How to treat allergies using anti-BET V1 antibodies

Abstract

The present disclosure provides methods for treating, preventing, or ameliorating one or more symptoms of birch allergy or allergic disease in a subject by administering to the subject an antibody or antigen-binding fragment thereof that binds Bet v 1, or a cocktail of antibodies or antigen-binding fragments thereof that bind Bet v 1. In another aspect, a cocktail comprising two or more anti-Bet v 1 antibodies or antigen-binding fragments thereof is provided for use in a method of treating a subject with seasonal or perennial allergy associated with birch and cross-reactive pollen. In some embodiments, the method comprises administering a cocktail disclosed herein to a subject in need thereof (e.g., a subject with seasonal or perennial allergy associated with birch and cross-reactive pollen).

Description

Technical Field

[0001] The present disclosure relates to the use of human antibodies that bind to Bet v 1 for performing the same in a subject that requires treatment or prevention of an allergic reaction and an allergic disease.

[0002] Sequence Listing The sequence listing was submitted electronically via EFS-Web as a sequence listing in ASCII format simultaneously with this specification, the file name is "10821WO01_Sequence_Listing_ST25", the creation date is June 30, 2021, and the size is about 32 kilobytes. The sequence listing contained in this ASCII format written document is part of this specification and is incorporated herein by reference in its entirety.

Background Art

[0003] Allergic rhinitis (AR) is an allergic disease characterized by one or more symptoms including sneezing, itching, nasal congestion, and rhinorrhea, and often includes symptoms such as fatigue, malaise, hypersensitivity, and in some cases neurocognitive deficits. Conjunctivitis is also common and has been reported in about 65% of people with AR (Rosario and Bielory, Current Opinion in Allergy and Clinical Immunology 2011, 11:471-476). Allergic rhinitis affects approximately 100 million people in Europe and its incidence is increasing (Wallace et al, Journal of Allergy and Clinical Immunology 2008, 122:S1-84). People with AR can be allergic to pollen, mold, house dust mites, and / or animal dander. The diagnosis of AR is confirmed by a positive history and evidence of immunoglobulin E (IgE)-mediated inflammation by either an allergen-specific skin prick test (SPT) and / or an allergen-specific IgE antibody test.

[0004] In Europe and the United States, clinically significant sensitization to birch affects approximately 20–30% of the allergic population (see, e.g., Pablos et al, Current Allergy and Asthma Reports 2016, 16:31). Birch pollen contains a mixture of allergenic and non-allergenic proteins, with Bet v 1 being the most abundant allergenic pollen protein (Erler et al, Proteomics 2011 11:1486-1498, Schenk et al, Journal of Proteomics 2011, 74:1290-1300). The sensitization rate to Bet v 1 in individuals allergic to birch reaches over 95%. Because the pollen grains are large (over 5 μm) and primarily affect nasal and ocular symptoms, they are generally not observed to enter the lower respiratory tract. However, cough and wheezing are not uncommon and are associated with breathable concentrations of Bet v 1 detectable in the air, suggesting that pollen fragments may enter the lower respiratory tract and exacerbate asthma (see, e.g., Taylor et al, Clinical and Experimental Allergy 2004, 34:1591-1596; Taylor et al, International Archives of Allergy and Immunology 2007, 144:162-170). Furthermore, mid-spring pollen exposure is associated with increased use of commercially available rhinitis medications and a peak in asthma-related emergency visits. The strongest association is found in children aged 5–17 years with allergies to birch, oak, beech, and ash pollen (Ito et al, Environmental Health: A Global Access Science Source 2015, 14:71). Up to 70% of people with pollen allergies also experience oral allergy syndrome (OAS), also known as pollen-food allergy, which involves an oral reaction to certain fresh fruits, vegetables, and nuts. Oral allergy syndrome is not a separate food allergy but occurs due to cross-reactive epitopes present in pollen and related foods.The symptoms of oral allergy syndrome typically manifest as itching of the lips, mouth, and throat, but may also be accompanied by swelling and angioedema of the lips and tongue (Bucher et al, Allergy 2004, 59:1272-1276), which causes people to avoid these fresh fruits, vegetables, and nuts. Recommendations for the treatment of AR include allergen avoidance, medications that provide short-term relief of symptoms such as antihistamines and intranasal corticosteroids, and allergen-specific immunotherapy (SIT). Antihistamines and intranasal corticosteroids are widely used, but approximately half of AR patients report poor or only partial symptom control and short-term relief (see, e.g., Wallace et al, Current Opinion in Allergy and Clinical Immunology 2017, 17:286-294; Wei, Biomedicine & Pharmacotherapy 2016, 83:989-997). Specific immunotherapy is indicated when moderate to severe symptoms of AR persist despite the use of antihistamines and intranasal corticosteroids. The use of allergen-specific SIT is well established, but there are numerous limitations to SIT in both subcutaneous immunotherapy (SCIT) and sublingual immunotherapy (SLIT) tablets or eye drops. For example, the efficacy of SIT is moderate, resulting in an improvement of approximately 20% to 30% of allergic symptoms in people who have completed treatment compared to those receiving placebo, and it may take at least 3 years to induce immune tolerance (see, e.g., Nolte et al, Journal of Allergy and Clinical Immunology 2016, 138:1631-1638). Furthermore, the efficacy of SIT requires high compliance to prevent symptoms of rhinitis upon allergen exposure. Side effects of SCIT occur in 40% to 50% of patients, ranging from mild reactions (e.g., swelling, injection site reactions, de novo allergic responses, and urticaria) to life-threatening reactions (e.g., asthma exacerbation and anaphylaxis) (see, e.g., Frew, Journal of Allergy and Clinical Immunology 2010, 125:S306-313). Additionally, asthma is a major risk factor for life-threatening systemic reactions to SCIT, and thus moderate to severe asthma is a contraindication to SCIT. Therefore, there remains a need for a safe and effective therapy for treating allergic individuals. [Primary Technology Documents]

Non-licensed literature

[0005] [Non-licensed document 1] Rosario and Bielory,Current Opinion in Allergy and Clinical Immunology 2011,11:471-476 [Non-licensed document 2] Wallace et al,Journal of Allergy and Clinical Immunology 2008,122:S1-84 [Non-licensed document 3] Pablos et al,Current Allergy and Asthma Reports 2016,16:31

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[0006] In one embodiment, a method for treating birch allergy in a subject is provided. In some embodiments, this method is (a) A first anti-Bet v 1 antibody or its antigen-binding fragment, wherein the first anti-Bet v 1 antibody comprises a heavy chain complementarity-determining region (HCDR) 1 containing the amino acid sequence of SEQ ID NO: 2, an HCDR2 containing the amino acid sequence of SEQ ID NO: 3, an HCDR3 containing the amino acid sequence of SEQ ID NO: 4, a light chain complementarity-determining region (LCDR) 1 containing the amino acid sequence of SEQ ID NO: 6, an LCDR2 containing the amino acid sequence of SEQ ID NO: 7, and an LCDR3 containing the amino acid sequence of SEQ ID NO: 8, and / or (b) A second anti-Bet v 1 antibody or its antigen-binding fragment, wherein the second anti-Bet v 1 antibody comprises HCDR1 containing the amino acid sequence of SEQ ID NO: 12, HCDR2 containing the amino acid sequence of SEQ ID NO: 13, HCDR3 containing the amino acid sequence of SEQ ID NO: 14, LCDR1 containing the amino acid sequence of SEQ ID NO: 16, LCDR2 containing the amino acid sequence of SEQ ID NO: 17, and LCDR3 containing the amino acid sequence of SEQ ID NO: 18, and / or (c) The third anti-Bet v 1 antibody or antigen-binding fragment thereof, comprising administering to one or more pharmaceutical compositions comprising the third anti-Bet v 1 antibody or antigen-binding fragment thereof, wherein the third anti-Bet v 1 antibody comprises HCDR1 containing the amino acid sequence of SEQ ID NO: 22, HCDR2 containing the amino acid sequence of SEQ ID NO: 23, HCDR3 containing the amino acid sequence of SEQ ID NO: 24, LCDR1 containing the amino acid sequence of SEQ ID NO: 26, LCDR2 containing the amino acid sequence of SEQ ID NO: 27, and LCDR3 containing the amino acid sequence of SEQ ID NO: 28.

[0007] In another embodiment, a method is provided for reducing one or more symptoms of an allergic reaction to a Fagales allergen in a subject. In some embodiments, the method is (a) A first anti-Bet v 1 antibody or its antigen-binding fragment, wherein the first anti-Bet v 1 antibody comprises HCDR1 containing the amino acid sequence of SEQ ID NO: 2, HCDR2 containing the amino acid sequence of SEQ ID NO: 3, HCDR3 containing the amino acid sequence of SEQ ID NO: 4, LCDR1 containing the amino acid sequence of SEQ ID NO: 6, LCDR2 containing the amino acid sequence of SEQ ID NO: 7, and LCDR3 containing the amino acid sequence of SEQ ID NO: 8, and / or (b) A second anti-Bet v 1 antibody or its antigen-binding fragment, wherein the second anti-Bet v 1 antibody comprises HCDR1 containing the amino acid sequence of SEQ ID NO: 12, HCDR2 containing the amino acid sequence of SEQ ID NO: 13, HCDR3 containing the amino acid sequence of SEQ ID NO: 14, LCDR1 containing the amino acid sequence of SEQ ID NO: 16, LCDR2 containing the amino acid sequence of SEQ ID NO: 17, and LCDR3 containing the amino acid sequence of SEQ ID NO: 18, and / or (c) The third anti-Bet v 1 antibody or antigen-binding fragment thereof, comprising administering to one or more pharmaceutical compositions comprising the third anti-Bet v 1 antibody or antigen-binding fragment thereof, wherein the third anti-Bet v 1 antibody comprises HCDR1 containing the amino acid sequence of SEQ ID NO: 22, HCDR2 containing the amino acid sequence of SEQ ID NO: 23, HCDR3 containing the amino acid sequence of SEQ ID NO: 24, LCDR1 containing the amino acid sequence of SEQ ID NO: 26, LCDR2 containing the amino acid sequence of SEQ ID NO: 27, and LCDR3 containing the amino acid sequence of SEQ ID NO: 28.

[0008] In some embodiments, the Fagales allergen is Bet v 1. In some embodiments, the subject is sensitized to Bet v 1 and at least one other Fagales allergen. In some embodiments, the at least one other Fagales allergen is alder, hazel, oak, hornbeam, Japanese jasmine, beech, horse chestnut, hazelnut, or apple.

[0009] In another embodiment, a method is provided for treating subjects having seasonal or perennial allergies related to birch and cross-reactive pollen. In some embodiments, the subjects have moderate to severe seasonal or moderate to severe perennial allergies. In some embodiments, the method is (a) A first anti-Bet v 1 antibody or its antigen-binding fragment, wherein the first anti-Bet v 1 antibody comprises HCDR1 containing the amino acid sequence of SEQ ID NO: 2, HCDR2 containing the amino acid sequence of SEQ ID NO: 3, HCDR3 containing the amino acid sequence of SEQ ID NO: 4, LCDR1 containing the amino acid sequence of SEQ ID NO: 6, LCDR2 containing the amino acid sequence of SEQ ID NO: 7, and LCDR3 containing the amino acid sequence of SEQ ID NO: 8, and / or (b) A second anti-Bet v 1 antibody or its antigen-binding fragment, wherein the second anti-Bet v 1 antibody comprises HCDR1 containing the amino acid sequence of SEQ ID NO: 12, HCDR2 containing the amino acid sequence of SEQ ID NO: 13, HCDR3 containing the amino acid sequence of SEQ ID NO: 14, LCDR1 containing the amino acid sequence of SEQ ID NO: 16, LCDR2 containing the amino acid sequence of SEQ ID NO: 17, and LCDR3 containing the amino acid sequence of SEQ ID NO: 18, and / or (c) The third anti-Bet v 1 antibody or antigen-binding fragment thereof, comprising administering to one or more pharmaceutical compositions comprising the third anti-Bet v 1 antibody or antigen-binding fragment thereof, wherein the third anti-Bet v 1 antibody comprises HCDR1 containing the amino acid sequence of SEQ ID NO: 22, HCDR2 containing the amino acid sequence of SEQ ID NO: 23, HCDR3 containing the amino acid sequence of SEQ ID NO: 24, LCDR1 containing the amino acid sequence of SEQ ID NO: 26, LCDR2 containing the amino acid sequence of SEQ ID NO: 27, and LCDR3 containing the amino acid sequence of SEQ ID NO: 28.

[0010] In some embodiments of the methods disclosed herein, the pharmaceutical composition comprises a first anti-Bet v 1 antibody. In some embodiments, the pharmaceutical composition comprises a second anti-Bet v 1 antibody. In some embodiments, the pharmaceutical composition comprises a third anti-Bet v 1 antibody. In some embodiments, the pharmaceutical composition comprises a first anti-Bet v 1 antibody and a second anti-Bet v 1 antibody. In some embodiments, the pharmaceutical composition comprises a first anti-Bet v 1 antibody and a third anti-Bet v 1 antibody. In some embodiments, the pharmaceutical composition comprises a second anti-Bet v 1 antibody and a third anti-Bet v 1 antibody. In some embodiments, the pharmaceutical composition comprises a first anti-Bet v 1 antibody, a second anti-Bet v 1 antibody, and a third anti-Bet v 1 antibody.

[0011] In some embodiments, the anti-Bet v 1 antibody is provided in a single pharmaceutical composition. In some embodiments, the anti-Bet v 1 antibody is provided in two or more pharmaceutical compositions, for example, in each anti-Bet v 1 antibody in separate pharmaceutical compositions.

[0012] In some embodiments, the pharmaceutical composition contains an anti-Bet v 1 antibody (e.g., each of a first anti-Bet v 1 antibody, a second anti-Bet v 1 antibody, and a third anti-Bet v 1 antibody) in an independently selected dose ranging from about 5 mg to about 500 mg. In some embodiments, the pharmaceutical composition contains an anti-Bet v 1 antibody (e.g., each of a first anti-Bet v 1 antibody, a second anti-Bet v 1 antibody, and a third anti-Bet v 1 antibody) in an independently selected dose ranging from about 50 mg, about 100 mg, about 150 mg, about 200 mg, about 250 mg, about 300 mg, about 350 mg, about 400 mg, about 450 mg, or about 500 mg. In some embodiments, the pharmaceutical composition contains each of a first anti-Bet v 1 antibody, a second anti-Bet v 1 antibody, and a third anti-Bet v 1 antibody in a dose of about 50 mg. In some embodiments, the pharmaceutical composition contains each of the first anti-Bet v 1 antibody, the second anti-Bet v 1 antibody, and the third anti-Bet v 1 antibody in a dose of about 150 mg. In some embodiments, the pharmaceutical composition contains each of the first anti-Bet v 1 antibody, the second anti-Bet v 1 antibody, and the third anti-Bet v 1 antibody in a dose of about 300 mg.

[0013] In some embodiments, the anti-Bet v 1 antibody or the pharmaceutical composition containing the anti-Bet v 1 antibody is administered subcutaneously. In some embodiments, the anti-Bet v 1 antibody or the pharmaceutical composition containing the anti-Bet v 1 antibody is administered intravenously.

[0014] In some embodiments, a single dose of an anti-Bet v 1 antibody or a pharmaceutical composition containing an anti-Bet v 1 antibody is administered. In some embodiments, the anti-Bet v 1 antibody or the pharmaceutical composition containing an anti-Bet v 1 antibody is administered once before the start of the pollen season.

[0015] In some embodiments, the first anti-Bet v 1 antibody comprises a heavy chain variable region (HCVR) containing the amino acid sequence of SEQ ID NO: 1 and a light chain variable region (LCVR) containing the amino acid sequence of SEQ ID NO: 5. In some embodiments, the first anti-Bet v 1 antibody comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 9 and a light chain containing the amino acid sequence of SEQ ID NO: 10.

[0016] In some embodiments, the second anti-Bet v 1 antibody comprises an HCVR containing the amino acid sequence of SEQ ID NO: 11 and an LCVR containing the amino acid sequence of SEQ ID NO: 15. In some embodiments, the second anti-Bet v 1 antibody comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 19 and a light chain containing the amino acid sequence of SEQ ID NO: 20.

[0017] In some embodiments, the third anti-Bet v 1 antibody comprises an HCVR containing the amino acid sequence of SEQ ID NO: 21 and an LCVR containing the amino acid sequence of SEQ ID NO: 25. In some embodiments, the third anti-Bet v 1 antibody comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 29 and a light chain containing the amino acid sequence of SEQ ID NO: 30.

[0018] In some embodiments, treatment using a pharmaceutical composition is To reduce the total nasal symptom score (TNSS) of the target group, To reduce the Total Ocular Symptom Score (TOSS) of the target group, To reduce the target's total symptom score (TSS), To reduce the target's daily medication score (DMS), To reduce the target complex symptom medication score (CSMS), To reduce the mean wheal diameter of the target birch skin prick test (SPT), and / or The goal is to increase the number of good days for subjects who do not require emergency medication, whose TSS score is 2 or less out of 18, and who do not require emergency medication.

[0019] In some embodiments, TNSS, TOSS, TSS, DMS, CSMS, SPT mean wheal diameter, and / or good days are measured over a period of at least 28, 57, 85, or 113 days. In some embodiments, TNSS, TOSS, TSS, DMS, CSMS, SPT mean wheal diameter, and / or good days are measured over a period of birch pollen season.

[0020] In some embodiments, treatment with the pharmaceutical composition reduces allergic rhinitis symptoms in the subject. In some embodiments, treatment with the pharmaceutical composition reduces the subject's total nasal symptom score (TNSS) AUC (0-1 hour) after nasal allergen induction (NAC) compared to the subject's baseline TNSS AUC (0-1 hour) after NAC prior to the initiation of treatment (e.g., by at least approximately 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, or more), and / or reduces the subject's peak TNSS after NAC compared to the subject's baseline peak TNSS after NAC prior to the initiation of treatment (e.g., by at least approximately 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, or more).

[0021] In some embodiments, a single dose of the pharmaceutical composition reduces the subject's TNSS AUC (0-1 hour) after NAC by at least about 20% for at least two months after administration of the pharmaceutical composition, and / or reduces the subject's peak TNSS after NAC by at least about 25% for at least two months after administration of the pharmaceutical composition.

[0022] In some embodiments, treatment with the pharmaceutical composition reduces allergic conjunctivitis symptoms in the subject. In some embodiments, treatment with the pharmaceutical composition reduces the subject's total ocular symptom score (TOSS) compared to the subject's baseline TOSS value before the initiation of treatment (e.g., by at least approximately 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, or more) and / or reduces the subject's TOSS AUC (0-1 hour) after NAC compared to the subject's baseline TOSS AUC (0-1 hour) after NAC before the initiation of treatment (e.g., by at least approximately 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, or more).

[0023] In some embodiments, treatment with a pharmaceutical composition reduces the target's CSMS during the birch pollen season compared to the target's baseline composite symptom medication score (CSMS) before the initiation of treatment or a control CSMS (e.g., by at least about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, or more).

[0024] In some embodiments, treatment with the pharmaceutical composition improves the subject's peak nasal inspiratory flow rate (PNIF) compared to the subject's baseline PNIF value before the start of treatment (e.g., by at least about 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, or more).

[0025] In some embodiments, treatment with the pharmaceutical composition reduces birch sensitization in a subject as measured by a skin prick test (SPT) using birch allergen extract (e.g., by at least about 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more). In some embodiments, a single dose of the pharmaceutical composition reduces birch sensitization in a subject by at least about 60% for at least two months after administration of the pharmaceutical composition (e.g., at least three months, at least four months, at least five months, or at least six months).

[0026] In some embodiments, the subject to be treated has a baseline serum allergen-specific IgE level of 0.35 kUa / L or higher in response to the allergen (e.g., birch pollen, Bet v 1 allergen, or Fagales allergen). In some embodiments, the subject to be treated is baseline positive on SPT using the allergen (e.g., birch allergen extract or Fagales allergen).

[0027] In another embodiment, a cocktail comprising two or more anti-Bet v 1 antibodies or their antigen-binding fragments is provided for use in a method of treating birch allergy in a subject. In some embodiments, the method comprises administering the cocktail disclosed herein to a subject in need (e.g., a subject having a birch allergy).

[0028] In another embodiment, the use of a cocktail comprising two or more anti-Bet v 1 antibodies or their antigen-binding fragments is provided in the manufacture of a drug for use in a method of treating birch allergy in a subject. In some embodiments, the method comprises administering the cocktail disclosed herein to a subject in need (e.g., a subject having a birch allergy).

[0029] In another embodiment, a cocktail comprising two or more anti-Bet v 1 antibodies or their antigen-binding fragments is provided for use in a method of reducing one or more symptoms of an allergic reaction to a Fagales allergen in a subject. In some embodiments, the method comprises administering the cocktail disclosed herein to a subject in need (for example, a subject having one or more symptoms of an allergic reaction to a Fagales allergen).

[0030] In another embodiment, the use of a cocktail comprising two or more anti-Bet v 1 antibodies or their antigen-binding fragments is provided in the manufacture of a drug for use in reducing one or more symptoms of an allergic reaction to a Fagales allergen in a subject. In some embodiments, the method comprises administering the cocktail disclosed herein to a subject in need (e.g., a subject having one or more symptoms of an allergic reaction to a Fagales allergen).

[0031] In another embodiment, a cocktail comprising two or more anti-Bet v 1 antibodies or their antigen-binding fragments is provided for use in a method of treating subjects with seasonal or perennial allergies related to birch and cross-reactive pollen. In some embodiments, the method comprises administering the cocktail disclosed herein to a subject in need (e.g., a subject with seasonal or perennial allergies related to birch and cross-reactive pollen).

[0032] In another embodiment, the use of a cocktail comprising two or more anti-Bet v 1 antibodies or their antigen-binding fragments is provided in the manufacture of a drug for use in a method of treating subjects with seasonal or perennial allergies related to birch and cross-reactive pollen. In some embodiments, the method comprises administering the cocktail disclosed herein to a subject in need (e.g., a subject with seasonal or perennial allergies related to birch and cross-reactive pollen).

[0033] Other embodiments will become apparent from the detailed description that follows. In embodiments of the present invention, for example, the following items are provided. (Item 1) A method for treating birch allergy in a subject, (a) A first anti-Bet v 1 antibody or its antigen-binding fragment, wherein the first anti-Bet v 1 antibody comprises a heavy chain complementarity-determining region (HCDR) 1 containing the amino acid sequence of SEQ ID NO: 2, an HCDR2 containing the amino acid sequence of SEQ ID NO: 3, an HCDR3 containing the amino acid sequence of SEQ ID NO: 4, a light chain complementarity-determining region (LCDR) 1 containing the amino acid sequence of SEQ ID NO: 6, an LCDR2 containing the amino acid sequence of SEQ ID NO: 7, and an LCDR3 containing the amino acid sequence of SEQ ID NO: 8, and / or (b) A second anti-Bet v 1 antibody or antigen-binding fragment thereof, wherein the second anti-Bet v 1 antibody comprises HCDR1 containing the amino acid sequence of SEQ ID NO: 12, HCDR2 containing the amino acid sequence of SEQ ID NO: 13, HCDR3 containing the amino acid sequence of SEQ ID NO: 14, LCDR1 containing the amino acid sequence of SEQ ID NO: 16, LCDR2 containing the amino acid sequence of SEQ ID NO: 17, and LCDR3 containing the amino acid sequence of SEQ ID NO: 18, and / or (c) A method comprising administering to a subject a pharmaceutical composition comprising a third anti-Bet v 1 antibody or antigen-binding fragment thereof, wherein the third anti-Bet v 1 antibody comprises HCDR1 comprising the amino acid sequence of SEQ ID NO: 22, HCDR2 comprising the amino acid sequence of SEQ ID NO: 23, HCDR3 comprising the amino acid sequence of SEQ ID NO: 24, LCDR1 comprising the amino acid sequence of SEQ ID NO: 26, LCDR2 comprising the amino acid sequence of SEQ ID NO: 27, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 28. (Item 2) The method according to item 1, wherein the pharmaceutical composition comprises the first anti-Bet v 1 antibody, the second anti-Bet v 1 antibody, and the third anti-Bet v 1 antibody. (Item 3) The method according to item 2, wherein the pharmaceutical composition comprises each of the first anti-Bet v 1 antibody, the second anti-Bet v 1 antibody, and the third anti-Bet v 1 antibody in a dose of about 5 mg to about 500 mg. (Item 4) The method according to item 3, wherein the pharmaceutical composition comprises, in a dose of approximately 50 mg, each of the first anti-Bet v 1 antibody, the second anti-Bet v 1 antibody, and the third anti-Bet v 1 antibody. (Item 5) The method according to item 3, wherein the pharmaceutical composition comprises, in a dose of approximately 150 mg, each of the first anti-Bet v 1 antibody, the second anti-Bet v 1 antibody, and the third anti-Bet v 1 antibody. (Item 6) The method according to item 3, wherein the pharmaceutical composition comprises, in a dose of approximately 300 mg, each of the first anti-Bet v 1 antibody, the second anti-Bet v 1 antibody, and the third anti-Bet v 1 antibody. (Item 7) The method according to any one of items 1 to 6, wherein the pharmaceutical composition is administered subcutaneously. (Item 8) The method according to any one of items 1 to 6, wherein the pharmaceutical composition is administered intravenously. (Item 9) The method according to any one of items 1 to 8, wherein a single dose of the pharmaceutical composition is administered before the start of the birch pollen season. (Item 10) The first anti-Bet v 1 antibody contains a heavy chain variable region including the amino acid sequence of SEQ ID NO: 1. The method according to any one of items 1 to 9, comprising a light chain variable region (LCVR) containing the amino acid sequence of HCVR and SEQ ID NO: 5. (Item 11) The method according to item 10, wherein the first anti-Bet v 1 antibody comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 9 and a light chain containing the amino acid sequence of SEQ ID NO: 10. (Item 12) The method according to any one of items 1 to 9, wherein the second anti-Bet v 1 antibody comprises HCVR containing the amino acid sequence of SEQ ID NO: 11 and LCVR containing the amino acid sequence of SEQ ID NO: 15. (Item 13) The method according to item 12, wherein the second anti-Bet v 1 antibody comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 19 and a light chain containing the amino acid sequence of SEQ ID NO: 20. (Item 14) The method according to any one of items 1 to 9, wherein the third anti-Bet v 1 antibody comprises HCVR containing the amino acid sequence of SEQ ID NO: 21 and LCVR containing the amino acid sequence of SEQ ID NO: 25. (Item 15) The method according to item 14, wherein the third anti-Bet v 1 antibody comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 29 and a light chain containing the amino acid sequence of SEQ ID NO: 30. (Item 16) Treatment using the aforementioned pharmaceutical composition To reduce the total nasal symptom score (TNSS) of the target group, To reduce the Total Ocular Symptom Score (TOSS) of the target group, To reduce the target's total symptom score (TSS), To reduce the target's daily medication score (DMS), To reduce the target complex symptom medication score (CSMS), To reduce the mean wheal diameter of the target birch skin prick test (SPT), and / or The method according to any one of items 1 to 15, wherein emergency medication is not used, the subject's TSS is 2 or less out of 18, and the number of good days for the subject is increased. (Item 17) The method according to item 16, wherein treatment using the aforementioned pharmaceutical composition reduces the target TNSS, TOSS, TSS, DMS, and / or CSMS during the birch pollen season. (Item 18) The method according to any one of items 1 to 17, wherein treatment using the pharmaceutical composition reduces allergic rhinitis symptoms in the subject. (Item 19) Treatment using the aforementioned pharmaceutical composition The total nasal symptom score (TNSS) AUC (0-1 hour) of the subject after nasal allergen induction (NAC) is reduced by at least approximately 20% compared to the baseline TNSS AUC (0-1 hour) value of the subject after NAC prior to the initiation of the treatment, and / or The method according to item 18, wherein the peak TNSS of the subject after NAC is reduced by at least about 25% compared to the baseline peak TNSS value of the subject after NAC before the initiation of the treatment. (Item 20) The administration of a single dose of the aforementioned pharmaceutical composition The pharmaceutical composition reduces the subject's TNSS AUC (0-1 hour) after NAC by at least about 20% for at least two months after administration, and / or The method according to item 19, which reduces the subject's peak TNSS after NAC by at least about 25% for at least two months after administration of the pharmaceutical composition. (Item 21) The method according to any one of items 1 to 20, wherein treatment using the pharmaceutical composition reduces allergic conjunctivitis symptoms in the subject. (Item 22) The method according to item 21, wherein treatment using the pharmaceutical composition reduces the subject's TOSS AUC (0-1 hour) after NAC compared to the baseline TOSS AUC (0-1 hour) value of the subject after NAC before the start of the treatment. (Item 23) The method according to any one of items 1 to 22, wherein treatment using the pharmaceutical composition improves the peak nasal inspiratory flow rate (PNIF) in the subject by at least about 30% compared to the baseline PNIF value of the subject before the start of treatment. (Item 24) The method according to any one of items 1 to 23, wherein treatment using the pharmaceutical composition reduces birch sensitization in the subject by at least about 50% as measured by a skin prick test (SPT) using birch allergen extract. (Item 25) The method according to item 24, wherein a single dose of the pharmaceutical composition reduces birch sensitization in the subject by at least about 60% for at least two months after administration of the pharmaceutical composition. (Item 26) The method according to any one of items 1 to 25, wherein the subject to be treated has a baseline serum allergen-specific IgE level of 0.35 kUa / L or higher in response to birch pollen and / or Bet v 1 allergen. (Item 27) The method according to any one of items 1 to 26, wherein the subject to be treated is baseline positive on SPT using birch allergen extract. (Item 28) A method for reducing one or more symptoms of an allergic reaction to a Fagales allergen in a subject, (a) A first anti-Bet v 1 antibody or its antigen-binding fragment, wherein the first anti-Bet v 1 antibody comprises a heavy chain complementarity-determining region (HCDR) 1 containing the amino acid sequence of SEQ ID NO: 2, an HCDR2 containing the amino acid sequence of SEQ ID NO: 3, an HCDR3 containing the amino acid sequence of SEQ ID NO: 4, a light chain complementarity-determining region (LCDR) 1 containing the amino acid sequence of SEQ ID NO: 6, an LCDR2 containing the amino acid sequence of SEQ ID NO: 7, and an LCDR3 containing the amino acid sequence of SEQ ID NO: 8, and / or (b) A second anti-Bet v 1 antibody or antigen-binding fragment thereof, wherein the second anti-Bet v 1 antibody comprises HCDR1 containing the amino acid sequence of SEQ ID NO: 12, HCDR2 containing the amino acid sequence of SEQ ID NO: 13, HCDR3 containing the amino acid sequence of SEQ ID NO: 14, LCDR1 containing the amino acid sequence of SEQ ID NO: 16, LCDR2 containing the amino acid sequence of SEQ ID NO: 17, and LCDR3 containing the amino acid sequence of SEQ ID NO: 18, and / or (c) A method comprising administering to a subject a pharmaceutical composition comprising a third anti-Bet v 1 antibody or antigen-binding fragment thereof, wherein the third anti-Bet v 1 antibody comprises HCDR1 comprising the amino acid sequence of SEQ ID NO: 22, HCDR2 comprising the amino acid sequence of SEQ ID NO: 23, HCDR3 comprising the amino acid sequence of SEQ ID NO: 24, LCDR1 comprising the amino acid sequence of SEQ ID NO: 26, LCDR2 comprising the amino acid sequence of SEQ ID NO: 27, and LCDR3 comprising the amino acid sequence of SEQ ID NO: 28. (Item 29) The method according to item 28, wherein the Fagales allergen is Bet v 1. (Item 30) The method according to item 28 or 29, wherein the subject is sensitized to Bet v 1 and at least one other Fagales allergen. (Item 31) The method according to any one of items 28 to 30, wherein the pharmaceutical composition comprises the first anti-Bet v 1 antibody, the second anti-Bet v 1 antibody, and the third anti-Bet v 1 antibody. (Item 32) The method according to item 31, wherein the pharmaceutical composition comprises each of the first anti-Bet v 1 antibody, the second anti-Bet v 1 antibody, and the third anti-Bet v 1 antibody in a dose of about 5 mg to about 500 mg. (Item 33) The method according to item 32, wherein the pharmaceutical composition comprises, in a dose of approximately 50 mg, each of the first anti-Bet v 1 antibody, the second anti-Bet v 1 antibody, and the third anti-Bet v 1 antibody. (Item 34) The method according to item 32, wherein the pharmaceutical composition comprises, in a dose of approximately 150 mg, each of the first anti-Bet v 1 antibody, the second anti-Bet v 1 antibody, and the third anti-Bet v 1 antibody. (Item 35) The method according to item 32, wherein the pharmaceutical composition comprises, in a dose of approximately 300 mg, each of the first anti-Bet v 1 antibody, the second anti-Bet v 1 antibody, and the third anti-Bet v 1 antibody. (Item 36) The method according to any one of items 28 to 35, wherein the pharmaceutical composition is administered subcutaneously. (Item 37) The method according to any one of items 28 to 35, wherein the pharmaceutical composition is administered intravenously. (Item 38) The method according to any one of items 28 to 37, wherein the first anti-Bet v 1 antibody comprises a heavy chain variable region (HCVR) containing the amino acid sequence of SEQ ID NO: 1 and a light chain variable region (LCVR) containing the amino acid sequence of SEQ ID NO: 5. (Item 39) The method according to item 38, wherein the first anti-Bet v 1 antibody comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 9 and a light chain containing the amino acid sequence of SEQ ID NO: 10. (Item 40) The method according to any one of items 28 to 37, wherein the second anti-Bet v 1 antibody comprises HCVR containing the amino acid sequence of SEQ ID NO: 11 and LCVR containing the amino acid sequence of SEQ ID NO: 15. (Item 41) The method according to item 40, wherein the second anti-Bet v 1 antibody comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 19 and a light chain containing the amino acid sequence of SEQ ID NO: 20. (Item 42) The method according to any one of items 28 to 37, wherein the third anti-Bet v 1 antibody comprises HCVR containing the amino acid sequence of SEQ ID NO: 21 and LCVR containing the amino acid sequence of SEQ ID NO: 25. (Item 43) The method according to item 42, wherein the third anti-Bet v 1 antibody comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 29 and a light chain containing the amino acid sequence of SEQ ID NO: 30. (Item 44) Treatment using the aforementioned pharmaceutical composition reduces allergic rhinitis symptoms in the subject. The method described in any one of items 28-43. (Item 45) The method according to any one of items 28 to 44, wherein treatment using the pharmaceutical composition reduces allergic conjunctivitis symptoms in the subject. (Item 46) The method according to any one of items 28 to 45, wherein treatment using the pharmaceutical composition reduces sensitization to the Fagales allergen in the subject. (Item 47) The method according to any one of items 28 to 46, wherein the subject to be treated has a baseline serum allergen-specific IgE level of 0.35 kUa / L or higher in response to the Fagales allergen. (Item 48) The method according to any one of items 28 to 47, wherein the subject to be treated is baseline positive for the Fagales allergen by SPT. [Brief explanation of the drawing]

[0034] [Figure 1]This study shows that a single dose of the anti-Bet v 1 antibody cocktail reduced the total nasal symptom score (TNSS) AUC (0-1 hour) after nasal allergen induction (NAC) at 8 days (Δ-1.2, p=0.001), 29 days (Δ-1.2, p=0.001), and 57 days (Δ-0.9, p=0.024) compared to placebo. Least squares mean change in presented AUC, adjusted for baseline AUC. Circles = placebo, squares = anti-Bet v 1 antibody cocktail. [Figure 2] When measured as a percentage change, a single dose of the anti-Bet v 1 antibody cocktail reduced the TNSS AUC (0-1 hour) after NAC at day 8 (Δ-32%, p=0.002), day 29 (Δ-27%, p=0.003), and day 57 (Δ-19%, p=0.053) compared to placebo. Least squares mean percentage change of presented AUC, adjusted for baseline AUC. Circles = placebo, squares = anti-Bet v 1 antibody cocktail. [Figure 3] This study shows that a single dose of the anti-Bet v 1 antibody cocktail reduced post-NAC peak TNSS compared to placebo at day 8 (Δ-35%, p=<0.001), day 29 (Δ-29%, p=<0.001), and day 57 (Δ-26%, p=0.003). TNSS in the subjects was assessed at each induction time point using the specific eligible dose of the allergen required for patients to achieve a TNSS of 7 or higher in baseline screening induction. Circles = placebo, squares = anti-Bet v 1 antibody cocktail. [Figure 4A] The single dose of the anti-Bet v 1 antibody cocktail showed a sustained response in reducing birch sensitization compared to placebo, as measured by the percentage change from baseline in the AUC of the mean wheal diameter in skin prick tests at day 8 (Δ-72%, p<0.001), day 29 (Δ-66%, p<0.001), day 57 (Δ-78%, p<0.001), and day 113 (Δ-74%, p<0.001). Circles = placebo, squares = anti-Bet v 1 antibody cocktail. [Figure 4B]The single dose of the anti-Bet v 1 antibody cocktail showed a sustained response in reducing alder sensitization, compared to placebo, as measured by the percentage change from baseline in the AUC of the mean wheal diameter in skin prick tests at day 8 (Δ-45%, p<0.001), day 29 (Δ-46%, p<0.001), day 57 (Δ-53%, p<0.001), and day 113 (Δ-50%, p<0.001). Circles = placebo, squares = anti-Bet v 1 antibody cocktail. [Figure 5A] This study shows that a single dose of the anti-Bet v 1 antibody cocktail reduced the total ocular symptom score (TOSS) AUC (0-1 hour) after NAC at day 8 (Δ-0.48, p=0.013), day 29 (Δ-0.59, p=0.007), and day 57 (Δ-0.30, p=0.191) compared to placebo. Least squares mean change in presented AUC, adjusted for baseline AUC. Circles = placebo, squares = anti-Bet v 1 antibody cocktail. [Figure 5B] When measured as a percentage change, a single dose of the anti-Bet v 1 antibody cocktail reduced the TOSS AUC (0-1 hour) after NAC at day 8 (Δ-42%, p=0.01), day 29 (Δ-48%, p=0.05), and day 57 (Δ-37%) compared to placebo. Least squares mean percentage change of presented AUC, adjusted for baseline AUC. Circles = placebo, squares = anti-Bet v 1 antibody cocktail. [Figure 6] A single dose of the anti-Bet v 1 antibody cocktail improved peak nasal inspiratory flow (PNIF) compared to placebo at day 8 (Δ32%, p=0.092), day 29 (Δ47%, p=0.017), and day 57 (Δ35%, p=0.121), with statistical significance achieved at day 29. Least squares mean percentage change in presented AUC, adjusted for baseline AUC. Circles = placebo, squares = anti-Bet v 1 antibody cocktail. [Figure 7A]This study shows that, compared to the placebo group, patients treated with the anti-Bet v 1 antibody cocktail showed a significant suppression of basophilic responses to birch, alder, hazelnut, and apple pollen, while no suppression of basophilic responses was detected in grass allergen-negative controls. (A) Basophilic responses to birch pollen extract were measured by EC50. When measured by EC values, a significant suppression of basophilic responses to birch allergen was observed in the treatment group (n=13) compared to placebo (days 8 and 57, all p<0.001; day 113, p<0.01). (B) Basophilic responses to alder pollen extract were measured by EC50. (C) Basophilic response to hazelnut pollen extract was measured by EC50. When measured by the percentage change in EC50 from baseline, a significant suppression of the basophilic response to alder allergen was observed in the treatment group (n=13) compared to placebo (n=13) (days 8, 57, and 113, all p<0.001). (D) Basophilic response to apple pollen extract was measured by EC50. When measured by the percentage change in EC50 from baseline, a significant suppression of the basophilic response to hazelnut allergen was observed in the treatment group (n=13) compared to placebo (n=13) (day 8 p<0.001, day 57 p=0.01). (D) Basophilic response to apple pollen extract was measured by EC50. When measured by the percentage change in EC50 from baseline, a significant suppression of the basophilic response to apple allergen was observed in the treatment group (n=13) compared to placebo (n=13) (p<0.01 on day 8, p<0.05 on days 57 and 113). (E) Basophilic response to grass pollen extract was measured by EC50. [Figure 7B] Same as above. [Figure 7C] Same as above. [Figure 7D] Same as above. [Figure 7E] Same as above. [Figure 8]This study shows a correlation between suppression of basophil activation (EC50 value) and reduction of skin prick test (SPT) in patients treated with an anti-Bet v 1 antibody cocktail. An inverse correlation was observed between the percentage change in birch BAT EC50 and birch SPT (mean wheal diameter) AUC at day 8 (r=-0.6, p=0.04) and day 57 (r=-0.57, p=0.04). No significant correlation was observed in the placebo-treated subjects. [Figure 9A] This study shows the correlation between suppression of basophil activation (EC50 value) and TNSS response (percentage change in TNSS (0-1 hour) AUC) in patients treated with an anti-Bet v 1 antibody cocktail. (A) On day 8 (r=-0.71, p=0.0095), a statistically significant inverse correlation was observed between EC50 value and the percentage improvement in TNSS (0-1 hour) AUC. No significant correlation was observed in the placebo-treated group. (B) Responder analysis of the active treatment group (TNSS (0-1 hour) AUC reduction of ≥60% vs. <60%). [Figure 9B] Same as above. [Modes for carrying out the invention]

[0035] Definition of Terms Before describing the present invention, it should be understood that the invention is not limited to the specific methods and experimental conditions described, and therefore the methods and conditions may vary. Since the scope of the invention is limited only by the appended claims, it should also be understood that the terms used herein are for the purpose of describing only specific embodiments and are not intended to be limiting.

[0036] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art to which the present invention pertains.

[0037] As used herein, the term “about” means that, when used in reference to a specific numerical value mentioned, the value may vary by up to 1% from the value mentioned. For example, as used herein, the expression “about 100” includes 99 and 101, as well as all values ​​in between (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

[0038] As used herein, terms such as “to treat” and “to treat” mean to alleviate symptoms, to eliminate the cause of symptoms, whether temporarily or permanently, or to prevent or delay the onset of symptoms of a specified disorder or condition.

[0039] As used herein, the term “Bet v 1” refers to the naturally occurring or recombinant Bet v 1 protein. The naturally occurring Bet v 1 protein is approximately 17 kD and exists as seven antiparallel β-sheets (β1-β7) in a chain, two short α-helices connecting Bet1 and Bet2, a long C-terminal α-helix (α3), and a glycine-rich loop motif between β2 and β3 (Kofler et al., J.Mol.Biol.2012, 422(1):109-123). In some embodiments, the Bet v 1 protein contains the amino acid sequence of SEQ ID NO: 31. In some embodiments, the Bet v 1 protein contains the naturally occurring or recombinantly produced form, which includes one or more amino acid substitutions, deletions, or additions to SEQ ID NO: 31. For example, in some embodiments, the Bet v 1 protein contains the amino acid sequence of SEQ ID NO: 32 (the Bet v 1 amino acid sequence from Uniprot:P15494).

[0040] As used herein, the term "Bet v 1 fragment" refers to a polypeptide having at least one antigenic site of Bet v 1. In some embodiments, the Bet v 1 fragment is a polypeptide having at least two antigenic sites of Bet v 1. In some embodiments, the antigenic sites are covalently linked. In some embodiments, the antigenic sites are linked by at least one peptide bond. In one embodiment, the two antigenic sites are linked by at least one peptide bond and a spacer between the antigenic sites. Exemplary Bet v 1 fragments are disclosed in WO2018 / 222854, which is incorporated herein by reference.

[0041] As used herein, the term "antibody" refers to an antigen-binding molecule or molecular complex that includes a set of complementarity-determining regions (CDRs) that specifically bind to or interact with a particular antigen (e.g., Bet v 1). As used herein, the term "antibody" includes immunoglobulin molecules, which include four polypeptide chains, two heavy (H) chains and two light (L) chains interconnected by disulfide bonds, and multimers thereof (e.g., IgM). In a typical antibody, each heavy chain includes a heavy-chain variable region (referred to herein as HCVR or V H and omitted) and a heavy-chain constant region. The heavy-chain constant region includes three domains, C H 1, C H 2, and C H 3. Each light chain includes a light-chain variable region (referred to herein as LCVR or V L and omitted) and a light-chain constant region. The light-chain constant region includes one domain (C L 1). The V H region and the V L region can be further subdivided into hypervariable regions called complementarity-determining regions (CDRs), with relatively conserved regions called framework regions (FRs) disposed therebetween. Each V H and V LIt consists of three CDRs and four FRs arranged from the amino terminus to the carboxyl terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. In some embodiments, the FRs of the antibody (or its antigen-binding portion) may be identical to the human germline sequence or may be naturally or artificially modified. The amino acid consensus sequence may be defined based on a parallel analysis of two or more CDRs.

[0042] As used herein, the term “antibody” also includes the antigen-binding fragment of a complete antibody molecule. Terms such as “antigen-binding portion” and “antigen-binding fragment” of an antibody, as used herein, include any natural, enzymatically obtainable, synthetic, or genetically engineered polypeptide or glycoprotein that specifically binds to an antigen to form a complex. Antigen-binding fragments of antibodies may originate from a complete antibody molecule using any suitable standard method, such as proteolytic digestion or recombinant genetic engineering techniques, involving the manipulation and expression of DNA encoding an antibody variable region and optionally a constant domain. Such DNA is known and / or readily available, for example, from commercially available sources, DNA libraries (including, for example, phage antibody libraries), or can be synthesized. DNA can be sequenced and manipulated, for example, by chemical or molecular biological techniques, to position one or more variable and / or constant domains in a suitable configuration, introduce codons, create cysteine ​​residues, modify, add, or delete amino acids.

[0043] Non-limiting examples of antigen-binding fragments include (i) Fab fragments, (ii) F(ab')2 fragments, (iii) Fd fragments, (iv) Fv fragments, (v) single-chain Fv(scFv) molecules, (vi) dAb fragments, and (vii) minimal recognition units consisting of amino acid residues mimicking the hypervariable region of an antibody (e.g., isolated complementarity-determining regions (CDRs) such as the CDR3 peptide) or the restricted FR3-CDR3-FR4 peptide. Other manipulated molecules such as domain-specific antibodies, single-domain antibodies, domain-deleted antibodies, chimeric antibodies, CDR-implanted antibodies, diabodies, triabodies, tetrabodies, minibodies, nanobodies (e.g., monovalent nanobodies, bivalent nanobodies, etc.), small modular immunotherapies (SMIPs), and shark variable IgNAR domains are also included in the expression “antigen-binding fragment” as used herein.

[0044] The antigen-binding fragment of an antibody typically contains at least one variable domain. The variable domain may be of any size or amino acid composition and generally contains at least one CDR adjacent to or in-framed to one or more framework sequences. L V associating with the domain H In an antigen-binding fragment having a domain, V H Domain and V L Domains can be arranged relative to each other in any suitable configuration. For example, the variable region is a dimer, V H -V H , V H -V L or V L -V L It may contain a dimer. Alternatively, the antigen-binding fragment of the antibody may be a monomer V H or V L It may contain a domain.

[0045] In certain embodiments, the antigen-binding fragment of the antibody may contain at least one variable domain covalently bound to at least one constant domain. Non-limiting exemplary configurations of variable and constant domains that may be found within the antigen-binding fragment of the antibody include (i)V H-C H 1. (ii)V H -C H 2, (iii)V H -C H 3, (iv)V H -C H 1-C H 2. (v)V H -C H 1-C H 2-C H 3. (vi)V H -C H 2-C H 3. (vii)V H -C L、 (viii)V L -C H 1. (ix)V L -C H 2, (x)V L -C H 3. (xi)V L -C H 1-C H 2. (xii)V L -C H 1-C H 2-C H 3. (xiii)V L -C H 2-C H 3, and (xiv)V L -C L Examples include: In any configuration of a variable domain and a constant domain, including any of the exemplary configurations listed above, the variable domain and the constant domain may be directly linked to each other, or they may be linked by a complete or partial hinge or linker region. The hinge region may consist of at least two (e.g., 5, 10, 15, 20, 40, 60 or more) amino acids that result in a mobile or semi-mobile chain between adjacent variable domains and / or constant domains in a single polypeptide molecule. Furthermore, the antigen-binding fragment of the antibody may consist of homodimers or heterodimers (or other polymers) of any of the variable and constant domain configurations listed above, linked to each other, and / or one or more monomers V H Domain or V L The domain may be included non-covalently (for example, by a disulfide bond).

[0046] As used herein, the term “antibody” also includes multispecific (e.g., bispecific) antibodies. A multispecific antibody or antigen-binding fragment of an antibody typically comprises at least two distinct variable domains, each of which can specifically bind to a separate antigen or to a different epitope on the same antigen. Any multispecific antibody format can be adapted for use relating to the antibody or antigen-binding fragment of an antibody of this disclosure using conventional techniques available in the art. For example, this disclosure includes a method comprising the use of a bispecific antibody, where one arm of the immunoglobulin is specific to Bet v 1 or a fragment thereof, and the other arm of the immunoglobulin is specific to a second therapeutic target or is conjugated to a therapeutic portion. Exemplary bispecificity formats that may be used in the context of this disclosure include, for example, scFv-based or diabody bispecificity formats, IgG-scFv fusions, dual variable domain (DVD)-Ig, quadromas, knob-into-holes, common light chains (e.g., common light chains with knob-into-holes), CrossMab, CrossFab, (SEED) bodies, leucine zippers, duobodies, IgG1 / IgG2, dual-acting Fab(DAF)-IgG, and Mab 2 Bispecificity formats include, but are not limited to, Klein et al. 2012, mAbs 4:6, 1-11, and the references cited in this document, for an overview of the above formats. Bispecificity antibodies may also be constructed using peptide / nucleic acid conjugations. For example, site-specific antibody-oligonucleotide conjugates may be generated using non-natural amino acids with orthogonal chemical reactivity, and then self-assembled into a multimeric complex with a specified composition, titer, and structure. (See, for example, Kazane et al., J.Am.Chem.Soc. [Epub:Dec.4,2012]).

[0047] As used herein, the term “human antibody” is intended to include antibodies having variable and constant regions derived from human germline immunoglobulin sequences. Nevertheless, the human antibodies of this disclosure may include amino acid residues not encoded by human germline immunoglobulin sequences, for example, in the CDR, particularly in CDR3 (mutations introduced, for example, by random or site-directed mutagenesis in vitro, or by somatic mutation in vivo). However, as used herein, the term “human antibody” is not intended to include antibodies in which germline CDR sequences of another mammalian species, such as mouse, are transplanted onto human framework sequences.

[0048] As used herein, the term “recombinant human antibody” is intended to include all human antibodies prepared, expressed, created, or isolated by genetic recombination means, such as antibodies expressed using a recombinant expression vector transduced into host cells (as further described below), antibodies isolated from recombinants, combinatorial human antibody libraries (as further described below), antibodies isolated from animals transgenic for human immunoglobulin genes (e.g., mice) (see, for example, Taylor et al. (1992) Nucl. Acids Res. 20:6287-6295), or antibodies prepared, expressed, created, or isolated by any other means, including splicing human immunoglobulin gene sequences to other DNA sequences. Such recombinant human antibodies have variable and constant regions derived from human germline immunoglobulin sequences. However, in certain embodiments, such recombinant human antibodies are subjected to in vitro mutagenesis (or in vivo somatic mutagenesis if transgenic animals for human Ig sequences are used), so the V of the recombinant antibody H Region and V L The amino acid sequence of the region is human germline V H Array and V L While derived from and related to the sequence, it is a sequence that cannot naturally exist in the in vivo human antibody germline repertoire.

[0049] "Isolated antibody" means an antibody identified, isolated, and / or recovered from at least one component of its natural environment. For example, an antibody isolated or removed from at least one component of an organism, or from a tissue or cell in which such antibody naturally exists or is naturally produced, is an "isolated antibody." Isolated antibodies also include antibodies in situ within recombinant cells. An isolated antibody is an antibody that has undergone at least one purification or isolation step. According to certain embodiments, an isolated antibody may substantially contain no other cellular material and / or chemical substances.

[0050] Terms such as "specifically bind" mean that an antibody or its antigen-binding fragment forms a complex with an antigen that is relatively stable under physiological conditions. Specific binding is at least about 1 × 10⁻⁶. -6 It can be characterized by an equilibrium dissociation constant less than or equal to M (for example, a smaller K). D (This indicates a tighter binding.) Methods for determining whether an antibody specifically binds to an antigen are well known in the art, and include, for example, equilibrium dialysis and surface plasmon resonance assays. In some embodiments, specific binding is measured by a surface plasmon resonance assay. An isolated antibody that specifically binds to an antigen from one species may or may not cross-react with other antigens, such as ortholog antigens from other species.

[0051] When used herein, "K D The term "equilibrium dissociation constant" refers to the equilibrium dissociation constant of a particular antibody-antigen interaction.

[0052] As used herein, the term “surface plasmon resonance” refers to an optical phenomenon that enables real-time analysis of biomolecular interactions by detecting changes in protein concentration within a biosensor matrix, for example, using BIACORE® systems (Pharmacia Biosensor AB, Uppsala, Sweden and Piscataway, NJ).

[0053] As used herein, the term “epitope” refers to an antigenic determinant that interacts with a specific antigen-binding site in a variable region of an antibody molecule, known as a paratope. A single antigen may have two or more epitopes. Thus, different antibodies may bind to different regions on an antigen and have different biological effects. The term “epitope” also refers to the site on an antigen to which B cells and / or T cells respond. This term also refers to the region of the antigen to which an antibody binds. Epitopes may be linear or discontinuous (e.g., steric). Linear epitopes are epitopes produced by adjacent amino acid residues in a polypeptide chain. Steric epitopes are produced by spatially juxtaposed amino acids from different segments of a linear polypeptide chain. In certain embodiments, epitopes may include determinants that are chemically active surface groups of molecules, such as amino acids, sugar side chains, phosphoryl groups, or sulfonyl groups, and in certain embodiments, they may have specific three-dimensional structural features and / or specific charge-to-mass features. Epitopes may also be defined as structural or functional. Functional epitopes are generally a subset of structural epitopes and contain residues that are directly involved in the affinity of the interaction. Epitopes formed from sequential amino acids are typically retained even when exposed to denaturing solvents, whereas epitopes formed by three-dimensional folding are typically lost upon treatment with denaturing solvents. Epitopes typically contain at least three, more commonly at least five, or eight to ten amino acids in their intrinsic spatial structure.

[0054] The terms “substantially identical” or “substantially identical,” when used in relation to nucleic acids or fragments thereof, indicate that, when optimally aligned with appropriate nucleotide insertions or deletions with another nucleic acid (or its complementary strand), there is nucleotide sequence identity in at least about 90% of the nucleotide bases, e.g., at least 91%, 92%, 93%, 94%, about 95%, 96%, 97%, 98%, or 99%, as measured by any well-known algorithm of sequence identity such as FASTA, BLAST, or GAP, as discussed below. A nucleic acid molecule having substantial identity with a reference nucleic acid molecule may, in certain particular cases, encode a polypeptide having the same or substantially similar amino acid sequence as the polypeptide encoded by the reference nucleic acid molecule.

[0055] As applied to polypeptides, the terms “substantial identity” and “substantially identical” mean that two peptide sequences, when optimally aligned, share at least about 90% sequence identity, e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity. In some embodiments, non-identical residue positions are distinguished by different conserved amino acid substitutions. A “conservative amino acid substitution” is when an amino acid residue is replaced by another amino acid residue having a side chain (R group) with similar chemical properties (e.g., charge or hydrophobicity). Generally, conservative amino acid substitutions do not substantially alter the functional properties of the protein.

[0056] Sequence similarity for polypeptides is typically measured using sequence analysis software. Protein analysis software matches similar sequences using similarity measurements assigned to various substitutions, deletions, and other modifications, including conserved amino acid substitutions. For example, GCG software includes programs such as GAP and BESTFIT, which can be used with default parameters to determine sequence homology or sequence identity between closely related polypeptides, such as homologous polypeptides from different species, or between wild-type proteins and their mutant proteins. See, for example, GCG version 6.1. Polypeptide sequences can also be compared using FASTA, a program in GCG version 6.1 with default or recommended parameters. FASTA (e.g., FASTA2 and FASTA3) provides alignment of best overlap regions and sequence identity percentage between query sequences and search sequences (Pearson, 2000, above). Another preferred algorithm for comparing the sequences of this disclosure with databases containing numerous sequences from different organisms is the computer program BLAST, particularly BLASTP or TBLASTN, using default parameters. (See, for example, Altschul et al., 1990, J.Mol.Biol.215:403-410 and 1997 Nucleic Acids Res.25:3389-3402).

[0057] As used herein, terms such as “allergic response,” “allergic reaction,” and “allergic symptoms” include one or more signs or symptoms selected from the group consisting of urticaria (e.g., hives), angioedema, rhinitis, asthma, vomiting, sneezing, runny nose, sinusitis, watery eyes, wheezing, bronchospasm, decreased peak expiratory flow (PEF), gastrointestinal disturbances, flushing, lip swelling, tongue swelling, hypotension, anaphylaxis, and organ dysfunction / failure. “Allergic response,” “allergic reaction,” and “allergic symptoms” also include immune responses and reactions such as increased IgE production and / or increased allergen-specific immunoglobulin production.

[0058] The term "allergen" refers to a substance, chemical, particle, or composition that can stimulate an allergic response in a susceptible individual. Allergens may be found in or derived from food items such as dairy products (e.g., cow's milk), eggs, celery, sesame, wheat, soy, fish, shellfish, sugars (e.g., sugars found in meat such as alpha-galactose), peanuts, other legumes (e.g., beans, peas, soybeans, etc.), and tree nuts. Alternatively, allergens may be contained in or derived from non-food items such as dust (e.g., house dust mites), pollen, insect toxins (e.g., toxins from honeybees, wasps, mosquitoes, fire ants, etc.), mold, animal hair, animal dander, latex, metals (e.g., nickel), household cleaning agents, detergents, pharmaceuticals, cosmetics (e.g., perfumes, etc.), drugs (e.g., penicillin, sulfonamides, salicylates, etc.), therapeutic monoclonal antibodies (e.g., cetuximab), ragweed, grasses, and birch. In some embodiments, the allergen is birch pollen, or contained in or derived from birch, for example, the Bet v 1 protein. The terms “allergen” and “antigen” are used interchangeably throughout this disclosure.

[0059] As used herein, the term “subject requiring it” means a human or non-human mammal that (i) exhibits signs of one or more symptoms or signs of an allergy (e.g., birch allergy), (ii) has been diagnosed with an allergy to an allergen (e.g., birch pollen allergen), and / or (iii) is at high risk of developing an allergy or allergic response to an allergen (e.g., birch allergy or allergic response). In certain embodiments, this term includes subjects that exhibit allergen sensitization to one or more allergens (e.g., birch allergen or its components, e.g., Bet v 1 protein). In some embodiments, a subject is sensitized to an allergen (e.g., birch allergen or Bet v 1 protein) if the subject exhibits an allergen-specific IgE level of 0.35 kU / L or higher for the allergen. In certain embodiments, subjects requiring treatment by the methods of the Disclosure are subjects having elevated levels of one or more serum biomarkers, including but not limited to total IgE, allergen-specific IgE (e.g., birch pollen IgE or Bet v 1 IgE), thymic and activation-regulated chemokines (TARCs), and eotaxin. For example, in some embodiments, the methods of the Disclosure include administering an anti-Bet v 1 antibody or antibody cocktail to a patient having elevated levels of allergen-specific IgE (e.g., a subject having birch pollen or Bet v 1 IgE levels of 0.35 kU / L or higher). The terms “subject” and “patient” are used interchangeably herein.

[0060] The term “subjects requiring treatment” may also include, for example, subjects with concomitant allergies or other conditions. For example, in some embodiments, a subject with birch allergy may also have oral allergy syndrome. In some embodiments, the subject to be treated is a subject with birch allergy and allergies to one or more other Fagales allergens. As used herein, Fagales allergens or "Fagales allergens" include, but are not limited to, birch pollen (Bet v 1), alder pollen (Aln g1 and Aln g4), hazel pollen (Cor a1, Cor a2, Cor a8, Cor a9, Cor a10, Cor a11, Cor a12, Cor a13, and Cor a14), hornbeam pollen (Car b1), Japanese oak pollen (Ost c1), horse chestnut pollen (Cas s1, Cas s5, Cas s8, and Cas s9), beech pollen (Fag s1), and white oak pollen (Que a1 and Que a2). Those skilled in the art will recognize that Bet v 1-related allergens (also known as "Fagales group 1" allergens or "PR-10 allergens") are also found in foods such as apples (Mal d 1), apricots (Pru ar 1), carrots (Dau c 1), celery (Api g 1), cherries (Pru av 1), horse chestnuts (Cas s 1), hazelnuts (Cor 1), kiwis (Act c 8, Act d 8, and Act d 11), mung beans (Vig r 1), peanuts (Ara h 8), pears (Pyr c 1), raspberries (Rubi 1), soybeans (Gly m 4), strawberries (Fra a 1), tomatoes (Sola l 4), and walnuts (Jug r 5). See Carlson, Annals of Allergy, Asthma & Immunology 2019, 123: pp. 359-365. Therefore, the term "Fagales allergen" includes not only pollen allergens but also food allergens.In some embodiments, the subject has an elevated allergen-specific IgE level (e.g., an allergen-specific IgE level of 0.35 kU / L or higher) in response to birch pollen (e.g., birch pollen extract) or a Bet v 1 allergen, and one or more other Fagales allergens.

[0061] Introduction As described herein, a monoclonal antibody cocktail has been developed against Bet v 1, the major birch tree allergen. High-affinity allergen-specific monoclonal IgG antibodies are hypothesized to be administered in a form that provides “passive immunity” to the allergen. Preclinical studies have demonstrated that the anti-Bet v 1 antibodies REGN5713, REGN5714, and REGN5715 bind independently and non-competitively to the Bet v 1 allergen. The data also show that optimal inhibition of Bet v 1 binding to human polyclonal IgE occurs when all three antibodies are administered together, resulting in in vitro effector cell degranulation and a reduction in subsequent type 1 hypersensitivity reactions. While not bound by any specific theory, this activity of the anti-Bet v 1 antibodies is expected to prevent the resulting inflammatory cascade that induces allergic reactions in response to birch tree allergens and potentially cross-reactive allergens.

[0062] The anti-Bet v 1 antibody cocktail disclosed herein is advantageous in at least some aspects over currently available therapies such as allergen-specific immunotherapy (SIT). For example, blocking the anti-antibody cocktail is expected to be safer because allergy patients are not exposed to natural allergens, to provide more predictable efficacy, to offer greater convenience as a single dose can prevent allergic symptoms across the entire birch group and birch homolog group during allergy season, to broaden the pool of patients who can receive treatment (e.g., asthma patients who have previously been contraindicated to receiving SIT), and to have a faster onset of action. As described herein, the anti-Bet v 1 antibody results in a rapid and sustained reduction of allergic symptoms in patients with birch allergy.

[0063] Treatment method In one embodiment, a method is provided for treating birch allergy in a subject, or for treating, preventing, or improving one or more symptoms of birch allergy. In another embodiment, a method is provided for treating, preventing, or improving seasonal or perennial allergy to birch and / or birch cross-reactive pollen (e.g., moderate to severe seasonal or perennial allergy). In some embodiments, the method comprises administering to a subject one or more doses of an anti-Bet v 1 antibody or a cocktail of anti-Bet v 1 antibodies (e.g., a pharmaceutical composition comprising one or more anti-Bet v 1 antibodies).

[0064] In some embodiments, the subjects to be treated have a history of allergic rhinitis symptoms induced by birch pollen, with or without conjunctivitis. In some embodiments, the subjects to be treated are diagnosed as positive by a skin prick test (SPT) using birch pollen extract. In some embodiments, the subjects are positive on SPT with an average wheal diameter at least 5 mm larger than that of the negative control. In some embodiments, the subjects to be treated are diagnosed as positive for birch pollen (e.g., birch pollen extract) and / or Bet v 1 antigen in an allergen-specific IgE test with a level of 0.35 kU / L or higher.

[0065] In some embodiments, the subject being treated is an adult. In some embodiments, the subject has a concomitant disease or condition. Non-limiting examples of concomitant diseases or conditions include allergies (e.g., allergies to one or more food allergens and / or allergies to one or more non-food allergens such as airborne allergens), oral allergy syndrome, and asthma. In some embodiments, the subject has asthma. In some embodiments, the subject has birch-induced asthma. In some embodiments, the subject has an allergy to birch allergen and one or more tree homologs (e.g., Fagales allergen).

[0066] In some embodiments, the subjects to be treated have a history of allergic rhinitis symptoms induced by birch pollen, with or without asthma. In some embodiments, the subjects to be treated have a history of allergic rhinitis symptoms induced by birch pollen, with or without conjunctivitis, with or without asthma.

[0067] In some embodiments, the subject being treated has altered levels of one or more biomarkers of allergic rhinitis. In some embodiments, the biomarker is related to type 2 immunoactivity and / or is an allergen-specific biomarker. In some embodiments, the biomarker is a serum biomarker. In some embodiments, the biomarker is total IgE, allergen-specific IgG4, or thymic and activating regulatory chemokines (TARCs).

[0068] In some embodiments, treatment with one or more anti-Bet v 1 antibodies disclosed herein results in improvement of one or more symptoms of birch allergy or conditions associated with birch allergy. In some embodiments, treatment by the methods disclosed herein improves one or more symptoms of allergic rhinitis in a subject. As used herein, “improving allergic rhinitis symptoms” includes reducing or eliminating the severity or duration of one or more symptoms of allergic rhinitis in a subject, including but not limited to sneezing, itching (of the nose, eyes, ears, or palate), runny nose, postnasal drip, congestion, osmotic dysphoria, headache, earache, tearing, conjunctivitis, ocular swelling, and fatigue. In some embodiments, the reduction of allergic rhinitis symptoms is measured by the Total Nasal Symptom Score (TNSS). The TNSS is a patient-reported composite symptom assessment of congestion, itching, rhinorrhea, and sneezing. The patient-assessed symptom score is assigned to each category at a given time point using a 4-point scale (0-3), where 0 indicates no symptoms, 1 is used for mild symptoms that are easily tolerable, 2 is used for bothersome but tolerable symptoms, and 3 is used for severe symptoms that are difficult to tolerate and interfere with daily activities. The TNSS is calculated by summing the scores for each symptom out of 12 points. In some embodiments, the TNSS score is measured after nasal allergen induction (NAC) using an allergen. In some embodiments, a baseline TNSS score is measured for the subject (e.g., during a pre-treatment screening visit).

[0069] In some embodiments, the treatment results in an improvement in TNSS compared to a baseline or control value (e.g., the subject's baseline TNS score before the initiation of treatment) during the birch pollen season (e.g., over at least 28, 57, 85, or 113 days of the birch pollen season, or over the entire birch pollen season). In some embodiments, the treatment results in a reduction of at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, or more in TNSS compared to a baseline or control value.

[0070] In some embodiments, the treatment results in an improvement in TNSS after NAC (e.g., using birch pollen extract), and the improvement includes a reduction in the score of one or more of (i) congestion, (ii) itching, (iii) runny nose, or (iv) sneezing, and / or the total TNSS score, compared to the baseline score of the subject. In some embodiments, the treatment results in a reduction of at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, or more in TNSS compared to the baseline score of the subject. In some embodiments, the treatment results in a reduction of 1, 2, 3, 4, 5, or more points in TNSS score compared to the baseline score of the subject.

[0071] In some embodiments, treatment by the methods disclosed herein (e.g., administration of the anti-Bet v 1 antibody or anti-Bet v 1 antibody cocktail disclosed herein) reduces the subject's TNSS AUC (0-1 hour) by at least about 15%, 20%, 25%, 30%, 35%, or more compared to the subject's baseline TNSS AUC (0-1 hour) (e.g., the subject's baseline TNSS AUC (0-1 hour) before the start of treatment). In some embodiments, the TNSS AUC (0-1 hour) is measured after NAC. In some embodiments, the treatment reduces the subject's peak TNSS by at least about 15%, 20%, 25%, 30%, 35%, or more compared to the subject's baseline peak TNSS (e.g., the subject's baseline peak TNSS before the start of treatment). In some embodiments, the peak TNSS is measured after NAC. In some embodiments, baseline peak TNSS is assessed by determining the dose of an allergen (e.g., Bet v 1 allergen or birch extract) that achieves a TNSS of 7 or higher in a subject before treatment initiation, and post-treatment peak TNSS is assessed by administering the same dose of the allergen that achieved a baseline TNSS of 7 or higher to the subject.

[0072] In some embodiments, treatment by the methods disclosed herein improves one or more symptoms of allergic conjunctivitis in a subject. As used herein, “improving allergic conjunctivitis symptoms” includes reducing or eliminating the severity or duration of one or more symptoms of allergic conjunctivitis in a subject, including but not limited to itching, redness, tearing, or puffiness of the eye. In some embodiments, the reduction of allergic conjunctivitis symptoms is measured by the Total Ocular Symptom Score (TOSS), which is a composite symptom assessment of patient-reported ocular symptoms.

[0073] In some embodiments, the TOSS ranges from 0 to 6 and is based on two symptoms, namely itching / conjunctival congestion / foreign body sensation and lacrimation / watery eyes. Each of the two symptoms is graded by the patient as 0 (none), 1 (mild), 2 (moderate), or 3 (severe). In other embodiments, the TOSS ranges from 0 to 12 and is based on four items: itching / burning sensation, conjunctival congestion, watery eyes and lacrimation, and puffiness and swelling. The symptom score assessed by the patient is assigned to each category for a given time point using a 4-point scale (0-3), where 0 indicates no symptoms, a score of 1 is used for mild symptoms that are easily tolerable, 2 is used for the recognition of bothersome but tolerable symptoms, and 3 is used for severe symptoms that are difficult to tolerate and interfere with daily activities.

[0074] In some embodiments, the TOSS score is measured after NAC using the allergen. In some embodiments, the baseline TOSS score is measured for the subject (e.g., during a pre-treatment screening visit). In some embodiments, the treatment results in an improvement in TOSS after NAC (e.g., using birch pollen extract), and the improvement includes a reduction in the subject's baseline score of one or more of the following: (i) pruritus / burning sensation, (ii) hyperemia, (iii) watery eyes and tearing, or (iv) puffiness and swelling, and / or a reduction in the total TOSS score.

[0075] In some embodiments, the treatment results in a reduction of at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, or more in the TOSS score compared to the baseline score of the subject. In some embodiments, the treatment results in a reduction of 1, 2, 3, 4, 5, or more points in the TOSS score compared to the baseline score of the subject. In some embodiments, the treatment reduces the subject's TOSS AUC(0-1 hour) by at least about 15%, 20%, 25%, 30%, 35%, or more compared to the subject's baseline TOSS AUC(0-1 hour) (e.g., the subject's baseline TOSS AUC(0-1 hour) before the start of treatment). In some embodiments, the TOSS AUC(0-1 hour) is measured after NAC.

[0076] In some embodiments, treatment by the methods disclosed herein results in an improvement (i.e., reduction) of the subject's Total Symptom Score (TSS). The TSS is calculated by summing the subject's TNS (ranging from 0 to 12) and TOSS (ranging from 0 to 6) to a composite TNSS of 0 to 18. In some embodiments, a baseline TSS score is measured for the subject (e.g., during a screening visit prior to the initiation of treatment). In some embodiments, the treatment results in an improvement in TSS compared to a baseline or control value (e.g., the subject's baseline TSS score before the initiation of treatment) during the birch pollen season (e.g., over at least 28, 57, 85, or 113 days of the birch pollen season, or over the entire birch pollen season). In some embodiments, the treatment results in a reduction of at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, or more TSS compared to a baseline or control value. In some embodiments, the treatment results in a reduction of 1, 2, 3, 4, 5, or more points in the TSS score compared to the baseline score of the subject.

[0077] In some embodiments, treatment by the method disclosed herein results in an improvement (i.e., reduction) in the subject's daily medication score (DMS). To calculate the DMS, the subject records the use of daily emergency medications, including which medications and the amounts of these pre-specified medications. Using this information, the DMS is calculated as follows: desloratadine 5 mg, 6 points / dose; maximum score of 6 points per day; olopatadine 1 mg / mL / drop, 1.5 points / drop; maximum score of 6 points per day; mometasone phlomate 50 μg / dose, 2.0 points / spray; maximum score of 8 points per day. The maximum DMS score is 20. See Calderon et al., Clin Exp Allergy 2014;44(10):1228-39. In some embodiments, a baseline DMS score is measured for the subject (e.g., during a pre-treatment screening visit). In some embodiments, the treatment results in an improvement in DMS compared to a baseline or control value (e.g., the subject's baseline TSS score before the start of treatment) during the birch pollen season (e.g., over at least 28, 57, 85, or 113 days of the birch pollen season, or over the entire birch pollen season). In some embodiments, the treatment results in a reduction of at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, or more DMS compared to a baseline or control value. In some embodiments, the treatment results in a reduction of 1, 2, 3, 4, 5, or more points in the DMS score compared to the subject's baseline score.

[0078] In some embodiments, treatment by the methods disclosed herein results in an improvement (i.e., reduction) of the subject's composite symptom medication score (CSMS). The CSMS is calculated by summing the subject's DMS (ranging from 0 to 20) and TSS (ranging from 0 to 18) to a composite CSMS of 0 to 38. In some embodiments, a baseline CSMS score is measured for the subject (e.g., during a pre-treatment screening visit). In some embodiments, treatment results in an improvement in CSMS compared to a baseline or control value (e.g., the subject's baseline CSMS score before treatment initiation) during the birch pollen season (e.g., over at least 28, 57, 85, or 113 days of the birch pollen season, or over the entire birch pollen season). In some embodiments, treatment results in a reduction of at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, or more in CSMS compared to a baseline or control value. In some embodiments, the treatment results in a reduction of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more points in the CSMS score compared to the baseline score of the subject.

[0079] In some embodiments, treatment by the method disclosed herein improves the subject's peak nasal inspiratory flow rate (PNIF) compared to a baseline value (e.g., the subject's baseline PNIF before the initiation of treatment). In some embodiments, PNIF is measured after NAC. In some embodiments, the treatment increases the subject's PNIF by at least about 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, or more compared to the subject's baseline PNIF (e.g., the subject's baseline PNIF before the initiation of treatment).

[0080] In some embodiments, treatment by the methods disclosed herein reduces the target allergen sensitization (e.g., sensitization to birch allergen) compared to a baseline value (e.g., the target's sensitization level before treatment initiation). In some embodiments, the treatment reduces the target allergen sensitization (e.g., birch sensitization) by at least about 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, or more compared to the target's sensitization level before treatment initiation. In some embodiments, the sensitization level is measured using a skin prick test with the allergen (e.g., birch allergen extract). In some embodiments, the sensitization level is assessed by measuring serum antibodies (e.g., allergen-specific IgE levels such as Bet v 1 or birch pollen IgE).

[0081] In some embodiments, treatment by the methods disclosed herein results in an increase in the number of good days for a subject during the birch pollen season. As used herein, “good day” is defined as a day in which the subject’s TSS is 2 or less without the use of anti-allergic emergency medication.

[0082] In some embodiments, treatment by the methods disclosed herein improves one or more symptoms of oral allergy syndrome. Symptoms of oral allergy syndrome typically include itching of the lips, mouth, and throat, and may also include swelling of the lips and tongue and angioedema. In some embodiments, a method is provided for treating birch pollen-related oral allergy syndrome by administering an anti-Bet v 1 antibody or anti-Bet v 1 antibody cocktail disclosed herein.

[0083] In some embodiments, treatment by the methods disclosed herein (for example, administering the anti-Bet v 1 antibody or anti-Bet v 1 antibody cocktail disclosed herein) results in one or more of the above-mentioned improvements over a long period, for example, at least one month, at least two months, at least three months, at least four months, or longer.

[0084] Anti-Bet v 1 antibody and its antigen-binding fragment According to certain embodiments of the present disclosure, the anti-Bet v 1 antibody or its antigen-binding fragment comprises a heavy chain variable region (HCVR), a light chain variable region (LCVR), and / or a complementarity-determining region (CDR) comprising any of the amino acid sequences of the anti-Bet v 1 antibody described in WO 2018 / 222854, which is incorporated herein by reference.

[0085] In some embodiments, the anti-Bet v 1 antibody or its antigen-binding fragment includes a heavy chain complementarity-determining region (HCDR) of a heavy chain variable region (HCVR) containing the amino acid sequence of SEQ ID NO: 1, and a light chain complementarity-determining region (LCDR) of a light chain variable region (LCVR) containing the amino acid sequence of SEQ ID NO: 5. In some embodiments, the anti-Bet v 1 antibody or its antigen-binding fragment includes three HCDRs (HCDR1, HCDR2, and HCDR3) and three LCDRs (LCDR1, LCDR2, and LCDR3), wherein HCDR1 contains the amino acid sequence of SEQ ID NO: 2, HCDR2 contains the amino acid sequence of SEQ ID NO: 3, HCDR3 contains the amino acid sequence of SEQ ID NO: 4, LCDR1 contains the amino acid sequence of SEQ ID NO: 6, LCDR2 contains the amino acid sequence of SEQ ID NO: 7, and LCDR3 contains the amino acid sequence of SEQ ID NO: 8. In some embodiments, the anti-Bet v 1 antibody or its antigen-binding fragment comprises HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 of SEQ ID NOs. 2, 3, 4, 6, 7, and 8, respectively, and further comprises an HCVR having at least 85% sequence identity to the amino acid sequence of SEQ ID NO. 1 (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity), and an LCVR having at least 85% sequence identity to the amino acid sequence of SEQ ID NO. 5 (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity). In some embodiments, the anti-Bet v 1 antibody or its antigen-binding fragment comprises an HCVR containing SEQ ID NO. 1 and an LCVR containing SEQ ID NO. 5. In some embodiments, the anti-Bet v 1 antibody or its antigen-binding fragment comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 9. In some embodiments, the anti-Bet v 1 antibody or its antigen-binding fragment comprises a light chain containing the amino acid sequence of SEQ ID NO: 10.

[0086] In some embodiments, the anti-Bet v 1 antibody or its antigen-binding fragment comprises an HCDR of HCVR containing the HCVR amino acid sequence of SEQ ID NO: 11, and an LCDR of LCVR containing the amino acid sequence of SEQ ID NO: 15. In some embodiments, the anti-Bet v 1 antibody or its antigen-binding fragment comprises three HCDRs (HCDR1, HCDR2, and HCDR3) and three LCDrs (LCDR1, LCDR2, and LCDR3), where HCDR1 contains the amino acid sequence of SEQ ID NO: 12, HCDR2 contains the amino acid sequence of SEQ ID NO: 13, HCDR3 contains the amino acid sequence of SEQ ID NO: 14, LCDR1 contains the amino acid sequence of SEQ ID NO: 16, LCDR2 contains the amino acid sequence of SEQ ID NO: 17, and LCDR3 contains the amino acid sequence of SEQ ID NO: 18. In some embodiments, the anti-Bet v 1 antibody or its antigen-binding fragment comprises HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 of SEQ ID NOs. 12, 13, 14, 16, 17, and 18, respectively, and further comprises an HCVR having at least 85% sequence identity to the amino acid sequence of SEQ ID NO. 11 (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity), and an LCVR having at least 85% sequence identity to the amino acid sequence of SEQ ID NO. 15 (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity). In some embodiments, the anti-Bet v 1 antibody or its antigen-binding fragment comprises an HCVR containing SEQ ID NO. 11 and an LCVR containing SEQ ID NO. 15. In some embodiments, the anti-Bet v 1 antibody or its antigen-binding fragment comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 19. In some embodiments, the anti-Bet v 1 antibody or its antigen-binding fragment comprises a light chain containing the amino acid sequence of SEQ ID NO: 20.

[0087] In some embodiments, the anti-Bet v 1 antibody or its antigen-binding fragment comprises an HCDR of HCVR containing the HCVR amino acid sequence of SEQ ID NO: 21, and an LCDR of LCVR containing the amino acid sequence of SEQ ID NO: 25. In some embodiments, the anti-Bet v 1 antibody or its antigen-binding fragment comprises three HCDRs (HCDR1, HCDR2, and HCDR3) and three LCDrs (LCDR1, LCDR2, and LCDR3), where HCDR1 contains the amino acid sequence of SEQ ID NO: 22, HCDR2 contains the amino acid sequence of SEQ ID NO: 23, HCDR3 contains the amino acid sequence of SEQ ID NO: 24, LCDR1 contains the amino acid sequence of SEQ ID NO: 26, LCDR2 contains the amino acid sequence of SEQ ID NO: 27, and LCDR3 contains the amino acid sequence of SEQ ID NO: 28. In some embodiments, the anti-Bet v 1 antibody or its antigen-binding fragment comprises HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 of SEQ ID NOs. 22, 23, 24, 26, 27, and 28, respectively, and further comprises an HCVR having at least 85% sequence identity to the amino acid sequence of SEQ ID NO 21 (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity), and an LCVR having at least 85% sequence identity to the amino acid sequence of SEQ ID NO 25 (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity). In some embodiments, the anti-Bet v 1 antibody or its antigen-binding fragment comprises an HCVR containing SEQ ID NO 21 and an LCVR containing SEQ ID NO 25. In some embodiments, the anti-Bet v 1 antibody or its antigen-binding fragment comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 29. In some embodiments, the anti-Bet v 1 antibody or its antigen-binding fragment comprises a light chain containing the amino acid sequence of SEQ ID NO: 30.

[0088] In some embodiments, the anti-Bet v 1 antibody is a bioequivalent of the antibody disclosed herein (e.g., a bioequivalent of REGN5713, REGN5714, or REGN5715). As used herein, the term “bioequivalent” refers to an anti-Bet v 1 antibody that, under similar experimental conditions and at the same molar dose, when administered in single or multiple doses, exhibits no significant difference in absorption rate and / or degree compared to that of the reference antibody (e.g., REGN5713, REGN5714, or REGN5715). In some embodiments, the term refers to an anti-Bet v 1 antibody that has no clinically significant difference in safety, purity, and / or potency compared to the anti-Bet v 1 antibodies of this disclosure (e.g., REGN5713, REGN5714, or REGN5715).

[0089] In some embodiments, the anti-Bet v 1 antibody is an IgG1 or IgG4 antibody. In some embodiments, the anti-Bet v 1 antibody comprises a heavy chain constant region of a human IgG1 or IgG4 isotype, the constant region having one or more amino acid modifications (e.g., substitutions or deletions), e.g., hinges, C H 2, or C H Includes amino acid modifications in 3 regions.

[0090] In some embodiments, the anti-Bet v 1 antibody used in the methods of the present disclosure may have pH-dependent binding properties. For example, the anti-Bet v 1 antibody for use in the methods of the present disclosure may exhibit reduced binding to Bet v 1 at acidic pH compared to neutral pH. Alternatively, the anti-Bet v 1 antibody of the present disclosure may exhibit enhanced binding to its antigen at acidic pH compared to neutral pH. The term "acidic pH" includes pH values ​​less than about 6.2, for example, about 6.0, 5.95, 5.9, 5.85, 5.8, 5.75, 5.7, 5.65, 5.6, 5.55, 5.5, 5.45, 5.4, 5.35, 5.3, 5.25, 5.2, 5.15, 5.1, 5.05, and 5.0. As used herein, the term "neutral pH" means a pH of about 7.0 to about 7.4. The term "neutral pH" includes pH values ​​of approximately 7.0, 7.05, 7.1, 7.15, 7.2, 7.25, 7.3, 7.35, and 7.4.

[0091] Antibodies with pH-dependent binding properties can be obtained, for example, by screening an antibody sample for decreased (or increased) binding to a specific antigen at acidic pH compared to neutral pH. In addition, modification of the antigen-binding domain at amino acid concentration can produce antibodies with pH-dependent properties. For example, by substituting one or more amino acids in the antigen-binding domain (e.g., within the CDR) with histidine residues, antibodies with decreased antigen binding at acidic pH compared to neutral pH can be obtained.

[0092] In some embodiments, the therapeutic methods disclosed herein involve the use of a pharmaceutical composition comprising two or more anti-Bet v 1 antibodies disclosed herein, for example, two or three anti-Bet v 1 antibodies disclosed herein.

[0093] In some embodiments, the combination or pharmaceutical composition is (a) A first anti-Bet v 1 antibody or its antigen-binding fragment, wherein the first anti-Bet v 1 antibody comprises a heavy chain complementarity-determining region (HCDR) 1 containing the amino acid sequence of SEQ ID NO: 2, an HCDR2 containing the amino acid sequence of SEQ ID NO: 3, an HCDR3 containing the amino acid sequence of SEQ ID NO: 4, a light chain complementarity-determining region (LCDR) 1 containing the amino acid sequence of SEQ ID NO: 6, an LCDR2 containing the amino acid sequence of SEQ ID NO: 7, and an LCDR3 containing the amino acid sequence of SEQ ID NO: 8, and / or (b) A second anti-Bet v 1 antibody or its antigen-binding fragment, wherein the second anti-Bet v 1 antibody comprises HCDR1 containing the amino acid sequence of SEQ ID NO: 12, HCDR2 containing the amino acid sequence of SEQ ID NO: 13, HCDR3 containing the amino acid sequence of SEQ ID NO: 14, LCDR1 containing the amino acid sequence of SEQ ID NO: 16, LCDR2 containing the amino acid sequence of SEQ ID NO: 17, and LCDR3 containing the amino acid sequence of SEQ ID NO: 18, and / or (c) A third anti-Bet v 1 antibody or its antigen-binding fragment, wherein the third anti-Bet v 1 antibody comprises HCDR1 containing the amino acid sequence of SEQ ID NO: 22, HCDR2 containing the amino acid sequence of SEQ ID NO: 23, HCDR3 containing the amino acid sequence of SEQ ID NO: 24, LCDR1 containing the amino acid sequence of SEQ ID NO: 26, LCDR2 containing the amino acid sequence of SEQ ID NO: 27, and LCDR3 containing the amino acid sequence of SEQ ID NO: 28.

[0094] In some embodiments, the pharmaceutical composition comprises (a) and (b). In some embodiments, the pharmaceutical composition comprises (a) and (c). In some embodiments, the pharmaceutical composition comprises (b) and (c). In some embodiments, the pharmaceutical composition comprises (a), (b), and (c).

[0095] Preparation of human antibodies Methods for generating human antibodies in transgenic mice are known in the art. Any such known method may be used in the context of this disclosure to produce human antibodies that specifically bind to the Bet v 1 protein.

[0096] A high-affinity chimeric antibody against IL-4R, possessing a human variable region and a mouse constant region, is first isolated using VELOCIMMUNE® technology (see, for example, US 6,596,541, Regeneron Pharmaceuticals) or any other known method for generating monoclonal antibodies. VELOCIMMUNE® technology involves generating a transgenic mouse having a genome containing a human heavy chain variable region and a human light chain variable region operably ligated to an endogenous mouse constant region locus, so that the mouse produces an antibody containing the human variable region and the mouse constant region in response to antigen stimulation. The DNA encoding the variable regions of the antibody's heavy chain and light chain is isolated and operably ligated to the DNA encoding the human heavy chain constant region and the human light chain constant region. The DNA is then expressed in cells capable of expressing a fully human antibody.

[0097] Generally, VELOCIMMUNE® mice are loaded with the target antigen, and lymphocytes (such as B cells) are recovered from mice that express antibodies. Immortalized hybridoma cell lines can be prepared by fusing these lymphocytes with myeloma cell lines, and such hybridoma cell lines are screened and selected to identify hybridoma cell lines that produce antibodies specific to the target antigen. DNA encoding the variable regions of the heavy and light chains may be isolated and bound to the desired isotype constant regions of the heavy and light chains. Such antibody proteins may be produced in cells such as CHO cells. Alternatively, antigen-specific chimeric antibodies or DNA encoding the variable domains of the light and heavy chains may be isolated directly from antigen-specific lymphocytes.

[0098] First, a high-affinity chimeric antibody having a human variable region and a mouse constant region is isolated. The antibody is characterized and selected for desirable properties, including affinity, selectivity, and epitope, using standard procedures known to those skilled in the art. The mouse constant region is replaced with a desired human constant region to produce the fully human antibody of this disclosure, e.g., wild-type or modified IgG1 or IgG4. While the selected constant region may vary depending on the specific application, the high-affinity antigen-binding and target-specific characteristics reside in the variable region.

[0099] Generally, antibodies that can be used in the methods of this disclosure exhibit high affinity as described above when measured by binding to an antigen immobilized on either a solid or solution phase. The mouse constant region is replaced with a desired human constant region to produce the fully human antibody of this disclosure. While the selected constant region may vary depending on the specific application, the high affinity antigen-binding and target specificity characteristics reside in the variable region.

[0100] Methods and techniques for identifying CDRs within HCVR amino acid sequences and LCVR amino acid sequences are well known in the art and can be used to identify CDRs within specific HCVR amino acid sequences and / or LCVR amino acid sequences disclosed herein. Exemplary rules that can be used to identify CDR boundaries include, for example, the Kabat definition, the Chothia definition, and the AbM definition. Under general conditions, the Kabat definition is based on sequence variability, the Chothia definition is based on the location of structural loop regions, and the AbM definition is a compromise between the Kabat and Chothia methods. See, for example, Kabat, “Sequences of Proteins of Immunological Interest,” National Institutes of Health, Bethesda, Md. (1991), Al-Lazikani et al., J.Mol.Biol.273:927-948 (1997), and Martin et al., Proc.Natl.Acad.Sci.USA 86:9268-9272 (1989). Public databases are also available for identifying CDR sequences within antibodies.

[0101] Pharmaceutical composition In one embodiment, the disclosure provides a method comprising administering one or more anti-Bet v 1 antibodies, the one or more anti-Bet v 1 antibodies contained within a pharmaceutical composition comprising one or more pharmaceutically acceptable vehicles, carriers, and / or excipients. In some embodiments, the antibodies are intended for use in the treatment of birch allergy or conditions associated with birch allergy (e.g., allergic rhinitis or oral allergy syndrome).

[0102] In some embodiments, the pharmaceutical composition comprises one anti-Bet v 1 antibody or its antigen-binding fragment (for example, antibodies comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 of SEQ ID NOs. 2, 3, 4, 6, 7, and 8, respectively; antibodies comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 of SEQ ID NOs. 12, 13, 14, 16, 17, and 18, respectively; or antibodies comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 of SEQ ID NOs. 22, 23, 24, 26, 27, and 28, respectively).

[0103] In some embodiments, the pharmaceutical composition comprises two anti-Bet vs. 1 antibodies or their antigen-binding fragments. For example, in some embodiments, the pharmaceutical composition comprises antibodies comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 of SEQ ID NOs. 2, 3, 4, 6, 7, and 8, respectively, and antibodies comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 of SEQ ID NOs. In some embodiments, the pharmaceutical composition comprises antibodies comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 of SEQ ID NOs. 2, 3, 4, 6, 7, and 8, respectively, and antibodies comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 of SEQ ID NOs. 22, 23, 24, 26, 27, and 28, respectively. In some embodiments, the pharmaceutical composition includes antibodies comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 of SEQ ID NOs. 12, 13, 14, 16, 17, and 18, respectively, and antibodies comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 of SEQ ID NOs. 22, 23, 24, 26, 27, and 28, respectively.

[0104] In some embodiments, the pharmaceutical composition comprises three anti-Bet vs. 1 antibodies or their antigen-binding fragments. For example, in some embodiments, the pharmaceutical composition comprises antibodies comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 of SEQ ID NOs. 2, 3, 4, 6, 7, and 8, respectively; antibodies comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 of SEQ ID NOs. 12, 13, 14, 16, 17, and 18, respectively; and antibodies comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 of SEQ ID NOs. 22, 23, 24, 26, 27, and 28, respectively.

[0105] Various pharmaceutically acceptable carriers and excipients are well known in the art. See, for example, Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, PA. In some embodiments, the carriers are suitable for intravenous, intramuscular, oral, intraperitoneal, intrathecal, transdermal, topical, or subcutaneous administration.

[0106] In some embodiments, the pharmaceutical composition includes injectable preparations such as intravenous, subcutaneous, intradermal, and intramuscular injections and infusions. These injectable preparations may be prepared by known methods. For example, an injectable preparation may be prepared by dissolving, suspending, or emulsifying the antibody or a salt thereof in a sterile aqueous or oily medium conventionally used for injection. Examples of aqueous media for injection include physiological saline, isotonic solutions containing glucose, and other adjuvants, which may be used in combination with suitable solubilizers such as alcohol (e.g., ethanol), polyalcohols (e.g., propylene glycol, polyethylene glycol), and nonionic surfactants [e.g., polysorbate 80, HCO-50 (polyoxyethylene (50 mol) adduct of hydrogenated castor oil)]. Examples of oily media include sesame oil and soybean oil, which may be used in combination with solubilizers such as benzyl benzoate and benzyl alcohol. The injection solution thus prepared may be filled into a suitable ampoule.

[0107] The dose of one or more antibodies administered to a patient according to the methods of this disclosure may vary depending on the patient's age and size, symptoms, condition, and route of administration. The dose is typically calculated according to body weight or body surface area. The frequency and duration of treatment may be adjusted depending on the severity of the condition. Effective doses and schedules for administering a pharmaceutical composition containing anti-Bet v 1 antibody may be determined empirically; for example, patient progression may be monitored by periodic assessments, and the dose may be adjusted accordingly. Furthermore, interspecies scaling of doses may be performed using methods well known in the art (e.g., Mordenti et al., 1991, Pharmaceut. Res. 8:1351). Specific exemplary doses of anti-Bet v 1 antibody and associated administration regimens that may be used in the context of this disclosure are disclosed elsewhere in this specification.

[0108] For example, various delivery systems are known and can be used to administer pharmaceutical compositions, such as liposomes, microparticles, microcapsule encapsulation, recombinant cells capable of expressing mutant viruses, and receptor-mediated endocytosis (see, for example, Wu et al., 1987, J. Biol. Chem. 262:4429-4432). Methods of administration include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes. The compositions can be administered by any convenient route, for example, by injection or bolus injection, by absorption through the epithelial or mucocutaneous linings (e.g., oral mucosa, rectal mucosa, and intestinal mucosa), and can be administered together with other bioactive agents. In some embodiments, the pharmaceutical compositions disclosed herein are administered intravenously. In some embodiments, the pharmaceutical compositions disclosed herein are administered subcutaneously.

[0109] In some embodiments, the pharmaceutical compositions of this disclosure are contained in a container. Thus, in another embodiment, a container containing the pharmaceutical compositions disclosed herein is provided. For example, in some embodiments, the pharmaceutical compositions are contained in a container selected from the group consisting of glass vials, syringes, pen-type delivery devices, and automatic injectors.

[0110] In some embodiments, the pharmaceutical compositions of the Disclosure are delivered, for example, subcutaneously or intravenously, using a standard needle and syringe. In some embodiments, the syringe is a pre-filled syringe. In some embodiments, a pen-type delivery device or autoinjector is used to deliver the pharmaceutical compositions of the Disclosure (for example, for subcutaneous delivery). Such pen-type delivery devices may be reusable or disposable. Reusable pen-type delivery devices generally utilize a replaceable cartridge containing the pharmaceutical composition. Once all of the pharmaceutical composition inside the cartridge has been administered and the cartridge is empty, the empty cartridge can be easily discarded and replaced with a new cartridge containing the pharmaceutical composition. The pen-type delivery device can then be reused. Disposable pen-type delivery devices do not have replaceable cartridges. Rather, disposable pen-type delivery devices are sold pre-filled with the pharmaceutical composition held in a reservoir inside the device. Once the pharmaceutical composition is emptied from the reservoir, the entire device is discarded.

[0111] Examples of suitable pen-type and auto-injector delivery devices include AUTOPEN® (Owen Mumford, Inc., Woodstock, UK), DISETRONIC® pen (Disetronic Medical Systems, Bergdorf, Switzerland), HUMALOG MIX 75 / 25® pen, HUMALOG® pen, HUMALIN 70 / 30® pen (Eli Lilly and Co., Indianapolis, IN), NOVOPEN® I, II, and III (Novo Nordisk, Copenhagen, Denmark), NOVOPEN JUNIOR® (Novo Nordisk, Copenhagen, Denmark), BD® pen (Becton Dickinson, Franklin Lakes, NJ), OPTIPEN®, OPTIPEN PRO®, and OPTIPEN®. Examples of disposable pen-type delivery devices that find applications in the subcutaneous delivery of the pharmaceutical compositions of this disclosure include, but are not limited to, STARLET® and OPTICLIK® (Sanofi-Aventis, Frankfurt, Germany).

[0112] In some embodiments, the pharmaceutical composition may be delivered using a controlled-release system. In one embodiment, a pump may be used (see Langer; Sefton, 1987, CRC Crit.Ref. Biomed.Eng. 14:201 above). In another embodiment, a multimeric material may be used (see Medical Applications of Controlled Release, Langer and Wise (eds.), 1974, CRC Press, Boca Raton, Florida). In yet another embodiment, the controlled-release system may be positioned near the target of the composition, and therefore only a fraction of the systemic dose may be required (see, for example, Goodson, 1984, Medical Applications of Controlled Release, above, vol.2, pp.115-138). Other controlled-release systems are discussed in the review by Langer, 1990, Science 249:1527-1533.

[0113] In some embodiments, the pharmaceutical compositions for the above-described use are prepared into dosage forms in unit doses suitable for adapting the dose of the active ingredient. Such dosage forms in unit doses include, for example, tablets, pills, capsules, injections (ampoules), suppositories, and the like.

[0114] Dosage and administration regimen Typically, the amount of anti-Bet v 1 antibody administered to a subject according to the methods disclosed herein is a therapeutically effective dose. As used herein, the term “therapeutically effective dose” means an amount of anti-Bet v 1 antibody (or a combination of anti-Bet v 1 antibodies) that results in one or more of the following: (a) a reduction in the severity or duration of one or more symptoms of birch allergy; (b) prevention or reduction of an allergic reaction to a birch allergen (e.g., birch pollen extract or Bet v 1 protein); (c) a reduction in induced allergic rhinitis symptoms following nasal allergen induction; (d) a reduction in the level of one or more markers of type 2 immunoactivity (e.g., serum TARC or total IgE); and (e) a reduction in the use or need for conventional allergy therapy (e.g., a reduction or elimination of the use of antihistamines, decongestants, nasal or inhaled steroids, anti-IgE therapy, epinephrine, etc.).

[0115] For anti-Bet v 1 antibodies (e.g., antibodies containing the CDR and / or HCVR and LCVR sequences of REGN5713, REGN5714, or REGN5715 disclosed herein), the therapeutically effective dose is anti-Bet v Approximately 0.05 mg to 600 mg of one antibody, approximately 10 mg to 600 mg, or approximately 10 mg to 300 mg, approximately 25 mg to 500 mg, or approximately 50 mg to 300 mg, for example, approximately 0.05 mg, approximately 0.1 mg, approximately 1.0 mg, approximately 1.5 mg, approximately 2.0 mg, approximately 10 mg, approximately 20 mg, approximately 30 mg, approximately 40 mg, approximately 50 mg, approximately 60 mg, approximately 70 mg, approximately 80 mg, approximately 90 mg, approximately 100 mg, approximately 110 mg, approximately 120 mg, approximately 130 mg, approximately 140 mg, approximately 150 mg, approximately 160 mg, approximately 170 mg, approximately 180 mg, approximately 190 mg, approximately 200 mg, approximately 210 mg, approximately 220 mg, approximately 230 mg It may be approximately 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, 350 mg, 360 mg, 370 mg, 380 mg, 390 mg, 400 mg, 410 mg, 420 mg, 430 mg, 440 mg, 450 mg, 460 mg, 470 mg, 480 mg, 490 mg, 500 mg, 510 mg, 520 mg, 530 mg, 540 mg, 550 mg, 560 mg, 570 mg, 580 mg, 590 mg, or 600 mg. In certain embodiments, approximately 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, or 300 mg of anti-Bet v 1 antibody is administered to the subject.

[0116] In some embodiments, a therapeutically effective dose of each of two or more anti-Bet v 1 antibodies disclosed herein is administered to the subject. For example, in some embodiments, a therapeutically effective dose of each of REGN5713 and REGN5714, or each of REGN5713 and REGN5715, or each of REGN5714 and REGN5715 is administered to the subject. In some embodiments, a therapeutically effective dose of each of REGN5713, REGN5714, and REGN5715 is administered to the subject. In some embodiments, anti-Bet v Each antibody is available in doses of approximately 0.05 mg to 600 mg, 10 mg to 600 mg, 10 mg to 300 mg, 25 mg to 500 mg, or 50 mg to 300 mg, for example, approximately 0.05 mg, 0.1 mg, 1.0 mg, 1.5 mg, 2.0 mg, 10 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, and 230 mg. It is administered in amounts of approximately 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, 350 mg, 360 mg, 370 mg, 380 mg, 390 mg, 400 mg, 410 mg, 420 mg, 430 mg, 440 mg, 450 mg, 460 mg, 470 mg, 480 mg, 490 mg, 500 mg, 510 mg, 520 mg, 530 mg, 540 mg, 550 mg, 560 mg, 570 mg, 580 mg, 590 mg, or 600 mg. In some embodiments, each of two or more anti-Bet v 1 antibodies (e.g., two or more of REGN5713, REGN5714, and REGN5715) is administered in amounts of approximately 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, or 300 mg. In some embodiments, the two or more anti-Bet v 1 antibodies are administered in equal amounts. In some embodiments, the two or more anti-Bet v 1 antibodies are administered in different amounts.In some embodiments in which three anti-Bet v 1 antibodies are administered, at least one antibody is administered in a different amount than the other antibodies. In some embodiments in which three anti-Bet v 1 antibodies are administered, all three antibodies are administered in different amounts. In some embodiments in which three anti-Bet v 1 antibodies are administered, all three antibodies are administered in equal amounts.

[0117] In some embodiments, the anti-Bet v 1 antibody (e.g., antibodies comprising the CDR and / or HCVR and LCVR sequences of REGN5713, REGN5714, and / or REGN5715 disclosed herein) is administered in a total dose of about 50 mg to about 1500 mg, for example, about 100 mg to about 1500 mg, about 100 mg to about 1000 mg, or about 300 mg to about 1000 mg. In some embodiments, the anti-Bet v One antibody is available in approximately 50mg, 75mg, 100mg, 125mg, 150mg, 175mg, 200mg, 225mg, 250mg, 275mg, 300mg, 325mg, 350mg, 375mg, 400mg, 425mg, 450mg, 475mg, 500mg, 525mg, 550mg, 575mg, 600mg, 625mg, 650mg, 675mg, 700mg, 725mg, 750mg, 775mg, 800mg, and approximately It is administered in a total dose of 825 mg, approximately 850 mg, approximately 875 mg, approximately 900 mg, approximately 925 mg, approximately 950 mg, approximately 975 mg, approximately 1000 mg, approximately 1025 mg, approximately 1050 mg, approximately 1075 mg, approximately 1100 mg, approximately 1125 mg, approximately 1150 mg, approximately 1175 mg, approximately 1200 mg, approximately 1225 mg, approximately 1250 mg, approximately 1275 mg, approximately 1300 mg, approximately 1325 mg, approximately 1350 mg, approximately 1375 mg, approximately 1400 mg, approximately 1450 mg, approximately 1475 mg, or approximately 1500 mg.

[0118] The amount of anti-Bet v 1 antibody contained in each dose may be expressed in milligrams (i.e., mg / kg) of the activator (e.g., antibody) per kilogram of the subject's body weight. For example, anti-Bet v 1 antibody may be administered to a patient in doses of approximately 0.0001 to approximately 10 mg / kg of patient's body weight, approximately 1 mg / kg to approximately 10 mg / kg, or approximately 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, or 10 mg / kg.

[0119] In some embodiments, an anti-Bet v 1 antibody, or a pharmaceutical composition comprising one or more anti-Bet v 1 antibodies, is administered to a subject at a frequency of approximately four times a week, twice a week, once a week, once every two weeks, once every three weeks, once every four weeks, once every five weeks, once every six weeks, once every eight weeks, once every twelve weeks, or less, as long as a therapeutic response is achieved. In some embodiments, the anti-Bet v 1 antibody is administered once every three months, once every four months, once every five months, once every six months, once every seven months, once every eight months, once every nine months, once every ten months, once every eleven months, or once every twelve months. In some embodiments, the anti-Bet v 1 antibody is administered once or twice a year. In some embodiments, the anti-Bet v 1 antibody is administered once or twice a year before the start of the allergy season (for example, before the birch pollen season).

[0120] In certain embodiments involving the administration of a pharmaceutical composition containing an anti-Bet v 1 antibody at the administration frequencies described herein, each dose is administered in amounts ranging from approximately 50 mg to approximately 600 mg, for example, approximately 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, or 300 mg, or in embodiments where two or more anti-Bet v 1 antibodies are administered, each anti-Bet v 1 antibody is administered in amounts ranging from approximately 50 mg to approximately 600 mg, for example, approximately 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, or 300 mg, for each dose administered. In certain embodiments where two anti-Bet v 1 antibodies are administered, the anti-Bet v 1 antibodies are administered in a 1:1 ratio for each dose administered. In certain embodiments where three anti-Bet v 1 antibodies are administered, the anti-Bet v 1 antibodies are administered in a 1:1:1 ratio for each dose administered.

[0121] In some embodiments, multiple doses of anti-Bet v 1 antibody or a cocktail of anti-Bet v 1 antibodies are administered to a subject over a defined time course. In some embodiments, the method of the Disclosure includes administering multiple doses of anti-Bet v 1 antibody sequentially to a subject. As used herein, “sequential administration” means that each dose of anti-Bet v 1 antibody is administered to the subject at different points in time, such as on different days separated by a predetermined interval (e.g., hours, days, weeks, or months). In some embodiments, the method of the Disclosure includes administering a single initial dose of anti-Bet v 1 antibody, followed by one or more secondary doses of anti-Bet v 1 antibody, and optionally followed by one or more tertiary doses of anti-Bet v 1 antibody, sequentially to a patient. The terms “initial dose,” “secondary dose,” and “tertiary dose” refer to the temporal sequence of administration of anti-Bet v 1 antibodies. Therefore, the "initial dose" is the dose administered at the start of the treatment regimen (also called the "loading dose"), the "secondary dose" is the dose administered after the initial dose, and the "tertiary dose" is the dose administered after the secondary dose.

[0122] In some embodiments, the initial, secondary, and tertiary doses all contain the same amount of anti-Bet v 1 antibody, but differ in their administration frequency. In other embodiments, the amounts of anti-Bet v 1 antibody contained in the initial, secondary, and / or tertiary doses differ from each other during the course of treatment (e.g., adjusted up or down as appropriate). In certain embodiments, one or more doses (e.g., 1, 2, 3, 4, or 5) are administered as a “loading dose” at the start of the treatment regimen, with subsequent doses administered on a lower frequency basis (e.g., “maintenance doses”). For example, the first amount of anti-Bet v 1 antibody or anti-Bet v 1 antibody cocktail may be 1.5 times, 2 times, 2.5 times, 3 times, 3.5 times, 4 times, or 5 times or more than the second amount of anti-Bet v 1 antibody or anti-Bet v 1 antibody cocktail.

[0123] In some embodiments, each secondary and / or tertiary dose is 1 to 30 of the immediately preceding dose (e.g., 1, 1 1 / 2, 2, 2 1 / 2, 3, 3 1 / 2, 4, 4 1 / 2, 5, 5 1 / 2, 6, 6 1 / 2, 7, 7 1 / 2, 8, 8 1 / 2, 9, 9 1 / 2, 10, 10 1 / 2, 11, 11 1 / 2, 12, 12 1 / 2, 13, 13 1 / 2, 14, 14 1 / 2, 15, 15 1 / 2, 16, 16 1 / 2, 17, 17 1 / 2, 18, 18 1 / 2, 19, 19 1 / 2, 20, 20 1 / 2, 21, 21 1 / 2, 22, 22 1 / 2, 23, 23 1 / 2, 24, 24) It is administered after 1 / 2, 25, 25 1 / 2, 26, 26 1 / 2, 27, 27 1 / 2, 28, 28 1 / 2, 29, 29 1 / 2, 30, or more weeks. As used herein, the phrase "immediately preceding dose" means, in a sequence of multiple doses, an administration of anti-Bet v 1 antibody administered to the patient immediately before the next dose in that sequence, without any interim doses.

[0124] The methods of the present disclosure may include administering to a patient any number of secondary and / or tertiary doses of an anti-Bet v 1 antibody or a cocktail of anti-Bet v 1 antibodies. For example, in one particular embodiment, the patient is administered only a single secondary dose. In another embodiment, the patient is administered two or more (e.g., 2, 3, 4, 5, 6, 7, 8, or more) secondary doses. Similarly, in one particular embodiment, the patient is administered only a single tertiary dose. In another embodiment, the patient is administered two or more (e.g., 2, 3, 4, 5, 6, 7, 8, or more) tertiary doses.

[0125] In some embodiments involving multiple secondary doses, each secondary dose is administered at the same frequency as the other secondary doses. Similarly, in some embodiments involving multiple tertiary doses, each tertiary dose is administered at the same frequency as the other tertiary doses. Alternatively, the frequency with which secondary and / or tertiary doses are administered to a patient may change throughout the course of the treatment regimen. The frequency of administration may be adjusted by the physician during the course of treatment according to the individual patient's needs after clinical examinations.

[0126] In some embodiments, a pharmaceutical composition comprising each of the first anti-Bet v 1 antibody, the second anti-Bet v 1 antibody, and the third anti-Bet v 1 antibody disclosed herein is administered to a subject at a dose of approximately 50 mg per antibody (total dose of approximately 150 mg of the three antibodies). In some embodiments, the pharmaceutical composition is administered to the subject subcutaneously or intravenously.

[0127] In some embodiments, a pharmaceutical composition comprising each of the first anti-Bet v 1 antibody, the second anti-Bet v 1 antibody, and the third anti-Bet v 1 antibody disclosed herein is administered to a subject at a dose of approximately 150 mg per antibody (total dose of approximately 450 mg of the three antibodies). In some embodiments, the pharmaceutical composition is administered to the subject subcutaneously or intravenously.

[0128] In some embodiments, a pharmaceutical composition comprising each of the first anti-Bet v 1 antibody, the second anti-Bet v 1 antibody, and the third anti-Bet v 1 antibody disclosed herein is administered to a subject at a dose of approximately 300 mg per antibody (total dose of approximately 900 mg of the three antibodies). In some embodiments, the pharmaceutical composition is administered to the subject subcutaneously or intravenously.

[0129] In some embodiments in which two or more anti-Bet v 1 antibodies are administered, the anti-Bet v 1 antibodies are in separate pharmaceutical compositions. In some embodiments, the pharmaceutical compositions are administered simultaneously (for example, by combining the compositions in a solution before administration). In some embodiments, the pharmaceutical compositions are administered separately (for example, by sequential administration).

[0130] Combination therapy In some embodiments, the methods of the present disclosure include administering one or more additional therapeutic agents to a target in combination with the anti-Bet v 1 antibody or cocktail of anti-Bet v 1 antibodies disclosed herein. Where used herein, the expression “in combination with” means that the additional therapeutic agent is administered before, after, or concurrently with the anti-Bet v 1 antibody or pharmaceutical composition comprising the anti-Bet v 1 antibody. The term “in combination with” also includes sequential or concurrent administration of the anti-Bet v 1 antibody and the second therapeutic agent or therapy.

[0131] In some embodiments, additional therapeutic agents are steroids, antihistamines, decongestants, anti-IgE agents, or agents that deplete plasma cells and / or B cells. In some embodiments, additional therapeutic agents are steroids (e.g., corticosteroids such as inhaled corticosteroids (ICS)). In some embodiments, additional therapeutic agents are antihistamines (e.g., loratadine, fexofenadine, cetirizine, diphenhydramine, promethazine, carbinoxamine, desloratadine, hydroxyzine, levocetirizine, triprolizine, brompheniramine, or chlorpheniramine). In some embodiments, additional therapeutic agents are decongestants (e.g., pseudoephedrine or phenylephrine). In some embodiments, additional therapeutic agents are anti-IgE agents (e.g., omalizumab). In some embodiments, additional therapeutic agents are agents that deplete plasma cells and / or B cells, such as BCMA-targeting agents (e.g., anti-BCMA antibodies conjugated to therapeutic agents such as cytotoxic drugs ("BCMA ADC" or "anti-BCMA ADC"), chimeric antigen receptors (CARs) that specifically bind to BCMA ("BCMA CAR" or "anti-BCMA CAR"), and anti-BCMA / anti-3BCMA bispecific antibodies as disclosed in WO 2020 / 018820). [Examples]

[0132] The following examples are provided to give a complete disclosure and explanation of how the methods and compositions of this disclosure are prepared and used, and are not intended to limit the scope of what the inventors consider to be their invention. Efforts have been made to ensure accuracy of the figures used (e.g., quantities, temperatures, etc.), but some experimental error and deviation should be taken into account. Unless otherwise indicated, parts are parts by weight, molecular weight is average molecular weight, temperature is Celsius, and pressure is atmospheric pressure or near atmospheric pressure.

[0133] Example 1: Clinical trial to investigate the safety, tolerability, pharmacokinetics, and pharmacodynamic effects of a single dose of REGN5713-5714-5715 in healthy adults. Study design and objectives This example describes a Phase 1, 2-part, randomized, double-blind, placebo-controlled study of the safety, tolerability, pharmacokinetics (PK), and pharmacodynamic (PD) effects of a single dose of REGN5713-5714-5715 in healthy adults (NCT03969849). Part A evaluated the safety, tolerability, and PK of single escalating doses of 150 mg subcutaneously (SC), 450 mg SC, 900 mg SC, and 900 mg intravenously (IV) in healthy adults. Part B evaluated the safety, tolerability, PK, and PD of a single dose of REGN5713-5714-5715 in healthy adults with birch pollen allergy.

[0134] REGN5713 is a fully human anti-Bet v 1 antibody containing HCVR (SEQ ID NO: 1), HCDR1 (SEQ ID NO: 2), HCDR2 (SEQ ID NO: 3), HCDR3 (SEQ ID NO: 4), LCVR (SEQ ID NO: 5), LCDR1 (SEQ ID NO: 6), LCDR2 (SEQ ID NO: 7), and LCDR3 (SEQ ID NO: 8).

[0135] REGN5714 is a fully human anti-Bet v 1 antibody containing HCVR (SEQ ID NO: 11), HCDR1 (SEQ ID NO: 12), HCDR2 (SEQ ID NO: 13), HCDR3 (SEQ ID NO: 14), LCVR (SEQ ID NO: 15), LCDR1 (SEQ ID NO: 16), LCDR2 (SEQ ID NO: 17), and LCDR3 (SEQ ID NO: 18).

[0136] REGN5715 is a fully human anti-Bet v 1 antibody containing HCVR (SEQ ID NO: 21), HCDR1 (SEQ ID NO: 22), HCDR2 (SEQ ID NO: 23), HCDR3 (SEQ ID NO: 24), LCVR (SEQ ID NO: 25), LCDR1 (SEQ ID NO: 26), LCDR2 (SEQ ID NO: 27), and LCDR3 (SEQ ID NO: 28).

[0137] The primary objective of this study was to evaluate the safety and tolerability of REGN5713-5714-5715 in healthy adults. The secondary objectives of this study were to characterize the concentration-time profile of a single dose of REGN5713-5714-5715 in healthy adults (Parts A and B), to evaluate the immunogenicity of a single dose of REGN5713-5714-5715 (Parts A and B), to evaluate the inhibition of allergic symptoms, as measured by the total nasal symptom score (TNSS) induced by nasal allergen induction (NAC) with birch allergens, after a single SC administration of REGN5713-5714-5715 in birch-sensitized allergy subjects (Part B), and to evaluate the responsiveness of skin tests induced by a skin prick test (SPT) using serial birch allergen titration after a single 900 mg SC administration of REGN5713-5714-5715 (Part B). Regarding Part B, additional preliminary objectives are to evaluate the inhibition of allergic symptoms, as measured by TOSS induced by birch allergen NAC, in birch-sensitized allergy subjects after a single SC administration of REGN5713-5714-5715, and to evaluate the inhibition of the allergic response, as measured by PNIF, in birch-sensitized AR subjects intranasally induced with birch pollen extract (NAC) after a single SC administration of REGN5713-5714-5715. The objectives were to evaluate the allergic response, as measured by RNA sequencing of nasal mucosal tissue, and the inhibition of chemokines / cytokines in nasal fluid after a single SC administration of REGN5713-5714-5715 in birch-sensitized AR subjects induced intranasally with birch pollen extract (NAC), and to assess the relationship between the clinical response to REGN5713-5714-5715 and the ability of REGN5713-5714-5715 and its components to inhibit endogenous serum IgE binding to Bet V 1 at baseline prior to drug exposure.

[0138] In Part A, after obtaining informed consent, subjects were evaluated for eligibility. Eligible subjects participated in a pre-baseline visit and were hospitalized during that period. The investigational drug was administered on day 1 of the study (randomization), the following day. Subjects were monitored at the clinic for at least 48 hours after SC administration or at least 24 hours after IV administration. After safety evaluation, subjects were discharged on day 3 after SC administration or day 2 after IV administration, respectively. Subjects returned to the clinic for safety evaluation on days 8, 15, 22, 29, 43, 57, 85, 99, and 113 (for the SC administration cohort), or on days 4, 8, 15, 22, 29, 43, 57, 85, 99, and 113 (for the IV administration cohort).

[0139] Part B was a randomized, placebo-controlled, double-blind, single-dose nasal allergen-induced (NAC) trial to evaluate the time-course safety, tolerability, and drug concentration of 900 mg of SC REGN5713-5714-5715, as well as the PD effect, in healthy adults with birch pollen allergy. Enrollment for Part B took place outside of the birch allergy season in birch endemic areas. After obtaining informed consent, subjects were evaluated for eligibility over three screening periods of up to 12 weeks. During screening visit 1, subjects with a history of AR to birch pollen underwent a medical history, physical examination, birch SPT, and blood tests for birch pollen and Bet v 1 specific IgE. If a subject tested positive for birch, birch pollen, and Bet v 1-specific IgE in a SPT (Special Test) according to the inclusion / exclusion criteria, and met the criteria, they were invited to Screening Visit 2. At Screening Visit 2, subjects underwent serum pregnancy testing (if applicable), spirometry, ECG, serological testing for chronic viral infections (human immunodeficiency virus infection [HIV] and hepatitis B and C), hematological testing, chemistry testing, and urinalysis, and were evaluated for the remaining study eligibility criteria. Screening Visits 1 and 2 could be combined into a single visit if the patient had previously tested positive for birch SPT or birch IgE within the past 12 months. Prior to the NAC (Non-Acceptance Test), subjects had to be observed for at least 10 minutes to achieve a resting / baseline of 2 or less, indicating that they did not have active nasal symptoms at rest (due to viral infection, sinusitis, allergies, etc.). Before initiating NAC, SPT was performed using serial allergen titration to evaluate the immediate phase reaction (EPR). After completing the EPR evaluation, NAC proceeded as follows. ●NAC was performed using increasing doses of birch extract every 10 minutes for up to 1 hour (up-titration stage), or until a TNSS of 7 or higher was achieved. ● Peak TNSS was recorded. ●The birch extract concentrations used to achieve a TNSS of 7 or higher were recorded. ●The subjects were observed for the following hour, and the TNSS was recorded at approximately 5 minutes, 15 minutes, 30 minutes, 45 minutes, and 1 hour, and then every hour up to 6 hours after the peak TNSS. ●TNSS, TOSS, PNIF (nasal congestion, measured in L / min), and sneezing were measured during the NAC procedure and the 6-hour observation period.

[0140] Subjects with birch allergy were eligible for registration based on having a TNSS of 2 or less prior to the screening NAC (visit 3) and a TNSS of 7 or more within the first hour of the up titration step. Furthermore, for eligibility, subjects had to experience a decrease of more than 20% in PNIF or three or more sneezes between the initial non-zero dose and approximately 10 minutes after the maximum / peak dose.

[0141] The investigational drug was administered on day 1 of the trial. Participants were monitored at the clinic for at least 8 hours after SC administration. After safety assessment, participants were discharged at least 8 hours after SC administration. Participants returned to the clinic for safety assessment on days 4, 8, 29, 57, 85, 99, and 113. Trial visits ended approximately 16 weeks after investigational drug administration. Participants participated in the trial for approximately 28 weeks, including the screening period.

[0142] This study was conducted in accordance with the provisions of the Declaration of Helsinki, the guidelines of the International Conference on Harmonization Good Clinical Practices, and applicable regulatory requirements. The protocol was reviewed and approved by the Institutional Review Board / Ethics Committee at all sites. Written informed consent was obtained from all adult patients.

[0143] Patient group This study enrolled healthy adult men and women aged 18-60 years (including those aged 18-60) at the time of their screening visit.

[0144] Inclusion Criteria: Patients had to meet the following criteria to be eligible for inclusion in this study: (1) Male or female aged 18-60 years (including) at the time of screening; (2) Weight 18-31 kg / m² at the time of screening. 2 (including) Body Mass Index (BMI), (3) Based on medical history, physical examination, vital signs measurement, and ECG performed prior to administration of the investigational drug, the subject is judged by the principal investigator to be in good health, (4) Based on clinical safety tests obtained during a screening visit prior to administration of the investigational drug, the subject is in good health, (5) The subject is willing and able to comply with clinic visits and study-related procedures, and (6) Informed consent is provided signed by the subject or their legal representative. Additional inclusion criteria for Part B: (7) able to understand and complete study-related questionnaires; (8) having a history of birch pollen-induced AR symptoms with or without conjunctivitis (over at least two seasons), based on subject recall; (9) testing positive on SPT with birch pollen extract during the screening period (mean wheal diameter at least 5 mm larger than negative controls); (10) testing positive on allergen-specific IgE (sIgE) tests for birch pollen and Bet v 1 (≥0.35 kUa / L) during the screening period; (11) testing positive on NAC with birch extract at the screening visit with 7 or more peak TNSS out of 12; and (12) experiencing a decrease of more than 20% in PNIF and / or 3 or more sneezes between the initial non-zero dose and approximately 10 minutes after the highest dose of NAC at the screening visit.

[0145] Exclusion Criteria: The following were the exclusion criteria for this study: (1) A history of clinically significant cardiovascular, respiratory, hepatic, renal, gastrointestinal, endocrine, hematological, psychiatric, or neurological disease as assessed by the principal investigator that could interfere with the results of the study or pose an additional risk to the subject by participating in the study; (2) Any physical examination findings and / or any medical history that, in the opinion of the principal investigator, could interfere with the results of the study or pose an additional risk to the subject by participating in the study; (3) Hospitalization for any reason (more than 24 hours) within 60 days of screening visit 1; (4) Current smokers or former smokers (cigarettes or e-cigarettes) who quit smoking within 3 months of screening visit 1; 5) History of drug or alcohol abuse within one year prior to the first screening visit (Note: Drug and alcohol screening must be negative before administration of the investigational drug), (6) Serologically positive for HIV, hepatitis B, or hepatitis C at the time of screening or within three months prior to administration, excluding false-positive screening tests demonstrated by polymerase chain reaction or Western blot, (7) Any malignant tumor within the past five years, excluding basal cell or squamous cell carcinoma of excised skin, or carcinoma in situ of the cervix or anus, with no evidence of local recurrence or metastatic disease for three years, (8) Blood pressure at 60 mL / min / 1.73 m² at the time of screening. 2(9) Having an estimated glomerular filtration rate of less than 100% (using the Modification of Diet in Renal Disease study equation or the Chronic Kidney Disease Epidemiology Collaboration equation), or clinically significant ECG abnormalities as assessed by the principal investigator at screening or before administration, or confirmed by repeated measurements with abnormal intervals (QTcF >470ms for males, >480ms for females), (10) a history of acute hypersensitivity and / or anaphylaxis to components of protein therapies or preparations, or allergies that, in the opinion of the principal investigator, could pose a substantial risk to the subject, (11) participation in any clinical research study evaluating another investigational drug or therapy within 90 days or at least 5 half-lives (whichever is longer) for investigational biological products, or at least 28 days for other investigational drugs, or within 6 months for immunotherapy, prior to the screening visit for the current study, (12) any permission or prohibition for this study (13) Members of the study team at the study site and / or their close relatives, unless otherwise approved by the sponsor; (14) Known hypersensitivity to doxycycline or a similar compound (i.e., tetracycline); (15) Pregnant or lactating women; (16) Women of childbearing potential who have a positive pregnancy test result at baseline or have no pregnancy test result; (17) Women of childbearing potential who do not intend to use highly effective contraception before the initiation of the first treatment, during the study, and for at least six months after the last dose; (18) Sexually active men who do not intend to use any form of medically acceptable birth control during the follow-up period of the investigational drug and for six months after the last dose of the investigational drug: vasectomy with a medical evaluation of the success of the surgery, or consistent use of condoms.Additional exclusion criteria for Part B: (19) Receipt of investigational drug REGN5713-5714-5715 in Part A; (20) Experiencing systemic symptoms in response to NAC at visit 3, including but not limited to wheezing, throat tightness, or shortness of breath; (21) Significant rhinitis or sinusitis outside of birch pollen season or due to daily exposure to other symptomatic allergens, as expected to be consistent with the study NAC assessment as assessed by the Principal Investigator; (22) Prediction of a significant change in allergen exposure in the home or work environment, as expected to be consistent with the study NAC assessment as assessed by the Principal Investigator; (23) Abnormal lung function as determined by the Principal Investigator, with an FEV1 less than 80% of the prediction; (24) Regular exposure for more than 6 months per year. (25) A history of asthma requiring long-term medication such as inhaled corticosteroids; (26) A history of asthma with two or more asthma exacerbations requiring hospitalization or systemic corticosteroids in the previous year; (27) A history of severe recurrent sinusitis defined as at least three episodes requiring antibiotic treatment per year in the past two years; (28) A history of chronic obstructive pulmonary disease (COPD); (29) A history of birch allergy immunotherapy (SCIT, sublingual immunotherapy, or oral immunotherapy) in the five years prior to screening; (30) Use of anti-IgE or other biological therapy within the six months prior to screening; (31) Allergen-specific immunotherapy using any allergen within the six months prior to screening; (32) Screening for an ACT of less than 20 at any screening visit.

[0146] Investigational drug REGN5713, REGN5714, and REGN5715 were supplied for this study at the following concentrations for each drug. ● 265 mg of lyophilized REGN5713, REGN5714, and REGN5715 pharmaceuticals in 20 mL vials. ●Liquid-dried REGN5713 / REGN5714 / REGN5715 placebo product in a 20mL vial

[0147] The reconstituted drug was 150 mg / mL for each drug: REGN5713, REGN5714, and REGN5715. Therefore, the total injectable drug volume (SC) was 1 mL for the total dose of 150 mg of REGN5713, REGN5714, and REGN5715 (Cohort 1), 3 mL for the total dose of 450 mg of REGN5713, REGN5714, and REGN5715 (Cohort 2), and 6 mL for the total dose of 900 mg of REGN5713, REGN5714, and REGN5715 (Cohort 3). Multiple syringes could be used for SC injection. For example, three syringes (2 mL per syringe) were used to inject the SC drugs into Cohort 3 and Part B. The preferred site for SC injection was the abdomen, but injections could also be administered to the abdomen, thigh, or upper arm.

[0148] For Part A, there were the following four cohorts: [Table 1]

[0149] For Part B, REGN5713-5714-5715 at 900 mg SC (300 mg / mAb) or placebo was administered on day 1.

[0150] Emergency Treatment: If necessary, subjects experiencing an allergic reaction were treated with emergency anti-allergic treatment at the discretion of the principal investigator. Subjects were also able to take oral antihistamines as needed for AR symptoms during the study period. However, oral antihistamines were prohibited within 5 days prior to or during visits for NAC or skin prick tests.

[0151] Evaluation of Results For both Part A and Part B, the primary endpoints were the incidence and severity of treatment-induced adverse events (TEAEs) as reported by subjects or observed by investigators, and other safety variables in subjects receiving SC or IV-administered REGN5713-5714-5715 compared to the placebo group.

[0152] The secondary endpoints for Parts A and B were the total concentrations of REGN5713, REGN5714, and REGN5715 in serum at the sample collection time specified in the clinic schedule, and the incidence of anti-drug antibodies developed under treatment against REGN5713, REGN5714, and REGN5715 in the subject.

[0153] The secondary endpoint for Part B only was the change and percent change in the area under the curve (AUC) of TNSS (0 - 1 hour after peak TNSS) from the pre-treatment baseline TNSS AUC of REGN5713 - 5714 - 5715 900 mg SC at 8, 29, and 57 days and the TNSS (0 - 1 hour after peak TNSS) in response to NAC compared to placebo, and the change and percent change in the mean wheal diameter of skin prick tests using serial Amb a 1 allergen titrations of REGN5713 - 5714 - 5715 900 mg SC from the pre-treatment baseline at 8, 29, and 57 days compared to placebo.

[0154] Procedures for evaluating efficacy are described below.

[0155] Total Nasal Symptom Score (TNSS): The total nasal symptom score (TNSS) measured on a scale of 0 - 12 is a composite symptom evaluation of congestion, itching, and rhinorrhea (each evaluated on a scale of 0 - 3, where 3 is severe), and sneezing (where 3 is more than 5 sneezes). TNSS is recorded in response to NAC at 8 days, 29 days, and 57 days from the pre-treatment baseline TNSS to evaluate the efficacy of REGN5713 - 5714 - 5715 against AR symptoms induced by NAC using Amb a 1 allergen.

[0156] Total Ocular Symptom Score (TOSS): The total ocular symptom score (TOSS) measured on a scale of 0 - 12 is a composite symptom evaluation of ocular symptoms (itching / burning, injection, epiphora and tearing, and swelling and edema). The TOSS score is recorded during NAC evaluation.

[0157] Peak nasal inspiratory flow (PNIF): Peak nasal inspiratory flow (PNIF, L / min) is a measure of nasal patency or nasal obstruction. PNIF is measured and recorded during NAC evaluation.

[0158] Safety variables Safety was evaluated throughout the entire study (Parts A and B) by investigating adverse events (AEs), vital signs, 12-lead electrocardiogram (ECG), physical examinations, and routine clinical safety tests. Further, for Part B, spirometry (performed in accordance with the American Thoracic Society [ATS] / European Respiratory Society [ERS]) including measurements of forced vital capacity (FVC) (L), forced expiratory volume in 1 second (FEV1) (L), FEV1 / FVC (%), peak expiratory flow (L / s), forced expiratory flow between 25% and 75% (L / s), and asthma control test (ACT) was performed at screening to exclude subjects with abnormal lung function and / or inadequately controlled asthma. ACT was performed in all subjects prior to all NAC procedures, and spirometry was measured prior to NAC in any subject with a history of asthma. FEV1 must be greater than 80% of predicted in any subject with a history of asthma in order to perform NAC.

[0159] Pharmacokinetic variables, pharmacodynamic variables, and other biomarker variables Pharmacokinetic variables include the following: Concentrations of REGN5713, REGN5714, and REGN5715 in serum at each specified time point.

[0160] In Part B, a standard SPT using serial allergen titration against birch allergen extract and at least one other birch homologous tree allergen (e.g., alder) will be performed at screening visit 1 and on designated test days (e.g., days 8, 29, 57, and 113) to assess sensitization status. Skin prick test data may be used to determine the relationship between baseline birch sensitization and the PD effect of REGN5713-5714-5715 for reducing nasal symptoms during NAC. Serial allergen titration and titration SPT with birch allergen extract will be performed at screening and on day 29 to assess the PD effect of REGN5713-5714-5715 on the skin wheal size response (mediated by mast cell degranulation).

[0161] Serum antibodies are evaluated as follows: ● Allergen-specific IgE levels at screening visit 1 (V1) and baseline (birch pollen, Bet v 1) ● Screening and baseline allergen-specific IgE levels (Bet v 1 and other common allergens) to assess sensitization status and evaluate the relationship between response to REGN5713-5714-5715 and poly / mono sensitization. ● Allergen-specific IgE levels (Bet v 1 and other common allergens) on days 8, 29, and 57, using blood samples collected before NAC. ● Additional subclasses of serum antibodies (e.g., IgG and IgG4 against birch pollen and Bet v 1) can be measured at screening, baseline, and on days 8, 29, and 57 using blood samples collected before NAC.

[0162] Furthermore, to better understand the effects of REGN5713-5714-5715 on birch allergy, blood may be obtained for additional preliminary research studies. This may include evaluating effective competition between REGN5713-5714-5715 and endogenous serum IgE for allergen binding in in vitro assays. Serum IgE for other related tree allergies will also be measured to correlate with symptom improvement after treatment with REGN5713-5714-5715. Peripheral blood samples (whole blood, peripheral blood mononuclear cells [PBMCs]) will also be collected for preliminary biomarker studies to investigate changes in ex vivo allergic responses to birch pollen and other allergens after treatment. Nasal brushing may be collected from subjects with birch allergy approximately 6 hours after NAC. Changes in the genetic signature of type 2 inflammatory response after NAC will be analyzed by RNA-seq.

[0163] Nasal fluid may be obtained from subjects with birch allergy up to approximately 6 hours after NAC. Type II cytokines and chemokines in the nasal fluid of subjects with birch pollen allergy are determined, and levels of type II cytokines (e.g., IL-4, IL-13) and chemokines (e.g., TARC, eotaxin) are measured up to 6 hours after NAC to evaluate the suppression of the allergic response to birch pollen after treatment with REGN5713-5714-5715.

[0164] result Test subjects In Part A, a total of 32 participants were randomized (24 to REGN5713-5714-5715 and 8 to placebo). In Part B, a total of 64 participants were randomized (32 to REGN5713-5714-5715 and 32 to placebo). All participants randomized in Parts A and B received the intended investigational drug and completed the trial until the end-of-trial visit on day 113.

[0165] Baseline demographics of age, race, ethnicity, and BMI were generally similar across the treatment groups in Part A (Table 1). Among the subjects in Part A (REGN 5713-5714-5715), 16 subjects (66.7%) were female, with mean heights of 169.2 cm and 67.5 kg, compared to 2 female subjects (25%) in the placebo group and mean heights of 177.9 cm and 75.6 kg. In Part B, baseline demographics of age, sex, race, ethnicity, height, weight, and BMI were generally similar across the treatment groups (Table 2).

[0166] safety REGN5713-5714-5715 were generally well-tolerated when administered via the SC and IV routes. There were no SAEs, deaths, or TEAEs leading to study discontinuation in Part A or Part B (Tables 3 and 4). The overall frequency of TEAEs was similar between REGN5713-5714-5715 (Part A: 88.9%, Part B: 81.3%) and placebo (Part A: 100%, Part B: 93.8%) (Tables 3 and 4). There was no dose-dependent trend in any of the TEAEs. The overall frequency of treatment-related TEAEs was also similar between REGN5713-5714-5715 (Part A: 25.0%, Part B: 21.9%) and placebo (Part A: 12.5%, Part B: 25.0%) (Tables 3 and 4).

[0167] In Part A, two injection site reactions were reported (1 [16.7%] in the 450 mg SC REGN5713-5714-5715 group and 1 [16.7%] in the 900 mg SC group), and in Part B, 11 were reported (5 [15.6%] in the 900 mg SC group versus 6 [18.8%] in the placebo group). The severity of all injection site reactions was mild. There was no dose-dependent trend in any of the TEAEs. In Part A of the REGN5713-5714-5715 IV+SC group, the most frequently observed TEAEs were headache (29.2% in those receiving REGN5713-5714-5715 vs. 50.0% in those receiving placebo), nasopharyngitis (20.8% vs. 62.5%), and abdominal pain (16.7% vs. 25.0%). In Part B, headache (34.4% vs. 28.1%), nasopharyngitis (15.6% vs. 34.4%), and sinusitis (6.3% vs. 15.6%) were the most frequent TEAEs in the REGN5713-5714-5715 group. [Table 2] [Table 3-1] [Table 3-2] [Table 4] [Table 5]

[0168] Pharmacokinetics In Part A, after SC administration, the PK profile showed a T of approximately 8.5 days for all three administered antibodies. maxshowed an initial absorption phase followed by a single-phase elimination phase. The half-lives of all three antibodies were similar, with mean half-lives of 25.9 ± 5.0 days for REGN5713 across three SC doses, 31.7 ± 4.3 days for REGN5714 across three SC doses, and 34.7 ± 5.9 days for REGN5715 across three SC doses. REGN5713-5714-5715 concentrations increased in a dose-proportional manner, showing dose-proportional increases in inf and max , consistent with linear PK for all three antibodies. After IV administration, a rapid initial distribution phase was observed, followed by a single-phase elimination phase. The half-lives of all three antibodies were consistent with SC administration, being 26.7 ± 5.0 days, 29.9 ± 4.7 days, and 32.1 ± 5.1 days for REGN5713, 5714, and 5715, respectively. REGN5713 showed slightly higher clearance and lower serum concentrations and a slightly shorter half-life than REGN5714 and REGN5715, but all three antibodies generally showed similar PK characteristics. Administration of the IV dose took into account the calculation of the bioavailability (F) from the mean inf . Among the three antibodies, F ranged from 70 to 93% at various dose levels, with mean F being 72% for REGN5713, 79% for REGN5714, and 88% for REGN5715 across all SC doses.

[0169] In Part B, administration of REGN5713-5714-5715 SC in Bet v 1-sensitive patients showed a concentration-time profile similar to that in healthy subjects after linear PK. Visual inspection of the concentration-time profiles and graphical overlay showed little difference between Bet v 1-sensitive patients and healthy volunteers.

[0170] Effectiveness The primary efficacy endpoints assessed in Part B were met. As shown in Figure 1 and Table 5, a single dose of REGN5713-5714-5715 significantly reduced the TNSS AUC (0-1 hour) after NAC compared to placebo at 8, 29, and 57 days after the single dose (placebo-adjusted change in TNSS AUC from baseline NAC: -1.17 [p=0.001], -1.18 [p=0.001], and -0.85 [p=0.024] for days 8, 29, and 57 of the study, respectively). The percentage change from baseline yielded similar results, with a significant reduction in TNSS AUC (0-1 hour) compared to baseline NAC up to day 57 (placebo-adjusted changes in TNSS AUC from baseline NAC of -31.57% [p=0.002], -27.35% [p=0.003], and -19.00% [p=0.053] for days 8, 29, and 57, respectively). See Figure 2 and Table 5. Treatment with the anti-Bet v 1 cocktail consistently reduced nasal symptoms over two months, with eight subjects showing a 100% reduction in TNSS AUC after two months. The relative decrease in therapeutic effect of REGN5713-5714-5715 on day 57 was associated with an increased placebo effect (percentage reductions in TNSS AUC (0-1 hour) of -31.39%, -31.15%, and -41.96% for days 8, 29, and 57, respectively). The therapeutic effect of REGN5714-5714-5715 remained consistently around 60% across all trial days (percentage reductions in TNSS AUC (0-1 hour) of -62.96%, -58.50%, and -60.96% for days 8, 29, and 57, respectively). Treatment with a single dose of REGN5713-5714-5715 also significantly reduced peak TNSS after NAC compared to placebo on days 8, 29, and 57. See Figure 3 and Table 5.

[0171] The mean wheal diameter at baseline was similar in the REGN5713-5714-5715 group and the placebo group (data not shown), with similar AUCs of 4.14 and 3.93, respectively, observed at baseline visits (Table 2). REGN5713-5714-5715 significantly reduced the AUC of mean wheal diameter at SPT compared to placebo up to day 113 (percentage change from baseline SPT adjusted for placebo: -72.39% [p<0.001], -65.61% [p<0.001], -77.90% [p<0.001], and -73.56% [p<0.001] at days 8, 29, 57, and 113, respectively). See Figure 4A and Table 5. Therefore, a single dose of the anti-Bet v 1 cocktail showed a sustained response that reduced birch sensitization, even 3.5 months after administration. [Table 6]

[0172] At baseline, NAC-induced ocular symptoms were minimal in the subjects, with mean TOSS AUCs of 1.3 and 2.1 in the REGN5713-5714-5715 group and the placebo group, respectively (Table 2). As shown in Table 5 and Figures 5A-5B, a single dose of REGN5713-5714-5715 significantly reduced TOSS AUC (0-1 hour) after NAC compared to placebo at 8 and 29 days post-administration, and this trend was also present at 57 days post-administration (placebo-adjusted change in TOSS AUC from baseline NAC: -0.48 [p=0.013], -0.59 [p=0.007], and -0.30 [p=0.191] for days 8, 29, and 57 of the study, respectively). Two (6.3%) of the placebo group and five (15.6%) of the REGN5713-5714-5715 group were symptom-free at baseline and had an AUC of 0. Furthermore, peak nasal inspiratory flow rate (PNIF) improved by at least 30% with the anti-Bet v 1 cocktail compared to placebo, with statistical significance achieved on day 29 (32% [p=0.092], 47% [p=0.017], and 35% [p=0.121] of placebo-adjusted percentage change from baseline NAC at days 8, 29, and 57, respectively). See Figure 6 and Table 5.

[0173] Titration skin prick test for birch homologous allergens The mean wheal diameter AUC of the serial alder titration SPT at baseline was similar between the REGN5713-5714-5715 (5.3) group and the placebo (4.9) group (Table 2). REGN5713-5714-5715 significantly reduced the mean wheal diameter AUC of the alder SPT up to day 113 compared to placebo. The placebo-adjusted percentage change from baseline SPT was -44.7% (day 8, P<.001), -46.3% (day 29, P<.001), -52.8% (day 57, P<.001), and -49.8% (day 113, P<.001). See Figure 4B.

[0174] Basophil activation A biomarker subtest was conducted on a subset of patients from Part B. A single-center basophil activation assay was performed on 26 subjects (13 treated with REGN5713-5714-5715, 13 receiving placebo). Basophil activation was characterized by CD63 surface expression, and the percentage of CD63+ basophils was measured, for example, by flow cytometry. Basophils in the whole blood of patients were stimulated ex vivo with increased concentrations of birch allergen extract. Basophil responsiveness to allergen stimulation was measured by EC50, the concentration of birch allergen required to achieve 50% of maximum basophil activation. Higher EC50 values ​​indicate decreased basophil responsiveness to allergen stimulation (i.e., increased suppression of basophil responsiveness). Percentage changes from baseline EC50 were compared between the treatment and placebo groups using non-parametric tests. The EC50 of birch tree correlated with clinical responses, including the total nasal symptom score (TNSS), after NAC and SPT.

[0175] Basophil activation studies (BAT) were performed using the Buhlmann's Flow CAST® kit (Buhlmann Laboratories AG, Switzerland). Dose-response curves were generated after ex vivo stimulation of basophils in patient whole blood using seven increasing concentrations of allergen extracts (0.0055–22.72 ng / mL), followed by basophil detection using CCR3 and CD63. Basophil activation was measured by flow cytometry analysis, specifically by the percentage of basophils positive for CD63 surface expression. Basophil responsiveness to extracts of birch, alder, hazel, apple, and a mixture of grasses (allergen sources: Buhlmann Laboratories AG, Switzerland, or Biomay AG, Vienna) was measured by EC50.

[0176] Significant suppression of basophilic responses to birch, alder, hazelnut, and apple pollen extracts was observed in the treatment group (n=13) versus placebo (n=13), measured by the percentage change in EC50 from baseline (birch and alder: days 8, 57, and 113, all p<0.001; hazelnut: day 8, p<0.001; day 57, p=0.01; apple: day 8, p<0.01; days 57 and 113, p<0.05). The highest EC50 was detected on day 8. See Figures 7A-7D. No suppression of basophilic responses to the grass allergen mixture (negative control) was detected in the REGN5713-5714-5715 treatment (Figure 7E).

[0177] The relationship between basophil suppression and reduction in skin prick test (SPT) was evaluated in patients treated with REGN5713-5714-5715. SPT was performed by pricking the forearm skin of patients with increasing concentrations of birch allergen. Mean wheal diameter (MWD) induced at each birch allergen concentration was used for AUC analysis. In the treatment group, an inverse correlation was observed between the percentage change in birch basophil activation assay (BAT) EC50 and birch SPT (mean wheal diameter) AUC at day 8 (r=-0.6, p=0.04) and day 57 (r=-0.57, p=0.04). See Figure 8.

[0178] The relationship between basophil suppression and TNSS response was also evaluated. Analysis of the active treatment group administered REGN5713-5714-5715 also showed that the TNSS response to birch allergen was maximally suppressed on day 8, when the basophil response to birch stimulation was most effectively suppressed. (0~1時間) An inverse correlation was observed between the percentage change in AUC and EC50 (r=-0.71, p=0.095). See Figures 9A-9B. One subject did not achieve any clinical improvement, and no suppression of basophil responsiveness was detected in that subject's blood.

[0179] conclusion REGN5713-5714-5715 was generally well-tolerated when administered via IV or SC route. There were no deaths or TEAEs leading to treatment or study discontinuation. The concentration-time profile of REGN5713-5714-5715 showed linear PK. A single dose of REGN5713-5714-5715 reduced allergic symptoms as measured by TNSS within 8 days and for at least 2 months, and resulted in a sustained response to reduce birch sensitization up to day 113 of the study.

[0180] Treatment with REGN5713-5714-5715, rather than placebo, significantly reduced the basophilic response to birch pollen extract. In particular, the EC50 of the birch allergen correlated with improved clinical response, indicating that the lower the basophilic sensitivity to birch pollen (i.e., the higher the EC50), the greater the clinical benefit achievable by patients. This study suggests that the clinical efficacy of anti-Bet v1 monoclonal antibodies in patients with birch allergy may be achieved in part by suppressing basophilic and mast cell-mediated allergic responses.

[0181] Following treatment with REGN5713-5714-5715, there was a significant suppression of basophilic responses to alder (r Aln g 1), hazel (r Cor a 1) pollen extracts, and apple (r Mal d 1). These data were also consistent with SPT results, indicating that REGN5713-5714-5715 significantly reduced the mean wheal diameter after SPT using alder pollen extract. The marked inhibitory effect on basophilic responses to homologous allergens suggests that REGN5713-5714-5715 as a cocktail may improve allergic responses to related tree allergens beyond birch and treat birch pollen-related OAS.

[0182] In conclusion, a single subcutaneous administration of REGN5713-5714-5715 is well-tolerated and results in a rapid and sustained reduction of allergic symptoms following birch allergen induction. The suppression of allergic responses to other cross-reactive allergens by the novel Bet v 1 mAb cocktail may provide a convenient, rapid, and long-acting therapy for the prevention and treatment of seasonal birch and related allergies. The three-antibody cocktail targeting Bet v 1, the major birch allergen, results in substantial clinical improvement in most patients.

[0183] Example 2: A clinical trial investigating the efficacy of anti-Bet v 1 monoclonal antibody in reducing symptoms of seasonal allergic rhinitis. Study design and objectives This example describes a phase 3, multicenter, randomized, double-blind, placebo-controlled, parallel-group comparative trial to evaluate the efficacy of an anti-Bet v 1 monoclonal antibody for reducing symptoms of allergic rhinitis and conjunctivitis, and the use of emergency medication during birch pollen season (NCT04709575).

[0184] The primary objective is to evaluate the reduction of allergic symptoms, as measured by the Combined Symptom Medication Score (CSMS), during the birch pollen season, after a single dose of REGN5713-5714-5715 versus placebo. Secondary objectives: (1) to evaluate the reduction of allergic symptoms and the use of allergy relief medications after a single dose of REGN5713-5714-5715 versus placebo, as measured by the Total Symptom Score (TSS), Total Nasal Symptom Score (TNSS), Total Ocular Symptom Score (TOSS), and Daily Medication Score (DMS); (2) to evaluate the safety and tolerability of REGN5713-5714-5715, including the incidence of hypersensitivity reactions and local injection site reactions; and (3) to evaluate the reduction of allergic symptoms, as measured by the mean wheal diameter of the skin prick test (SPT), compared to placebo. (4) assess the reduction of the early allergic response to birch allergen after a single dose of (5) REGN5713, REGN5714, and REGN5715 in serum, determine the systemic exposure of total antibodies (i.e., free and antigen-bound), (5) assess the immunogenicity of subjects to REGN5713, REGN5714, and REGN5715 after a single dose of REGN5713-5714-5715, and (6) assess the number of "good days" (defined as the number of days on which the TSS is 2 or less without the use of anti-allergic emergency drugs).

[0185] The total trial period is approximately 28 weeks, including screening, and depends on the start and end dates of the local birch pollen season. The length of the birch pollen season varies by region. Approximately 300 subjects with birch allergies will be randomized in a 1:1 ratio to receive REGN5713-5714-5715 or placebo.

[0186] Patient group This trial will enroll generally healthy adult men and women with birch allergies.

[0187] Selection Criteria: To be eligible for this study, participants must meet the following criteria: (1) generally healthy men and women aged 18 years or older at the time of screening; (2) documented or self-reported history of birch pollen-induced allergic rhinitis symptoms, with or without conjunctivitis, for at least two years; (3) positive SPT with birch pollen extract during the screening period (i.e., mean wheal diameter at least 5 mm larger than the negative control); (4) positive sIgE test for birch pollen and Bet v 1 (i.e., ≥0.7 kUa / L); (5) willingness and ability to attend clinic visits and comply with study-related procedures; (6) provide informed consent signed by the study participant or their legal representative; and (7) be able to understand and complete study-related questionnaires.

[0188] Exclusion Criteria: Participants meeting any of the following criteria will be excluded from this study: (1) prior participation in the REGN5713-5714-5715 clinical trial; (2) recurrent or chronic rhinitis or sinusitis, as assessed by the Principal Investigator, due to daily exposure to other allergens that are not related to the birch pollen season or are expected to cause symptoms consistent with the birch pollen season; (3) at the discretion of the Principal Investigator, who anticipate a significant change in allergen exposure in the home or work environment that is expected to be consistent with the study assessment; (4) persistent chronic or recurrent acute infection, or any untreated respiratory infection, requiring treatment with antibiotics, antivirals, or antifungals within four weeks prior to screening. Participants may be reassessed for eligibility after symptoms have subsided.(5) Documentary evidence of active SARS-CoV-2 infection, (6) Abnormal lung function as determined by the principal investigator, with an FEV1 less than 70% of the predicted value at screening or randomization, (7) History of asthma with two or more asthma exacerbations requiring hospitalization or systemic corticosteroids in the previous year, (8) History of severe recurrent sinusitis defined as at least three episodes requiring antibiotic treatment per year in the past two years prior to screening, (9) History of nasal polyps, (10) Active lung disease other than asthma, (11) History of immunotherapy for birch or related tree allergy (SCIT, SLIT, or oral immunotherapy) within five years prior to screening, (12) Use of anti-IgE or other biological therapies modifying type 2 inflammation within six months prior to screening, (13) Six months prior to screening (14) A history of clinically significant cardiovascular, respiratory, hepatic, renal, gastrointestinal, endocrine, hematological, psychiatric, or neurological disease that, in the opinion of the principal investigator, could interfere with the results of the study or present further risk to the subject by participation in the study; (15) Any physical examination findings and / or any medical history that, in the opinion of the principal investigator, could interfere with the results of the study or present further risk to the subject by participation in the study; (16) Any laboratory findings, as determined by the principal investigator at the time of screening, that show evidence of organ dysfunction or clinically significant deviation from the normal range, including but not limited to: (a) clinically significant / active underlying hepatobiliary disease; or (b) 1.5 × 10. 3 Neutrophils less than / μL or 100,000 cells / mm³ 3(17) Abnormal laboratory values ​​at screening, such as platelets less than 100%, (18) History of drug or alcohol abuse within the year prior to screening, (19) Any malignant tumor within the past 5 years, excluding basal cell or squamous cell carcinoma of excised skin, or carcinoma in situ of the cervix or anus, with no evidence of local recurrence or metastatic disease for 3 years, (10) Clinically significant ECG abnormalities during the screening period as assessed by the principal investigator, (21) History of acute hypersensitivity and / or anaphylaxis to excipients in the therapeutic agent, or allergies that, in the opinion of the principal investigator, may pose a substantial risk to the subject, (22) Within 2 months, or at least within 5 half-lives (if known), whichever is longer (22) persons who are not willing or unable to comply with the permitted and prohibited drug specifications for this study, (23) members of the study team at the study site and / or their close relatives, unless otherwise approved by the sponsor, (24) pregnant or breastfeeding women, (25) women of childbearing potential (WOCBP)* who are not willing to use highly effective contraception before, during, and for at least six months after administration of the study, (26) sexually active men who are not willing to use any form of medically acceptable birth control during the follow-up period for the study and for six months after administration of the study: vasectomy with a medical evaluation of the success of the procedure, or consistent use of condoms.

[0189] Investigational drug REGN5713, REGN5714, and REGN5715 are supplied individually in open-label carton vials. Each 20 mL vial contains 265 mg of lyophilized protein. The corresponding placebo is supplied as lyophilized powder in a 20 mL open-label carton vial. Instructions for preparing the following therapeutic agents are provided in the pharmacy manual. ●REGN5713-5714-5715 900mg single subcutaneous (SC) dose (300mg per mAb) ●Single-dose SC of the corresponding placebo to replace REGN5713-5714-5715

[0190] The subjects will be provided with the following medications to treat allergic symptoms during the trial. ● Desloratadine 5mg (second-generation antihistamine) ● Olopatadine 1 mg / mL (antihistamine eye drops) ● Mometasone fluate 50 ug / dose (intranasal steroid)

[0191] From the time of investigational drug administration through the entire birch pollen season, participants will be required to use an electronic diary to record daily medication use, including information on which medications were used and the pre-specified amounts used. Emergency medication use should be initiated when a participant reaches a symptom threshold of approximately 4 / 18 or higher for TSS. Participants will be trained to understand the severity of symptoms associated with 4 / 18 or higher TSS. Participants should be instructed not to use emergency medication in anticipation of birch pollen season. Participants should also be instructed not to use antihistamines (i.e., desloratadine or olopatadine) for 5 days prior to the end of their trial visits.

[0192] Participants will be randomized in a 1:1 ratio to receive either REGN5713-5714-5715 or the corresponding placebo. Randomization will be stratified based on the following criteria: ● Serum-specific birch pollen IgE levels at screening (<17.5 kUa / L vs. ≥17.5 kUa / L) ● North America only: Serum-specific oak pollen IgE levels at screening (<0.7 kUa / L vs. ≥0.7 kUa / L) ●Geographical regions (North America vs. Europe)

[0193] Validation Procedure The procedure for evaluating effectiveness is described below.

[0194] The Total Nasal Symptom Score (TNSS) ranges from 0 to 12 and is based on an assessment of four nasal symptoms, including congestion, itchiness, rhinorrhea, and sneezing, graded on a Likert scale from 0 (none) to 3 (severe). The TNSS is recorded using an electronic diary.

[0195] Total Ocular Symptom Score (TOSS): The TOSS ranges from 0 to 6 and is based on two symptoms: itching / conjunctival congestion / foreign body sensation and lacrimation / watery eyes. Each of the two symptoms is graded on a scale of 0 (none), 1 (mild), 2 (moderate), or 3 (severe). The TOSS is recorded using an electronic diary.

[0196] Total Symptom Score (TSS): The TSS is calculated by summing the TNS and TOSS scores to arrive at a composite TSS of 0 to 18.

[0197] Daily Medication Score: Participants are required to use an electronic diary to record their daily emergency medication use, including which medications are used and the amounts of these pre-specified medications. Using this information, the DMS is calculated as follows: Desloratadine 5 mg, 6 points / dose; maximum daily score of 6 points; Olopatadine 1 mg / mL / drop, 1.5 points / drop; maximum daily score of 6 points; Mometasone phlomate 50 μg / dose, 2.0 points / spray; maximum daily score of 8 points. The maximum DMS score is 20.

[0198] The Combined Symptom Medication Score (CSMS) is calculated by summing the DMS and TSS scores, with the score ranging from 0 to 38.

[0199] The Asthma Control Questionnaire (ACQ) measures the adequacy of asthma control and changes in asthma control that occur spontaneously or as a result of treatment. The ACQ-5 consists of five patient-reported items that clinicians consider most important for assessing control: (1) nighttime awakenings due to symptoms, (2) morning symptoms, (3) limitation of daily activities, (4) shortness of breath, and (5) wheezing. The total score ranges from 0 to 6, with higher scores indicating less asthma control. A score of 1.5 or higher is considered poorly controlled asthma. The ACQ-5 is recorded using an electronic diary.

[0200] The Standardized Quality of Life Questionnaire for Rhinitis and Conjunctivitis (RQLQ(S)) consists of 28 questions across seven domains: activity limitations, sleep problems, nasal symptoms, ocular symptoms, non-nasal / ocular symptoms, practical problems, and emotional functioning. The activity domain includes three subject-specific questions that ask subjects to select the three activities most limited by rhinitis and conjunctivitis. Subjects recall how much they suffered from rhinitis and conjunctivitis in the previous week and answer each question on a 7-point scale (0 [no impairment] to 6 [severe impairment]). The overall RQLQ(S) score is the average of all 28 responses, and individual domain scores are the average of the items within those domains. The RQLQ(S) is recorded using an electronic journal.

[0201] Pollen and Food Allergy Questionnaire (PFASQ): The PFASQ is to be administered at the designated time to determine the types of foods that cause allergic reactions, the type of reaction, and how quickly the reaction occurs.

[0202] Patient's Overall Severity Impression (PGI-S): The PGI-S assesses the severity of seasonal allergy symptoms over the past week. Symptom severity ranges from 0 (no symptoms), 1 (mild), 2 (moderate), to 3 (severe). [Table 7-1] [Table 7-2] [Table 7-3]

[0203] The present invention is not limited to the scope of the specific embodiments described herein. In fact, various modifications of the present invention will be apparent to those skilled in the art from the above description and accompanying drawings, in addition to those described herein. Such modifications are intended to fall within the scope of the appended claims. All patent and non-patent literature disclosures referenced herein are expressly incorporated in their entirety by reference.

Claims

1. A pharmaceutical composition for treating birch allergy in a subject, wherein the pharmaceutical composition is (a) A first anti-Bet v 1 antibody, wherein the first anti-Bet v 1 antibody comprises a heavy chain complementarity determining region (HCDR) 1 containing the amino acid sequence of SEQ ID NO: 2, an HCDR 2 containing the amino acid sequence of SEQ ID NO: 3, an HCDR 3 containing the amino acid sequence of SEQ ID NO: 4, a light chain complementarity determining region (LCDR) 1 containing the amino acid sequence of SEQ ID NO: 6, an LCDR 2 containing the amino acid sequence of SEQ ID NO: 7, and an LCDR 3 containing the amino acid sequence of SEQ ID NO:

8. (b) A second anti-Bet v 1 antibody, wherein the second anti-Bet v 1 antibody comprises HCDR1 containing the amino acid sequence of SEQ ID NO: 12, HCDR2 containing the amino acid sequence of SEQ ID NO: 13, HCDR3 containing the amino acid sequence of SEQ ID NO: 14, LCDR1 containing the amino acid sequence of SEQ ID NO: 16, LCDR2 containing the amino acid sequence of SEQ ID NO: 17, and LCDR3 containing the amino acid sequence of SEQ ID NO: 18, and (c) A third anti-Bet v 1 antibody, wherein the third anti-Bet v 1 antibody comprises HCDR1 containing the amino acid sequence of SEQ ID NO: 22, HCDR2 containing the amino acid sequence of SEQ ID NO: 23, HCDR3 containing the amino acid sequence of SEQ ID NO: 24, LCDR1 containing the amino acid sequence of SEQ ID NO: 26, LCDR2 containing the amino acid sequence of SEQ ID NO: 27, and LCDR3 containing the amino acid sequence of SEQ ID NO:

28. Includes, The pharmaceutical composition comprises, in a dose of approximately 300 mg per antibody, each of the first anti-Bet v 1 antibody, the second anti-Bet v 1 antibody, and the third anti-Bet v 1 antibody. A pharmaceutical composition wherein a single dose of the pharmaceutical composition is administered subcutaneously or intravenously before the start of the birch pollen season.

2. The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition is administered subcutaneously.

3. The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition is administered intravenously.

4. The first anti-Bet v 1 antibody described above, (a) a heavy chain variable region (HCVR) containing the amino acid sequence of SEQ ID NO: 1 and a light chain variable region (LCVR) containing the amino acid sequence of SEQ ID NO: 5, and / or (b) Heavy chain containing the amino acid sequence of SEQ ID NO: 9 and light chain containing the amino acid sequence of SEQ ID NO: 10 A pharmaceutical composition according to any one of claims 1 to 3, comprising:

5. The second anti-Bet v 1 antibody described above, (a) HCVR containing the amino acid sequence of SEQ ID NO: 11 and LCVR containing the amino acid sequence of SEQ ID NO: 15, and / or (b) Heavy chain containing the amino acid sequence of SEQ ID NO: 19 and light chain containing the amino acid sequence of SEQ ID NO: 20 A pharmaceutical composition according to any one of claims 1 to 3, comprising:

6. The third anti-Bet v 1 antibody described above, (a) HCVR containing the amino acid sequence of SEQ ID NO: 21 and LCVR containing the amino acid sequence of SEQ ID NO: 25, and / or (b) Heavy chain containing the amino acid sequence of SEQ ID NO: 29 and light chain containing the amino acid sequence of SEQ ID NO: 30 A pharmaceutical composition according to any one of claims 1 to 3, comprising:

7. (a) The first anti-Bet v 1 antibody comprises an HCVR containing the amino acid sequence of SEQ ID NO: 1 and an LCVR containing the amino acid sequence of SEQ ID NO: 5 (b) The second anti-Bet v 1 antibody comprises an HCVR containing the amino acid sequence of SEQ ID NO: 11 and an LCVR containing the amino acid sequence of SEQ ID NO: 15, (c) The third anti-Bet v 1 antibody comprises an HCVR containing the amino acid sequence of SEQ ID NO: 21 and an LCVR containing the amino acid sequence of SEQ ID NO:

25. A pharmaceutical composition according to any one of claims 1 to 3.

8. (a) The first anti-Bet v 1 antibody comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 9 and a light chain containing the amino acid sequence of SEQ ID NO: 10, (b) The second anti-Bet v 1 antibody comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 19 and a light chain containing the amino acid sequence of SEQ ID NO: 20, (c) The third anti-Bet v 1 antibody comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 29 and a light chain containing the amino acid sequence of SEQ ID NO:

30. A pharmaceutical composition according to any one of claims 1 to 3.

9. Treatment using the aforementioned pharmaceutical composition To reduce the total nasal symptom score (TNSS) of the target group, To reduce the Total Ocular Symptom Score (TOSS) of the target group, To reduce the target's Total Symptom Score (TSS), To reduce the target's daily medication score (DMS), To reduce the target complex symptom medication score (CSMS), To reduce the mean wheal diameter of the target birch skin prick test (SPT), and / or A pharmaceutical composition according to any one of claims 1 to 8, wherein emergency medicine is not used, the TSS of the subject is 2 or less out of 18, and the number of good days of the subject is increased.

10. The pharmaceutical composition according to any one of claims 1 to 9, wherein treatment using the pharmaceutical composition reduces allergic rhinitis symptoms in the subject.

11. The pharmaceutical composition according to any one of claims 1 to 10, wherein treatment using the pharmaceutical composition reduces allergic conjunctivitis symptoms in the subject.

12. The pharmaceutical composition according to any one of claims 1 to 11, wherein the subject to be treated has a baseline serum allergen-specific IgE level of 0.35 kUa / L or higher in relation to birch pollen and / or Bet v 1 allergen.

13. The pharmaceutical composition according to any one of claims 1 to 12, wherein the subject to be treated exhibits baseline positive SPT using birch allergen extract.

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