used in the allergic asthma experiment, respectively. Briefly, an anti-OVA Th2-driven immune response was induced by inoculation with OVA / alum on days 0 and 14 to induce allergic asthma by sensitization to ovalbumin (OVA). On days 23, 25, and 27, mice were treated with anti-mouse CD6Sc domain 1 antibody (mCD6D1), which is a mouse substitute for EQ001 that binds to domain 1 of CD6 and has similar characteristics to the anti-human CD6 antibody itolizumab. Subsequently, sensitized mice were intranasally attacked with OVA on days 25-27, and then terminated on day 28 to evaluate cells and cytokines in the lungs.

[0213] CD6 blockade during the challenge resulted in decreased levels of Th2 cytokines IL-4, IL-5, and IL-13 in bronchiolar lavage fluid (BALF) ( Figure 5 C), accompanied by a modest decrease in pneumocytes, indicating an inhibitory effect of CD6 blockade on Th2 responses.

[0214] To further support the ability of CD6 blockade to influence Th2 responses, we tested the blocking effect of CD6 blockade in the classical Th2 model of classical OVA vaccination. Figure 6 A and Figure 6 B shows the experimental groups and experimental scheme used in this experiment, respectively. Briefly, mice were vaccinated with OVA / alum on days 0 and 14 and treated with anti-CD6 twice a week from day -1 to day 16. On day 19, mice were sacrificed to examine anti-OVA antibody responses. Prophylactic CD6 blockade inhibited the production of OVA-specific IgE, demonstrating the influence of the CD6 pathway on Th2 responses ( Figure 6 C).

[0215] Therefore, these data combined suggest that CD6 blockade is a viable treatment for Th2-mediated allergic asthma, both acutely and preventively. Moreover, the data provided herein combined with the known ability of anti-CD6 antibodies (e.g., EQ001) to inhibit Th1 / Th17-type T cell activation and migration support that CD6 inhibition (e.g., using EQ001) will be able to treat severe and allergic asthma.

[0216] Example 4

[0217] Efficacy of anti-CD6 antibodies in a severe asthma model

[0218] Mice were sensitized intranasally with 25 μg of HDM on days 1, 3, and 5. The mice were then left for 5 days before a 3-challenge setup involving 3 consecutive challenges with 25 μg of HDM, with a 4-day rest between challenge setups. 25 μg of HDM was used only in the last two attacks. Dexamethasone (Dex) positive treatment control or anti-CD6 antibody test product was administered intraperitoneally at a concentration of 4 mg / kg starting on the first day of the challenge, and then Dex was repeated every three days and anti-CD6 antibody was administered every other day.

[0219] Example 5

[0220] Efficacy of anti-CD6 antibodies in asthma models

[0221] Due to the ability of CD6 blockade to target multiple T cell subtypes, it is expected that CD6 inhibition will also show efficacy in many asthma models, including HDM-induced, HDM+LPS-induced, cockroach-induced, and Alternaria alternata-induced asthma models, as well as the generation of asthma models capable of having a Th1 / Th17 phenotype in STAT6- / - mice. Therefore, several models were used to test doses of anti-CD6 antibodies ranging from 600ug down to 10ug (Table 3).

[0222] Table 3: Additional mouse models

[0223]

[0224]

[0225] The results of these experiments will demonstrate that CD6 inhibition is effective in inhibiting: Th1 / Th17 forms of asthma; Th2, and T2 / Th17 forms of asthma, etc. Thus, the present invention supports the development of anti-CD6 antibodies for the treatment of severe and allergic asthma.

[0226] Example 6

[0227] Analysis of tissue samples

[0228] Tissue samples were collected after completing the protocols in Examples 4 and 5 and analyzed according to the following procedures.

[0229] Body weights and clinical scores of all mice were collected daily according to protocols approved by the Institutional Animal Care and Use Committee (IACUC).

[0230] Twenty-four hours after the last allergen challenge, mice were anesthetized and bronchoalveolar lavage (BAL) was performed. The left bronchus of each mouse was ligated and the right lobe was lavaged with 0.7 ml of sterile PBS to obtain BAL fluid. The total BAL cell count was determined using trypan blue staining and standard light microscopy, and then 750,000 BAL fluid cells were centrifuged onto a clean slide and differentiated cell counts were performed using Giemsa staining.

[0231] BAL fluid was centrifuged to separate cells from the supernatant. The supernatant was analyzed using standard cytokine assays to identify specific cell markers and / or cytokines, and the cells were analyzed by standard flow cytometry methods. Lavaged lungs were placed in fixative for 48 hours and then transferred to 70% ethanol until paraffin embedding for periodic acid–Schiff (PAS) staining, hematoxylin and eosin staining, and / or immunohistochemical or immunofluorescent staining for T cell markers (e.g., CD3, CD4, CD8), CD6, ALCAM, and other known or potential ligands for CD6.

[0232] The various embodiments described above can be combined to provide other embodiments. All U.S. patents, U.S. patent application publications, U.S. patent applications, foreign patents, foreign patent applications, and non-patent publications cited in this specification or listed in any application data sheet are fully incorporated herein by reference. If it is necessary to adopt the concepts of various patents, applications, and publications to provide other embodiments, various aspects of the embodiments can be modified.

[0233] These and other changes can be made to the embodiments in light of the above detailed description. In general, the terms used in the claims should not be interpreted as limiting the claims to the specific embodiments disclosed in the specification and claims, but should be interpreted as including all possible embodiments, and all equivalent ranges of the protection scope of the claims. Therefore, the claims are not limited by what is disclosed. Sequence Listing <110> Balance Biotech Co., Ltd. Stephen Connolly Cherie NG <120> Anti-CD6 antibody for treating severe asthma <130> EQIL-006 / 01WO 330372-2026 <150> US 62 / 636,092 <151> 2018-02-27 <160> 10 <170> PatentIn version 3.5 <210> 1 <211> 119 <212> PRT <213> Artificial Sequence <220> <223> Made in Lab - humanized T1h VH sequence <400> 1 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Lys Phe Ser Arg Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Arg Leu Glu Trp Val 35 40 45 Ala Thr Ile Ser Ser Gly Gly Ser Tyr Ile Tyr Tyr Pro Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Val Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Ser Ser Leu Arg Ser Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Arg Arg Asp Tyr Asp Leu Asp Tyr Phe Asp Ser Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 2 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> Made in Lab - humanized T1h VL sequence <400> 2 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Arg Asp Ile Arg Ser Tyr 20 25 30 Leu Thr Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Thr Leu Ile 35 40 45 Tyr Tyr Ala Thr Ser Leu Ala Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Gln Asp Tyr Ser Leu Thr Ile Ser Ser Leu Glu Ser 65 70 75 80 Asp Asp Thr Ala Thr Tyr Tyr Cys Leu Gln His Gly Glu Ser Pro Phe 85 90 95 Thr Leu Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 3 <211> 357 <212> DNA <213> Artificial Sequence <220> <223> Made in Lab - humanized T1h VH nucleic acid sequence <400> 3 gaagtgcagc tggtggagtc tgggggaggc ttagtgaagc ctggagggtc cctgaaactc 60 tcctgtgcag cctctggatt caagtttagt agatatgcca tgtcttgggt tcgccaggct 120 ccggggaaga ggctggagtg ggtcgcaacc attagtagtg gtggtagtta catctactat 180 ccagacagtg tgaagggtcg attcaccatc tccagagaca atgtcaagaa caccctgtat 240 ctgcaaatga gcagtctgag gtctgaggac acggccatgt attactgtgc aagacgagat 300 tacgacctgg actactttga ctcctggggc caaggcaccc ttgtcaccgt ctcctca 357 <210> 4 <211> 321 <212> DNA <213> Artificial Sequence <220> <223> Made in Lab - humanized T1h VL nucleic acid sequence <400> 4 gacatccaga tgacccagtc tccatcctcc ctgtctgcat cggtgggaga cagagtcact 60 atcacttgca aggcgagtcg ggacattaga agctatttaa cctggtacca gcagaaacca 120 gggaaagctc ctaagaccct gatctattat gcaacaagct tggcagatgg ggtcccgtcg 180 agattcagtg gcagtggatc tgggcaagat tattctctca ccatcagcag cctggagtct 240 gacgatacag caacttacta ctgtctacaa catggtgaga gtccattcac gctcggctcg 300 gggaccaagc tggaaatcaa a 321 <210> 5 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Made in Lab - humanized T1h VH CDR1 <400> 5 Gly Phe Lys Phe Ser Arg Tyr Ala Met Ser 1 5 10 <210> 6 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> Made in Lab - humanized T1h VH CDR2 <400> 6 Thr Ile Ser Ser Gly Gly Ser Tyr Ile Tyr Tyr Pro Asp Ser Val Lys 1 5 10 15 Gly <210> 7 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Made in Lab - humanized T1h VH CDR3 <400> 7 Arg Asp Tyr Asp Leu Asp Tyr Phe Asp Ser 1 5 10 <210> 8 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Made in Lab - humanized T1h VL CDR1 <400> 8 Lys Ala Ser Arg Asp Ile Arg Ser Tyr Leu Thr 1 5 10 <210> 9 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Made in Lab - humanized T1h VL CDR2 <400> 9 Tyr Ala Thr Ser Leu Ala Asp 1 5 <210> 10 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Made in Lab - humanized T1h VL CDR3 <400> 10 Leu Gln His Gly Glu Ser Pro 1 5

Claims

1. Use of an anti-CD6 antibody or an antigen-binding fragment thereof in the preparation of a medicament for treating non-allergic severe asthma, characterized in that: The CDR sequences of the anti-CD6 antibody or antigen-binding fragment thereof are shown in SEQ ID NOs: 5-10, and the non-allergic severe asthma is characterized by: - No blood eosinophils or blood eosinophil count ≤ 300 cells / μl, - High levels of Th1, Th17, or both Th1 and Th17 T cell types in T cells, - is refractory to steroid therapy, and - is agranulocytic.

2. The use according to claim 1, characterized in that: The anti-CD6 antibody or antigen-binding fragment thereof is Itolizumab, wherein the heavy chain and light chain variable regions comprise the amino acid sequences shown in SEQ ID NO: 1 and SEQ ID NO:

2.

3. The use according to any one of claims 1-2, characterized in that: The treatment may also include administering one or more additional agents that are capable of treating, preventing or alleviating one or more of the asthma symptoms.

4. The use according to claim 3, characterized in that: The additional agent includes ipratropium.

5. The use according to claim 4, characterized in that: The ipratropium is administered in a spray form.

6. The use according to any one of claims 1-2, characterized in that: Such treatment also includes administration of intubation, mechanical ventilation and / or oxygen therapy.

7. The use according to any one of claims 1-2, characterized in that: The anti-CD6 antibody or antigen-binding fragment thereof is administered as a pharmaceutical composition comprising one or more pharmaceutically acceptable salts, excipients or vehicles.

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