Treatment of peanut allergy with tolerant nanoparticles

Abstract

The present disclosure generally relates to methods for treating peanut allergy using nanoparticles encapsulating peanut antigens to induce antigen-specific tolerance.

Description

Cross - Reference to Related Applications

[0001] This application claims the priority benefit of U.S. Provisional Patent Application No. 63 / 380,173, filed on October 19, 2022, and U.S. Provisional Patent Application No. 63 / 486,812, filed on February 24, 2023, which are hereby incorporated by reference in their entirety. Technical Field

[0002] The present disclosure relates to methods of treating peanut allergy using tolerogenic immunomodulatory nanoparticles encapsulating a purified peanut extract containing allergenic peanut proteins or antigenic fragments thereof. Description of Text Files Submitted Electronically

[0003] The content of the text file submitted electronically with this application is hereby incorporated by reference in its entirety: computer-readable form copy of the sequence listing (filename: 58512_SeqListing.XML, created on October 18, 2023, file size: 27,434 bytes). Background Art

[0004] Peanut allergy is one of the most common food allergies, affecting nearly 1.2% of the total U.S. population and 2.5% of the pediatric population, and its incidence has been on the rise in the past decade (Cannon HE. Am J ManagCare. 2018;24(19 Suppl):S428 - s433). Peanut allergy is driven by a pathologic hyperimmune response, in which exposure to peanuts can lead to mild to severe symptoms such as nausea, vomiting, rash, weakened breathing, decreased blood pressure, and even death.

[0005] Normal individuals with a healthy immune system are able to maintain non-responsiveness to antigens encountered in common foods such as peanuts; however, in peanut-allergic subjects, the loss of immune tolerance to peanut antigens prompts a pathologic hyperimmune response. To date, eight peanut proteins (Ara h1 to Ara h8) have been identified as the major antigenic peanut proteins that prompt allergic hyperimmune responses (Keet et al., Journal of Allergy and Clinical Immunology: In Practice (J Allergy Clin Immunol Pract.) 2013;1(1):101-103). Additional peanut allergens have recently been described, reporting up to 18 allergenic peanut proteins. Ozias-Akins et al., Allergy May 2019; 74(5): 888-898). The allergic immune response to peanut antigen proteins is mediated by T cell-dependent mechanisms that involve upregulation of the production of type 2 T helper (Th2) cytokines (e.g., IL-4, IL-5, IL-9, and IL-13) and B cell class switching, resulting in the production of IgE antibodies and the degranulation of mast cells and basophils (Sampath et al., Journal of Clinical Investigation (J Clin Invest.) 2019;129(4):1431-1440).

[0006] Currently, there is no cure for peanut allergy, and strict avoidance of peanut antigens and management of allergic reactions are the only options available to patients. Immunotolerance therapies that can induce T cell tolerance to peanut antigens are considered the gold standard for treating peanut allergy but have been elusive to date. Several attempts have been made to develop immunotolerance therapies using oral immunotherapy (OIT), subcutaneous immunotherapy (SCIT), epicutaneous immunotherapy (EPIT), and sublingual immunotherapy (SLIT) methods (Feuille et al., Allergy Asthma Immunol Res. 2018;10(3):189-206). The success of these therapies has been variable, and desensitization to peanut proteins has been reported to only protect against accidental exposure but not cure (Chinthrajah et al., Lancet. 2019;394(10207):1437-1449; Vickery et al., New England Journal of Medicine (N Engl J Med.) 2018;379(21):1991-2001; Fleischer et al., Journal of the American Medical Association (J. Am Med Assoc.) 2019;321(10):946-955). Summary of the Invention

[0007] Tolerogenic immunomodulatory particles (TIMPs) containing one or more antigens have previously been described for the treatment of immune-mediated disorders (e.g., autoimmune diseases and allergies) by inducing antigen-specific immune tolerance (WO20131319253 and WO 2015023796, incorporated herein by reference). In several preclinical models of autoimmune diseases and allergies, TIMPs have been shown to be efficacious in inducing T cell tolerance. The use of TIMPs encapsulating purified peanut extract (PPE) and / or containing peanut extract or one or more peanut proteins or antigenic fragments thereof (including Ara h1, Ara h2, Ara h3, Ara h5, Ara h6, Ara h7, Ara h8, Ara h9, Ara h10, Ara h11, Ara h12, Ara h13, Arah14, Ara h15, Ara h16, Ara h17, and Ara h18) to induce antigen-specific T cell tolerance to peanut antigens may improve or may cure peanut allergy (PA).

[0008] The present disclosure provides a method of treating a subject having a peanut allergy, the method comprising administering TIMP-PPE to the subject, wherein the TIMP-PPE is administered at a dose of from 0.001 mg / kg to 12 mg / kg. The present disclosure also provides a method of reducing an allergic immune response to peanut antigen in a subject having PA, the method comprising administering TIMP-PPE to the subject, wherein the TIMP-PPE is administered at a dose of from about 0.001 mg / kg to 12 mg / kg. In various embodiments, the TIMP-PPE is administered at a dose of from about 0.001 mg / kg to 10 mg / kg, from about 0.005 mg / kg to 12 mg / kg, from about 0.01 mg / kg to 12 mg / kg, from about 0.05 mg / kg to 12 mg / kg, from about 0.1 mg / kg to 12 mg / kg, from about 0.5 mg / kg to 10 mg / kg, from about 1 mg / kg to 8 mg / kg, from about 1.5 mg / kg to 10 mg / kg, from about 2 mg / kg to 12 mg / kg, from about 2 mg / kg to 10 mg / kg, from about 3 mg / kg to 10 mg / kg, from about 4 mg / kg to 10 mg / kg, from about 4 mg / kg to 12 mg / kg, or from about 5 mg / kg to 12 mg / kg. In various embodiments, the TIMP-PPE is administered at a dose of about 0.001 mg / kg, 0.0025 mg / kg, 0.005 mg / kg, 0.01 mg / kg, 0.025 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.25 mg / kg, 0.5 mg / kg, 1.0 mg / kg, 2.0 mg / kg, 3.0 mg / kg, 4.0 mg / kg, 5 mg / kg, 6 mg / kg, 8.0 mg / kg, 10 mg / kg, or 12 mg / kg. In various embodiments, the TIMP-PPE is administered at a dose of about 0.1 mg, 0.25 mg, 0.5 mg, 1 mg, 2 mg, 2.5 mg, 5 mg, 10 mg, 25 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 325 mg, 350 mg, 400 mg, 425 mg, 450 mg, 475 mg, 500 mg, 525 mg, 550 mg, 575 mg, 600 mg, 625 mg, 650 mg, 675 mg, 700 mg, 725 mg, 750 mg, 775 mg, or 800 mg.

[0009] In various embodiments, TIMP-PPE is administered at a concentration between about 0.0005 mg / mL and about 50 mg / mL. In various embodiments, TIMP-PPE is administered at a concentration of about 0.0005 mg / mL, 0.001 mg / mL, 0.005 mg / mL, 0.01 mg / mL, 0.05 mg / mL, 0.1 mg / mL, 0.5 mg / mL, 1 mg / mL, 2 mg / mL, 3 mg / mL, 4 mg / mL, 5 mg / mL, 6 mg / mL, 7 mg / mL, 8 mg / mL, 9 mg / mL, 10 mg / mL, 11 mg / mL, 12.5 mg / mL, 15 mg / mL, 17.5 mg / mL, 20 mg / mL, 25 mg / mL, 30 mg / mL, 40 mg / mL or 50 mg / mL. In various embodiments, TIMP-PPE is administered by intravenous infusion for about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 10, 12, 18 or 20 hours.

[0010] In various embodiments, TIMP-PPE is administered in a single dose or multiple doses. In various embodiments, TIMP-PPE is administered in two doses spaced one week apart. In various embodiments, TIMP-PPE is administered once a week, once every two weeks, once every three weeks, once every 4 weeks, once every two months, once every three months, once every 6 months or once a year.

[0011] In various embodiments, a booster dose of TIMP-PPE is administered, i.e., TIMP-PPE is re-administered in a single dose or multiple doses after the initial or first administration of TIMP-PPE. In various embodiments, the booster dose of TIMP-PPE is administered once a week, once every two weeks, once every three weeks, once every 4 weeks, once every two months, once every three months, once every 6 months or once a year. In various embodiments, TIMP-PPE is administered in two doses spaced one week apart and then the booster dose of TIMP-PPE is re-administered in a single dose every three months.

[0012] In various embodiments, the booster dose of TIMP-PPE is re-administered at a dose of from 0.001 mg / kg to 12 mg / kg. In various embodiments, the booster dose of TIMP-PPE is re-administered at a dose of about 0.001 mg / kg to 10 mg / kg, about 0.005 mg / kg to 12 mg / kg, about 0.01 mg / kg to 12 mg / kg, about 0.05 mg / kg to 12 mg / kg, about 0.1 mg / kg to 12 mg / kg, about 0.5 mg / kg to 10 mg / kg, about 1 mg / kg to 8 mg / kg, about 1.5 mg / kg to 10 mg / kg, about 2 mg / kg to 12 mg / kg, about 2 mg / kg to 10 mg / kg, about 3 mg / kg to 10 mg / kg, about 4 mg / kg to 10 mg / kg, about 4 mg / kg to 12 mg / kg, or about 5 mg / kg to 12 mg / kg, or at a dose of 0.001 mg / kg, 0.0025 mg / kg, 0.005 mg / kg, 0.01 mg / kg, 0.025 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.25 mg / kg, 0.5 mg / kg, 1.0 mg / kg, 2.0 mg / kg, 4.0 mg / kg, 6 mg / kg, 8.0 mg / kg, 10 mg / kg, or 12 mg / kg. In various embodiments, the booster dose of TIMP-PPE is re-administered at a dose of about 0.1 mg, 0.25 mg, 0.5 mg, 1 mg, 2 mg, 2.5 mg, 5 mg, 10 mg, 25 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 325 mg, 350 mg, 400 mg, 425 mg, 450 mg, 475 mg, 500 mg, 525 mg, 550 mg, 575 mg, 600 mg, 625 mg, 650 mg, 675 mg, 700 mg, 725 mg, 750 mg, 775 mg, or 800 mg. In various embodiments, the booster dose of TIMP-PPE is administered at a concentration between about 0.0005 mg / mL and about 50 mg / mL.In various embodiments, the booster dose of TIMP-PPE is re-administered at a concentration of about 0.0005 mg / mL, 0.001 mg / mL, 0.005 mg / mL, 0.01 mg / mL, 0.05 mg / mL, 0.1 mg / mL, 0.5 mg / mL, 1 mg / mL, 2 mg / mL, 3 mg / mL, 4 mg / mL, 5 mg / mL, 6 mg / mL, 7 mg / mL, 8 mg / mL, 9 mg / mL, 10 mg / mL, 11 mg / mL, 12.5 mg / mL, 15 mg / mL, 17.5 mg / mL, 20 mg / mL, 25 mg / mL, 30 mg / mL, 40 mg / mL or 50 mg / mL.

[0013] In various embodiments, the booster dose of TIMP-PPE is administered by intravenous infusion for about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 10, 12, 18 or 20 hours.

[0014] In various embodiments, TIMP-PPE encapsulates peanut proteins Ara h1, Ara h2, Ara h3, Ara h5, Arah6, Ara h7, Ara h8, Ara h9, Ara h10, Ara h11, Ara h12, Ara h13, Ara h14, Ara h15, Arah16, Ara h17 or Ara h18 and / or antigenic fragments of peanut proteins. In various embodiments, TIMP-PPE encapsulates peanut proteins Ara h1, Ara h2, Ara h3, Ara h5, Ara h6, Ara h7, Ara h8, Ara h9, Ara h10, Ara h11, Arah12, Ara h13, Ara h14, Ara h15, Ara h16, Ara h17, Ara h18 or combinations thereof and / or antigenic fragments of peanut proteins. In various embodiments, TIMP-PPE contains peanut extract or one or more peanut proteins or antigenic fragments thereof selected from the group consisting of: Ara h1, Ara h2, Ara h3, Ara h5, Ara h6, Ara h7, Ara h8, Ara h9, Ara h10, Ara h11, Ara h12, Ara h13, Ara h14, Ara h15, Ara h16, Ara h17 and Ara h18.

[0015] In various embodiments, the TIMP-PPE consists of poly(lactic-co-glycolic acid) (PLGA) particles encapsulating one or more peanut antigens and a suitable buffer or excipient. In various embodiments, the TIMP-PPE particles are surface-functionalized. In various embodiments, the TIMP-PPE particles are surface-functionalized by carboxylation. In various embodiments, the TIMP-PPE particles have a negative ζ potential. In various embodiments, the negative ζ potential of the TIMP-PPE particles is between about -100 mV and about 0 mV. In various embodiments, the ζ potential of the particles is from about -100 mV to about -25 mV, about -100 mV to about -30 mV, about -80 mV to about -30 mV, about -75 mV to about -30 mV, about -70 mV to about -30 mV, about -75 mV to about -35 mV, about -70 mV to about -25 mV, about -60 mV to about -30 mV, about -60 mV to about -35 mV, or about -50 mV to about -30 mV. In various embodiments, the ζ potential is about -25 mV, -30 mV, -35 mV, -40 mV, -45 mV, -50 mV, -55 mV, -60 mV, -65 mV, -70 mV, -75 mV, -80 mV, -85 mV, -90 mV, -95 mV, or -100 mV.

[0016] In various embodiments, the size or diameter of the TIMP-PPE particles ranges from 0.05 µm to about 10 µm. In various embodiments, the diameter of the TIMP-PPE particles ranges from 0.1 µm to about 10 µm. In various embodiments, the diameter of the TIMP-PPE particles ranges from 0.1 µm to about 5 µm. In various embodiments, the diameter of the TIMP-PPE particles ranges from 0.1 µm to about 3 µm. In various embodiments, the diameter of the TIMP-PPE particles ranges from 0.3 µm to about 5 µm. In various embodiments, the diameter of the TIMP-PPE particles ranges from about 0.3 µm to about 3 µm. In various embodiments, the diameter of the TIMP-PPE particles ranges from about 0.3 µm to about 1 µm. In various embodiments, the diameter of the TIMP-PPE particles ranges from about 0.4 µm to about 1 µm. In various embodiments, the diameter of the TIMP-PPE particles ranges from about 100 nm to 10000 nm, from about 100 nm to 5000 nm, from about 100 nm to 3000 nm, from about 100 nm to 2000 nm, from about 300 nm to 5000 nm, from about 300 nm to 3000 nm, from about 300 nm to 1000 nm, from about 300 nm to 800 nm, from about 400 to 800 nm, or from about 200 nm to 700 nm. In various embodiments, the diameter of the TIMP-PPE particles is about 50 nm, 100 nm, 200 nm, 300 nm, 400 nm, 500 nm, 600 nm, 700 nm, 800 nm, 900 nm, 1000 nm, 1100 nm, 1200 nm, 1300 nm, 1400 nm, 1500 nm, or 2000 nm. In various embodiments, the diameter of the negatively charged particles ranges from 400 nm to 800 nm.

[0017] In various embodiments, TIMP-PPE is administered intravenously, subcutaneously, intramuscularly, intraperitoneally, intranasally, or orally.

[0018] In various embodiments, TIMP-PPE is administered alone or in combination with one or more additional therapeutic agents. In various embodiments, the present disclosure provides a method of treating peanut allergy or peanut allergy-related symptoms in a subject in need thereof, the method comprising administering to the subject a composition comprising TIMP-PPE alone or in combination with a therapeutic agent useful for treating peanut allergy.

[0019] In various embodiments, a therapeutic agent useful for treating peanut allergy induces regulatory T cells (Tregs). In various embodiments, a therapeutic agent useful for treating peanut allergy increases the frequency and / or number of Tregs. In various embodiments, a therapeutic agent useful for treating peanut allergy is an IL-2 therapy that induces Tregs. In various embodiments, the IL-2 therapy is low-dose IL-2, an IL-2 mutant protein engineered to expand Tregs, an IL-2 variant engineered to expand Tregs, an IL-2 molecule engineered to be selective for the high-affinity IL-2 receptor, PEGylated IL-2, an IL-2 complex, or an IL-2 / CD25 fusion protein. In various embodiments, additional therapeutic agents are IgE inhibitors, basophil activation inhibitors, mast cell activation inhibitors, antihistamines, or small molecule or biologic therapeutic agents. In various embodiments, additional therapeutic agents are IgE inhibitors, competitors of IgE for allergen binding sites, basophil activation inhibitors, mast cell activation inhibitors, antihistamines, cytokine inhibitors, microbiome therapies, small molecule or biologic therapeutic agents. In various embodiments, an additional therapeutic agent inhibits IgE. In various embodiments, an additional therapeutic agent inhibits IgE antibodies. In various embodiments, an additional therapeutic agent inhibits basophil activation. In various embodiments, an additional therapeutic agent inhibits mast cell activation. In various embodiments, an additional therapeutic agent is a biologic agent or a small molecule. In various embodiments, an additional therapeutic agent is an anti-IgE antibody, an anti-IL-4Rα antibody, an anti-IL13 antibody, an anti-IL-33 antibody, an antihistamine, a steroid, a corticosteroid, a leukotriene modifier, or a non-steroidal anti-inflammatory drug (NSAID).

[0020] In various embodiments, a therapeutic agent useful for treating peanut allergy is an IgE inhibitor, a competitor of IgE for allergen binding sites, a basophil activation inhibitor, a mast cell activation inhibitor, an antihistamine, a cytokine inhibitor, a microbiome therapy, a small molecule or a biologic therapeutic agent. In various embodiments, a therapeutic agent useful for treating peanut allergy inhibits IgE. In various embodiments, a therapeutic agent useful for treating peanut allergy inhibits IgE antibodies. In various embodiments, a therapeutic agent useful for treating peanut allergy inhibits basophil activation. In various embodiments, a therapeutic agent useful for treating peanut allergy inhibits mast cell activation. In various embodiments, a therapeutic agent useful for treating peanut allergy is a biologic agent or a small molecule. In various embodiments, a therapeutic agent useful for treating peanut allergy is an anti-IgE antibody, an anti-IL-4Rα antibody, an anti-IL13 antibody, an anti-IL-33 antibody, an antihistamine, a steroid, a corticosteroid, a leukotriene modifier, or a non-steroidal anti-inflammatory drug (NSAID).

[0021] In various embodiments, the additional therapeutic agent is an antihistamine. In various embodiments, the therapeutic agent useful for treating peanut allergy is an antihistamine. In various embodiments, the antihistamine is a first-generation antihistamine. In various embodiments, the antihistamine is a second-generation antihistamine. In various embodiments, the antihistamine is selected from the group consisting of brompheniramine, carbinoxamine maleate, chlorpheniramine, clemastine, diphenhydramine, hydroxyzine, triprolidine, azelastine, cetirizine, desloratadine, fexofenadine, levocetrizine, doxylamine, ebastine, embramine, epinephrine, fexofenadine, loratadine, and olopatadine.

[0022] In various embodiments, the therapeutic agent administered in combination with TIMP-PPE is an anti-IgE antibody, an anti-IL-4Rα antibody, an anti-IL13 antibody, an anti-IL-33 antibody, an antihistamine, a steroid, a corticosteroid, a leukotriene modifier, low-dose IL-2, an IL-2 mutant protein engineered to expand Tregs, an IL-2 variant engineered to expand Tregs, an IL-2 molecule engineered to be selective for the high-affinity IL-2 receptor, PEGylated IL-2, an IL-2 complex, an IL-2 / CD25 fusion protein, a prebiotic, a probiotic, a histone deacetylase inhibitor, short-chain fatty acids (e.g., acetate, butyrate, propionate, butyrate polymers), an IgE inhibitor, a competitor of IgE for the allergen-binding site, a basophil activation inhibitor, a mast cell activation inhibitor, a cytokine inhibitor, a microbiome therapy, a small molecule or a biotherapeutic agent or a non-steroidal anti-inflammatory drug (NSAID).

[0023] In various embodiments, the additional therapeutic agent is a steroid. In various embodiments, the therapeutic agent useful for treating peanut allergy is a steroid. In various embodiments, the steroid is selected from the group consisting of beclomethasone, ciclesonide, fluticasone furoate, mometasone, budenoside, fluticasone, triamcinolone, and loteprednol.

[0024] In various embodiments, the additional therapeutic agent is a corticosteroid. In various embodiments, the therapeutic agent useful for treating peanut allergy is a corticosteroid. In various embodiments, the corticosteroid is selected from the group consisting of cortisone, prednisone, prednisolone, methylprednisolone, dexamethasone, betamethasone, and hydrocortisone.

[0025] In various embodiments, the additional therapeutic agent is a non-steroidal anti-inflammatory drug (NSAID). In various embodiments, the therapeutic agent useful for treating peanut allergy is a non-steroidal anti-inflammatory drug (NSAID). In various embodiments, the NSAID is a non-selective NSAID. In various embodiments, the NSAID is a COX-2 selective NSAID. In various embodiments, the NSAID is a COX-1 selective NSAID. In various embodiments, the NSAID is a prostaglandin synthase inhibitor. In various embodiments, the NSAID is selected from the group consisting of: diclofenac, diclofenac potassium, diclofenac sodium, diflunisal, etodolac, flurbiprofen, fenoprofen, fenoprofen calcium, ketorolac, ketorolac tromethamine, ketoprofen, tolmetin, tolmetin sodium, acetylsalicylic acid, aspirin, ibuprofen, naproxen, indomethacin, indomethacin sodium, sulindac, felbinac, piroxicam, mefenamic acid, meclofenamate sodium, meloxicam, nabumetone, oxaprozin, piroxicam, celecoxib, etodolac, etoricoxib, lumiracoxib, rofecoxib, and valdecoxib.

[0026] In various embodiments, the additional therapeutic agent is a leukotriene modifier. In various embodiments, the therapeutic agent useful for treating peanut allergy is a leukotriene modifier. In various embodiments, the leukotriene modifier is an anti-leukotriene. In various embodiments, the leukotriene modifier is a leukotriene receptor antagonist. In various embodiments, the leukotriene modifier is a leukotriene synthesis inhibitor. In various embodiments, the leukotriene modifier is selected from the group consisting of montelukast, zileuton, and zafirlukast.

[0027] In various embodiments, the biologic is an antibody. In various embodiments, the antibody is an anti-IgE antibody, an anti-IL-4Rα antibody, an anti-IL-13 antibody, or an anti-IL-33 antibody. In various embodiments, the anti-IgE antibody is omalizumab (XOLAIR®). In various embodiments, the anti-IL-4Rα antibody is dupilumab (DUPIXENT®). In various embodiments, the anti-IL-33 antibody is etokinumab. In various embodiments, the additional therapeutic agent is administered before, during, or after the administration of TIMP-PPE. In various embodiments, the additional therapeutic agent is administered intravenously, subcutaneously, intramuscularly, intraperitoneally, intranasally, or orally.

[0028] In various embodiments, the therapeutic agent useful for treating peanut allergy is administered before, during, or after the administration of TIMP-PPE. In various embodiments, the therapeutic agent useful for treating peanut allergy is administered intravenously, subcutaneously, intramuscularly, intraperitoneally, intranasally, or orally.

[0029] In various embodiments, the therapeutic agent is administered before, concomitantly with, or immediately following the administration of TIMP-PPE. In various embodiments, the therapeutic agent is administered 1, 2, 3, 4, 5, 6, or 7 days before the administration of TIMP-PPE. In various embodiments, the therapeutic agent is administered 1, 2, 3, or 4 weeks before the administration of TIMP-PPE. In various embodiments, the therapeutic agent is administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months before the administration of TIMP-PPE. In various embodiments, the therapeutic agent is administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 years before the administration of TIMP-PPE. In various embodiments, the therapeutic agent is administered 1, 2, 3, 4, 5, 6, or 7 days immediately following the administration of TIMP-PPE. In various embodiments, the therapeutic agent is administered 1, 2, 3, or 4 weeks immediately following the administration of TIMP-PPE. In various embodiments, the therapeutic agent is administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months immediately following the administration of TIMP-PPE. In various embodiments, the therapeutic agent is administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 years immediately following the administration of TIMP-PPE.

[0030] The present disclosure describes a method of treating peanut allergy in a subject, the method comprising administering to the subject a combination of TIMP-PPE and an anti-IgE antibody, wherein TIMP-PPE is administered at a dose of about 0.001 mg / kg to 12 mg / kg, and wherein the anti-IgE antibody is administered at a dose of about 10 mg to about 500 mg. In various embodiments, the anti-IgE antibody is omalizumab (XOLAIR®). In various embodiments, TIMP-PPE is administered at a dose of about 0.001 mg / kg to 10 mg / kg, about 0.005 mg / kg to 12 mg / kg, about 0.01 mg / kg to 12 mg / kg, about 0.05 mg / kg to 12 mg / kg, about 0.1 mg / kg to 12 mg / kg, about 0.5 mg / kg to 10 mg / kg, about 1 mg / kg to 8 mg / kg, about 1.5 mg / kg to 10 mg / kg, about 2 mg / kg to 12 mg / kg, about 2 mg / kg to 10 mg / kg, about 3 mg / kg to 10 mg / kg, about 4 mg / kg to 10 mg / kg, about 4 mg / kg to 12 mg / kg or about 5 mg / kg to 12 mg / kg or at a dose of about 0.001 mg / kg, 0.0025 mg / kg, 0.005 mg / kg, 0.01 mg / kg, 0.025 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.25, 0.5 mg / kg, 1.0 mg / kg, 2.0 mg / kg, 4.0 mg / kg, 6 mg / kg, 8.0 mg / kg, 10 mg / kg or 12 mg / kg. In various embodiments, TIMP-PPE is administered at a dose of about 0.1 mg, 0.25 mg, 0.5 mg, 1 mg, 2 mg, 2.5 mg, 5 mg, 10 mg, 25 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 325 mg, 350 mg, 400 mg, 425 mg, 450 mg, 475 mg, 500 mg, 525 mg, 550 mg, 575 mg, 600 mg, 625 mg, 650 mg, 675 mg, 700 mg, 725 mg, 750 mg, 775 mg or 800 mg.In various embodiments, the anti-IgE antibody administered in combination with TIMP-PPE is administered at a dose of about 10 mg, 25 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 375 mg, 400 mg, 425 mg, 450 mg, 475 mg or 500 mg. In various embodiments, the dose level of the anti-IgE antibody is determined based on the serum IgE level. In various embodiments, the serum IgE level is 30 - 100 IU / mL, 100 - 200 IU / mL, 200 - 300 IU / mL, 300 - 400 IU / mL, 400 - 500 IU / mL, 500 - 600 IU / mL, 600 - 700 IU / mL, 700 - 800 IU / mL, 800 - 900 IU / mL, 900 - 1000 IU / mL, 1000 - 1100 IU / mL, 1100 - 1200 IU / mL, 1200 - 1300 IU / mL, 1300 - 1400 IU / mL or 1400 - 1500 IU / mL. In various embodiments, the dose level of the anti-IgE antibody is determined based on the weight of the subject. In various embodiments, the weight of the subject is 30 - 40 kg, 40 - 50 kg, 50 - 60 kg, 60 - 70 kg, 70 - 80 kg, 80 - 90 kg, 90 - 125 kg or 125 - 150 kg.

[0031] In various embodiments, the anti-IgE antibody is administered in a single dose or multiple doses. In various embodiments, the anti-IgE antibody is administered once a week, once every two weeks, once every three weeks or once every four weeks. In various embodiments, the anti-IgE antibody is administered before, concomitantly with or after the administration of TIMP-PPE. In various embodiments, the anti-IgE antibody is administered one week, two weeks, three weeks, four weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months or 12 months before the administration of TIMP-PPE. In various embodiments, the anti-IgE antibody is administered one week, two weeks, three weeks or four weeks after the administration of TIMP-PPE.

[0032] The present disclosure provides methods for treating peanut allergy, the methods comprising administering to a subject a combination of TIMP-PPE and an anti-IgE antibody, wherein the anti-IgE antibody is administered to the subject once a week for two weeks or once a week for four weeks prior to administering TIMP-PPE, wherein TIMP-PPE is administered in two dose intervals one week apart at a dose level between 0.001 mg / kg and 12 mg / kg, and wherein the anti-IgE antibody is administered at a dose level between about 50 mg and 500 mg.

[0033] The present disclosure provides methods for treating peanut allergy, the methods comprising administering to a subject a combination of TIMP-PPE and an anti-IgE antibody, wherein the anti-IgE antibody is administered to the subject once a week for two weeks or once a week for four weeks prior to administering TIMP-PPE, wherein TIMP-PPE is administered in two dose intervals one week apart at a dose level between 0.1 mg and 800 mg, and wherein the anti-IgE antibody is administered at a dose level between about 50 mg and 500 mg.

[0034] The present disclosure describes a method of treating peanut allergy in a subject, the method comprising administering to the subject a combination of TIMP-PPE and an anti-IL-4Rα antibody, wherein TIMP-PPE is administered at a dose of about 0.001 mg / kg to 12 mg / kg, and wherein the anti-IL-4Rα antibody is administered at a dose of about 10 mg to about 500 mg. In various embodiments, the anti-IL-4Rα antibody is dupilumab (DUPIXENT®). In various embodiments, TIMP-PPE is administered at a dose of about 0.001 mg / kg to 10 mg / kg, about 0.005 mg / kg to 12 mg / kg, about 0.01 mg / kg to 12 mg / kg, about 0.05 mg / kg to 12 mg / kg, about 0.1 mg / kg to 12 mg / kg, 0.5 mg / kg to 10 mg / kg, about 1 mg / kg to 8 mg / kg, about 1.5 mg / kg to 10 mg / kg, about 2 mg / kg to 12 mg / kg, about 2 mg / kg to 10 mg / kg, about 3 mg / kg to 10 mg / kg, about 4 mg / kg to 10 mg / kg, about 4 mg / kg to 12 mg / kg or about 5 mg / kg to 12 mg / kg or at a dose of about 0.001 mg / kg, 0.0025 mg / kg, 0.005 mg / kg, 0.01 mg / kg, 0.025 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.25, 0.5 mg / kg, 1.0 mg / kg, 2.0 mg / kg, 4.0 mg / kg, 6 mg / kg, 8.0 mg / kg, 10 mg / kg or 12 mg / kg. In various embodiments, TIMP-PPE is administered at a dose of about 0.1 mg, 0.25 mg, 0.5 mg, 1 mg, 2 mg, 2.5 mg, 5 mg, 10 mg, 25 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 325 mg, 350 mg, 400 mg, 425 mg, 450 mg, 475 mg, 500 mg, 525 mg, 550 mg, 575 mg, 600 mg, 625 mg, 650 mg, 675 mg, 700 mg, 725 mg, 750 mg, 775 mg or 800 mg.In various embodiments, the anti-IL-4Rα antibody is administered at a dose of about 10 mg, 25 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 375 mg, 400 mg, 425 mg, 450 mg, 475 mg, 500 mg, 525 mg, 550 mg, 575 mg or 600 mg. In various embodiments, the dose level of the anti-IL-4Rα antibody is determined based on the serum IgE level. In various embodiments, the serum IgE level is 30 - 100 IU / mL, 100 - 200 IU / mL, 200 - 300 IU / mL, 300 - 400 IU / mL, 400 - 500 IU / mL, 500 - 600 IU / mL, 600 - 700 IU / mL, 700 - 800 IU / mL, 800 - 900 IU / mL, 900 - 1000 IU / mL, 1000 - 1100 IU / mL, 1100 - 1200 IU / mL, 1200 - 1300 IU / mL, 1300 - 1400 IU / mL or 1400 - 1500 IU / mL. In various embodiments, the dose level of the anti-IL-4Rα antibody is determined based on the weight of the subject. In various embodiments, the weight of the subject is 30 - 40 kg, 40 - 50 kg, 50 - 60 kg, 60 - 70 kg, 70 - 80 kg, 80 - 90 kg, 90 - 125 kg or 125 - 150 kg. In various embodiments, the anti-IL-4Rα antibody is administered as a single dose or multiple doses. In various embodiments, the anti-IL-4Rα antibody is administered once a week, once every two weeks, once every three weeks or once every four weeks. In various embodiments, the anti-IL-4Rα antibody is administered before, concomitantly with or after the administration of TIMP-PPE. In various embodiments, the anti-IL-4Rα antibody is administered one week, two weeks, three weeks or four weeks before the administration of TIMP-PPE. In various embodiments, the anti-IL-4Rα antibody is administered one week, two weeks, three weeks or four weeks after the administration of TIMP-PPE. In various embodiments, the anti-IL-4Rα antibody is administered in two doses at an initial dose level between 400 mg and 600 mg, followed by a maintenance dose level between 200 mg and 300 mg for subsequent doses.

[0035] In various embodiments, the antihistamine administered in combination with TIMP-PPE is administered in a dose of from about 0.05 mg to 2000 mg. In various embodiments, the antihistamine is administered in a dose of about 0.05 mg, 0.1 mg, 0.5 mg, 1 mg, 2 mg, 4 mg, 5 mg, 10 mg, 25 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 375 mg, 400 mg, 425 mg, 450 mg, 475 mg, 500 mg, 1000 mg, 1500 mg or 2000 mg. In various embodiments, the antihistamine is administered in a single dose or multiple doses. In various embodiments, the antihistamine is administered once a day, once a week, once every two weeks, once every three weeks or once every four weeks. In various embodiments, the antihistamine is administered two, three, four, five or six times a day. In various embodiments, the antihistamine is administered before, concomitantly with or after the administration of TIMP-PPE. In various embodiments, the antihistamine is administered one, two, three, four, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months or 12 months before the administration of TIMP-PPE. In various embodiments, the antihistamine is administered one, two, three, four, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months or 12 months after the administration of TIMP-PPE. In various embodiments, the antihistamine is administered 5, 10, 15, 30, 45 or 60 minutes before the administration of TIMP-PPE. In various embodiments, the antihistamine is administered 5, 10, 15, 30, 45 or 60 minutes after the administration of TIMP-PPE. In various embodiments, the antihistamine is administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 36, 48, 72 or 96 hours before the administration of TIMP-PPE. In various embodiments, the antihistamine is administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 36, 48, 72 or 96 hours after the administration of TIMP-PPE. In various embodiments, the antihistamine is administered 1, 2, 3, 4, 5, 6 or 7 days before the administration of TIMP-PPE. In various embodiments, the antihistamine is administered 1, 2, 3, 4, 5, 6 or 7 days after the administration of TIMP-PPE.

[0036] In various embodiments, the steroid administered in combination with TIMP-PPE is administered at a dose of from about 0.05 mg to 2000 mg. In various embodiments, the steroid is administered at a dose of about 0.05 mg, 0.1 mg, 0.5 mg, 1 mg, 2 mg, 4 mg, 5 mg, 10 mg, 25 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 375 mg, 400 mg, 425 mg, 450 mg, 475 mg, 500 mg, 1000 mg, 1500 mg or 2000 mg. In various embodiments, the steroid is administered in a single dose or multiple doses. In various embodiments, the steroid is administered once a day, once a week, once every two weeks, once every three weeks or once every four weeks. In various embodiments, the steroid is administered two, three, four, five or six times a day. In various embodiments, the steroid is administered before, concomitantly with or after the administration of TIMP-PPE. In various embodiments, the steroid is administered one, two, three, four, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months before the administration of TIMP-PPE. In various embodiments, the steroid is administered one, two, three, four, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months after the administration of TIMP-PPE. In various embodiments, the steroid is administered 5, 10, 15, 30, 45 or 60 minutes before the administration of TIMP-PPE. In various embodiments, the steroid is administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 36, 48, 72 or 96 hours before the administration of TIMP-PPE. In various embodiments, the steroid is administered 1, 2, 3, 4, 5, 6 or 7 days before the administration of TIMP-PPE.

[0037] In various embodiments, the corticosteroid administered in combination with TIMP-PPE is administered in a dose of from about 0.05 mg to 2000 mg. In various embodiments, the corticosteroid is administered in a dose of about 0.05 mg, 0.1 mg, 0.5 mg, 1 mg, 2 mg, 4 mg, 5 mg, 10 mg, 25 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 375 mg, 400 mg, 425 mg, 450 mg, 475 mg, 500 mg, 1000 mg, 1500 mg, or 2000 mg. In various embodiments, the corticosteroid is administered in a single dose or multiple doses. In various embodiments, the corticosteroid is administered once a day, once a week, once every two weeks, once every three weeks, or once every four weeks. In various embodiments, the corticosteroid is administered two, three, four, five, or six times a day. In various embodiments, the corticosteroid is administered before, concomitantly with, or after administering TIMP-PPE. In various embodiments, the corticosteroid is administered one, two, three, four, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months before administering TIMP-PPE. In various embodiments, the corticosteroid is administered one, two, three, four, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months after administering TIMP-PPE. In various embodiments, the corticosteroid is administered 5, 10, 15, 30, 45, or 60 minutes before administering TIMP-PPE. In various embodiments, the corticosteroid is administered 5, 10, 15, 30, 45, or 60 minutes after administering TIMP-PPE. In various embodiments, the corticosteroid is administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 36, 48, 72, or 96 hours before administering TIMP-PPE. In various embodiments, the corticosteroid is administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 36, 48, 72, or 96 hours after administering TIMP-PPE. In various embodiments, the corticosteroid is administered 1, 2, 3, 4, 5, 6, or 7 days before administering TIMP-PPE. In various embodiments, the corticosteroid is administered 1, 2, 3, 4, 5, 6, or 7 days after administering TIMP-PPE.

[0038] In various embodiments, the NSAID administered in combination with TIMP-PPE is administered in a dose of from about 0.05 mg to 2000 mg. In various embodiments, the NSAID is administered in a dose of about 0.05 mg, 0.1 mg, 0.5 mg, 1 mg, 2 mg, 4 mg, 5 mg, 10 mg, 25 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 325 mg, 350 mg, 375 mg, 400 mg, 425 mg, 450 mg, 475 mg, 500 mg, 1000 mg, 1500 mg or 2000 mg. In various embodiments, the NSAID is administered in a single dose or multiple doses. In various embodiments, the NSAID is administered once a day, once a week, once every two weeks, once every three weeks or once every four weeks. In various embodiments, the NSAID is administered two, three, four, five or six times a day. In various embodiments, the NSAID is administered before, concomitantly with or after the administration of TIMP-PPE. In various embodiments, the NSAID is administered one, two, three, four, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months or 12 months before the administration of TIMP-PPE. In various embodiments, the NSAID is administered one, two, three, four, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months or 12 months after the administration of TIMP-PPE. In various embodiments, the NSAID is administered 5, 10, 15, 30, 45 or 60 minutes before the administration of TIMP-PPE. In various embodiments, the NSAID is administered 5, 10, 15, 30, 45 or 60 minutes after the administration of TIMP-PPE. In various embodiments, the NSAID is administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 36, 48, 72 or 96 hours before the administration of TIMP-PPE. In various embodiments, the NSAID is administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 36, 48, 72 or 96 hours after the administration of TIMP-PPE. In various embodiments, the NSAID is administered 1, 2, 3, 4, 5, 6 or 7 days before the administration of TIMP-PPE. In various embodiments, the NSAID is administered 1, 2, 3, 4, 5, 6 or 7 days after the administration of TIMP-PPE.

[0039] In various embodiments, the leukotriene regulator administered in combination with TIMP-PPE is administered in a dose of about 0.05 mg to 2000 mg. In various embodiments, the leukotriene regulator is administered in a dose of about 0.05 mg, 0.1 mg, 0.5 mg, 1 mg, 2 mg, 4 mg, 5 mg, 10 mg, 25 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 375 mg, 400 mg, 425 mg, 450 mg, 475 mg, 500 mg, 1000 mg, 1500 mg, or 2000 mg. In various embodiments, the leukotriene regulator is administered in a single dose or multiple doses. In various embodiments, the leukotriene regulator is administered once a day, once a week, once every two weeks, once every three weeks, or once every four weeks. In various embodiments, the leukotriene regulator is administered two, three, four, five, or six times a day. In various embodiments, the leukotriene regulator is administered before, concomitantly with, or after the administration of TIMP-PPE. In various embodiments, the leukotriene regulator is administered one, two, three, four, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, or 12 months before the administration of TIMP-PPE. In various embodiments, the leukotriene regulator is administered one, two, three, four, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, or 12 months after the administration of TIMP-PPE. In various embodiments, the leukotriene regulator is administered 5, 10, 15, 30, 45, or 60 minutes before the administration of TIMP-PPE. In various embodiments, the leukotriene regulator is administered 5, 10, 15, 30, 45, or 60 minutes after the administration of TIMP-PPE. In various embodiments, the leukotriene regulator is administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 36, 48, 72, or 96 hours before the administration of TIMP-PPE. In various embodiments, the leukotriene regulator is administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 36, 48, 72, or 96 hours before and after the administration of TIMP-PPE. In various embodiments, the leukotriene regulator is administered 1, 2, 3, 4, 5, 6, or 7 days before the administration of TIMP-PPE. In various embodiments, the leukotriene regulator is administered 1, 2, 3, 4, 5, 6, or 7 days after the administration of TIMP-PPE.

[0040] It is contemplated that when TIMP-PPE is administered in combination with one or more additional therapeutic agents as described herein, TIMP-PPE is administered in a single dose or multiple doses. In various embodiments, TIMP-PPE is administered in two doses one week apart.

[0041] It is contemplated that when TIMP-PPE is administered in combination with one or more therapeutic agents useful for treating peanut allergy as described herein, TIMP-PPE is administered in a single dose or multiple doses. In various embodiments, TIMP-PPE is administered in two doses one week apart. In various embodiments, TIMP-PPE is administered in two doses one week apart and then a booster dose of TIMP-PPE is administered in a single dose every three months.

[0042] In various embodiments, administering TIMP-PPE alone or in combination with one or more additional therapeutic agents to a subject in need thereof alleviates one or more symptoms of peanut allergy. In various embodiments, the symptoms of peanut allergy are selected from the group consisting of: skin reactions, hives, skin redness, skin swelling, itching, tightness in the throat, difficulty breathing, shortness of breath, and anaphylaxis.

[0043] In various embodiments, administering TIMP-PPE alone or in combination with one or more therapeutic agents useful for treating peanut allergy to a subject in need thereof alleviates one or more symptoms of peanut allergy. In various embodiments, the symptoms of peanut allergy are selected from the group consisting of: skin reactions, hives, skin redness, skin swelling, itching, tightness in the throat, difficulty breathing, shortness of breath, digestive problems such as diarrhea, stomach cramps, nausea or vomiting, drop in blood pressure, and anaphylaxis.

[0044] In various embodiments, administering TIMP-PPE alone or in combination with one or more additional therapeutic agents to a subject in need thereof shortens the duration and reduces the severity of the allergic immune response to peanut protein. In various embodiments, administering TIMP-PPE alone or in combination with one or more additional therapeutic agents to a subject in need thereof shortens the duration and reduces the severity of the allergic immune response following exposure to peanut protein. In various embodiments, the allergic immune response is a Th2 T cell response, B cell activation, basophil activation, eosinophil activation, mast cell activation, and / or IgE induction.

[0045] In various embodiments, the efficacy of TIMP-PPE in alleviating one or more symptoms of peanut allergy and / or shortening the duration and reducing the severity of the allergic immune response to peanut protein is determined by the assay of one or more biological samples of a subject. In various embodiments, the biological sample is selected from the group consisting of: whole blood, peripheral blood, peripheral blood mononuclear cells (PBMCs), serum, plasma, urine, cerebrospinal fluid (CSF), feces, tissue biopsy, and / or bone marrow biopsy. In various embodiments, the efficacy of TIMP-PPE in alleviating one or more symptoms of peanut allergy and / or shortening the duration and reducing the severity of the allergic immune response to peanut protein is determined by double-blind placebo-controlled food challenge (DBPCFC). In various embodiments, the efficacy of TIMP-PPE in alleviating one or more symptoms of peanut allergy and / or shortening the duration and reducing the severity of the allergic immune response to peanut protein is determined by skin prick test (SPT).

[0046] In various embodiments, administering TIMP-PPE alone or in combination with another therapeutic agent to a subject reduces the proportion of Th2a+ T cells present in the total T cell population in peripheral blood.

[0047] In various embodiments, administering TIMP-PPE alone or in combination with a therapeutic agent to a subject reduces the proportion of Th2a+ T cells present in the total T cell population in peripheral blood.

[0048] In various embodiments, administering TIMP-PPE alone or in combination with another therapeutic agent to a subject reduces the ratio of activated peanut protein-specific T cells to unactivated peanut protein-specific T cells in peripheral blood.

[0049] In various embodiments, administering TIMP-PPE alone or in combination with a therapeutic agent to a subject reduces the proportion of activated peanut protein-specific T cells in peripheral blood.

[0050] In various embodiments, administering TIMP-PPE alone or in combination with another therapeutic agent to a subject increases the level of peanut protein-specific Treg cells in the blood. In various embodiments, administering TIMP-PPE alone or in combination with a therapeutic agent to a subject increases the level of peanut protein-specific Treg cells in the blood.

[0051] In various embodiments, administering TIMP-PPE alone or in combination with another therapeutic agent to a subject reduces basophil activation. In various embodiments, administering TIMP-PPE alone or in combination with a therapeutic agent to a subject reduces basophil activation.

[0052] In various embodiments, administering to a subject a TIMP-PPE alone or in combination with another therapeutic agent reduces the level of peanut protein-specific IgE in the blood. In various embodiments, administering to a subject a TIMP-PPE alone or in combination with a therapeutic agent reduces the level of peanut protein-specific IgE in the blood.

[0053] In various embodiments, administering to a subject a TIMP-PPE alone or in combination with another therapeutic agent reduces the ratio of peanut protein-specific IgE to IgG levels in the blood. In various embodiments, administering to a subject a TIMP-PPE alone or in combination with a therapeutic agent reduces the ratio of peanut protein-specific IgE to IgG levels in the blood.

[0054] In various embodiments, administering to a subject a TIMP-PPE alone or in combination with another therapeutic agent reduces the level of Th2 cytokine levels in the blood. In various embodiments, administering to a subject a TIMP-PPE alone or in combination with a therapeutic agent reduces the level of Th2 cytokine levels in the blood. In various embodiments, the Th2 cytokines are selected from the group consisting of: IL-4, IL-5, IL-9, and IL-13.

[0055] In various embodiments, administering to a subject a TIMP-PPE alone or in combination with another therapeutic agent increases tolerance to peanut protein. In various embodiments, administering to a subject a TIMP-PPE alone or in combination with a therapeutic agent increases tolerance to peanut protein.

[0056] Also contemplated are compositions comprising a TIMP-PPE for treating peanut allergy as described herein. In various embodiments, the present disclosure provides the use of a composition comprising a TIMP-PPE as described herein in the preparation of a medicament for treating peanut allergy.

[0057] It should be understood that each feature or embodiment or combination described herein is a non-limiting illustrative example of any aspect of the present invention and is thus intended to be combinable with any other feature or embodiment or combination described herein. For example, in the case of describing features in language such as "one embodiment", "some embodiments", "certain embodiments", "further embodiments", "specific exemplary embodiments" and / or "another embodiment", each of these types of embodiments is a non-limiting example of a feature intended to be combinable with any other feature or combination of features described herein, without necessarily listing every possible combination. Such features or combinations of features apply to any aspect of the present invention. In the case of disclosing examples of values within a range, any of these examples is contemplated as a possible endpoint of the range, any and all numerical values between such endpoints are contemplated, and any and all combinations of the upper and lower endpoints are envisioned.

[0058] The title of this article is for the convenience of the reader and is not restrictive. Further aspects, embodiments and variations of the present invention will become apparent in accordance with the detailed description and / or the drawings and / or the claims. Description of the Drawings

[0059] Figure 1A . Schedule of activities for subjects participating in Part A of the human study of CNP-201 particles. Figure 1B . Schedule of activities for subjects participating in Part B of the human study of CNP-201 particles. Figure 1C . Schedule of activities for subjects participating in the low starting dose study of CNP-201 particles.

[0060] Figure 2 . Summary of results for patients receiving low-dose CNP-201 particles in peanut allergy: Patients received approximately one dose of 1 mg or 25 mg of CNP-201 intravenously. The BAT threshold increased in patients receiving low-dose CNP-201, the ratio of peanut-specific IgE to IgG decreased, and a reduction in antigen-specific Tregs and / or pathogenic peanut-specific T cell subsets (Th2a, TFH, B cell plasmablasts) was induced.

[0061] Figure 3 . Basophil sensitivity test using crude peanut extract: Peanut-allergic patients receiving one dose of 25 mg CNP-201 intravenously showed an increase in the EC50 (peanut allergen concentration) required for increased expression of activation markers on the surface of basophils after stimulation with peanut allergen.

[0062] Figure 4. At a CNP-201 dose of up to 25 mg, the shift of IgE to IgG phenotype: Peanut-allergic patients receiving an intravenous dose of up to 25 mg of CNP-201 showed an increase in peanut-specific IgG and a decrease in the peanut-specific IgE / IgG ratio on days 15 and 38 after dosing compared to placebo.

[0063] Figure 5 . Reduction of immune cell subsets related to peanut allergy. Peanut-allergic patients receiving an intravenous dose of 25 mg of CNP-201 showed a reduction in the pro-allergic subsets T helper cell 2A (Th2A), T follicular helper cell (TFH), terminally differentiated effector memory cell (TEMRA), and B cell plasmablasts.

[0064] Figure 6 . After a single dose of 25 mg of CNP-201, fewer pathogenic activated peanut-specific CD4+ T cells were observed. Compared to placebo, the peanut-specific activated T cell subsets CD4+CD25+, CD4+CD69+, and CD4+PD-1+ were reduced in patients treated with low-dose CNP-201.

[0065] Figure 7 . Induction of antigen-specific regulatory T cells (Tregs): Peanut-allergic patients receiving an intravenous dose of 25 mg of CNP-201 showed an increase in peanut-specific Tregs compared to placebo.

[0066] Figure 8 . Activity schedule of subjects participating in the 1 mg dose study of CNP-201 particles. Detailed implementation

[0067] The present disclosure provides methods for monitoring the induction and maintenance of immune tolerance in a subject after receiving immunotherapy. Definitions

[0068] Unless otherwise specified, the following terms used in this application (including the specification and claims) have the definitions given below.

[0069] As used in the specification and the appended claims, unless the context clearly dictates otherwise, the indefinite articles "a / an" and the definite article "the" include plural as well as singular referents.

[0070] The term "about" or "approximately" means an acceptable error of a particular value as determined by a person of ordinary skill in the art, which depends in part on how the value is measured or determined. In certain embodiments, the term "about" or "approximately" means within 1, 2, 3, or 4 standard deviations. In certain embodiments, the term "about" or "approximately" means within 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.05% of a given value or range. Whenever the term "about" or "approximately" precedes the first of a series of two or more numerical values, it is understood that the term "about" or "approximately" applies to each numerical value in that series.

[0071] As used herein, "particle" refers to any non-tissue-derived substance composition, which can be a sphere or sphere-like entity, bead, or liposome. The terms "particle", "immunomodified particle", "carrier particle", and "bead" may be used interchangeably depending on the context. Additionally, the term "particle" is used to encompass beads and spheres.

[0072] As used herein, "negatively charged particle" refers to a particle that has been modified to have a net surface charge less than zero.

[0073] "Carboxylated particle" or "carboxylated bead" or "carboxylated sphere" includes any particle that has been modified to contain carboxyl groups on its surface. In some embodiments, the addition of carboxyl groups, for example, enhances the uptake of particles from the circulation by phagocytes / monocytes through interaction with scavenger receptors such as MARCO. Carboxylation of particles can be achieved using any compound that adds carboxyl groups.

[0074] As used herein, "TIMP-PPE" refers to a negatively charged tolerogenic immunomodified particle (TIMP) that contains peanut extract or one or more peanut proteins or antigenic fragments thereof (including Ara h1, Ara h2, Ara h3, Ara h5, Ara h6, Ara h7, Ara h8, Ara h9, Ara h10, Arah11, Ara h12, Ara h13, Ara h14, Ara h15, Ara h16, Ara h17, and Ara h18).

[0075] As used herein, the term "Th cell" or "helper T cell" refers to CD4+ cells. CD4+ T cells assist other white blood cells in immune processes, including the maturation of B cells into plasma cells and memory B cells, and the activation of cytotoxic T cells and macrophages. These cells are activated when peptide antigens are presented to T cells by MHC class II molecules expressed on the surface of antigen-presenting cells (APCs).

[0076] As used herein, the term "Th1 cell" refers to a subset of Th cells that produce pro-inflammatory mediators. Th1 cells secrete cytokines to promote immune responses and play a role in host defense against pathogens by mediating, to some extent, the recruitment of neutrophils and macrophages to infected tissues. Th1 cells secrete cytokines, including IFN-γ, IL-2, IL-10, and TNF-α / β, to coordinate the defense against intracellular pathogens such as viruses and some bacteria.

[0077] As used herein, the term "Th2 cell" refers to a subset of Th cells that mediate the activation and maintenance of antibody-mediated immune responses against extracellular parasites, bacteria, allergens, and toxins. Th2 cells mediate these functions by producing various cytokines such as IL-4, IL-5, IL-6, IL-9, IL-13, and IL-17E (IL-25), which are responsible for antibody production, eosinophil activation, and inhibition of several macrophage functions, thereby providing a protective response independent of phagocytes.

[0078] "Polypeptide" and "protein" refer to polymers composed of amino acid residues linked by peptide bonds or peptide bond equivalents, related naturally occurring structural variants, and synthetic non-naturally occurring analogs. Synthetic polypeptides can be synthesized, for example, using an automated polypeptide synthesizer. The terms "polypeptide" and "protein" are not limited to the minimum length of the product. The term "protein" generally refers to large polypeptides. The term "peptide" generally refers to short polypeptides. Thus, peptides, oligopeptides, dimers, multimers, etc. are all included in the definition. The definition encompasses full-length proteins and their fragments. The terms "polypeptide" and "protein" also include post-expression modifications of the polypeptide or protein, such as glycosylation, acetylation, phosphorylation, etc. In addition, for the purposes of this disclosure, "polypeptide" can include "modifications" of the native sequence, such as deletions, additions, substitutions (which can be conservative in nature or can include substitutions with any of the 20 amino acids commonly found in human proteins or any other natural or non-natural or non-canonical amino acids), and chemical modifications (e.g., addition or substitution with peptidomimetics). These modifications may be intentional, such as by site-directed mutagenesis or by chemical modification of amino acids to remove or attach chemical moieties, or may be accidental, such as mutations caused by the host cell producing the protein or errors due to PCR amplification prior to host cell transfection.

[0079] "Antigenic portion" or "antigen" as used herein refers to any portion recognized by the host immune system, such as a peptide. Examples of antigenic portions include, but are not limited to, autoantigens, allergens, enzymes, and / or bacterial or viral proteins, peptides, drugs, or components).

[0080] "Pharmaceutically acceptable carrier" means any standard pharmaceutical carrier, buffer, etc., such as phosphate buffered saline solution, 5% aqueous glucose solution, and emulsions (e.g., oil / water or water / oil emulsions). Non-limiting examples of excipients include adjuvants, binders, fillers, diluents, disintegrants, emulsifiers, wetting agents, lubricants, thickeners, sweeteners, flavoring agents, and coloring agents. Suitable pharmaceutical carriers, excipients, and diluents are described in Remington's Pharmaceutical Sciences, 19th Edition (Mack Publishing Co., Easton, 1995). Preferred pharmaceutical carriers depend on the intended mode of administration of the active agent. Typical modes of administration include enteral (e.g., oral) or parenteral (e.g., subcutaneous injection, intramuscular injection, intravenous injection, or intraperitoneal injection; or topical administration, transdermal administration, or transmucosal administration) or administration by inhalation.

[0081] "Pharmaceutically acceptable" or "pharmacologically acceptable" means a material that is not biologically or otherwise undesirable, i.e., the material can be administered to an individual without causing any adverse biological effects and does not interact in a harmful manner with any component of the composition containing the material or with any component present on or in the individual.

[0082] As used herein, the term "subject" encompasses mammals and non-mammals. Examples of mammals include, but are not limited to, any member of the following mammalian classes: humans, non-human primates such as chimpanzees and other anthropoid apes and monkey species; farm animals such as cows, horses, sheep, goats, pigs; domestic animals such as rabbits, dogs, and cats; laboratory animals, including rodents such as rats, mice, and guinea pigs. Examples of non-mammals include, but are not limited to, birds, fish, etc. The term does not denote a particular age or sex.

[0083] The term "epitope" means a portion of any molecule that can be recognized and bound by a selective binding agent at one or more antigen-binding regions in an antigen-binding domain. Epitopes typically consist of the chemically reactive surface groups of a molecule, such as amino acid or carbohydrate side chains, and have specific three-dimensional structural characteristics as well as specific charge characteristics. As used herein, an epitope can be continuous or discontinuous. In addition, an epitope can be a mimetic (mimotope) in that it contains the same three-dimensional structure as the epitope used to generate an antibody, but does not contain or contains only some of the amino acid residues seen in the target that are used to stimulate an antibody immune response. As used herein, a mimotope is not considered a different antigen from the epitope that binds to the selective binding agent; the selective binding agent recognizes the same three-dimensional structure of the epitope and the mimotope.

[0084] The term "therapeutically effective amount" is used herein to indicate the amount of the antigen - specific composition of the present disclosure that is effective to ameliorate or reduce the symptoms or signs of a disease to be treated.

[0085] As used herein with respect to methods, the terms "treat", "treated", "treating" and "treatment" mean to temporarily or permanently, partially or completely eliminate, reduce, suppress or ameliorate the clinical symptoms, manifestations or progression of an event, disease or condition. Such treatment is not necessarily curative.

[0086] As used herein, the term "symptom" means any physical or observable manifestation of a disorder, whether or not it is a general characteristic of the disorder. The term "symptom" can mean all such manifestations or any subset thereof.

[0087] As used herein, a "booster dose" refers to the readministration of TIMP after an initial or primary administration. The administration or readministration of a booster dose enhances the tolerogenic immune response elicited by the initial administration. The booster dose can be greater than, equal to, or less than the initial dose of TIMP particles administered to the subject. Subjects treated with tolerogenic therapy are monitored to confirm the maintenance of immune tolerance. Based on the observation of changes, attenuation or loss of immune tolerance, the decision to administer a booster dose is made. Methods for monitoring the immune tolerance status of subjects treated with tolerogenic therapy have been previously described (see International Patent Publication WO 2022 / 221622, which is incorporated herein by reference). The booster dose of TIMP - PPE is administered in the presence of the same, different or no combination therapy as the combination therapy described herein that is capable of being administered with the primary TIMP - PPE dose. Particle

[0088] The size and charge of the particles are important for tolerance induction. Although the size and charge of the particles will vary based on the antigen encapsulated therein, generally, when the particles described herein are between about 100 nanometers and about 1500 nanometers and have a charge between 0 mV and about -100 mV, they are effective in inducing tolerance. In various embodiments, the particles have a diameter of 400 - 800 nanometers and have a charge between about -25 mV and -70 mV. In various embodiments, the particles have a diameter of 400 - 800 nanometers and have a charge between about -30 mV and -60 mV. In various embodiments, the particles have a diameter of 400 - 800 nanometers and have a charge between about -30 mV and -80 mV. The average particle size and charge may change slightly during the lyophilization process, and thus, the post-synthesis average values and the post-lyophilization average values are described. As used herein, the terms "post-synthesis size" and "post-synthesis charge" refer to the size and charge of the particles before lyophilization. "Post-lyophilization size" and "post-lyophilization charge" refer to the size and charge of the particles after lyophilization.

[0089] In some embodiments, the particles are non-metallic. In these embodiments, the particles can be formed from a polymer. In preferred embodiments, the particles are biodegradable in an individual. In this embodiment, the particles can be provided to the individual across multiple doses without accumulating in the individual. Examples of suitable particles include polystyrene particles, PLGA particles, PLURONICS-stabilized polyphenylene sulfide particles, and diamond particles.

[0090] Preferably, the particle surface is composed of a material that minimizes non-specific or unwanted biological interactions. The interaction between the particle surface and the stroma may be a factor in lymphatic uptake. The particle surface can be coated with a material that prevents or reduces non-specific interactions. As demonstrated by the improved lymphatic uptake after subcutaneous injection, steric hindrance stabilization by coating the particles with a hydrophilic layer such as polyethylene glycol (PEG) and its copolymers (such as PLURONICS®) (including copolymers of polyethylene glycol - bl - polypropylene glycol - bl - polyethylene glycol) can reduce non-specific interactions with stromal proteins. All of these facts suggest the relevance of the physical properties of the particles in lymphatic uptake. Biodegradable polymers can be used to fabricate all or part of the polymer and / or particles and / or layers. Biodegradable polymers, for example, may undergo degradation due to the reaction of functional groups with water in the solution. As used herein, the term "degradation" refers to becoming soluble by a decrease in molecular weight or by the conversion of hydrophobic groups to hydrophilic groups. Polymers having ester groups (e.g., poly(lactide) and poly(glycolide)) typically undergo spontaneous hydrolysis.

[0091] The particles disclosed herein may also contain additional components. For example, the carrier may have an imaging agent incorporated or conjugated to the carrier. An example of a carrier nanosphere with a currently commercially available imaging agent is Kodak X-sight nanospheres. Inorganic quantum-confined luminescent nanocrystals called quantum dots (QDs) are ideal donors in FRET applications: their high quantum yields and tunable size-dependent Stokes Shifts allow different sizes to emit blue to infrared light when excited at a single ultraviolet wavelength. (Bruchez et al., Science, 1998, 281, 2013; Niemeyer, C. M Angew. Chem. Int. Ed., 2003, 42, 5796; Waggoner, A. Methods Enzymol., 1995, 246, 362; Brus, L. E. J. Chem. Phys., 1993, 79, 5566). Quantum dots, such as hybrid organic / inorganic quantum dots based on a class of polymers called dendrimers, can be used in bio-labeling, imaging, and optical biosensing systems. (Lemon et al., J. Am. Chem. Soc., 2000, 122, 12886). Unlike the synthesis of traditional inorganic quantum dots, the synthesis of these hybrid quantum dot nanoparticles does not require high temperatures or highly toxic and unstable reagents. (Etienne et al., Appl. Phys. Lett., 87, 181913, 2005).

[0092] The particles can be formed from a wide variety of materials. The particles are preferably composed of materials suitable for biological use. For example, the particles can be composed of glass, silica, polyesters of hydroxycarboxylic acids, polyanhydrides of dicarboxylic acids, or copolymers of hydroxycarboxylic acids and dicarboxylic acids. More generally, the carrier particles can be composed of: linear or branched, substituted or unsubstituted, saturated or unsaturated, linear or crosslinked alkyl, haloalkyl, thioalkyl, aminoalkyl, aryl, aralkyl, alkenyl, aralkenyl, heteroaryl or alkoxyhydroxy acid polyesters or linear or branched, substituted or unsubstituted, saturated or unsaturated, linear or crosslinked alkyl, haloalkyl, thioalkyl, aminoalkyl, aryl, aralkyl, alkenyl, aralkenyl, heteroaryl or alkoxydicarboxylic acid polyanhydrides. Additionally, the carrier particles can be quantum dots, or composed of quantum dots such as quantum dot polystyrene particles (Joumaa et al. (2006) Langmuir 22: 1810-6). Carrier particles comprising a mixture of ester and anhydride bonds (e.g., a copolymer of glycolic acid and sebacic acid) can also be employed. For example, the carrier particles can comprise materials including: polyglycolic acid (PGA) polymers, polylactic acid (PLA) polymers, polysebacic acid (PSA) polymers, poly(lactic-co-glycolic acid) (PLGA or PLG; the terms are interchangeable) copolymers, poly(lactic-co-sebacic acid) (PLSA) copolymers, poly(glycolic-co-sebacic acid) (PGSA) copolymers, polyphenylene sulfide polymers, polycaprolactone, chitosan, and the like. Other biocompatible, biodegradable polymers useful in the present invention include polymers or copolymers of caprolactone, carbonates, amides, amino acids, orthoesters, acetals, cyanoacrylates, and degradable polyurethanes, as well as their copolymers with linear or branched, substituted or unsubstituted alkyl, haloalkyl, thioalkyl, aminoalkyl, alkenyl or aromatic hydroxyacids or dicarboxylic acids. Additionally, biologically important amino acids having reactive side chain groups, such as lysine, arginine, aspartic acid, glutamic acid, serine, threonine, tyrosine, and cysteine or their enantiomers, can be included in copolymers with any of the above materials to provide reactive groups for conjugation with antigenic peptides and proteins or conjugate moieties. Biodegradable materials suitable for the present invention include diamond, PLA, PGA, polyphenylene sulfide, and PLGA polymers. Biocompatible but non-biodegradable materials can also be used in the carrier particles of the present invention. For example, acrylate, ethylene-vinyl acetate, acyl-substituted cellulose acetate, non-degradable polyurethane, styrene, vinyl chloride, vinyl fluoride, vinyl imidazole, chlorosulfonated olefins, ethylene oxide, vinyl alcohol, TEFLON ® (DuPont, Wilmington, Del.) and non-biodegradable polymers of nylon.

[0093] In certain embodiments, the particles are copolymers having a molar ratio of from about 80:20 to about 100:0. Suitable copolymer ratios for the present immunomodified particles can be 25:75, 30:70, 35:65, 40:60, 45:55, 50:50, 55:45, 60:40, 65:35, 70:30, 75:25, 80:20, 81:19, 82:18, 83:17, 84:16, 85:15, 86:14, 87:13, 88:12, 89:11, 90:10, 91:9, 92:8, 93:7, 94:6, 95:5, 96:4, 97:3, 98:2, 99:1 or 100:0. In certain embodiments, the particles are PLURONICS-stabilized polyphenylene sulfide particles, polyglycolic acid (PGA) particles, polylactic acid (PLA) particles, or poly(lactic-co-glycolic acid) particles. In certain embodiments, the copolymer ratio of the particles is polylactic acid / polyglycolic acid 80:20, polylactic acid / polyglycolic acid 90:10, or polylactic acid:polyglycolic acid 50:50. In various embodiments, the particles are poly(lactic-co-glycolic acid) particles, and the copolymer ratio of polylactic acid:polyglycolic acid is about 50:50.

[0094] It is contemplated that the particles may further comprise a surfactant. The surfactant can be anionic, cationic, or nonionic. Surfactants from the Pluronic and Poloxamine families are commonly used in particle synthesis. Surfactants that can be used include, but are not limited to, PEG, Tween-80, gelatin, dextran, pluronic L-63, PVA, PAA, methylcellulose, lecithin, DMAB, and PEMA. Additionally, biodegradable and biocompatible surfactants, including but not limited to vitamin E TPGS (D-α-tocopherol polyethylene glycol 1000 succinate), polyamino acids (e.g., polymers of lysine, arginine, aspartic acid, glutamic acid, serine, threonine, tyrosine, and cysteine or their enantiomers), and sulfate polymers. In certain embodiments, two surfactants are used. For example, if the particles are produced by the double emulsion method, the two surfactants can include a hydrophobic surfactant for the first emulsion and a hydrophobic surfactant for the second emulsion.

[0095] In some embodiments, the polypeptide antigen is encapsulated in particles by the single emulsion method. In further embodiments, the polypeptide antigen is more hydrophobic. Sometimes, the double emulsion method results in the formation of large particles, which may lead to leakage of hydrophilic active components and low retention efficiency. Coalescence and Ostwald ripening are two mechanisms that can destabilize double emulsion droplets, while diffusion of the hydrophilic active component through the organic phase is the main mechanism leading to low retention levels of the active component. In some embodiments, reducing the nanoparticle size may be beneficial. One strategy to achieve this is to apply a second strong shear rate. The leakage effect can be reduced by using a high polymer concentration and high polymer molar mass, which is accompanied by an increase in the viscosity of the internal aqueous phase and an increase in the surfactant molar mass. In certain embodiments, the particles encapsulating the antigen are manufactured by nanoprecipitation, coprecipitation, inert gas condensation, sputtering, microemulsion, sol-gel method, layer-by-layer technique, or ion gelation method. Several methods for manufacturing nanoparticles have been described in the literature and are incorporated herein by reference (Sánchez, Mejía, and Orozco 2020; Zielińska et al. 2020). antigen

[0096] An antigen refers to a discrete portion of a molecule, such as a polypeptide or peptide sequence, the 3-D structure formation of a polypeptide or peptide, a polysaccharide, or a polynucleotide that can be recognized by a host immune cell. Antigen specificity refers to the ability of the host cells of a subject to recognize and generate an immune response against a separate antigen or a molecule very similar to the antigen, such as an epitope or a mimotope.

[0097] "Anergy", "tolerance", or "antigen-specific tolerance" refers to the insensitivity of T cells to T cell receptor-mediated stimulation. This insensitivity is usually antigen-specific and persists after cessation of exposure to the antigenic peptide. For example, T cell anergy is characterized by a lack of cytokine production, such as IL-2. T cell anergy occurs when T cells are exposed to an antigen and receive a first signal (T cell receptor or CD-3-mediated signal) in the absence of a second signal (co-stimulatory signal). Under these conditions, re-exposure of the cells to the same antigen (even when re-exposure occurs in the presence of co-stimulatory molecules) results in the inability to produce cytokines and subsequent inability to proliferate. Thus, the inability to produce cytokines prevents proliferation. However, anergic T cells can proliferate if cultured with cytokines, such as IL-2.

[0098] It is contemplated that the tolerance therapies described herein are antigen - specific. For example, TIMP encapsulation administered in the form of a tolerance therapy encompasses one or more antigens associated with the tolerance therapy and the relevant disease or condition being treated. It is contemplated that TIMP for a tolerance therapy contains one or more peanut antigens. The one or more peanut antigens are derived from a peanut protein extract or may be peptides derived from known peanut proteins.

[0099] The WHO / IUIS Allergen Nomenclature Sub-Committee (www.allergen.org) has officially recognized more than 15 peanut allergens: Ara h1 to Ara h18, including Ara h1, Ara h2, Ara h3, Ara h5, Ara h6, Ara h7, Ara h8, Ara h9, Ara h10, Ara h11, Ara h12, Ara h13, Ara h14, Ara h15, Ara h16, Ara h17, and Ara h18. Based on Ara h1, Ara h2, Ara h3, Ara h5, Ara h6, and Ara h8, peanut allergens can be classified into different groups according to their structures (e.g., trimers, monomers, cupins, albumins, prolamins, protease inhibitors, oleosins, defensins, vicilins, and nonspecific lipid transfer proteins (nsLTPs)), and each of these groups has a different degree of allergenic potency (Ozias-Akins et al., Allergy 74:888-898, 2019). Known peanut allergens include allergens derived from Arachis hypogaea: Ara h1, Ara h2, Ara h3, Ara h5, Ara h6, Ara h7, Ara h8, and Ara h18. See, for example, UNIPROT database accession number E5G076 showing the Ara h1 polypeptide sequence (SEQ ID NO: 1), UNIPROT database accession number A0A445BYI5 for the Ara h2 polypeptide (SEQ ID NO: 2), UNIPROT database accession number E5G077 for the Ara h3 polypeptide (SEQ ID NO: 3) (see also UNIPROT database accession numbers O82580 (SEQ ID NO: 4) and Q9SQH7 (SEQ ID NO: 5) for Ara h3 isoallergen 1 and Arah3 isoallergen 2 (previously known as Ara h4), respectively), UNIPROT database accession number L7QH52 for the Ara h5 polypeptide (SEQ ID NO: 6), UNIPROT database accession number A5Z1R0 for the Ara h6 polypeptide (SEQ ID NO: 7), UNIPROT database accession number B4XID4 for the Ara h7 polypeptide (SEQ IDNO: 8), UNIPROT database accession number Q6VT83 for the Ara h8 polypeptide sequence (SEQ ID NO: 9); Ara h9 isoallergen 1 and Ara h9 isoallergen 2, UNIPROT database accession numbers B6CEX8 and B6CG41 (SEQ ID NO: 10 and 11), respectively;Allergens Ara h10 isoallergen 1 and Ara h10 isoallergen 2, with UNIPROT database accession numbers Q647G5 and Q647G4 respectively (SEQ ID NO: 12 and 13); Allergens Ara h11 isoallergen 1 and Ara h11 isoallergen 2, with UNIPROT database accession numbers Q45W87 and Q45W86 respectively (SEQ ID NO: 14 and 15); Allergen Ara h12, with UNIPROT database accession number B3EWP3 (SEQ ID NO: 16); Allergens Ara h13 isoallergen 1 and Ara h13 isoallergen 2, with UNIPROT database accession numbers B3EWP4 and C0HJZ1 respectively (SEQ ID NO: 17 and 18); Allergens Ara h14 isoallergen 1, Ara h14 isoallergen 2 and Ara h14 isoallergen 3, with UNIPROT database accession numbers Q9AXI1, Q9AXI0 and Q6J1J8 respectively (SEQ ID NO: 19 to 21); Allergen Ara h15, with UNIPROT database accession number Q647G3 (SEQ ID NO: 22); Allergen Ara h16, with UNIPROT database accession number A0A509ZX51 (SEQ ID NO: 23); Allergen Ara h17, with UNIPROT database accession number 0A510A9S3 (SEQ ID NO: 24); Allergen Ara h18, with UNIPROT database accession number A0A444XS96 (SEQ ID NO: 25).;

[0100] In various embodiments, the allergenic peanut proteins include one or more of the following: Ara h1, Ara h2, Arah3, Ara h5, Ara h6, Ara h7, Ara h8, Ara h9, Ara h10, Ara h11, Ara h12, Ara h13, Arah14, Ara h15, Ara h16, Ara h17 and Ara h18. In various embodiments, the peptides derived from peanut proteins comprise allergenic epitopes from one or more of the following: Ara h1, Ara h2, Ara h3, Ara h5, Ara h6, Ara h7, Ara h8, Ara h9, Ara h10, Ara h11, Ara h12, Ara h13, Ara h14, Ara h15, Ara h16, Ara h17 and Ara h18.

[0101] In certain embodiments, one, two, three, or more antigens or antigenic peptides are used in the TIMP. In certain embodiments, one or more peanut antigens are encapsulated in the TIMP by covalently attaching them to the inner surface of the particle (see, e.g., U.S. Patent Publication US20190282707, incorporated herein by reference). In certain embodiments, it is contemplated that the sequences of two or more peanut proteins, such as from Ara h1, Ara h2, Ara h3, Ara h5, Ara h6, Ara h7, and / or Ara h8, are ligated in a fusion protein and encapsulated in the TIMP described herein. In certain embodiments, it is contemplated that the sequences of two or more peanut proteins, such as from Ara h1, Ara h2, Ara h3, Ara h5, Ara h6, Ara h7, Ara h8, Ara h9, Ara h10, Arah11, Ara h12, Ara h13, Ara h14, Ara h15, Ara h16, Ara h17, and Ara h18, are ligated in a fusion protein and encapsulated in the TIMP described herein. Methods for preparing TIMPs with ligated epitopes are described in U.S. Patent Publication US20190365656, incorporated herein by reference. Method of Use

[0102] Peanut allergy is the most common food allergy in the United States, with up to 0.6% of adults and 0.8% of children diagnosed. Additionally, due to the high prevalence of peanut allergy in the population and the widespread use of peanut-derived products as fillers in many packaged foods, peanut allergy is the most common cause of fatal food-related allergic reactions. Thus, peanut allergy poses a significant health hazard in the United States.

[0103] The four major peanut allergen proteins are Ara h1, Ara h2, Ara h3, and Ara h6. Ara h1 and Ara h2 are recognized by more than 95% of peanut-allergic patients.

[0104] For a long time, dietary avoidance of peanuts has been the most effective treatment for peanut allergy. However, severe cases of peanut allergy can also be triggered by foods processed near peanuts and / or inhaling air near peanut products. When it is not possible to completely avoid ingesting peanut products and entering areas contaminated with peanut products, severe allergic reactions may occur, and patients need to receive medications such as epinephrine, antihistamines, and / or oral steroids.

[0105] Current therapies approved for the treatment of peanut allergy, such as PALFORZIA™, and therapies in clinical trials only prevent accidental exposure to peanuts by desensitizing the subject. The treatment regimen of PALFORZIA™ also includes a rather extensive daily dose escalation and maintenance schedule and may pose challenges to subject compliance and still requires the subject to maintain a peanut-free diet.

[0106] A treatment method that renders T cells tolerant to peanut proteins may cure peanut allergy, thereby eliminating the burden of strict avoidance of peanut products.

[0107] Provided herein is a method of treating peanut allergy in a subject, the method comprising administering TIMP-PPE to the subject, wherein the TIMP-PPE is administered at a dose level determined based on the weight of the subject. It is also contemplated that the TIMP-PPE may be administered at a fixed dose regardless of the weight of the subject. Provided herein is a method of treating peanut allergy in a subject, the method comprising administering TIMP-PPE to the subject, wherein the TIMP-PPE is administered at a dose of 0.1 mg / kg to 12 mg / kg. Provided herein is a method of treating peanut allergy in a subject, the method comprising administering TIMP-PPE to the subject, wherein the TIMP-PPE is administered at a dose of 0.001 mg / kg to 12 mg / kg. In various embodiments, a method of treating peanut allergy in a subject is contemplated, the method comprising administering TIMP-PPE to the subject, wherein the TIMP-PPE is administered at a dose of 0.001 mg / kg to 12 mg / kg based on the weight of the subject or at a fixed dose between 0.1 mg and 800 mg. Also provided herein is a method of reducing the allergic immune response of a subject with PA to peanut antigen, the method comprising administering TIMP-PPE to the subject, wherein the TIMP-PPE is administered at a dose of 0.001 mg / kg to 12 mg / kg. Further contemplated herein is a method of reducing the allergic immune response of a subject with PA to peanut antigen, the method comprising administering TIMP-PPE to the subject, wherein the TIMP-PPE is administered at a dose of 0.001 mg / kg to 12 mg / kg based on the weight of the subject or at a fixed dose between 0.1 mg and 800 mg.

[0108] It is also contemplated that TIMP-PPE is administered at a dose of about 0.001 mg / kg to 10 mg / kg, about 0.005 mg / kg to 12 mg / kg, about 0.01 mg / kg to 12 mg / kg, about 0.05 mg / kg to 12 mg / kg, about 0.1 mg / kg to 12 mg / kg, about 0.5 mg / kg to 10 mg / kg, about 1 mg / kg to 8 mg / kg, about 1.5 mg / kg to 10 mg / kg, about 2 mg / kg to 12 mg / kg, about 2 mg / kg to 10 mg / kg, about 3 mg / kg to 10 mg / kg, about 4 mg / kg to 10 mg / kg, about 4 mg / kg to 12 mg / kg or about 5 mg / kg to 12 mg / kg. Optionally, TIMP-PPE is administered at a dose of about 0.001 mg / kg, 0.0025 mg / kg, 0.005 mg / kg, 0.01 mg / kg, 0.025 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.25, 0.5 mg / kg, 1.0 mg / kg, 2.0 mg / kg, 4.0 mg / kg, 6 mg / kg, 8.0 mg / kg, 10 mg / kg or 12 mg / kg. Alternatively, TIMP-PPE is administered at a dose of about 0.1 mg, 0.25 mg, 0.5 mg, 1 mg, 2 mg, 2.5 mg, 5 mg, 10 mg, 25 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 325 mg, 350 mg, 400 mg, 425 mg, 450 mg, 475 mg, 500 mg, 525 mg, 550 mg, 575 mg, 600 mg, 625 mg, 650 mg, 675 mg, 700 mg, 725 mg, 750 mg, 775 mg or 800 mg.In another embodiment, TIMP-PPE is administered at a concentration between about 0.0005 mg / mL and about 50 mg / mL, optionally about 0.0005 mg / mL, 0.001 mg / mL, 0.005 mg / mL, 0.01 mg / mL, 0.05 mg / mL, 0.1 mg / mL, 0.5 mg / mL, 1 mg / mL, 2 mg / mL, 3 mg / mL, 4 mg / mL, 5 mg / mL, 6 mg / mL, 7 mg / mL, 8 mg / mL, 9 mg / mL, 10 mg / mL, 11 mg / mL, 12.5 mg / mL, 15 mg / mL, 17.5 mg / mL, 20 mg / mL, 25 mg / mL, 30 mg / mL, 40 mg / mL, or 50 mg / mL.

[0109] It is contemplated that TIMP-PPE is administered in a single dose or multiple doses. In various embodiments, TIMP-PPE is administered once a week, once every two weeks, once every three weeks, once every 4 weeks, once every two months, once every three months, once every 6 months, or once a year. In certain embodiments, TIMP-PPE is administered in two doses one week apart.

[0110] In various embodiments, TIMP-PPE is administered intravenously, subcutaneously, intramuscularly, intraperitoneally, intranasally, or orally. It is contemplated that if TIMP-PPE is administered intravenously, it can be administered by intravenous infusion for about 1, 2, 3, 4, 5, 6, 7, 8, 10, 12, 18, or 20 hours or by intravenous infusion for about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 10, 12, 18, or 20 hours.

[0111] Also provided herein is a method of enhancing the duration or efficacy of tolerance to peanut allergen induction by TIMP-PPE treatment, the method comprising administering a booster dose of TIMP-PPE. In various embodiments, the booster dose of TIMP-PPE is administered in a single dose or multiple doses after the initial or first administration of TIMP-PPE. In various embodiments, the booster dose of TIMP-PPE is administered once a week, once every two weeks, once every three weeks, once every 4 weeks, once every two months, once every three months, once every 6 months, or once a year. In various embodiments, the booster dose of TIMP-PPE is administered as a single dose every three months.

[0112] It is further contemplated that the TIMP-PPE is administered alone or in combination with one or more additional therapeutic agents. Additional exemplary therapeutic agents include, but are not limited to, IgE inhibitors, basophil activation inhibitors, mast cell activation inhibitors, antihistamines, non-steroidal anti-inflammatory drugs (NSAIDs), or small molecule or biologic therapeutic agents.

[0113] In various embodiments, the TIMP-PPE is administered alone or in combination with one or more therapeutic agents useful for treating peanut allergy. Exemplary therapeutic agents include, but are not limited to, IgE inhibitors, basophil activation inhibitors, mast cell activation inhibitors, antihistamines, non-steroidal anti-inflammatory drugs (NSAIDs), prebiotics, probiotics, histone deacetylase inhibitors, short-chain fatty acids (e.g., acetate, butyrate, propionate, butyrate polymers), competitors of IgE for allergen binding sites, cytokine inhibitors, microbiome therapies, steroids, corticosteroids, leukotriene modifiers, or small molecule or biologic therapeutic agents. Exemplary additional therapeutic agents or therapeutic agents useful for treating peanut allergy also include therapeutic agents that increase the number, frequency, or activity of Tregs.

[0114] In various embodiments, the biologic is an antibody. In various embodiments, the antibody is an anti-IgE antibody, an anti-IL-4Rα antibody, an anti-IL-13 antibody, or an anti-IL-33 antibody. In various embodiments, the anti-IgE antibody is omalizumab (XOLAIR®). Exemplary anti-IL-4Rα antibodies include dupilumab (DUPIXENT®), and the anti-IL-33 antibody includes etokinumab.

[0115] In various embodiments, the antihistamine is a first-generation antihistamine. In various embodiments, the antihistamine is a second-generation antihistamine. In various embodiments, the antihistamine is selected from the group consisting of: brompheniramine, carbinoxamine maleate, chlorpheniramine, clemastine, diphenhydramine, hydroxyzine, triprolidine, azelastine, cetirizine, desloratadine, fexofenadine, levocetrizine, loratadine, and olopatadine. In various embodiments, the additional therapeutic agent is a steroid. In various embodiments, the steroid is selected from the group consisting of: beclomethasone, ciclesonide, fluticasone furoate, mometasone, budenoside, fluticasone, triamcinolone, and loteprednol. In various embodiments, the additional therapeutic agent is a corticosteroid. In various embodiments, the corticosteroid is selected from the group consisting of: cortisone, prednisone, prednisolone, methylprednisolone, dexamethasone, betamethasone, and hydrocortisone.

[0116] In various embodiments, the additional therapeutic agent is a non-steroidal anti-inflammatory drug (NSAID). In various embodiments, the therapeutic agent useful for treating peanut allergy is a non-steroidal anti-inflammatory drug (NSAID). In various embodiments, the NSAID is a non-selective NSAID. In various embodiments, the NSAID is a selective NSAID. In various embodiments, the NSAID is a COX-2 selective NSAID. In various embodiments, the NSAID is a COX-1 selective NSAID. In various embodiments, the NSAID is a prostaglandin synthase inhibitor. In various embodiments, the NSAID is selected from the group consisting of: diclofenac, diclofenac potassium, diclofenac sodium, diflunisal, etodolac, flurbiprofen, fenoprofen, fenoprofen calcium, ketorolac, ketorolac tromethamine, ketoprofen, tolmetin, tolmetin sodium, aspirin, ibuprofen, naproxen, indomethacin, indomethacin sodium, sulindac, felbinac, piroxicam, mefenamic acid, meclofenamate sodium, meloxicam, nabumetone, oxaprozin, piroxicam, celecoxib, etodolac, etoricoxib, lumiracoxib, rofecoxib, and valdecoxib.

[0117] In various embodiments, the additional therapeutic agent is a leukotriene modifier. In various embodiments, the therapeutic agent useful for treating peanut allergy is a leukotriene modifier. In various embodiments, the leukotriene modifier is an anti-leukotriene. In various embodiments, the leukotriene modifier is a leukotriene receptor antagonist. In various embodiments, the leukotriene modifier is a leukotriene synthesis inhibitor. In various embodiments, the leukotriene modifier is selected from the group consisting of: montelukast, zileuton, and zafirlukast.

[0118] In various embodiments, the additional therapeutic agent is administered before, during, or after administration of TIMP-PPE. In various embodiments, the therapeutic agent useful for treating peanut allergy is administered before, during, or after administration of TIMP-PPE.

[0119] In various embodiments, the additional therapeutic agent is administered intravenously, subcutaneously, intramuscularly, intraperitoneally, intranasally, by inhalation, or orally. In various embodiments, the therapeutic agent useful for treating peanut allergy is administered intravenously, subcutaneously, intramuscularly, intraperitoneally, intranasally, by inhalation, or orally.

[0120] The present disclosure provides a method of treating peanut allergy in a subject, the method comprising administering to the subject a combination of TIMP-PPE and an anti-IgE antibody, wherein TIMP-PPE is administered at a dose of about 0.001 mg / kg to 12 mg / kg, and wherein the anti-IgE antibody is administered at a dose of about 10 mg to about 500 mg. In various embodiments, the anti-IgE antibody is omalizumab (XOLAIR®), quilizumab, or ligelizumab.

[0121] In various embodiments, the anti-IgE antibody administered in combination with TIMP-PPE is administered at a dose of about 10 mg, 25 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 375 mg, 400 mg, 425 mg, 450 mg, 475 mg, or 500 mg. The dose level of the anti-IgE antibody is determined based on the serum IgE level, which can be between about 30 - 100 IU / mL, 100 - 200 IU / mL, 200 - 300 IU / mL, 300 - 400 IU / mL, 400 - 500 IU / mL, 500 - 600 IU / mL, 600 - 700 IU / mL, 700 - 800 IU / mL, 800 - 900 IU / mL, 900 - 1000 IU / mL, 1000 - 1100 IU / mL, 1100 - 1200 IU / mL, 1200 - 1300 IU / mL, 1300 - 1400 IU / mL, or 1400 - 1500 IU / mL. Alternatively, the dose level of the anti-IgE antibody is determined based on the weight of the subject. In various embodiments, the weight of the subject is 30 - 40 kg, 40 - 50 kg, 50 - 60 kg, 60 - 70 kg, 70 - 80 kg, 80 - 90 kg, 90 - 125 kg, or 125 - 150 kg.

[0122] In various embodiments, the anti-IgE antibody is administered in a single dose or multiple doses. In various embodiments, the anti-IgE antibody is administered once a week, once every two weeks, once every three weeks, or once every four weeks. In various embodiments, the anti-IgE antibody is administered before, concomitantly with, or immediately before or after administering TIMP-PPE. In various embodiments, the anti-IgE antibody is administered one week, two weeks, three weeks, or four weeks before administering TIMP-PPE. In various embodiments, the anti-IgE antibody is administered one week, two weeks, three weeks, or four weeks after administering TIMP-PPE.

[0123] Concurrent or combined administration of two therapeutic agents does not require that the agents be administered at the same time or by the same route, provided that the time periods during which the agents exert their therapeutic effects overlap. Concurrent or sequential administration is contemplated, as is administration on different days or weeks. Further contemplated is that the therapeutic agents are administered as separate formulations and are combined or co-administered, where combination means agents administered within 30 minutes of each other. Pre-administration means administration of the therapeutic agent within the range of up to 30 minutes prior to administration of TIMP-PPE and within one week prior to treatment with TIMP-PPE. Concomitant administration is intended to describe administration from 30 minutes after TIMP-PPE treatment up to one week after administration of TIMP-PPE.

[0124] The present disclosure provides a method for treating peanut allergy, the method comprising administering to a subject a combination of TIMP-PPE and an anti-IgE antibody, wherein the anti-IgE antibody is administered to the subject once weekly for two weeks or once weekly for four weeks prior to administration of TIMP-PPE, wherein TIMP-PPE is administered in two dose intervals one week apart at a dose level between 0.001 mg / kg and 12 mg / kg, and wherein the anti-IgE antibody is administered at a dose level between about 50 mg and 500 mg.

[0125] Also provided is a method for treating peanut allergy in a subject, the method comprising administering to the subject a combination of TIMP-PPE and an anti-IL-4Rα antibody, wherein TIMP-PPE is administered at a dose of about 0.001 mg / kg to 12 mg / kg, and wherein the anti-IL-4Rα antibody is administered at a dose of about 10 mg to about 500 mg. In various embodiments, the anti-IL-4Rα antibody is dupilumab (DUPIXENT®).

[0126] In various embodiments, the anti-IL-4Rα antibody is administered at a dose of about 10 mg, 25 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 375 mg, 400 mg, 425 mg, 450 mg, 475 mg, 500 mg, 525 mg, 550 mg, 575 mg or 600 mg. In various embodiments, the dose level of the anti-IL-4Rα antibody is determined based on the serum IgE level, which can be about 30 - 100 IU / mL, 100 - 200 IU / mL, 200 - 300 IU / mL, 300 - 400 IU / mL, 400 - 500 IU / mL, 500 - 600 IU / mL, 600 - 700 IU / mL, 700 - 800 IU / mL, 800 - 900 IU / mL, 900 - 1000 IU / mL, 1000 - 1100 IU / mL, 1100 - 1200 IU / mL, 1200 - 1300 IU / mL, 1300 - 1400 IU / mL or 1400 - 1500 IU / mL. Alternatively, the dose level of the anti-IL-4Rα antibody is determined based on the weight of the subject. In various embodiments, the weight of the subject is 30 - 40 kg, 40 - 50 kg, 50 - 60 kg, 60 - 70 kg, 70 - 80 kg, 80 - 90 kg, 90 - 125 kg or 125 - 150 kg.

[0127] In various embodiments, the anti-IL-4Rα antibody is administered as a single dose or multiple doses. In various embodiments, the anti-IL-4Rα antibody is administered once a week, once every two weeks, once every three weeks or once every four weeks. In various embodiments, the anti-IL-4Rα antibody is administered before, concomitantly with or after the administration of TIMP-PPE. In various embodiments, the anti-IL-4Rα antibody is administered one week, two weeks, three weeks or four weeks before the administration of TIMP-PPE. In various embodiments, the anti-IL-4Rα antibody is administered one week, two weeks, three weeks or four weeks after the administration of TIMP-PPE. In various embodiments, the anti-IL-4Rα antibody is administered in two doses at an initial dose level between 400 mg and 600 mg, followed by a maintenance dose level between 200 mg and 300 mg for subsequent doses.

[0128] In various embodiments, the antihistamine administered in combination with TIMP-PPE is administered at a dose of about 0.05 mg to 2000 mg. In various embodiments, the antihistamine is administered at a dose of about 0.05 mg, 0.1 mg, 0.5 mg, 1 mg, 2 mg, 4 mg, 5 mg, 10 mg, 25 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 375 mg, 400 mg, 425 mg, 450 mg, 475 mg, 500 mg, 1000 mg, 1500 mg, or 2000 mg. In various embodiments, the antihistamine is administered as a single dose or multiple doses. In various embodiments, the antihistamine is administered once a day, once a week, once every two weeks, once every three weeks, or once every four weeks. In various embodiments, the antihistamine is administered two, three, four, five, or six times a day. In various embodiments, the antihistamine is administered before, concomitantly with, or after the administration of TIMP-PPE. In various embodiments, the antihistamine is administered one, two, three, four, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, or 12 months before the administration of TIMP-PPE. In various embodiments, the antihistamine is administered one, two, three, four, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, or 12 months after the administration of TIMP-PPE. In various embodiments, the antihistamine is administered 5, 10, 15, 30, 45, or 60 minutes before the administration of TIMP-PPE. In various embodiments, the antihistamine is administered 5, 10, 15, 30, 45, or 60 minutes after the administration of TIMP-PPE. In various embodiments, the antihistamine is administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 hours before the administration of TIMP-PPE. In various embodiments, the antihistamine is administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 hours after the administration of TIMP-PPE. In various embodiments, the antihistamine is administered 1, 2, 3, 4, 5, 6, or 7 days before the administration of TIMP-PPE. In various embodiments, the antihistamine is administered 1, 2, 3, 4, 5, 6, or 7 days after the administration of TIMP-PPE. In various embodiments, the antihistamine is a first-generation antihistamine or a second-generation antihistamine.In various embodiments, the antihistamine is selected from the group consisting of: brompheniramine, carbinoxamine maleate, chlorpheniramine, clemastine, diphenhydramine, hydroxyzine, triprolidine, azelastine, cetirizine, desloratadine, fexofenadine, levocetrizine, doxylamine, ebastine, embramine, epinephrine, fexofenadine, loratadine, and olopatadine.

[0129] In various embodiments, the steroid administered in combination with TIMP-PPE is administered at a dose of from about 0.05 mg to 2000 mg. In various embodiments, the steroid is administered at a dose of about 0.05 mg, 0.1 mg, 0.5 mg, 1 mg, 2 mg, 4 mg, 5 mg, 10 mg, 25 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 375 mg, 400 mg, 425 mg, 450 mg, 475 mg, 500 mg, 1000 mg, 1500 mg or 2000 mg. In various embodiments, the steroid is administered in a single dose or multiple doses. In various embodiments, the steroid is administered once a day, once a week, once every two weeks, once every three weeks or once every four weeks. In various embodiments, the steroid is administered two, three, four, five or six times a day. In various embodiments, the steroid is administered before, concomitantly with or after the administration of TIMP-PPE. In various embodiments, the steroid is administered one, two, three, four, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months or 12 months before the administration of TIMP-PPE. In various embodiments, the steroid is administered one, two, three, four, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months or 12 months after the administration of TIMP-PPE. In various embodiments, the steroid is administered 5, 10, 15, 30, 45 or 60 minutes before the administration of TIMP-PPE. In various embodiments, the steroid is administered 5, 10, 15, 30, 45 or 60 minutes after the administration of TIMP-PPE. In various embodiments, the steroid is administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 hours before the administration of TIMP-PPE. In various embodiments, the steroid is administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 hours after the administration of TIMP-PPE. In various embodiments, the steroid is administered 1, 2, 3, 4, 5, 6 or 7 days before the administration of TIMP-PPE. In various embodiments, the steroid is administered 1, 2, 3, 4, 5, 6 or 7 days after the administration of TIMP-PPE.In various embodiments, the steroid is selected from the group consisting of beclomethasone, ciclesonide, fluticasone furoate, mometasone, budenoside, fluticasone, triamcinolone, and loteprednol.

[0130] In various embodiments, the additional therapeutic agent is a corticosteroid. In various embodiments, the therapeutic agent useful for treating peanut allergy is a corticosteroid. In various embodiments, the corticosteroid administered in combination with TIMP-PPE is administered at a dose of about 0.05 mg to 2000 mg. In various embodiments, the corticosteroid is administered at a dose of about 0.05 mg, 0.1 mg, 0.5 mg, 1 mg, 2 mg, 4 mg, 5 mg, 10 mg, 25 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 375 mg, 400 mg, 425 mg, 450 mg, 475 mg, 500 mg, 1000 mg, 1500 mg, or 2000 mg. In various embodiments, the corticosteroid is administered in a single dose or multiple doses. In various embodiments, the corticosteroid is administered once a day, once a week, once every two weeks, once every three weeks, or once every four weeks. In various embodiments, the corticosteroid is administered two, three, four, five, or six times a day. In various embodiments, the corticosteroid is administered before, concomitantly with, or after the administration of TIMP-PPE. In various embodiments, the corticosteroid is administered one, two, three, four, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, or 12 months before the administration of TIMP-PPE. In various embodiments, the corticosteroid is administered one, two, three, four, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, or 12 months after the administration of TIMP-PPE. In various embodiments, the corticosteroid is administered 5, 10, 15, 30, 45, or 60 minutes before the administration of TIMP-PPE. In various embodiments, the corticosteroid is administered 5, 10, 15, 30, 45, or 60 minutes after the administration of TIMP-PPE. In various embodiments, the corticosteroid is administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 hours before the administration of TIMP-PPE. In various embodiments, the corticosteroid is administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 hours after the administration of TIMP-PPE. In various embodiments, the corticosteroid is administered 1, 2, 3, 4, 5, 6, or 7 days before the administration of TIMP-PPE.In various embodiments, a corticosteroid is administered 1, 2, 3, 4, 5, 6, or 7 days after administration of the TIMP-PPE. In various embodiments, the corticosteroid is selected from the group consisting of cortisone, prednisone, prednisolone, methylprednisolone, dexamethasone, betamethasone, and hydrocortisone.

[0131] In various embodiments, the NSAID administered in combination with TIMP-PPE is administered in a dose of about 0.05 mg to 2000 mg. In various embodiments, the NSAID is administered in a dose of about 0.05 mg, 0.1 mg, 0.5 mg, 1 mg, 2 mg, 4 mg, 5 mg, 10 mg, 25 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 375 mg, 400 mg, 425 mg, 450 mg, 475 mg, 500 mg, 1000 mg, 1500 mg or 2000 mg. In various embodiments, the NSAID is administered in a single dose or multiple doses. In various embodiments, the NSAID is administered once a day, once a week, once every two weeks, once every three weeks or once every four weeks. In various embodiments, the NSAID is administered two, three, four, five or six times a day. In various embodiments, the NSAID is administered before, concomitantly with or after the administration of TIMP-PPE. In various embodiments, the NSAID is administered one, two, three, four, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 months before the administration of TIMP-PPE. In various embodiments, the NSAID is administered one, two, three, four, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 months after the administration of TIMP-PPE. In various embodiments, the NSAID is administered 5, 10, 15, 30, 45 or 60 minutes before the administration of TIMP-PPE. In various embodiments, the NSAID is administered 5, 10, 15, 30, 45 or 60 minutes after the administration of TIMP-PPE. In various embodiments, the NSAID is administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 36, 48, 72 or 96 hours before the administration of TIMP-PPE. In various embodiments, the NSAID is administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 36, 48, 72 or 96 hours after the administration of TIMP-PPE. In various embodiments, the NSAID is administered 1, 2, 3, 4, 5, 6 or 7 days before the administration of TIMP-PPE. In various embodiments, the NSAID is administered 1, 2, 3, 4, 5, 6 or 7 days after the administration of TIMP-PPE.In various embodiments, the NSAID is a non-selective NSAID, a COX-2 selective NSAID, or a COX-1 selective NSAID. In various embodiments, the NSAID is a prostaglandin synthase inhibitor. In various embodiments, the NSAID is selected from the group consisting of: diclofenac, diclofenac potassium, diclofenac sodium, diflunisal, etodolac, flurbiprofen, fenoprofen, fenoprofen calcium, ketorolac, ketorolac tromethamine, ketoprofen, tolmetin, tolmetin sodium, acetylsalicylic acid, aspirin, ibuprofen, naproxen, indomethacin, indomethacin sodium, sulindac, biphenylacetic acid, piroxicam, mefenamic acid, mefenamic acid sodium, meloxicam, nabumetone, oxaprozin, piroxicam, celecoxib, etodolac, etoricoxib, lumiracoxib, rofecoxib, valdecoxib.

[0132] In various embodiments, the leukotriene regulator administered in combination with TIMP-PPE is administered at a dose of about 0.05 mg to 2000 mg. In various embodiments, the leukotriene regulator is administered at a dose of about 0.05 mg, 0.1 mg, 0.5 mg, 1 mg, 2 mg, 4 mg, 5 mg, 10 mg, 25 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 375 mg, 400 mg, 425 mg, 450 mg, 475 mg, 500 mg, 1000 mg, 1500 mg, or 2000 mg. In various embodiments, the leukotriene regulator is administered in a single dose or multiple doses. In various embodiments, the leukotriene regulator is administered once a day, once a week, once every two weeks, once every three weeks, or once every four weeks. In various embodiments, the leukotriene regulator is administered two, three, four, five, or six times a day. In various embodiments, the leukotriene regulator is administered before, concomitantly with, or after the administration of TIMP-PPE. In various embodiments, the leukotriene regulator is administered one, two, three, four, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, or 12 months before the administration of TIMP-PPE. In various embodiments, the leukotriene regulator is administered one, two, three, four, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, or 12 months after the administration of TIMP-PPE. In various embodiments, the leukotriene regulator is administered 5, 10, 15, 30, 45, or 60 minutes before the administration of TIMP-PPE. In various embodiments, the leukotriene regulator is administered 5, 10, 15, 30, 45, or 60 minutes after the administration of TIMP-PPE. In various embodiments, the leukotriene regulator is administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 36, 48, 72, or 96 hours before the administration of TIMP-PPE. In various embodiments, the leukotriene regulator is administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 36, 48, 72, or 96 hours after the administration of TIMP-PPE. In various embodiments, the leukotriene regulator is administered 1, 2, 3, 4, 5, 6, or 7 days before the administration of TIMP-PPE. In various embodiments, the leukotriene regulator is administered 1, 2, 3, 4, 5, 6, or 7 days after the administration of TIMP-PPE.In various embodiments, the leukotriene modifier is an anti-leukotriene, a leukotriene receptor antagonist, or a leukotriene synthesis inhibitor. In various embodiments, the leukotriene modifier is selected from the group consisting of montelukast, zileuton, and zafirlukast.

[0133] Provided is administering to a subject in need thereof a TIMP-PPE alone or in combination with one or more additional therapeutic agents to alleviate one or more symptoms of peanut allergy. Symptoms of peanut allergy include skin reactions, hives, skin redness, skin swelling, itching, tightness in the throat, difficulty breathing, shortness of breath, and anaphylaxis. Additional symptoms include digestive problems such as diarrhea, stomach cramps, nausea, or vomiting, and a drop in blood pressure.

[0134] Provided is administering to a subject in need thereof a TIMP-PPE alone or in combination with one or more therapeutic agents useful for treating peanut allergy to alleviate one or more symptoms of peanut allergy. Symptoms of peanut allergy include skin reactions, hives, skin redness, skin swelling, itching, tightness in the throat, difficulty breathing, shortness of breath, digestive problems such as diarrhea, stomach cramps, nausea, or vomiting, a drop in blood pressure, and anaphylaxis.

[0135] Also contemplated is administering to a subject in need thereof a TIMP-PPE alone or in combination with one or more additional therapeutic agents to shorten the duration and reduce the severity of an allergic immune response to peanut protein or upon exposure to peanut protein. The allergic immune responses contemplated herein include Th2 T cell responses, B cell activation, basophil activation, eosinophil activation, mast cell activation, and / or IgE induction. Additional immune responses include T cell-dependent mechanisms involving upregulation of the production of type 2 T helper (Th2) cytokines (e.g., IL-4, IL-5, IL-9, and IL-13), B cell class switching leading to the production of IgE antibodies, and / or the IgE:IgG ratio.

[0136] Also contemplated is administering to a subject in need thereof a TIMP-PPE alone or in combination with one or more therapeutic agents useful for treating peanut allergy to shorten the duration and reduce the severity of an allergic immune response to peanut protein or upon exposure to peanut protein. The allergic immune responses contemplated herein include T cell-dependent mechanisms involving upregulation of the production of type 2 T helper (Th2) cytokines (e.g., IL-4, IL-5, IL-9, and IL-13), B cell activation, B cell class switching leading to the production of IgE antibodies, basophil activation, eosinophil activation, mast cell activation, and / or the IgE:IgG ratio. Screening methods

[0137] It is contemplated to monitor the induction and maintenance of immune tolerance in a subject having a peanut allergy, the subject being treated or about to be treated with an antigen-specific tolerance therapy consisting of TIMPs encapsulating peanut allergens as described herein.

[0138] Methods for screening cell types, cytokines, or other tolerance metrics from subjects undergoing tolerance therapy as described herein are known in the art. Methods for assessing tolerance are accomplished using techniques such as flow cytometry, mass cytometry (CyTOF), ELISA, ELISPOT, in vitro / ex vivo cell stimulation assays (including but not limited to cell proliferation assays, basophil activation tests (BAT), macrophage stimulation assays), measuring autoantibodies, or measuring Ig serotypes (e.g., by ImmunoCap assay), among others.

[0139] In various embodiments, the immune tolerance status of a subject is determined based on assays of one or more biological samples from the subject. Biological samples include whole blood, peripheral blood, peripheral blood mononuclear cells (PBMC), serum, plasma, urine, cerebrospinal fluid (CSF), feces, tissue biopsy, and / or bone marrow biopsy. In various embodiments, assays of biological samples include analyzing the levels and / or presence or absence of cell surface proteins, extracellular proteins, intracellular proteins, nucleic acids, metabolites, and / or combinations thereof.

[0140] Cells assayed from biological samples include immune cells, non-immune cells, and / or combinations thereof. Immune cells include innate immune cells, adaptive immune cells, and / or combinations thereof. Innate immune cells assayed from biological samples include antigen-presenting cells (APC). Exemplary innate immune cells assayed from biological samples include monocytes, macrophages, neutrophils, granulocytes, dendritic cells, mast cells, eosinophils, basophils, and / or combinations thereof. Adaptive immune cells assayed from biological samples include effector immune cells such as CD4+ T cells, CD8+ T cells, B cells, NK cells, NK-T cells, and / or combinations thereof. In various embodiments, the T cells are Th1 cells, Th2a cells, Treg cells, and Tr1 cells.

[0141] In certain embodiments, the cells assayed from biological samples are epithelial cells, stromal cells, endothelial cells, fibroblasts, pericytes, adipocytes, mesenchymal stem cells, hematopoietic stem cells, hematopoietic progenitor cells, liver sinusoidal endothelial cells (LSEC), and / or Kupffer cells.

[0142] One aspect of the immune tolerance status and immune profile of a subject is determined by analyzing one or more proteins from one or more biological samples from the subject. In various embodiments, the protein is a cytokine and / or chemokine. In various embodiments, the protein is a cell signaling protein. In various embodiments, the cytokines and chemokines are selected from the group consisting of: IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-12p70, IL-13, IL-14, IL-15, IL-16, IL-17, IL-17, IL-18, IL-20, IL-21, IL-22, IL-23, IL-24, IL-25, IL-26, IL-27, IL-27b, IL-28, IL-29, IL-30, IL-31, IL-32, IL-33, IL-35, IL-36, CCL1, CCL2, CCL3, CCL4, CCL5, CCL6, CCL7, CCL8, CCL9, CCL10, CCL11, CCL12, CCL14, CCL15, CCL16, CCL17, CCL18, CCL19, CCL20, CCL21, CCL22, CCL23, CCL24, CCL25, CCL26, CCL27, CCL28, CXCL1, CXCL2 (MCP-1), CXCL3 (MIP-1α), CXCL4 (MIP-1β), CXCL5 (RANTES), CXCL6, CXCL7, CXCL8, CXCL9, CXCL10, CXCL11, CXCL12, CXCL13, CXCL14, CXCL15, CXCL16, CXCL17, GM-CSF, IFN-α, IFN-β, IFN-γ, TNF-α, TGF-β1, TGF-β2, TGF-β3, soluble CD14, and / or combinations thereof.

[0143] In various embodiments, the protein is a protease. In various embodiments, the protease is an aspartic protease, a cysteine protease, a metalloprotease, a serine protease, and / or a threonine protease. In various embodiments, the protease is selected from the group consisting of: ADAM1, ADAM2, ADAM7, ADAM8, ADAM9, ADAM10, ADAM11, ADAM12, ADAM15, ADAM17, ADAM18, ADAM19, ADAAM20, ADAM21, ADAM22, ADAM23, ADAM28, ADAM29, ADAM30, ADAM33, MMP1, MMP2, MMP3, MMP7, MMP8, MMP9, MMP10, MMP11, MMP12, MMP13, MMP14, MMP15, MMP16, MMP17, MMP18, MMP19, MMP20, MMP21, MMP23A, MMP23B, MMP24, MMP25, MMP26, MMP27, and MMP28. In various embodiments, the proteins associated with apoptosis are selected from the group consisting of: P53, caspase 1, caspase 2, caspase 3, caspase 4, caspase 5, caspase 6, caspase 7, caspase 8, caspase 9, caspase 10, caspase 11, caspase 12, caspase 13, caspase 14, BCL-2, BCL-XL, MCL-1, CED-9, A1, BFL1, BAX, BAK, DIVA, BCL-XS, BIK, BIM, BAD, BID, and EGL-1. Several methods for determining proteins in biological samples have been described in the literature, including enzyme-linked immunosorbent assay (ELISA), Western blotting, and mass spectrometry. In various embodiments, the protein is one or more immunoglobulins (Ig). In various embodiments, the Ig is selected from the group consisting of IgA, IgD, IgE, IgM, and / or variants thereof. In various embodiments, the immunoglobulin is antigen-specific. Several methods for detecting immunoglobulins in biological samples have been described in the literature, including ELISA and ImmunoCap.

[0144] Lists of human metabolites measured from biological samples can be found in the literature, including (Psychogios et al., 2011), (Wishart et al., HMDB: Human Metabolome Database, Nucleic Acids Res. January 2007; 35 (Database issue): D521-6, 2007), and the Human Metabolome Database (HMDB), and are hereby incorporated by reference.

[0145] Determining an aspect of the immune tolerance status and immune markers of a subject by analyzing one or more cell surface proteins from a biological sample. In various embodiments, the cell surface proteins include CD1c, CD2, CD3, CD4, CD5, CD8, CD9, CD10, CD11b, CD11c, CD14, CD15, CD16, CD18, CD19, CD20, CD21, CD22, CD23, CD24, TACI, CD25, CD27, CD28, CD30, CD30L, CD31, CD32, CD32b, CD34, CD33, CD38, CD39, CD40, CD40-L, CD41b, CD42a, CD42b, CD43, CD44, CD45, CD45RA, CD47, CD45RA, CD45RO, CD48, CD52, CD55, CD56, CD58, CD61, CD66b, CD69, CD70, CD72, CD79, CD68, CD84, CD86, CD93, CD94, CD95, CRACC, BLAME, BCMA, CD103, CD107, CD112, CD120a, CD120b, CD123, CD125, CD127, CD134, CD135, CD140a, CD141, CD154, CD155, CD160, CD161, CD163, CD172a, XCR1, CD203c, CD204, CD206, CD207, CD226, CD244, CD267, CD268, CD269, CD355, CD358, CRTH2, NKG2A, NKG2B, NKG2C, NKG2D, NKG2E, NKG2F, NKG2H, KIR2DL1, KIR2DL2, KIR2DL3, KIR2DL5A, KIR2DL5B, KIR3DL1, KIR3DL2, KIR3DL3, KIR3DL4, KIR2DS1, KIR2DS2, KIR2DS3, KIR2DS4, KIR2DS5, DAP12, KIR3DS, NKp44, NKp46, TCR, BCR, integrin, FcβεRI, MHC-I, MHC-II, IL-1R, IL-2Rα, IL-2Rβ, IL-2Rγ, IL-3Rα, CSF2RB, IL-4R, IL-5Rα, CSF2RB, IL-6Rα, gp130, IL-7Rα, IL-9R, IL-10R, IL-12Rβ1, IL-12Rβ2, IL-13Rα1, IL-13Rα2, IL-15Rα, IL-21R, IL-23R, IL-27Rα, IL-31Rα, OSMR, CSF-1R, cell surface IL-15,IL-10Rα, IL-10Rβ, IL-20Rα, IL-20Rβ, IL-22Rα1, IL-22Rα2, IL-22Rβ, IL-28RA, PD-1, PD-1H, BTLA, CTLA-4, PD-L1, PD-L2, 2B4, B7-1, B7-2, B7-H1, B7-H4, B7-DC, DR3, LIGHT, LAIR, LTα1β2, LTβR, TIM-1, TIM-3, TIM-4, TIGIT, LAG-3, ICOS, ICOS-L, SLAM, SLAMF2, OX-40, OX-40L, GITR, GITRL, TL1A, HVEM, 41-BB, 41BB-L, TL-1A, TRAF1, TRAF2, TRAF3, TRAF5, BAFF, BAFF-R, APRIL, TRAIL, RANK, AITR, TRAMP, CCR1, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, CCR10, CCR11, CXCR1, CXCR2, CXCR3, CXCR4, CXCR5, CXCR6, CXCR7, CLECL9a, DC-SIGN, IGSF4A, SIGLEC, EGFR, PDGFR, VEGFR, FAP, α-SMA, FAS, FAS-L, FC, ICAM-1, ICAM-2, ICAM-3, ICAM-4, ICAM-5, PECAM-1, MICA, MICB, UL16, ULBP1, ULBP2, ILBP3, ULBP4, ULBP5, ULBP6, MULT1, RAE1 α, β, γ, δ and ε, H60a, H60b, H60c, GPR15, ST2 and / or combinations thereof. Integrins include α1, α2, αIIb, α3, α4, α5, α6, α7, α8, α9, α10, α11, αD, αE, αL, αM, αV, αX, β1, β2, β3, β4, β5, β6, β7, β8 and / or combinations thereof. TCRs include α, β, γ, δ, ε, ζ chains and / or combinations thereof. Several methods for determining cell surface protein expression have been described in the literature, including flow cytometry and mass cytometry (CyTOF).

[0146] One aspect of the immune tolerance status and immune profile of a subject is determined by analyzing one or more metabolites from a biological sample. In various embodiments, the metabolite is an inflammatory metabolite. In various embodiments, the metabolite is an anti-inflammatory metabolite. In various embodiments, examples of inflammatory metabolites include acids, lipids, sugars, amino acids, lactate, trimethylamine N-oxide, O-acetylcarnitine, L-carnitine, choline, succinate, glutamine, fatty acids, cholesterol, 3-hydroxybutyric acid, 3'-sialyllactose, arachidonic acid, prostaglandins (G2 and H2), PGD2, PGE2, PGF2a, PGI2, TXA2, leukotrienes (A4, B4, C4, D4, E4), kynurenine, 3-hydroxykynurenine, lipoxin A4, and lipoxin B4. In various embodiments, examples of anti-inflammatory metabolites include 2-amino-3-carboxymuconic acid 6-semialdehyde, picolinic acid, anthranilic acid, 3-hydroxyanthranilic acid, glutaryl co-A, NAD+, quinolinic acid, arginine, butyrate, and adenosine. A list of human metabolites that can be assayed from biological samples can be found in the literature, including (Psychogios et al., 2011 PLoS One 6(2):e16957), (Wishart et al., HMDB: Human Metabolome Database Nucleic Acids Res. January 2007; 35(Database issue):D521-6, 2007), and the Human Metabolome Database (HMDB), each of which is incorporated herein by reference in its entirety.

[0147] In certain embodiments, the tolerance status of a subject is determined by analyzing nucleic acids from a biological sample. In various embodiments, the nucleic acids are DNA and / or RNA, including but not limited to single-stranded DNA, double-stranded DNA, mRNA, rRNA, tRNA, siRNA, miRNA, long non-coding RNA (long ncRNA, lncRNA), and non-coding RNA (ncRNA), mitochondrial RNA. In various embodiments, the immune tolerance status of a subject is determined by assaying gene expression from a biological sample. In various embodiments, the immune tolerance status is determined by assaying gene expression associated with immune function, antibodies, foreign body response, metabolism, apoptosis, cell death, necrosis, ferroptosis, autophagy, cell migration, endocytosis, phagocytosis, pinocytosis, tight junction regulation, cell adhesion, differentiation, and / or combinations thereof. In various embodiments, the immune tolerance status is determined by assaying gene expression associated with immunosuppression. In various embodiments, the immune tolerance status is determined by assaying gene expression associated with immune activation. In various embodiments, the immune tolerance status is determined by assaying gene expression associated with immune regulatory function. In various embodiments, nucleic acid analysis is used to generate immune tolerance markers. Several methods for high-throughput gene expression analysis have been described in the literature, including RNA sequencing (RNA-seq), single-cell RNA sequencing (scRNA-seq), exome sequencing, and microarray-based analysis.

[0148] After in vivo and / or ex vivo stimulation with one or more stimuli such as an antigen, an allergen, and one or more activators, a biological sample is optionally assayed. It is contemplated that the T cells, B cells, and immunoglobulins used in the assay are antigen-specific. Exemplary T cells include effector memory T cells, antigen-specific T cells, activated antigen-specific T cells, Th1 cells, pathogenic Th2a+ cells, Th17 cells, T follicular helper (TFH) cells, TH0 cells, or other antigen-specific T cells. B cells include effector B cells, memory B cells, plasma B cells, and regulatory B (Breg) cells. In certain embodiments, T cells or B cells are identified based on the expression of the proteins described in Table 1.

[0149] In various embodiments, the immune tolerance status of a subject is determined by obtaining one or more samples (e.g., whole blood) from the subject on the day of the first administration of TIMP-PPE (Day 1) before dosing, 14 days after the administration of the second dose, and then every 90 days after the second dose (e.g., Days 90, 180, 270, and 360 after the second dose). The whole blood can then be processed to isolate peripheral blood mononuclear cells (PBMCs), basophils, neutrophils, plasma, and serum for downstream analysis. Assays are performed on cells isolated from one or more samples collected from the subject and analyzed using the methods described below.

[0150] In various embodiments, the immune tolerance status of a subject is determined by obtaining one or more samples, such as whole blood, from the subject before dosing on the day of the first TIMP-PPE administration (Day 1), on Day 15 (14 days after the second dose), on Day 60, and then optionally every 90 days after the second dose (e.g., Days 90, 180, 270, and 360 after the second dose). The whole blood can then be processed to isolate peripheral blood mononuclear cells (PBMCs), basophils, neutrophils, plasma, and serum for downstream analysis. Assays and analysis are performed on cells isolated from one or more samples collected from the subject using such methods as described below.

[0151] In various embodiments, the immune tolerance status of a subject determined prior to the administration of TIMP-PPE serves as a baseline. In various embodiments, the baseline of a subject is determined by assaying one or more biological samples 1, 2, 3, 4, 5, 6, or 7 days before the administration of TIMP-PPE. In various embodiments, the baseline of a subject is determined by assaying one or more biological samples 1, 2, 3, or 4 weeks before the administration of TIMP-PPE. In various embodiments, the baseline of a subject is determined by assaying one or more biological samples 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months before the administration of TIMP-PPE.

[0152] In various embodiments, the immune tolerance status of a subject is determined after administration of TIMP-PPE. In various embodiments, the immune tolerance status of a subject is determined by assaying one or more biological samples at 1, 2, 3, 4, 5, 6, or 7 days after administration of TIMP-PPE. In various embodiments, immune tolerance is determined by assaying one or more biological samples at 1, 2, 3, or 4 weeks after administration of TIMP-PPE. In various embodiments, the immune tolerance of a subject is determined by assaying one or more biological samples at 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months after administration of TIMP-PPE. In various embodiments, the immune tolerance status of a subject determined after administration of TIMP-PPE is compared to a baseline. In various embodiments, the immune tolerance status of a subject determined after administration of TIMP-PPE is compared to the immune tolerance status of a healthy subject or a subject administered a placebo.

[0153] In various embodiments, the immune tolerance status of a subject is determined after administration of TIMP-PPE, whether a primary or initial dose of TIMP-PPE or a booster dose of TIMP-PPE.

[0154] The following assays are envisioned to track the immune status and tolerance induction of subjects receiving TIMP-PPE tolerance therapy.

[0155] For example, by flow cytometry, the proportion of peanut-specific Th2a+ cells (Th2a+ cells / total peanut-specific T cells) stimulated ex vivo with purified peanut antigen can be measured. Th2a+ cells are defined as CRTH2+ / CD161+ / CD154+ / CD27-. Total peanut-reactive cells are defined as CRTH2- / CD161+ / CD154+ / CD27-. In various embodiments, relative to placebo administration and / or one or more baseline measurements taken from the subject during treatment, administration of TIMP-PPE to the subject maintains or reduces peanut-specific Th2a+ cells by about 1% - 100% (e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95% or about 100%, including all values and ranges between these values), 10 - 95%, 15 - 90%, 20 - 85%, 25 - 75%, 30 - 70%, 35 - 65%, 40 - 60%, 45 - 55% or 50% or about 2 - 100-fold (e.g., about 2, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 or 100-fold, including all values and ranges between these values).

[0156] The proportion of activated peanut-specific T cells (activated peanut-specific T cells / total peanut-specific T cells) after ex vivo stimulation with peanut protein is determined by flow cytometry. Activated peanut-specific T cells are defined as CD154+ / CD38+. Unactivated peanut-specific T cells are defined as CD154+. In various embodiments, relative to placebo administration and / or one or more baseline measurements taken from the subject during treatment, administration of TIMP-PPE to the subject maintains or reduces activated peanut-specific T cells by about 1% - 100% (e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95% or about 100%, including all values and ranges between these values), 10 - 95%, 15 - 90%, 20 - 85%, 25 - 75%, 30 - 70%, 35 - 65%, 40 - 60%, 45 - 55% or 50% or about 2 - 100-fold (e.g., about 2, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 or 100-fold, including all values and ranges between these values).

[0157] The frequency of T regulatory cell populations (CD4+ / CD25+ / FoxP3+ / Helios+ / IL-10+) or (CD4+CD45RA 低 CD4+CD137+ CD25+ CD127 低 ) was determined by flow cytometry. Multicolor flow analysis was performed to provide the proportion of peanut-specific T regulatory cells (peanut-specific T regulatory cells / peanut-specific CD4+ effector memory cells). In various embodiments, administration of TIMP-PPE to a subject increased Treg cells by about 1% - 100% (e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95% or about 100%, including all values and ranges between these values), 10 - 95%, 15 - 90%, 20 - 85%, 25 - 75%, 30 - 70%, 35 - 65%, 40 - 60%, 45 - 55% or 50% or about 2 - 100-fold (e.g., about 2, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 or 100-fold, including all values and ranges between these values), relative to placebo administration and / or one or more baseline measurements taken from the subject during treatment.

[0158] The ratio of IL-5 to IFN-γ in the cell culture supernatant of PBMCs was measured, for example, as detected by Luminex 200.

[0159] The following indicators of the immunotolerant state can be examined by determining basophils isolated from one or more blood samples collected from a subject and stimulated ex vivo with purified antigenic peanut proteins: the proportion of CD203+ / CD63+ basophils activated after stimulation ex vivo with purified antigenic peanut proteins using the basophil activation test (BAT) (Santos and Lack 2016 Clin Transl Allergy. 6:10) and the effective concentration (EC50) at 50% maximum basophil activation after stimulation ex vivo with purified antigenic peanut proteins measured using the basophil activation test, where the activated basophils are CD203+ / CD63+ / -). Analysis was performed to provide the effective concentration at 50% maximum basophil activation (EC50).

[0160] The following indicators of the immune tolerance status can be examined by determining serum isolated from one or more blood samples obtained from a subject: the ratio of peanut-specific IgE to IgG measured by ImmunoCap assay. In various embodiments, relative to placebo administration and / or one or more baseline measurements taken from the subject during treatment, administration of TIMP-PPE to the subject reduces the peanut-specific IgE:IgG ratio by about 1% - 100% (e.g., about 1%, about 2%, about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95% or about 100%, including all values and ranges between these values), 10 - 95%, 15 - 90%, 20 - 85%, 25 - 75%, 30 - 70%, 35 - 65%, 40 - 60%, 45 - 55% or 50% or about 2 - 100-fold (e.g., about 2, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 or 100-fold, including all values and ranges between these values).

[0161] The change in peanut-specific IgE measured by ImmunoCap assay. In various embodiments, relative to placebo administration and / or one or more baseline measurements taken from the subject during treatment, administration of TIMP-PPE to the subject maintains or reduces the peanut-specific IgE level by about 1% - 100% (e.g., about 1%, about 2%, about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95% or about 100%, including all values and ranges between these values), 10 - 95%, 15 - 90%, 20 - 85%, 25 - 75%, 30 - 70%, 35 - 65%, 40 - 60%, 45 - 55% or 50% or about 2 - 100-fold (e.g., about 2, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 or 100-fold, including all values and ranges between these values).

[0162] The following indicators of the immunotolerant state can be examined by determining basophils isolated from one or more blood samples collected from a subject and stimulated ex vivo with purified antigenic peanut protein: the proportion of activated CD203+ / CD63+ basophils after stimulation ex vivo with purified antigenic peanut protein using the basophil activation test (BAT) (Santos and Lack 2016 Clin Transl Allergy 6: 10) and the effective concentration (EC50) at 50% of the maximum basophil activation after stimulation ex vivo with purified antigenic peanut protein measured using the basophil activation test, wherein the activated basophils are CD203+ / CD63+ / - . An analysis is performed to provide the effective concentration at 50% of the maximum basophil activation (EC50). In various embodiments, administration of TIMP-PPE reduces basophil activation. In various embodiments, administration of TIMP-PPE to a subject increases the EC50 of the maximum basophil activation by about 1% - 100% (e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95% or about 100%, including all values and ranges between these values), 10 - 95%, 15 - 90%, 20 - 85%, 25 - 75%, 30 - 70%, 35 - 65%, 40 - 60%, 45 - 55% or 50% or about 2 - 10,000 fold (e.g., about 2, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 1000 or 10,000 fold, including all values and ranges between these values), relative to placebo administration and / or one or more baseline measurements taken from the subject during treatment.

[0163] In certain embodiments, the efficacy of TIMP-PPE in alleviating one or more symptoms of peanut allergy, increasing tolerance to peanut protein, and / or shortening the duration and reducing the severity of the allergic immune response to peanut protein is determined by double-blind placebo-controlled food challenge (DBPCFC), changes in the cumulative tolerated dose of peanut protein administered during DBPCFC, and / or skin prick test (SPT). Procedures for performing DBPCFC and SPT have been previously described (Sampson et al., Journal of Allergy and Clinical Immunology 2012;130(6):1260-1274; Heinzerling et al., “The skin prick test - European standards,” Clinical and Translational Allergy 2013;3(1):3).

[0164] The efficacy of TIMP-PPE in alleviating one or more symptoms of peanut allergy and / or reducing the duration and severity of the allergic immune response to peanut protein is determined by assaying one or more biological samples from the subject. Biological samples include whole blood, peripheral blood, peripheral blood mononuclear cells (PBMC), serum, plasma, urine, cerebrospinal fluid (CSF), feces, tissue biopsy, and / or bone marrow biopsy. In various embodiments, the assay of the biological sample includes analyzing the levels and / or presence or absence of cell surface proteins, extracellular proteins, intracellular proteins, nucleic acids, metabolites, and / or combinations thereof.

[0165] One or more of the following parameters assayed in one or more biological samples obtained from the subject and stimulated in vivo and / or ex vivo are used to generate an immune tolerance marker for the subject: a. The proportion of effector T cells in the total T cell population; b. The proportion of Treg cells in the total T cell population; c. The proportion of effector B cells in the total B cell population; d. The levels of specific IgG, IgA, IgM, and / or IgE; e. The levels of inflammatory cytokines and chemokines; f. The levels of anti-inflammatory cytokines and chemokines; g. The levels of inflammatory metabolites; and h. The levels of anti-inflammatory metabolites.

[0166] If one, two, three, four, five, six, seven, or eight of the parameters listed in (a) through (h) above indicate maintenance of immune tolerance, the immune tolerance marker indicates maintenance of immune tolerance. In various embodiments, if at least 2 / 8 of the parameters listed in (a) through (h) above indicate maintenance of immune tolerance, the immune tolerance marker indicates maintenance of immune tolerance. In various embodiments, if one, two, three, four, five, six, seven, or eight of the parameters listed in (a) through (h) above indicate maintenance of immune tolerance, it is determined that the subject does not require treatment with TIMP. In various embodiments, if at least 3 / 8 of the parameters listed in (a) through (h) above indicate maintenance of immune tolerance, it is determined that the subject does not require treatment with TIMP.

[0167] If:

[0168] a. The proportion of effector T cells in the total T cell population is 5% - 100% (e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100%, including all values and the ranges between these values); and / or

[0169] b. The proportion of Treg cells in the total T cell population is 1 - 3%; and / or

[0170] c. The proportion of effector B cells in the total B cell population is 5% - 100% (e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100%, including all values and the ranges between these values); and / or

[0171] d. The levels of IgG, IgA, IgM, and / or IgE are increased by about 5% - 100% (e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100%, including all values and ranges between these values), 10 - 95%, 15 - 90%, 20 - 85%, 25 - 75%, 30 - 70%, 35 - 65%, 40 - 60%, 45 - 55%, or 50% or about 2 - 100 - fold (e.g., about 2, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 - fold, including all values and ranges between these values) relative to healthy subjects and / or one or more baseline measurements taken from the subject during treatment; and / or

[0172] e. The levels of inflammatory cytokines / chemokines are increased by about 5% - 100% (e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100%, including all values and ranges between these values), 10 - 95%, 15 - 90%, 20 - 85%, 25 - 75%, 30 - 70%, 35 - 65%, 40 - 60%, 45 - 55%, or 50% or about 2 - 100 - fold (e.g., about 2, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 - fold, including all values and ranges between these values) relative to healthy subjects and / or one or more baseline measurements taken from the subject during treatment; and / or

[0173] f. The levels of anti-inflammatory cytokines and chemokines are reduced by about 5% - 100% (e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95% or about 100%, including all values and ranges between these values), 10 - 95%, 15 - 90%, 20 - 85%, 25 - 75%, 30 - 70%, 35 - 65%, 40 - 60%, 45 - 55% or 50% or about 2 - 100-fold (e.g., about 2, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 or 100-fold, including all values and ranges between these values) relative to healthy subjects and / or one or more baseline measurements taken from the subject during treatment; and / or

[0174] g. The levels of inflammatory metabolites are increased by about 5% - 100% (e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95% or about 100%, including all values and ranges between these values), 10 - 95%, 15 - 90%, 20 - 85%, 25 - 75%, 30 - 70%, 35 - 65%, 40 - 60%, 45 - 55% or 50% or about 2 - 100-fold (e.g., about 2, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 or 100-fold, including all values and ranges between these values) relative to healthy subjects and / or one or more baseline measurements taken from the subject during treatment; and / or

[0175] h. The level of the anti-inflammatory metabolite is reduced by about 5% - 100% (e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95% or about 100%, including all values and ranges between these values), 10 - 95%, 15 - 90%, 20 - 85%, 25 - 75%, 30 - 70%, 35 - 65%, 40 - 60%, 45 - 55% or 50% or about 2 - 100-fold (e.g., about 2, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 or 100-fold, including all values and ranges between these values) relative to healthy subjects and / or one or more baseline measurements taken from the subject during treatment, then the immune tolerance markers of the subject generated using one or more of the parameters described herein indicate a weakening and / or absence of immune tolerance before or after treatment with TIMP-PPE.

[0176] It is assumed that the proportion of Th2a+ cells at the time point of Day 1 before dosing in subjects with peanut allergy is expected to be > 15%. Treatment with TIMP-PPE is expected to reduce the proportion of Th2a+ cells to < 15% 14 days after the second dose, which indicates the induction of immune tolerance. At any subsequent time point (e.g., Day 90, 180, 270 and 360 after dosing), an increase in the proportion of Th2a+ cells to > 15% will indicate a weakening of immune tolerance and require re-administration of TIMP-PPE to restore immune tolerance. Collectively, the results from the above analysis can be used to determine immune tolerance markers and whether the subject has maintained immune tolerance. If such analysis indicates a weakening and / or loss of immune tolerance, TIMP-PPE can be re-administered to the subject to restore immune tolerance. Drug formulation

[0177] Depending on the route of administration, the pharmaceutical compositions of the present disclosure containing the TIMP-PPE described herein as an active ingredient may contain pharmaceutically acceptable carriers or additives. Examples of such carriers or additives include water, pharmaceutically acceptable organic solvents, collagen, polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymer, sodium carboxymethylcellulose, sodium polyacrylate, sodium alginate, water-soluble dextran, sodium carboxymethyl starch, pectin, methylcellulose, ethylcellulose, xanthan gum, gum arabic, casein, gelatin, agar, diglycerol, glycerol, propylene glycol, polyethylene glycol, petrolatum, paraffin, stearyl alcohol, stearic acid, human serum albumin (HSA), mannitol, sorbitol, lactose, pharmaceutically acceptable surfactants, etc. The additives used are selected from, but not limited to, the above or combinations thereof, and depending on the dosage form of the present disclosure as appropriate.

[0178] The formulation of the pharmaceutical composition will vary depending on the selected route of administration (e.g., solution, emulsion). Suitable compositions containing the therapeutic agent to be administered can be prepared in a physiologically acceptable vehicle or carrier. For solutions or emulsions, suitable carriers include, for example, aqueous solutions or alcoholic / aqueous solutions, emulsions or suspensions, including saline and buffered media. Extrinsic media can include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's solution or non-volatile oils. Intravenous media can include various additives, preservatives or fluids, nutrient or electrolyte supplements.

[0179] Various aqueous carriers can be used, for example, sterile phosphate buffered saline solution, bacteriostatic water, water, buffered water, 0.4% saline, 0.3% glycine, etc., and may include other proteins for enhanced stability such as albumin, lipoprotein, globulin, etc. that have been mildly chemically modified, etc.

[0180] Therapeutic formulations of the inhibitor are prepared for storage by mixing an inhibitor having the desired degree of purity with optional physiologically acceptable carriers, excipients or stabilizers (Remington's Pharmaceutical Sciences, 16th Edition, Osol, A. Ed. (1980)) in the form of a lyophilized formulation or aqueous solution. Acceptable carriers, excipients or stabilizers are non-toxic to recipients at the dosages and concentrations employed and include: buffers, such as phosphates, citrates and other organic acids; antioxidants, including ascorbic acid and methionine; preservatives (such as octadecyl dimethyl benzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butanol or benzyl alcohol; alkyl parabens, such as methylparaben or propylparaben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin or immunoglobulins; hydrophilic polymers, such as polyvinylpyrrolidone; amino acids, such as glycine, glutamine, asparagine, histidine, arginine or lysine; monosaccharides, disaccharides and other carbohydrates, including glucose, mannose or dextrin; chelating agents, such as EDTA; sugars, such as sucrose, mannitol, trehalose or sorbitol; salt-forming counterions, such as sodium; metal complexes (e.g., zinc-protein complexes); and / or nonionic surfactants, such as TWEEN™, PLURONICS™ or polyethylene glycol (PEG).

[0181] Formulations for in vivo administration must be sterile. This is readily accomplished by filtration through sterile filtration membranes.

[0182] Aqueous suspensions may contain the active compounds mixed with excipients suitable for the manufacture of aqueous suspensions. Such excipients are suspending agents, for example, sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia; dispersing or wetting agents may be a naturally-occurring phosphatide (e.g., lecithin), or a condensation product of an alkylene oxide with a fatty acid (e.g., polyoxyethylene stearate), or a condensation product of ethylene oxide with a long chain aliphatic alcohol (e.g., heptadecaethyleneoxycetanol), or a condensation product of ethylene oxide with a partial ester derived from fatty acids and hexitol anhydrides (e.g., polyoxyethylene sorbitan monooleate), or a condensation product of ethylene oxide and a partial ester derived from fatty acids and hexitol anhydrides (e.g., polyoxyethylene sorbitan monooleate). Aqueous suspensions may also contain one or more preservatives, such as ethyl p-hydroxybenzoate or n-propyl p-hydroxybenzoate.

[0183] The TIMP-PPE described herein can be stored in a lyophilized form and reconstituted in a suitable carrier prior to use.

[0184] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the modified particles are mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dibasic calcium phosphate and / or (a) fillers or extenders such as starch, lactose, sucrose, glucose, mannitol, and silicic acid, (b) binders such as, for example, carboxymethylcellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and acacia, (c) humectants such as glycerin, (d) disintegrants such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, (e) solution blockers such as paraffin, (f) absorption promoters such as quaternary ammonium compounds, (g) wetting agents such as, for example, cetyl alcohol and glyceryl monostearate, (h) adsorbents such as kaolin and bentonite, and (i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may also contain buffering agents. Kit

[0185] As a further aspect, the present disclosure includes kits that contain one or more compounds or compositions packaged in a manner that facilitates their use in practicing the methods of the present disclosure. In one embodiment, such a kit includes a compound or composition described herein (e.g., a composition containing TIMP alone or in combination with a second reagent), packaged in a container (such as a sealed bottle or vessel), with a label attached to the container or included in the packaging that describes the use of the compound or composition in practicing the method. Preferably, the compound or composition is packaged in unit dosage form. The kit may further include a device suitable for administering the composition according to a particular route of administration or for performing a screening assay. Preferably, the kit contains a label that describes the use of the inhibitor composition.

[0186] Further aspects and details of the present disclosure will become apparent from the following examples, which are intended to be illustrative and not limiting. Example 1 - Phase I / II Trial of TIMP-PPE in Peanut Allergy

[0187] This example describes a two-part, randomized, double-blind, placebo-controlled Phase 1b / 2a study to evaluate the safety, tolerability, pharmacodynamics, and efficacy of TIMP-PPE (CNP-201) in peanut-allergic subjects aged 16 to 55 years.

[0188] CNP-201 consists of PLGA nanoparticles encapsulating a purified peanut extract. The average diameter of the CNP-201 particles is 400 - 800 nm, and the negative ζ potential is between -32 mV and -50 mV. The CNP-201 particles are provided in a lyophilized formulation. The CNP-201 particles are reconstituted in sterile water for injection and diluted with sterile saline (0.9% sodium chloride) before administration.

[0189] This study consists of Part A and Part B.

[0190] Part A: Part A is a randomized, double-blind, placebo-controlled study of the safety of CNP-201 at escalating dose levels for tolerance. Part A will enroll 3 cohorts to receive CNP-201 or placebo at multiple escalating dose levels. Part B follows a randomized, double-blind, placebo-controlled, repeated-dose study using the safe and tolerable dose level of CNP-201 determined from Part A.

[0191] Subjects who meet all inclusion and no exclusion criteria after the initial screening assessment undergo a skin prick test (SPT), followed by a baseline double-blind, placebo-controlled food challenge (DBPCFC) (peanut and placebo (oat) challenges, administered over two days) to confirm peanut allergy. The SPT and DBPCFC are performed by a study physician or a staff member trained in the management of on-site clinical emergencies in the immediate availability of emergency medications and equipment and near the hospital emergency department to provide rapid emergency care if needed. Subjects who continue to meet all inclusion and no exclusion criteria two days after completing the DBPCFC are eligible to participate in this study.

[0192] All subjects who continue to meet all I / E criteria after the DBPCFC receive subcutaneous injection of omalizumab (XOLAIR®). The dose of omalizumab (XOLAIR®) follows the product label specified in the protocol and is determined by the subject's body weight and serum IgE at the initial screening. Subjects are dosed every 2 weeks or every 4 weeks according to the product label.

[0193] Subjects who continue to meet the inclusion / exclusion criteria are randomized at a ratio of 2:1 (Part A) or 1:1 (Part B) on Day 1 to receive CNP-201 or placebo (0.9% sodium chloride USP) by intravenous (IV) infusion. On Day 1 and Day 8, the subjects are dosed with CNP-201 or placebo.

[0194] Peanut-allergic subjects eligible to participate in this study are defined based on the following inclusion criteria: 1. Males and non-pregnant females, aged between 16 and 55 years (inclusive). 2. Subjects with a body mass index (BMI) ≥ 18 and ≤ 32, a body weight > 30 kg and ≤ 150 kg at the time of screening. 3. Subjects with a serum IgE ≥ 30 IU / mL and ≤ 1500 IU / mL at the time of screening. 4. Subjects with a physician-diagnosed peanut allergy or a history of peanut allergy. 5. Subjects with a history of non-severe peanut allergic reactions (≤ grade 3) (including mild wheezing or non-hypoxic dyspnea). 6. Subjects with a peanut-specific IgE > 5 kU / L measured by ImmunoCAP at the time of screening. 7. Subjects who self-reported a peanut-free diet for at least 14 days prior to screening, had no suspected peanut exposure (including any peanut food challenges), and agreed to continue to restrict peanut exposure during the study (except for the study DBPCFC). 8. Subjects with a positive peanut skin prick test (SPT) and a change in wheal diameter > 3 mm compared to the negative control (50% glycerol) at the time of screening. 9. Subjects who are willing and able to provide a written informed consent approved by the Institutional Review Board (IRB). 10. Subjects with a positive peanut double-blind placebo-controlled food challenge (DBPCFC) (baseline DBPCFC) and an inducing dose ≥ 10 mg and ≤ 300 mg of peanut protein at the time of screening. 11. Subjects with peanut-specific Th2a+ T cells (peanut-specific Th2a+ cells / total peanut-specific T cells) ≥ 15% after in vitro stimulation of PBMCs at the time of screening.

[0195] Subjects who meet all inclusion and no exclusion criteria after completing two screening visits (including two sets of baseline DBPCFCs (peanut and placebo challenges, administered on two separate days)) are enrolled in one of three dose-escalation cohorts. Subjects are randomly assigned at a ratio of 2:1 on Day 1 and Day 8 to receive CNP-201 or placebo (0.9% sodium chloride injection) by 200 mL intravenous infusion.

[0196] The dose levels for the three cohorts are as follows: Cohort 1: 250 mg, Cohort 2: 450 mg, Cohort 3: 650 mg. The dosing interval for subjects within a dose cohort is at least 48 hours.

[0197] Subjects who meet all inclusion and no exclusion criteria after initial screening assessment will undergo skin prick testing (SPT), followed by baseline double-blind placebo-controlled food challenge (DBPCFC) (peanut and placebo (oat) challenges, administered on two separate days) to confirm peanut allergy. If a reaction occurs and treatment is administered, peanut and placebo challenges are performed at least 48 hours apart. The procedural schedule for this study is as Figure 1C shown.

[0198] After completion of the second set of DBPCFC and a 2-hour observation period, subjects who continue to meet all inclusion and no exclusion criteria receive the first subcutaneous injection of omalizumab. The dose of omalizumab (XOLAIR®) follows the product label and is determined by the subject's weight and serum IgE at screening, as described in Table 2. According to the product label, subjects are dosed every 2 weeks (days 29, 15, and 1) or every 4 weeks (days 29 and 1).

[0199] Any subject who experiences a severe hypersensitivity reaction to omalizumab is appropriately treated and the study is discontinued, and then replaced with another subject at the same dose level.

[0200] Subjects return to the outpatient clinic on day 1 for a final assessment of eligibility and collection of laboratory samples. Subjects who continue to meet all inclusion and no exclusion criteria are randomly assigned to the dose cohort that is open at that time. On day 1 and on day 8, subjects are administered CNP-201 or placebo. CNP-201 or placebo is administered by intravenous infusion using a progressive infusion rate over approximately 3 to 4 hours. Subjects receive medical observation for 4 hours for acute adverse events (AEs) (including infusion reactions (IRs)) at the outpatient clinic after the infusion. If an allergic reaction occurs, antihistamines / adrenaline are immediately available for treatment of anaphylaxis.

[0201] Subjects return to the outpatient clinic on days 3 and 10 (2 days after each infusion) for a clinic visit for collection of safety laboratory samples, drug review, and assessment of AEs, and are followed up by telephone visit daily between infusions (days 33 to 36) to assess and record any AEs and drug changes. During the post-dose period, subjects return to the outpatient clinic on day 15 for collection of safety laboratory samples, PD measurements, and assessment of AEs and drug changes.

[0202] After all subjects in a dose cohort have completed the Day 15 clinic visit (7 days after the second dose), the Data Monitoring Committee (DMC) is convened to review all available safety data and determine whether it is acceptable to proceed to the next escalating dose cohort, whether it is necessary to expand the cohort (randomizing at least three additional subjects in a 2:1 ratio to receive CNP-201 or placebo (0.9% sodium chloride USP)), or whether any other clinical recommendations should be made.

[0203] Subjects return to the outpatient clinic on Day 60 for collection of immunological safety laboratory samples, PD measurements, and a second SPT, followed by DBPCFC (to be completed on Day 61). At the end of the study visit, subjects return to the outpatient clinic on Day 90 for collection of safety laboratory samples, PD measurements, and a final assessment of AEs and medication changes. Once all subjects have completed the Day 15 visit, a recommendation will be made to continue with the safe and tolerable dose of CNP-201 identified in Part A into Part B.

[0204] Part B: Subjects in Part B are randomized in a 1:1 ratio to receive the safe and tolerable level of CNP-201 identified in Part A or placebo (0.9% sodium chloride USP). Subjects who meet all inclusion and no exclusion criteria after the initial screening assessment undergo a skin prick test (SPT), followed by a baseline double-blind placebo-controlled food challenge (DBPCFC) (peanut and placebo (oat) challenges, administered over two days) to confirm peanut allergy. If a reaction occurs and treatment is received, the peanut and placebo challenges are performed at least 48 hours apart. After completion of the second set of DBPCFC and a 2-hour observation period, subjects who continue to meet all inclusion and no exclusion criteria will receive the first subcutaneous injection of omalizumab.

[0205] The dose of omalizumab (XOLAIR®) follows the product label specified in the protocol and is determined by the subject's weight and serum IgE at screening and is described in Table 2. Subjects are dosed every 2 weeks or every 4 weeks according to the product label. Any subject with a severe hypersensitivity reaction to omalizumab is treated appropriately and withdrawn from the study. These subjects are replaced in Part B.

[0206] Subjects return to the outpatient clinic on Day 1 for a final assessment of eligibility and collection of laboratory samples. Subjects who continue to meet all inclusion and no exclusion criteria are randomized to receive CNP-201 or placebo.

[0207] Subjects will receive CNP-201 or placebo on Day 1 and Day 8. The study product is administered via IV infusion using a progressive infusion rate over approximately 3 to 4 hours. Subjects will receive medical observation for acute AEs for 4 hours in the outpatient setting after the infusion. In case of an allergic reaction, antihistamines / adrenaline will be immediately available for treatment of anaphylaxis. Subjects will be followed up daily by telephone interview between each infusion (Days 2 to 7 and Days 9 to 14) to assess and record any AEs and medication changes.

[0208] During the post-dose period, subjects will return to the outpatient setting on Day 15 for collection of safety laboratory samples, PD measurements, and assessment of AEs and medication changes. Subjects will also return to the outpatient setting on Day 60 for collection of immunological safety laboratory samples, PD measurements, and a second SPT, followed by DBPCFC (to be completed on Day 61). At the end of the study visit, subjects will return to the outpatient setting on Day 90 for collection of safety laboratory samples, PD measurements, and a final assessment of AEs and medication changes.

[0209] Emerging safety and tolerability data are monitored in Part B. The medical monitor is notified of any serious adverse event (SAE) and any > Grade 2 adverse event (CTCAE v.5.0) within 24 hours of becoming aware of such an event. The medical monitor may then convene an ad hoc DMC meeting to evaluate the safety and tolerability data to determine whether continued dosing is still acceptable and whether recommendations, including but not limited to continued dosing and stopping or suspending dosing of subjects, should be made. If the DMC determines that continued dosing poses a safety risk to the subjects, they may recommend stopping or suspending the study at any time during the study.

[0210] In Parts A and B of the study, subjects will receive CNP-201 via intravenous infusion over approximately 3 to 4 hours at the following progressive infusion rates: 20 mL / hour for the first 15 minutes, 40 mL / hour for the next 15 minutes, and 80 mL / hour for the remaining infusion time.

[0211] Study duration: 2 doses, 7 days apart (Parts A and B). The total study duration for an individual subject is approximately 134 days; 14 days for screening, 30 days for omalizumab administration, 60 days for IP administration, and 30 days for the follow-up period.

[0212] Primary endpoints (Parts A and B) include: frequency of adverse events (AE) and serious adverse events (SAE), MedDRA 23.0 (CTCAE v.5.0); laboratory safety assessments (hematology, serum chemistry, coagulation panel, urinalysis); physical examinations, including vital signs (blood pressure, heart rate, body temperature); 12-lead electrocardiogram (ECG); serum cytokines (TNF-α, IL-2, IL-6, IL-8, IL-1β, MCP-1, MIP-1β, MIP-1α, IFN-γ, IL-12p70); and the change in the proportion of peanut-specific Th2a + T cells (peanut-specific Th2a + cells / total peanut-specific T cells) between placebo and CNP-201 after ex vivo stimulation of PBMCs at baseline (Day 1 before dosing) and Day 15; and the change in the proportion of activated peanut-specific T cells among total peanut-specific T cells between placebo and CNP-201 after ex vivo stimulation of PBMCs at baseline (Day 1 before dosing) and Day 15.

[0213] Secondary endpoints (Parts A and B) include: the change in the IL-5 to IFN-γ ratio between placebo and CNP-201 after ex vivo stimulation of PBMCs at baseline (Day 1 before dosing) and Day 15.

[0214] Exploratory endpoints (Parts A and B) include: the change in the proportion of peanut-specific T regulatory cells (peanut-specific T regulatory cells / peanut-specific CD4+ effector memory cells) between placebo and CNP-201 after ex vivo stimulation of PBMCs at baseline (Day 1 before dosing) and Day 15; the change in the effective concentration at 50% maximal basophil activation (EC 50 ) measured by basophil activation test (CD203c+ / CD63+ / - basophil activation) between placebo and CNP-201 at baseline (Day -30 before DBPCFC) and Day 60; the change in the ratio of peanut-specific IgE to IgG measured by ImmunoCap between placebo and CNP-201 at baseline (Day -30 to -29) and Day 60 to 61; and the change in the cumulative tolerated dose (CTD) (mg) of peanut protein administered between placebo and CNP-201 during DBPCFC at baseline (Days [-30] to [-29]) and Days 60 to 61. The cell markers used to evaluate the exploratory endpoints are described in Table 3 below: Example 1A - TIMP - PPE Phase I / II Trial in Peanut Allergy

[0215] An alternative is provided for a two-part, randomized, double-blind, placebo-controlled Phase 1b / 2a study to evaluate the safety, tolerability, pharmacodynamics, and efficacy of TIMP-PPE (CNP-201) in peanut-allergic subjects aged 16 to 35 years.

[0216] CNP-201 consists of PLGA nanoparticles encapsulating purified peanut extract, with an average diameter of 400 - 800 nm and a negative ζ potential between -30 mV and -60 mV. The CNP-201 particles are provided in a lyophilized formulation. The CNP-201 particles are reconstituted in sterile water for injection and diluted with sterile saline (0.9% sodium chloride) prior to administration.

[0217] This study consists of Part A and Part B. Part A: Part A is a randomized, double-blind, placebo-controlled study of the safety of escalating dose levels of CNP-201 for tolerability. Part A will enroll 3 cohorts to receive CNP-201 or placebo at multiple escalating dose levels. Part B follows a randomized, double-blind, placebo-controlled, repeated-dose study using the safe and tolerable dose level of CNP-201 determined from Part A.

[0218] Subjects who meet all inclusion (except IgG and IgE results which may not be available at the 2nd visit) and no exclusion criteria after the initial screening assessment undergo a skin prick test (SPT), followed by a baseline double-blind placebo-controlled food challenge (DBPCFC) (peanut and placebo (oat) challenges, administered over two days) to confirm peanut allergy. The SPT and DBPCFC are performed by a study physician or a staff member trained in the management of on-site clinical emergencies, with immediate access to emergency medications and equipment and in close proximity to the hospital emergency department to provide rapid emergency care if needed. Subjects who continue to meet all inclusion and no exclusion criteria two days after completion of the DBPCFC are eligible to participate in this study.

[0219] All subjects who continue to meet all I / E criteria after the DBPCFC receive subcutaneous injection of omalizumab (XOLAIR). The dose of omalizumab (XOLAIR®) follows the product label specified in the protocol and is determined by the subject's body weight and serum IgE at the initial screening. Subjects are dosed every 2 weeks or every 4 weeks according to the product label.

[0220] Subjects who continue to meet all inclusion and no-exclusion criteria will be randomly assigned at a ratio of 2:1 (Part A) or 1:1 (Part B) on Day 1 to receive CNP-201 or placebo (0.9% sodium chloride USP) by intravenous (IV) infusion. CNP-201 or placebo will be administered to the subjects on Day 1 and Day 8. Unless infusion reactions, allergic reactions, or other adverse events require extended monitoring, the subjects will remain outpatient from Day 1 (prior to administration of CNP-201 or placebo) and Day 8 (prior to administration of CNP-201 or placebo) after admission until 4 hours after administration of the drug on the same day. If the safety parameters are acceptable to the investigator, the subject will be discharged.

[0221] Peanut-allergic subjects eligible for inclusion in this study are defined based on the following inclusion criteria: 1. Males and non-pregnant females, aged 16 to 35 years (inclusive). 2. Subjects with a body mass index (BMI) ≥ 18 and ≤ 32 and a weight > 30 kg and ≤ 150 kg at screening. Subjects outside this range may be included at the discretion of the investigator. 3. Subjects with a serum IgE ≥ 30 IU / mL and ≤ 1500 IU / mL at screening. Subjects outside this range may be included at the discretion of the investigator. 4. Subjects with a physician-diagnosed peanut allergy or a history of documented peanut allergy. 5. Subjects with a history of documented non-severe peanut allergic reactions (≤ grade 3) (including mild wheezing or dyspnea without hypoxia). 6. Subjects with a peanut-specific IgE > 2 kU / L measured by ImmunoCAP and / or positive for peanut skin prick test (SPT) with a change in wheal diameter > 3 mm compared to negative control subjects (50% glycerol) at screening. 7. Subjects who self-report being on a peanut-free diet for at least 14 days prior to screening, have no suspected peanut exposure (including any peanut food challenges), and agree to continue to restrict peanut exposure during the study (except for the study DBPCFC). 8. Female subjects and male subjects and their female spouses / partners who are willing to practice highly effective contraceptive methods, which may include but are not limited to abstinence, sexual relations only with the same sex, monogamous relationships with vasectomized partners, vasectomy, hysterectomy, bilateral tubal ligation, approved hormonal methods, intrauterine devices (IUDs), or the use of spermicides in combination with barrier methods (e.g., condoms, diaphragms) starting at screening and continuing throughout the study until Day 90 (EOS / ET). 9. Female subjects who agree not to breastfeed starting from initial screening and throughout the study period until Day 90 (EOS / ET). 10. Female subjects who agree not to donate eggs starting from initial screening and throughout the study period until Day 90 (EOS / ET). 11. Subjects who are willing and able to provide a written informed consent approved by the Institutional Review Board (IRB). 12. Subjects who are willing to perform and comply with all study procedures (including attending study visits as scheduled and completing two DBPCFCs). 13. Male subjects who agree not to donate sperm starting from screening and throughout the study period until Day 90 (EOS / ET).

[0222] Subjects must have a positive peanut DBPCFC at screening with an inducing dose ≥ 10 mg and ≤ 300 mg of peanut protein to be included in the statistical analysis of exploratory endpoints. Subjects who are tolerant to > 444 mg of peanut (cumulative tolerated dose) will be followed up for safety and evaluated separately.

[0223] Subjects who meet all inclusion and no exclusion criteria after completing two screening visits (including two baseline DBPCFCs (peanut and placebo challenges, administered on two separate days)) are randomized into one of three dose-escalation cohorts at a ratio of 2:1 on Day 1 and Day 8 to receive CNP-201 or placebo (0.9% sodium chloride injection) as a 200 mL intravenous infusion. The dose levels for the three cohorts are as follows: Cohort 1: 250 mg, Cohort 2: 450 mg, Cohort 3: 650 mg. The dosing interval for subjects within a dose cohort is at least 48 hours.

[0224] Subjects who meet all inclusion and no exclusion criteria after initial screening assessment will undergo a skin prick test (SPT), followed by a baseline double-blind placebo-controlled food challenge (DBPCFC) (peanut and placebo (oat) challenges, administered on two separate days) to confirm peanut allergy. If a reaction occurs and treatment is received, the peanut and placebo challenges are performed at least 48 hours apart.

[0225] Subjects who continue to meet all inclusion and no exclusion criteria after completing the second DBPCFC challenge and a 2-hour observation period will proceed with the first subcutaneous injection of XOLAIR®.

[0226] As described in Table 2, the dose and dosing frequency of XOLAIR® (every 2 weeks or every 4 weeks) are determined by the subject's serum IgE at screening and the body weight measured at XOLAIR® dose 1.

[0227] On Day 1, subjects returned to the outpatient clinic for a final assessment of eligibility and collection of laboratory samples. Subjects who continued to meet all inclusion and no exclusion criteria were randomly assigned to the dose cohort that was open at that time. On Day 1 and on Day 8, subjects were administered CNP-201 or placebo. CNP-201 or placebo was administered via intravenous infusion over approximately 3 to 4 hours using a progressive infusion rate. Subjects received medical observation for 4 hours for acute adverse events (AEs) (including infusion reactions (IRs)) in the outpatient clinic after the infusion. If an allergic reaction occurred, antihistamines / adrenaline would be immediately available for treatment of anaphylaxis.

[0228] If two identical Grade 3 adverse events (AEs) or one ≥ Grade 4 adverse event (CTCAE v.5.0 or CoFAR V.1 for allergy-related AEs) that were considered possibly related to CNP-201 occurred, dosing would be suspended and the DMC would convene a meeting to review all available safety data obtained to date. Grade 4 or 5 AEs that were considered serious AEs (SAEs) would be reported to the institution. After reviewing all available safety data, the DMC would make recommendations, including but not limited to stopping dosing, down-titrating dosing, continuing dosing, expanding the current cohort, adding a cohort in Part A, or determining the dose-limiting toxicity. The DMC could also convene an interim meeting based on the sponsor medical monitor's ongoing daily monitoring of safety data to address any safety issues that arose during dosing of any subject in Part A. The DMC would evaluate the available safety data, including but not limited to AEs, physical examinations, vital signs, 12-lead ECGs, and available laboratory results.

[0229] Subjects returned to the outpatient clinic for a clinic visit to collect safety laboratory samples, review the medication, and evaluate AEs on Days 3 and 10, 2 days after each infusion. Follow-up was conducted via telephone visit daily after each infusion (Days 4 to 7 and Days 11 to 14) to assess and record any AEs and medication changes. During the post-dosing period, subjects returned to the outpatient clinic 7 days after receiving the second dose of CNP-201 or placebo to collect safety laboratory samples, measure PD, and evaluate AEs and medication changes.

[0230] After all subjects in the dose cohort had completed the clinic visit on Day 15 (7 days after the second dose), the DMC was convened to review all available safety data and determine whether it was acceptable to proceed to the next escalating dose cohort, whether it was necessary to expand the cohort (randomly assign at least three additional subjects in a 2:1 ratio to receive CNP-201 or placebo), or whether any other clinical recommendations (such as expanding to Part B) should be made.

[0231] Subjects returned to the outpatient clinic on Day 60 for collection of immunological safety laboratory samples, PD measurements, and the second SPT, followed by post-dose DBPCFC, and at the end of the study visit, returned to the outpatient clinic on Day 90 for collection of safety laboratory samples, PD measurements, and final assessment of AEs and medication changes. Newly emerging safety data and tolerance data will be continuously monitored. The medical monitor is notified of any serious adverse event (SAE) and any adverse event of grade > 2 that may be related to the test product (CTCAE v.5.0 or CoFAR V.1) within 24 hours of becoming aware of it. The medical monitor may then convene an ad hoc DMC meeting to evaluate the safety data and tolerance data to determine whether continued dosing is still acceptable and whether recommendations are made, including but not limited to continued dosing and stopping or suspending dosing of the subject. If the DMC determines that continued dosing poses an unacceptable safety risk to the subject, they may recommend stopping or suspending the study at any time during the study. Once all subjects have completed the visit on Day 15, a recommendation will be made to continue with the safe and tolerable dose of CNP-201 identified in Part A into Part B.

[0232] Part B: Subjects in Part B will be randomized at a 1:1 ratio to receive CNP-201 or placebo at the safe and tolerable level identified in Part A. Subjects in Part A and Part B receive the same evaluations, and the only difference between the two parts is the dose escalation in Part A.

[0233] Subjects who meet all inclusion and no exclusion criteria after the initial screening assessment undergo a skin prick test (SPT), followed by a baseline double-blind placebo-controlled food challenge (DBPCFC) (peanut and placebo (oatmeal) challenges, administered over two days) to confirm peanut allergy. If a reaction occurs and treatment is received, the peanut and placebo challenges are performed at least 48 hours apart.

[0234] After completion of the second DBPCFC challenge and the 2-hour observation period, subjects who continue to meet all inclusion and no exclusion criteria proceed with the first subcutaneous injection of XOLAIR®. As described in Table 2, the dose and dosing frequency of XOLAIR® (every 2 weeks or every 4 weeks) are determined by the subject's serum IgE at screening and the body weight measured at XOLAIR® dose 1.

[0235] Subjects returned to the outpatient clinic on Day 1 for final assessment of eligibility and collection of laboratory samples. Subjects who continue to meet all inclusion and no exclusion criteria are randomized to receive CNP-201 or placebo.

[0236] On Day 1 and on Day 8, CNP-201 or placebo was administered to the subjects. CNP-201 or placebo was administered by intravenous infusion at a progressive infusion rate over approximately 3 to 4 hours. The subjects received medical observation for 4 hours of acute adverse events (AEs), including infusion reactions (IRs), in the outpatient setting after the infusion. If an allergic reaction occurred, antihistamines / adrenaline were immediately available for the treatment of anaphylaxis.

[0237] The subjects returned to the outpatient setting on Days 2 (Day 3 and Day 10) after each infusion for a clinic visit for the collection of safety laboratory samples, review of the drug, and assessment of AEs, and were followed up by telephone visit daily after each infusion (Days 4 to 7 and Days 11 to 14) to evaluate and record any AEs and drug changes. During the post-dose period, the subjects returned to the outpatient setting 7 days after the administration of the second dose of CNP-201 or placebo for the collection of safety laboratory samples, PD measurements, and assessment of AEs and drug changes.

[0238] The subjects returned to the outpatient setting on Day 60 for the collection of immunological safety laboratory samples, PD measurements, and the second SPT, followed by the post-dose DBPCFC, and at the end of the study visit, returned to the outpatient setting on Day 90 for the collection of safety laboratory samples, PD measurements, and the final assessment of AEs and drug changes. Newly emerging safety data and tolerance data were continuously monitored. The medical monitor was notified of any serious adverse event (SAE) and any adverse event of > Grade 2 (CTCAE v.5.0 or CoFAR V.1) that might be related to the test product within 24 hours of becoming aware of it. The medical monitor could then convene an ad hoc DMC meeting to evaluate the safety data and tolerance data to determine whether continued dosing was still acceptable and whether recommendations should be made, including but not limited to continuing dosing and stopping or suspending dosing of the subjects. If the DMC considered that continued dosing posed an unacceptable safety risk to the subjects, they could recommend stopping or suspending the study at any time during the study.

[0239] In Parts A and B of the study, the subjects received CNP-201 by intravenous infusion over approximately 3 to 4 hours at the following progressive infusion rates: 20 mL / hour for the first 15 minutes, 40 mL / hour for the next 15 minutes, and 80 mL / hour for the remaining infusion time.

[0240] In Parts A and B, the subjects randomized to the placebo group received an injection of 0.9% sodium chloride (normal saline [NS]). The placebo was administered as a 200 mL intravenous infusion on Day 1 and Day 8 at the following progressive infusion rates: 20 mL / hour for the first 15 minutes, 40 mL / hour for the next 15 minutes, and 80 mL / hour for the remaining infusion time.

[0241] Study duration: 2 doses, 7 days apart (Parts A and B). The total study duration for an individual subject is approximately 134 days; 14 days for screening, approximately 30 days for XOLAIR® administration, 60 days for test product administration, and approximately 30 days for follow-up after dosing.

[0242] Primary endpoints (Parts A and B) include: frequency of adverse events (AE) and serious adverse events (SAE), MedDRA 23.0 (CTCAE v.5.0 or CoFAR V.1 for allergy-related AEs); laboratory safety assessments (hematology, serum chemistry, coagulation panel, and urine analysis); physical examinations, including vital signs (blood pressure, heart rate, and body temperature); 12-lead electrocardiogram (ECG); serum cytokines (TNF-α, IL-2, IL-6, IL-8, IL-1β, MCP-1, MIP-1β, MIP-1α, IFN-γ, and IL-12p70).

[0243] Exploratory endpoints (Parts A and B) include: change in the proportion of peanut-specific Th2a+ T cells (peanut-specific Th2a+ cells / total peanut-specific T cells) after ex vivo stimulation of PBMC between placebo and CNP-201 at baseline (Day 1 before dosing) and Day 15; change in the effective concentration at 50% of the maximum basophil activation (EC50) measured by basophil activation test (CD203c+ / CD63+ / - basophil activation) between placebo and CNP-201 at baseline (pre-DBPCFC screening, 2nd visit) and 11th visit; change in peanut-specific IgE measured by ImmunoCap assay between placebo and CNP-201 at baseline (pre-DBPCFC screening, 2nd visit) and 11th visit; change in the ratio of peanut-specific IgE to IgG measured by ImmunoCap assay between placebo and CNP-201 at baseline (screening DBPCFC, 2nd and 3rd visits) and after dosing (post-dose DBPCFC, 11th and 12th visits); change in the cumulative tolerated dose (CTD) (mg) of peanut protein administered during DBPCFC between placebo and CNP-201 at baseline (Day 1 before dosing) and Day 15; change in the proportion of activated peanut-specific T cells (activated peanut-specific T cells / total peanut-specific T cells) after ex vivo stimulation of PBMC between placebo and CNP-201 at baseline (Day 1 before dosing) and Day 15. Example 2 - A Phase I / II Trial of TIMP-PPE Without an IgE Inhibitor for the Treatment of Peanut Allergy

[0244] This example describes a Phase 1b / 2a randomized, double-blind, placebo-controlled study to evaluate the safety, tolerability, and pharmacodynamics of TIMP-PPE (CNP-201) without IgE inhibitors in peanut-allergic subjects aged 16 to 55 years.

[0245] CNP-201 consists of PLGA nanoparticles encapsulating purified peanut extract, with an average diameter of 400 - 800 nm and a negative ζ potential between -30 mV and -60 mV. CNP-201 particles are provided in a lyophilized formulation. CNP-201 particles are reconstituted in sterile water for injection and diluted with sterile saline (0.9% sodium chloride) before administration.

[0246] This study is a randomized, double-blind, placebo-controlled study on the safety and tolerability of escalating dose levels of CNP-201. This study will enroll 3 cohorts to receive CNP-201 or placebo at multiple escalating dose levels.

[0247] Subjects who meet all inclusion and no exclusion criteria are eligible for this study. Subjects who continue to meet the inclusion / exclusion criteria are randomized at a ratio of 2:1 on Day 1 to receive CNP-201 or placebo (0.9% sodium chloride USP) by intravenous (IV) infusion. Subjects are dosed with CNP-201 or placebo on Day 1 and Day 8.

[0248] Subjects may receive pre-treatment with antihistamines (such as 10 mg IV cetirizine) and corticosteroids (such as 125 mg IV methylprednisolone) 30 minutes before each infusion of CNP-201 or placebo. Subjects will continue their current SoC during the study, excluding a 12-hour washout period (only for β-agonists, theophylline, and cromolyn) and a 7-day washout period for antihistamines before skin prick test (SPT). Subjects will resume their SoC regimen after SPT and continue with SoC during dosing / study.

[0249] Peanut-allergic subjects eligible for this study are defined based on the following inclusion criteria: 1. Males and non-pregnant females, aged 16 to 55 years (inclusive). 2. Subjects with a physician-diagnosed peanut allergy or a history of peanut allergy documented. 3. Subjects with a screening weight ≥ 31.25 kg. Subjects outside this range may be included at the discretion of the investigator. 4. Subjects with a history of non-severe peanut allergic reactions (≤ grade 3) (including mild wheezing or non-hypoxic dyspnea). 5. Subjects with peanut-specific IgE > 5 kU / L measured by ImmunoCAP at screening. 6. Subjects who self-reported a peanut-free diet for at least 14 days prior to screening, had no suspected peanut exposure (including any peanut food challenges), and agreed to continue to restrict peanut exposure during the study. 7. Subjects who had a positive skin prick test (SPT) to peanut with a wheal diameter change > 3 mm compared to the negative control (50% glycerol) at screening. 8. Female and male subjects and their female spouses / partners willing to practice highly effective contraceptive methods, which may include but are not limited to abstinence, sexual relations only with the same sex, monogamous relationships with vasectomized partners, vasectomy, hysterectomy, bilateral tubal ligation, approved hormonal methods, intrauterine devices (IUDs), or the use of spermicides in combination with barrier methods (e.g., condoms, diaphragms) starting at screening and continuing throughout the study until Day 38 (EOS / ET). 9. Female subjects who agreed not to breastfeed from the initial screening and throughout the study until Day 38 (EOS / ET). 10. Female subjects who agreed not to donate eggs from the initial screening and throughout the study until Day 38 (EOS / ET). 11. Subjects willing and able to provide a written informed consent approved by the Institutional Review Board (IRB). 12. Subjects willing to perform and comply with all study procedures. 13. Male subjects who agreed not to donate sperm from screening and throughout the study until Day 38 (EOS / ET).

[0250] Subjects who met all inclusion and no exclusion criteria after completing the screening visit were enrolled in one of three dose-escalation cohorts. Subjects were randomly assigned at a 2:1 ratio on Day 1 and Day 8 to receive CNP-201 or placebo (0.9% sodium chloride injection) as a 200 mL intravenous infusion. The dose levels for the three cohorts were as follows: Cohort 1: 250 mg, Cohort 2: 450 mg, Cohort 3: 650 mg. The dosing interval for subjects within a dose cohort was at least 48 hours.

[0251] Subjects who met all inclusion and no exclusion criteria at screening were enrolled in this study. Subjects will return to the outpatient clinic on Day 1 for a final assessment of eligibility and collection of laboratory samples. Subjects who continued to meet all inclusion and no exclusion criteria were randomly assigned to the dose cohorts that were open at that time. Subjects could receive premedication with antihistamines (such as 10 mg IV cetirizine) and corticosteroids (such as 125 mg IV methylprednisolone) 30 minutes before each infusion of CNP-201 or placebo on Days 1 and 8. CNP-201 or placebo was administered by intravenous infusion over approximately 3 to 4 hours using a progressive infusion rate. Subjects were observed medically for 4 hours for acute adverse events (AEs) (including infusion reactions (IRs)) after the infusion in the outpatient clinic. If an allergic reaction occurred, antihistamines / adrenaline (including intramuscular (IM) and intravenous (IV) adrenaline) were immediately available for the treatment of anaphylaxis.

[0252] Subjects were followed up by telephone visit daily after each infusion (Days 2 to 7 and Days 9 to 14) to assess and record any AEs and medication changes. During the post-dosing period, subjects returned to the outpatient clinic on Day 15 (7 days after the administration of the second dose of CNP-201 or placebo) for collection of safety laboratory samples, PD measurements, and assessment of AEs and medication changes.

[0253] After all subjects in the dose cohort completed the clinic visit on Day 15 (7 days after the second dose), the DMC was convened to review all available safety data and determine whether it was acceptable to proceed to the next escalating dose cohort.

[0254] At the end of the study visit, subjects returned to the outpatient clinic on Day 38 for collection of safety laboratory samples, PD measurements, and final assessment of AEs and medication changes, as well as a second SPT.

[0255] Newly emerging safety data and tolerance data will be continuously monitored. The medical monitor is notified of any serious adverse event (SAE) and any ≥ Grade 2 adverse event (CTCAE v.5.0 or CoFAR V.3.0) that may be related to the test product within 24 hours of becoming aware of such an event. The medical monitor may convene an ad hoc DMC meeting to evaluate the safety data and tolerance data to determine whether continued dosing is still acceptable and whether recommendations are made, including but not limited to continuing dosing and stopping or suspending dosing of the subject. If the DMC determines that continued dosing poses an unacceptable safety risk to the subject, they may recommend stopping or suspending this study at any time during the study.

[0256] Subjects received CNP-201 via an intravenous infusion lasting approximately 3 to 4 hours at the following escalating infusion rates: 20 mL / hour for the first 15 minutes, 40 mL / hour for the next 15 minutes, and 80 mL / hour for the remaining infusion time.

[0257] Study duration: 2 doses, 7 days apart. The total study duration for an individual subject was approximately 45 days; 7 days for screening; 8 days for test product administration; and 30 days for post-administration evaluation.

[0258] Primary endpoints included: the frequency of adverse events (AE) and serious adverse events (SAE), MedDRA 23.0 (CTCAE v.5.0 or CoFAR V.3.0 for allergy-related AE); laboratory safety assessments (hematology, serum chemistry, coagulation panel, and urine analysis); physical examinations, including vital signs (blood pressure, heart rate, and body temperature); 12-lead electrocardiogram (ECG); serum cytokines (TNF-α, IL-2, IL-6, IL-8, IL-1β, MCP-1, MIP-1β, MIP-1α, IFN-γ, and IL-12p70).

[0259] Exploratory endpoints included: the change in the ratio of peanut-specific IgE to IgG measured by ImmunoCap between placebo and CNP-201 at baseline and Day 38; the change in peanut-specific IgE measured by ImmunoCap between placebo and CNP-201 at baseline and Day 38.

[0260] This document describes the preferred embodiments of the present disclosure. Variations of these preferred embodiments may become apparent to those of ordinary skill in the art after reading the foregoing description. The inventors expect those skilled in the art to adopt such variations where appropriate, and the inventors intend to practice the present disclosure in a manner different from that specifically described herein. Accordingly, the present disclosure includes all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. In addition, any combination of the above elements in all possible variations thereof is included in the present disclosure unless otherwise indicated herein or otherwise clearly contradicted by the context. Example 3 - Phase I / II Trial of Low-Dose TIMP-PPE in Peanut Allergy

[0261] This example describes a Phase 1b / 2a randomized, double-blind, placebo-controlled study evaluating the safety, tolerability, and pharmacodynamics of TIMP-PPE (CNP-201) in peanut-allergic subjects aged 16 to 55 years.

[0262] CNP-201 consists of PLGA nanoparticles encapsulating purified peanut extract, with an average diameter of 400 - 800 nm and a negative ζ potential between -30 mV and -60 mV. CNP-201 particles are provided in a lyophilized formulation. CNP-201 particles are reconstituted in sterile water for injection and diluted with sterile saline (0.9% sodium chloride) before administration.

[0263] This study is a randomized, double-blind, placebo-controlled study on the safety and tolerance of escalating dose levels of CNP-201. This study will enroll 4 cohorts to receive CNP-201 or placebo at multiple escalating dose levels.

[0264] Subjects meeting all inclusion and no exclusion criteria are eligible for this study. Subjects who continue to meet the inclusion / exclusion criteria are randomly assigned at a ratio of 2:1 on Day 1 to receive CNP-201 or placebo (0.9% sodium chloride USP) by intravenous (IV) infusion. On Day 1 and Day 8, subjects are administered CNP-201 or placebo.

[0265] Subjects may receive pre-treatment with NSAIDs (such as oral 325 mg acetylsalicylic acid) daily for 2 days and 60 minutes before each infusion of CNP-201 or placebo. Subjects may receive pre-treatment and / or post-treatment with leukotriene modifiers (such as oral 10 mg montelukast) 12 hours before, 45 minutes before, and 12 hours after each infusion of CNP-201 or placebo. Subjects may receive pre-treatment with antihistamines (such as 50 mg IV diphenhydramine) and corticosteroids (such as 125 mg IV methylprednisolone) 30 minutes before each infusion of CNP-201 or placebo.

[0266] During the study, subjects will continue to receive their current standard of care (SoC), excluding a 12-hour washout period (only for beta-agonists, theophylline, and cromolyn) and a 7-day washout period for antihistamines before skin prick test (SPT). Subjects will resume their SoC regimen after SPT and continue to receive SoC during dosing / study.

[0267] Peanut-allergic subjects eligible for this study are defined based on the following inclusion criteria: 1. Males and non-pregnant females, aged 16 to 55 years (inclusive). 2. Subjects with a physician-diagnosed peanut allergy or a history of peanut allergy documented. 3. Subjects with a screening weight ≥ 31.25 kg. Subjects outside this range may be included at the discretion of the investigator. 4. Subjects with a documented history of non-severe peanut allergic reactions (⧠3), including mild wheezing or dyspnea without hypoxia. 5. Subjects with peanut-specific IgE ≥ 5 kU / L measured by ImmunoCAP at screening, unless previously treated for peanut allergy with OIT. Subjects previously treated for peanut allergy with OIT and those without peanut-specific IgE ≥ 5 kU / L measured by ImmunoCap at screening may be included at the discretion of the investigator, or subjects who are positive for peanut SPT with a wheal diameter change ≥ 3 mm compared to the negative control (50% glycerol) at screening. Subjects previously treated for peanut allergy with OIT and those without a positive peanut skin prick test (SPT) with a wheal diameter change ≥ 3 mm at screening may be included at the discretion of the investigator. 6. Subjects who self-report a peanut-free diet for at least 14 days prior to screening, have no suspected peanut exposure (including any peanut food challenges), and agree to continue to restrict peanut exposure during the study. 7. Subjects who are positive for peanut skin prick test (SPT) with a wheal diameter change > 3 mm compared to the negative control (50% glycerol) at screening. 8. Female and male subjects and their female spouses / partners who are willing to practice highly effective contraceptive methods, which may include but are not limited to abstinence, sexual relations only with the same sex, monogamous relationships with vasectomized partners, vasectomy, hysterectomy, bilateral tubal ligation, approved hormonal methods, intrauterine devices (IUDs), or the use of spermicides in combination with barrier methods (e.g., condoms, diaphragms) starting at screening and continuing throughout the study until Day 38 (EOS / ET). 9. Female subjects who agree not to breastfeed from initial screening and throughout the study until Day 38 (EOS / ET). 10. Female subjects who agree not to donate eggs from initial screening and throughout the study until Day 38 (EOS / ET). 11. Subjects who are willing and able to provide a written informed consent approved by the Institutional Review Board (IRB). 12. Subjects who are willing to perform and comply with all study procedures. 13. Male subjects who agree not to donate sperm from screening and throughout the study until Day 38 (EOS / ET).

[0268] Subjects who meet all inclusion and no exclusion criteria after the screening visit are assigned to one of the multiple dosing groups. Subjects are randomly assigned at a ratio of 2:1 on Day 1 and Day 8 to receive CNP-201 or placebo (0.9% sodium chloride injection) by 200 mL intravenous infusion. The dose level of the first group is 25 mg. The other dose levels of the remaining 3 groups decrease to 0.1 mg, 0.25 mg, 0.5 mg, 1 mg, 2 mg, 2.5 mg, 5 mg, 10 mg or may increase up to a maximum of 650 mg. The dosing interval for subjects in the dose groups is at least 48 hours.

[0269] Subjects who meet all inclusion and no exclusion criteria at screening are eligible for this study. Subjects will return to the outpatient clinic on Day 1 for a final assessment of eligibility and collection of laboratory samples. Subjects who continue to meet all inclusion and no exclusion criteria are randomly assigned to the dose groups that are open at that time. Subjects may receive pre-treatment with NSAIDs (such as oral 325 mg acetylsalicylic acid) daily for 2 days and 60 minutes before each infusion of CNP-201 or placebo on Day 1 and Day 8. Subjects may receive pre-treatment and / or post-treatment with a leukotriene modifier (such as oral 10 mg montelukast) 12 hours before, 45 minutes before, and 12 hours after each infusion of CNP-201 or placebo on Day 1 and Day 8. Subjects may receive pre-treatment with an antihistamine (such as 50 mg IV diphenhydramine) and a corticosteroid (such as 125 mg IV methylprednisolone) 30 minutes before each infusion of CNP-201 or placebo on Day 1 and Day 8.

[0270] CNP-201 or placebo is administered by intravenous infusion using a progressive infusion rate over approximately 3 to 4 hours. Subjects receive medical observation for 4 hours for acute adverse events (AEs) (including infusion reactions (IRs)) in the outpatient clinic after the infusion. If an allergic reaction occurs, antihistamines / adrenaline (including intramuscular injection (IM) and intravenous injection (IV) adrenaline) are immediately available for treatment of anaphylactic reactions.

[0271] Subjects are followed up daily by telephone visit after each infusion (Days 2 to 7 and Days 9 to 14) to assess and record any AEs and medication changes. During the post-dosing period, subjects return to the outpatient clinic 7 days (Day 15) after administration of the second dose of CNP-201 or placebo for collection of safety laboratory samples, PD measurements, and assessment of AEs and medication changes.

[0272] After all subjects in the dose groups complete the clinic visit on Day 15 (7 days after the second dose), the DMC is convened to review all available safety data and determine whether it is acceptable to proceed to the next escalating dose group.

[0273] At the end of the study visit, subjects return to the outpatient clinic on Day 38 for collection of safety laboratory samples and PD measurements, assessment of AEs and medication changes, and the second SPT. Subjects return to the outpatient clinic on Day 60 for safety laboratory samples, PD measurements, and DBPCFC. Subjects will optionally return to the outpatient clinic on Days 90 and 120 for collection of safety laboratory samples and PD measurements. Subjects will optionally return on Day 180 for the second DBPCFC, which consists of peanut and placebo (oatmeal) challenges, safety laboratory samples, PD measurements, and final assessment of AEs and medication changes.

[0274] Newly emerging safety and tolerability data will be continuously monitored. The medical monitor is notified of any serious adverse event (SAE) and any ≥ Grade 2 adverse event (CTCAE v.5.0 or CoFAR V.3.0) that may be related to the test product within 24 hours of becoming aware of such an event. The medical monitor may convene an ad hoc DMC meeting to evaluate the safety and tolerability data to determine whether continued dosing remains acceptable and whether recommendations, including but not limited to continued dosing and stopping or suspending dosing of subjects, should be made. If the DMC determines that continued dosing poses an unacceptable safety risk to the subjects, they may recommend stopping or suspending the study at any time during the study.

[0275] Subjects receive CNP-201 via intravenous infusion over approximately 3 to 4 hours at the following escalating infusion rates: 1 mL / hour for the first 10 minutes, 2 mL / hour for the next 10 minutes, 5 mL / hour for the next 10 minutes, 10 mL / hour for the next 10 minutes, 20 mL / hour for the next 15 minutes, 40 mL / hour for the next 15 minutes, and 80 mL / hour for the remaining infusion time.

[0276] Study duration: 2 doses, 7 days apart. The total study duration for an individual subject is approximately 67 days; 7 days for screening; 8 days for test product administration; and 52 days for post-administration evaluation. An additional 120-day evaluation after administration is optional.

[0277] Primary objectives include: safety and tolerability of CNP-201.

[0278] Exploratory objectives include: changes in response to DBPCFC in patients treated with CNP-201 or placebo, changes in the ratio of peanut-specific IgE to IgG in patients treated with CNP-201 or placebo, changes in peanut-specific IgE in patients treated with CNP-201 or placebo, changes in the proportion of peanut-specific Th2a+ T cells in patients treated with CNP-201 or placebo, changes in response to basophil activation test in patients treated with CNP-201 or placebo, changes in the proportion of peanut-specific T regulatory cells in patients treated with CNP-201 or placebo.

[0279] Primary endpoints include: the frequency of adverse events (AE) and serious adverse events (SAE), MedDRA 23.0 (CTCAE v.5.0 or CoFAR V.3.0 for allergy-related AE); laboratory safety assessments (hematology, serum chemistry, coagulation panel, and urinalysis); physical examinations, including vital signs (blood pressure, heart rate, and body temperature); 12-lead electrocardiogram (ECG); serum cytokines (TNF-α, IL-2, IL-6, IL-8, IL-1β, MCP-1, MIP-1β, MIP-1α, IFN-γ, and IL-12p70).

[0280] Exploratory endpoints include: the difference in the percentage of subjects passing DBPCFC (not reaching the 2000 mg dose level or previous provocation dose, cumulative 4043 mg) between placebo and CNP-201 at Day 60 and Day 180; the change in the cumulative tolerated dose (CTD) (mg) of peanut protein administered during DBPCFC between placebo and CNP-201 at Day 60 and Day 180; the change in the ratio of peanut-specific IgE to IgG measured by ImmunoCap between placebo and CNP-201 at baseline and Day 38; the change in peanut-specific IgE measured by ImmunoCap between placebo and CNP-201 at baseline and Day 38; the change in the proportion of peanut-specific Th2a+ T cells (peanut-specific Th2a+ cells / total peanut-specific T cells) after ex vivo stimulation of PBMC between placebo and CNP-201 at baseline and Day 15; the change in the effective concentration at 50% maximum basophil activation (EC50) measured by basophil activation test (CD203c+ / CD63+ / - basophil activation) between placebo and CNP-201 at baseline, Day 60, and Day 180; the change in the proportion of peanut-specific T regulatory cells (peanut-specific T regulatory cells / peanut-specific CD4+ effector memory cells) after ex vivo stimulation of PBMC between placebo and CNP-201 at baseline and Day 15. Example 4 - Interim Results of a Phase I / II Trial of Low-Dose TIMP-PPE in Peanut Allergy

[0281] This example describes the results of a Phase 1b / 2a randomized, double-blind, placebo-controlled study demonstrating the efficacy of administering low-dose CNP-201 in the treatment of peanut allergy.

[0282] Subjects received a single intravenous dose of 1 mg or 25 mg of CNP-201 on Day 1. Figure 2 Results are summarized for peanut-allergic subjects who received low-dose CNP-201 particles with respect to peanut allergy. Three out of four patients who received low-dose CNP-201 had an increased BAT threshold, with two patients having a duration extended up to 60 days. Peanut-allergic patients who received up to 25 mg of CNP-201 showed a decrease in the ratio of peanut-specific IgE to IgG on Days 15 and 38. Peanut-allergic patients who received a single dose of 25 mg of CNP-201 showed induction of antigen-specific Tregs and a decrease in pathogenic peanut-specific T cell subsets (Th2a, TFH, B cell plasmablasts).

[0283] Blood samples were taken before and after low-dose CNP-201 treatment for BAT testing. PBMCs isolated from the blood were stimulated with increasing concentrations of peanut allergen. Basophil degranulation was measured by flow cytometry through the expression of CD203 and CD63 positive markers on basophils. The activation threshold, i.e., the critical value for a positive response, was 50% CD203 + CD63 + expression. In Figure 3 , basophil activation was detected before treatment with 25 mg of CNP-201. Peanut-allergic subjects who received a single intravenous dose of 25 mg of CNP-201 showed an increase in the EC50 (peanut allergen concentration) required for increased expression of activation markers on the surface of basophils after stimulation with the allergen. After treatment with CNP-201, basophil degranulation was below the activation threshold on Days 15, 38, and 60. The increase in BAT indicates unresponsiveness to peanut allergen.

[0284] Peanut-specific IgE and IgG levels in the patient sera were determined. Peanut-allergic patients who received up to 25 mg of CNP-201 intravenously showed an increase in peanut-specific IgG, while peanut-specific IgE levels did not change ( Figure 4 ). Compared to placebo, a decrease in the peanut-specific IgE / IgG ratio was detected on Days 15 and 38 after dosing ( Figure 4 ).

[0285] PBMCs obtained from the blood of peanut-allergic patients who received a single dose of 25 mg CNP-201 were stimulated with whole peanut extract and CD40 ligand. Peanut-specific activated T cell subsets were further characterized by the expression of lineage-specific markers. Figure 5 It was shown that up to 100 days after administration of a single dose of 25 mg CNP-201, peanut allergy-related immune cell subsets were reduced. Peanut-allergic patients who received a single intravenous dose of 25 mg CNP-201 showed a reduction in the pro-allergic subsets T helper cell 2 (Th2A), T follicular helper cells (TFH), terminally differentiated effector memory cells (TEMRA), and B cell plasmablasts. Figure 6 It was shown that there were fewer pathogenic activated peanut-specific CD4 + T cells. Compared with placebo, peanut-specific activated T cell subsets CD4 + CD25 + 、CD4 + CD69 + and CD4 + PD-1 + were reduced in peanut-allergic patients treated with low-dose CNP-201. Peanut-allergic subjects who received a single intravenous dose of 25 mg CNP-201 showed an increase in peanut-specific Tregs (CD4 + CD137 + CD25 + CD127 lo ), compared with placebo ( Figure 7 ). Example 5: Phase I / II trial of 1 mg dose of TIMP-PPE in peanut allergy

[0286] This example describes a randomized, double-blind, placebo-controlled 1b / 2a-phase study to evaluate the safety, tolerability, and pharmacodynamics of TIMP-PPE (CNP-201) in peanut-allergic subjects aged 16 to 55 years.

[0287] CNP-201 consists of PLGA nanoparticles encapsulating purified peanut extract, with an average diameter of 400 - 800 nm and a negative ζ potential between -30 mV and -80 mV. CNP-201 particles are provided in a lyophilized formulation. CNP-201 particles are reconstituted in sterile water for injection and diluted with sterile saline (0.9% sodium chloride) before administration.

[0288] This study is a randomized, double-blind, placebo-controlled study of the safety, pharmacodynamics, and tolerability of multiple escalating dose levels of CNP-201 (escalation phase) with the goal of determining a safe and tolerable dose level for further evaluation in a large number of subjects (expansion phase). This study will enroll 4 cohorts to receive CNP-201 or placebo at multiple escalating dose levels.

[0289] During the study, subjects will continue their current SoC, excluding a 12-hour washout period (for beta-agonists, theophylline, and cromolyn only) and a 7-day washout period for antihistamines prior to skin prick testing (SPT). Subjects will resume their SoC regimen after SPT and continue on SoC during dosing / study.

[0290] Peanut-allergic subjects eligible for this study are defined based on the following inclusion criteria: 1. Males and non-pregnant females, aged 16 to 55 years (inclusive). 2. Subjects with a physician-diagnosed peanut allergy or a documented history of peanut allergy. 3. Subjects with a screening weight ≥ 31.25 kg. Subjects outside this range may be included at the discretion of the investigator. 4. Subjects with a documented history of non-severe peanut allergic reactions (≤ grade 3), including mild wheezing or dyspnea without hypoxia. 5. Subjects with a peanut-specific IgE ≥ 5 kU / L measured by ImmunoCAP at screening, unless previously treated for peanut allergy with OIT. Subjects previously treated for peanut allergy with OIT and those without a peanut-specific IgE ≥ 5 kU / L measured by ImmunoCap at screening may be included at the discretion of the investigator. 6. Subjects with a positive peanut SPT, with a change in wheal diameter ≥ 5 mm compared to the negative control (50% glycerol) at screening. Subjects previously treated for peanut allergy with OIT and those without a positive peanut skin prick test (SPT) with a change in wheal diameter ≥ 5 mm at screening may be included at the discretion of the investigator. 7. Subjects who self-report a peanut-free diet for at least 14 days prior to screening, no suspected peanut exposure (including any peanut food challenges), and agree to continue to restrict peanut exposure during the study. 8. Female and male subjects and their female spouses / partners who are willing to practice highly effective contraceptive methods, which may include but are not limited to abstinence, sexual relations only with the same sex, monogamous relationships with vasectomized partners, vasectomy, hysterectomy, bilateral tubal ligation, approved hormonal methods, intrauterine devices (IUDs), or spermicides in combination with barrier methods (e.g., condoms, diaphragms) starting from screening and continuing throughout the study until Day 38 (EOS / ET). 9. Female subjects who are willing to refrain from breastfeeding starting from initial screening and continuing until Day 60. 10. Female subjects who are willing to refrain from donating eggs starting from initial screening and continuing until Day 60. 11. Subjects who are willing and able to provide a written informed consent approved by the Institutional Review Board (IRB). 12. Subjects who are willing to perform and comply with all study procedures. 13. Male subjects who are willing to refrain from donating sperm starting from screening and continuing until Day 60.

[0291] Subjects who meet all inclusion and no exclusion criteria after completing the screening visit are enrolled in one of the multiple dosing groups. During the escalation period, subjects will receive CNP-201 by intravenous infusion on Day 1 and Day 8. The dose level for the first group is 1 mg. The other dose levels for the remaining 3 groups decrease to 0.1 mg, 0.25 mg, 0.5 mg, 1 mg, 2 mg, 2.5 mg, 5 mg, 10 mg or may increase to a maximum of 650 mg.

[0292] Subjects who meet all inclusion and no exclusion criteria at screening are enrolled in this study. CNP-201 or placebo is administered by intravenous infusion using a progressive infusion rate. The following progressive infusion rates will be used to administer CNP-201: 1 mL / hour for the first 10 minutes, 2 mL / hour for the next 10 minutes, 5 mL / hour for the next 10 minutes, 10 mL / hour for the next 10 minutes, and 20 mL / hour for the remaining infusion time.

[0293] Unless infusion reactions, allergic reactions, or other adverse events require extended monitoring time, subjects remain outpatient on Day 1 and Day 8 (before TP administration) after admission until 4 hours after the drug administration on the same day. If the safety parameters are acceptable to the researchers, the subjects are discharged.

[0294] Subjects are followed up by telephone visits daily after each infusion (Days 2 to 7 and Days 9 to 14) to assess and record any AEs and drug changes.

[0295] Seven days after the second administration of CNP-201 (Day 15), the subject must return to the outpatient clinic for collection of safety laboratory samples, assessment of AEs and medication changes, and collection of PD measurements. During the post-dose period, safety and tolerability follow-up of the subject continues.

[0296] On Day 60 (end of study), the subject returns to the outpatient clinic for collection of immune safety laboratory samples, PD measurements, and a double-blind placebo-controlled food challenge (DBPCFC) consisting of peanut and placebo (oat) challenges. The DBPCFC is conducted by a study physician or staff member trained in the management of on-site clinical emergencies, with immediate access to emergency medications and equipment, and near the hospital emergency department to provide rapid emergency care if needed.

[0297] At the end of the study visit, the subject then returns to the outpatient clinic on Day 60 for collection of safety laboratory samples, PD measurements, and final assessment of AEs and medication changes.

[0298] The total study duration for an individual subject is approximately 240 days; up to 180 days for screening; 8 days of CNP-201 or placebo administration and 52 days of post-administration evaluation ( Figure 8 ).

[0299] After all subjects in the dose cohort complete the clinic visit on Day 15 (7 days after the second dose), the DMC is convened to review all available safety data. At this time, the DMC determines whether it is acceptable to proceed to the next dose cohort and makes recommendations regarding the dose level of that cohort.

[0300] If a safe and effective dose is identified during the escalation phase, the study proceeds to the expansion phase. Subjects in the expansion phase are randomized at a 1:1 ratio to receive CNP-201 or placebo at the safe and tolerable dose level identified during the escalation phase. Subjects in the escalation and expansion phases will undergo the same evaluations.

[0301] The primary objectives include: safety and tolerability of CNP-201.

[0302] The exploratory objectives include: changes in response to DBPCFC in patients treated with CNP-201 or placebo, changes in the ratio of peanut-specific IgE to IgG in patients treated with CNP-201 or placebo, changes in peanut-specific IgE in patients treated with CNP-201 or placebo, changes in the proportion of peanut-specific Th2a+ T cells in patients treated with CNP-201 or placebo, changes in response to the basophil activation test in patients treated with CNP-201 or placebo, changes in the proportion of peanut-specific regulatory T cells in patients treated with CNP-201 or placebo.

[0303] The primary endpoints included: the frequencies of adverse events (AEs) and serious adverse events (SAEs), MedDRA 23.0 (CTCAE v.5.0 or CoFAR V.3.0 for allergy-related AEs); laboratory safety assessments (hematology, serum chemistry, coagulation panel, and urinalysis); physical examinations, including vital signs (blood pressure, heart rate, and body temperature); 12-lead electrocardiogram (ECG); serum cytokines (TNF-α, IL-2, IL-6, IL-8, IL-1β, MCP-1, MIP-1β, MIP-1α, IFN-γ, and IL-12p70).

[0304] The exploratory endpoints included: the difference in the percentage of subjects between placebo and CNP-201 at Day 60 by double-blind placebo-controlled food challenge (DBPCFC) (not reaching the 2000 mg dose level or previous provocative dose, cumulative 4043 mg); the change in the cumulative tolerated dose (CTD) (mg) of peanut protein administered during DBPCFC between placebo and CNP-201 at Day 60; the change in the ratio of peanut-specific IgE to IgG measured by ImmunoCap assay between placebo and CNP-201 at baseline and Day 60; the change in peanut-specific IgE measured by ImmunoCap assay between placebo and CNP-201 at baseline and Day 60; the change in the proportion of peanut-specific Th2a+ T cells (peanut-specific Th2a+ cells / total peanut-specific T cells) after ex vivo stimulation of PBMCs between placebo and CNP-201 at baseline and Day 15; the change in the effective concentration at 50% maximal basophil activation (EC50) measured by basophil activation test (CD203c+ / CD63+ / - basophil activation) between placebo and CNP-201 at baseline and Day 15 after ex vivo stimulation of PBMCs; the change in the proportion of peanut-specific regulatory T cells (peanut-specific regulatory T cells / peanut-specific CD4+ effector memory cells) after ex vivo stimulation of PBMCs between placebo and CNP-201 at baseline and Days 15 and 60.

[0305] This document describes preferred embodiments of the present disclosure. After reading the foregoing description, variations of those preferred embodiments may become apparent to those of ordinary skill in the art. The inventors expect that those skilled in the art may adopt such variations as appropriate, and the inventors intend for the present disclosure to be practiced in other ways different from those specifically described herein. Accordingly, to the extent permitted by applicable law, the present disclosure includes all modifications and equivalents of the subject matter recited in the appended claims. In addition, unless otherwise indicated herein or clearly contradicted by context, the present disclosure covers any combination of the above elements in all possible variations thereof.

Claims

1. A method of treating peanut allergy in a subject, the method comprising administering to the subject either a tolerogenic immunomodulatory particle (TIMP-PPE) encapsulating peanut protein alone or in combination with one or more therapeutic agents, wherein the TIMP-PPE is administered at a dose level between about 0.001 mg / kg and 12 mg / kg.

2. The method according to claim 1, wherein the TIMP-PPE particles have an average diameter between 100 nm and 1500 nm.

3. The method according to claim 1 or 2, wherein the TIMP-PPE particles have a negative ζ potential.

4. The method according to any one of the preceding claims, wherein the negative ζ potential of the particles is between -30 mV and -100 mV.

5. The method according to any one of the preceding claims, wherein the TIMP-PPE is administered at a concentration between about 0.0005 mg / mL and about 50 mg / mL, optionally 0.0005 mg / mL, 0.001 mg / mL, 0.005 mg / mL, 0.01 mg / mL, 0.05 mg / mL, 0.1 mg / mL, 0.5 mg / mL, 1 mg / mL, 2 mg / mL, 3 mg / mL, 4 mg / mL, 5 mg / mL, 6 mg / mL, 7 mg / mL, 8 mg / mL, 9 mg / mL, 10 mg / mL, 11 mg / mL, 12.5 mg / mL, 15 mg / mL, 17.5 mg / mL, 20 mg / mL, 25 mg / mL, 30 mg / mL, 40 mg / mL or 50 mg / mL.

6. The method according to any one of the preceding claims, wherein the TIMP-PPE is administered at a dose level of about 0.001 mg / kg, 0.0025 mg / kg, 0.005 mg / kg, 0.01 mg / kg, 0.025 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.25 mg / kg, 0.5 mg / kg, 1.0 mg / kg, 2.0 mg / kg, 4.0 mg / kg, 6 mg / kg, 8.0 mg / kg, 10 mg / kg or 12 mg / kg.

7. The method according to any one of the preceding claims, wherein the TIMP-PPE is administered at a dose level between about 0.1 mg and 800 mg.

8. The method according to any one of the preceding claims, wherein the TIMP-PPE is administered at a dose of about 0.1 mg, 0.25 mg, 0.5 mg, 1 mg, 2 mg, 2.5 mg, 5 mg, 10 mg, 25 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 325 mg, 350 mg, 400 mg, 425 mg, 450 mg, 475 mg, 500 mg, 525 mg, 550 mg, 575 mg, 600 mg, 625 mg, 650 mg, 675 mg, 700 mg, 725 mg, 750 mg, 775 mg or 800 mg.

9. The method according to any one of the preceding claims, wherein the TIMP-PPE is administered in a single dose or multiple doses.

10. The method according to any one of the preceding claims, wherein the TIMP-PPE is administered once a week, once every two weeks, once every three weeks, once every 4 weeks, once every two months, once every three months, once every 6 months or once a year.

11. The method according to any one of the preceding claims, wherein the TIMP-PPE is administered in two doses at one-week intervals.

12. The method according to any one of the preceding claims, wherein the TIMP-PPE is administered intravenously, subcutaneously, intramuscularly, intraperitoneally, intranasally or orally.

13. The method according to any one of the preceding claims, wherein the TIMP-PPE is administered at a concentration between 0.0005 mg / mL and 50 mg / mL.

14. The method according to any one of the preceding claims, wherein the TIMP-PPE is administered in two doses at one-week intervals and then a booster dose of TIMP-PPE is re-administered in a single dose every three months.

15. The method according to any one of claims 1 to 14, wherein the therapeutic agent administered in combination with TIMP-PPE is an anti-IgE antibody, an anti-IL-4Rα antibody, an anti-IL13 antibody, an anti-IL-33 antibody, an antihistamine, a steroid, a corticosteroid, a leukotriene regulator, low-dose IL-2, an IL-2 mutant protein engineered to expand Tregs, an IL-2 variant engineered to expand Tregs, an IL-2 molecule engineered to be selective for the high-affinity IL-2 receptor, PEGylated IL-2, an IL-2 complex, an IL-2 / CD25 fusion protein, a prebiotic, a probiotic, a histone deacetylase inhibitor, short-chain fatty acids (e.g., acetate, butyrate, propionate, butyrate polymers), an IgE inhibitor, a competitor of IgE for the allergen binding site, a basophil activation inhibitor, a mast cell activation inhibitor, a cytokine inhibitor, a microbiome therapy, a small molecule or a biotherapeutic agent or a non-steroidal anti-inflammatory drug (NSAID).

16. The method according to claim 15, wherein the anti-IgE antibody is omalizumab.

17. The method according to claim 15 or 16, wherein the anti-IgE antibody is administered subcutaneously.

18. The method according to any one of claims 15 to 17, wherein the anti-IgE antibody is administered in a single dose or multiple doses.

19. The method according to any one of claims 15 to 18, wherein the anti-IgE antibody is administered before, concomitantly with, or after TIMP-PPE.

20. The method according to any one of claims 15 to 19, wherein the anti-IgE antibody is administered in three doses two weeks apart or in two doses four weeks apart before TIMP-PPE.

21. The method according to any one of claims 15 to 20, wherein the anti-IgE antibody is administered at a dose between about 10 mg and 500 mg.

22. The method according to claim 21, wherein the anti-IgE antibody is administered at a dose of about 10 mg, 25 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 375 mg, 400 mg, 425 mg, 450 mg, 475 mg, or 500 mg.

23. The method according to claim 21 or 22, wherein the dose level of the anti-IgE antibody is determined based on the level of IgE in the blood of the subject.

24. The method according to claim 21 or 22, wherein the dose level of the anti-IgE antibody is determined based on the body weight of the subject.

25. The method according to any one of the preceding claims, wherein administering TIMP-PPE alone or in combination with another therapeutic agent to the subject reduces one or more symptoms of peanut allergy.

26. The method according to claim 19, wherein one or more symptoms of peanut allergy are selected from the group consisting of: skin reactions, urticaria, skin flushing, skin swelling, itching, tightness in the throat, difficulty breathing, shortness of breath, digestive problems such as diarrhea, stomach cramps, nausea or vomiting, drop in blood pressure, and anaphylactic reactions.

27. The method according to any one of the preceding claims, wherein administering to the subject the TIMP-PPE alone or in combination with a therapeutic agent shortens the duration and reduces the severity of the allergic immune response to peanut protein.

28. The method according to claim 27, wherein the allergic immune response is a Th2 cell response, B cell activation, basophil activation, eosinophil activation, mast cell activation, and / or IgE induction.

29. The method according to claim 28, wherein the Th2 cell response, B cell activation, basophil activation, eosinophil activation, mast cell activation, and / or IgE induction is determined in one or more biological samples obtained from the subject.

30. The method according to claim 29, wherein the biological sample is selected from the group consisting of: whole blood, peripheral blood, peripheral blood mononuclear cells (PBMC), serum, plasma, urine, cerebrospinal fluid (CSF), feces, tissue biopsy, and / or bone marrow biopsy.

31. The method according to claim 27, wherein administering to the subject the TIMP-PPE alone or in combination with a therapeutic agent reduces the proportion of Th2a+ T cells present in the total T cell population in peripheral blood.

32. The method according to claim 27, wherein administering to the subject the TIMP-PPE alone or in combination with an additional therapeutic agent reduces the proportion of activated peanut protein-specific T cells in peripheral blood.

33. The method according to claim 27, wherein administering to the subject the TIMP-PPE alone or in combination with a therapeutic agent increases the level of peanut protein-specific Treg cells in the blood.

34. The method according to claim 27, wherein administering to the subject the TIMP-PPE alone or in combination with a therapeutic agent reduces basophil activation.

35. The method according to claim 34, wherein basophil activation is determined by in vitro stimulation of basophils with peanut protein in a basophil activation test (BAT).

36. The method according to claim 27, wherein administering to the subject the TIMP-PPE alone or in combination with a therapeutic agent reduces the level of peanut protein-specific IgE in the blood.

37. The method according to claim 27, wherein administering to the subject the TIMP-PPE alone or in combination with a therapeutic agent reduces the ratio of peanut protein-specific IgE to IgG levels in the blood.

38. The method according to any one of claims 1 to 37, wherein administering to the subject the TIMP-PPE alone or in combination with a therapeutic agent reduces the level of Th2 cytokine levels in the blood.

39. The method according to claim 38, wherein the Th2 cytokines are selected from the group consisting of: IL-4, IL-5, IL-9, and IL-13.

40. The method according to any one of the preceding claims, wherein administering the TIMP-PPE alone or in combination with a therapeutic agent to the subject increases tolerance to peanut protein.

41. The method according to claim 40, wherein tolerance to peanut protein is determined by a double-blind placebo-controlled food challenge.

42. The method according to claim 40, wherein tolerance to peanut protein is determined by a skin prick test (SPT).

43. The method according to any one of the preceding claims, wherein the TIMP-PPE comprises a peanut extract or one or more peanut proteins or antigenic fragments thereof selected from the group consisting of: Ara h1, Ara h2, Ara h3, Ara h5, Ara h6, Ara h7, Ara h8, Ara h9, Ara h10, Ara h11, Ara h12, Ara h13, Ara h14, Ara h15, Ara h16, Ara h17, and Ara h18.

44. The method according to any one of the preceding claims, wherein the TIMP-PPE comprises one or more proteins or antigenic fragments thereof of Ara h1, Ara h2, Ara h3, Ara h5, Ara h6, Ara h7, or Ara h8, or a combination thereof.

45. The method according to any one of the preceding claims, wherein the subject is administered a combination of an antihistamine, a corticosteroid, an NSAID, and / or a leukotriene modifier with the TIMP-PPE.

46. The method according to claim 45, wherein the antihistamine is diphenhydramine.

47. The method according to claim 45 or 46, wherein the antihistamine is administered at a dose of about 0.05 mg, 0.1 mg, 0.5 mg, 1 mg, 2 mg, 4 mg, 5 mg, 10 mg, 25 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 375 mg, 400 mg, 425 mg, 450 mg, 475 mg, 500 mg, 1000 mg, 1500 mg, or 2000 mg.

48. The method according to any one of claims 45 to 47, wherein the antihistamine is administered intravenously.

49. The method according to any one of claims 45 to 48, wherein the antihistamine is administered in a single dose or multiple doses.

50. The method according to any one of claims 45 to 49, wherein the antihistamine is administered 30 minutes before administering the TIMP-PPE.

51. The method according to claim 45, wherein the corticosteroid is methylprednisolone.

52. The method according to any one of claims 45 to 51, wherein the corticosteroid is administered in a dose of about 0.05 mg, 0.1 mg, 0.5 mg, 1 mg, 2 mg, 4 mg, 5 mg, 10 mg, 25 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 375 mg, 400 mg, 425 mg, 450 mg, 475 mg, 500 mg, 1000 mg, 1500 mg or 2000 mg.

53. The method according to any one of claims 45 to 52, wherein the corticosteroid is administered intravenously.

54. The method according to any one of claims 45 to 53, wherein the corticosteroid is administered in a single dose or multiple doses.

55. The method according to any one of claims 45 to 54, wherein the corticosteroid is administered 30 minutes before administering TIMP-PPE.

56. The method according to claim 45, wherein the NSAID is acetylsalicylic acid.

57. The method according to any one of claims 45 and 56, wherein the NSAID is administered in a dose of about 0.05 mg, 0.1 mg, 0.5 mg, 1 mg, 2 mg, 4 mg, 5 mg, 10 mg, 25 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 325 mg, 375 mg, 400 mg, 425 mg, 450 mg, 475 mg, 500 mg, 1000 mg, 1500 mg or 2000 mg.

58. The method according to claim 45 or any one of claims 56 to 57, wherein the NSAID is administered orally.

59. The method according to claim 45 or any one of claims 56 to 58, wherein the NSAID is administered in a single dose or multiple doses.

60. The method according to claim 45 or any one of claims 56 to 59, wherein the NSAID is administered daily for two days and is administered 60 minutes before administering TIMP-PPE.

61. The method according to claim 45, wherein the leukotriene modifier is montelukast.

62. The method according to any one of claims 45 or 61, wherein the leukotriene regulator is administered at a dose of about 0.05 mg, 0.1 mg, 0.5 mg, 1 mg, 2 mg, 4 mg, 5 mg, 10 mg, 25 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 325 mg, 375 mg, 400 mg, 425 mg, 450 mg, 475 mg, 500 mg, 1000 mg, 1500 mg or 2000 mg.

63. The method according to any one of claims 45 or 61 to 62, wherein the leukotriene regulator is administered orally.

64. The method according to any one of claims 45 or 61 to 63, wherein the leukotriene regulator is administered in a single dose or multiple doses.

65. The method according to any one of claims 45 or 61 to 64, wherein the leukotriene regulator is administered 12 hours before, 45 minutes before and / or 12 hours after the administration of TIMP-PPE.

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