Treating peanut allergy with tolerizing nanoparticles

Abstract

Peanut allergy is one of the most common food allergies, affecting nearly 1.2% of the total US population and 2.5% of the pediatric population, and has been increasing in incidence over the past decade. The present disclosure generally relates to methods for treating peanut allergy using nanoparticles encapsulating peanut antigens to induce antigen-specific tolerance. A method for treating peanut allergy in a subject is provided, comprising administering to the subject tolerizing immune-modifying particles encapsulating peanut proteins (TIMP-PPE), wherein the TIMP-PPE is administered at a dose of 0.001 to 12 mg / kg.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 380,173, filed October 19, 2022, and U.S. Provisional Patent Application No. 63 / 486,812, filed February 24, 2023, which are incorporated by reference in their entireties.

[0002] The present disclosure relates to methods of treating peanut allergy using tolerizing, immunomodulating nanoparticles encapsulating purified peanut extracts containing allergenic peanut proteins or antigenic fragments thereof.

[0003] Description of electronically submitted text files The contents of the text file submitted electronically herein are incorporated herein by reference in their entirety: Computer-readable format copy of the Sequence Listing (Filename: 58512_SeqListing.xml, Creation Date: October 18, 2023, File Size: 27,434 bytes). [Background technology]

[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, with incidence rates increasing over the past decade (Cannon HE. Am J Manag Care. 2018;24(19 Suppl):S428-s433). Peanut allergy is caused by a pathological hyperimmune response, and exposure to peanuts can lead to mild to severe symptoms, including nausea, vomiting, rash, breathing problems, low blood pressure, and even death.

[0005] While normal individuals with healthy immune systems can maintain non-responsiveness to antigens encountered in common food substances such as peanuts, in peanut-allergic subjects, loss of immune tolerance to peanut antigens drives pathological hyperimmune responses. To date, eight peanut proteins (Ara h1–Ara h8) have been identified as the predominant antigenic peanut proteins that drive allergic hyperimmune responses (Keet et al., J Allergy Clin Immunol Pract. 2013;1(1):101–103). Additional peanut allergens have also been reported recently, bringing the total to 18 allergenic peanut proteins (Ozias-Akins et al., Allergy. 2019 May;74(5):888–898). Allergic immune responses to peanut antigenic proteins are mediated by T cell-dependent mechanisms, including upregulated T helper type 2 (Th2) cytokine production (e.g., IL-4, IL-5, IL-9, and IL-13), and B cell class switching, which leads to the production of IgE antibodies and degranulation of mast cells and basophils (Sampath et al., J Clin Invest. 2019;129(4):1431-1440).

[0006] Currently, there is no cure for peanut allergy, and strict avoidance of exposure to peanut antigens and management of anaphylaxis are the only options available to patients. Immune tolerization therapy, which can induce T cell tolerance to peanut antigens, is considered the gold standard for treating peanut allergy but has been difficult to achieve. Several attempts to develop immune tolerization therapy have been made using oral immunotherapy (OIT), subcutaneous immunotherapy (SCIT), epicutaneous immunotherapy (EPIT), and sublingual immunotherapy (SLIT) approaches (Feuille et al., Allergy Asthma Immunol Res. 2018;10(3):189-206). The success of these therapies is highly variable, with only desensitization to peanut proteins reported, which provides protection only against accidental exposure but is not a cure (Chinthrajah et al., Lancet. 2019;394(10207):1437-1449; Vickery et al., N Engl J Med. 2018;379(21):1991-2001; Fleischer et al., J. Am Med Assoc. 2019;321(10):946-955). Summary of the Invention

[0007] Tolerizing immune-modifying particles (TIMPs) containing one or more antigens have previously been described for the treatment of immune-mediated disorders (e.g., autoimmune diseases and allergies) through the induction of antigen-specific immune tolerance (WO2013 / 1319253 and WO2015 / 023796, which are incorporated herein by reference). In several preclinical models of autoimmune disease and allergy, TIMPs have demonstrated efficacy in inducing T cell tolerance. Induction of antigen-specific T cell tolerance to peanut antigens using TIMPs (TIMP-PPE) encapsulating purified peanut extract (PPE) and / or comprising 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, Ara h14, Ara h15, Ara h16, Ara h17, and Ara h18, may ameliorate or potentially cure peanut allergy (PA).

[0008] Provided herein are methods for treating peanut allergy in a subject, comprising administering TIMP-PPE to the subject, wherein the TIMP-PPE is administered at a dose of 0.001 to 12 mg / kg. Also provided herein are methods for reducing an allergic immune response to a peanut antigen in a subject suffering from PA, comprising administering TIMP-PPE to the subject, wherein the TIMP-PPE is administered at a dose of about 0.001 to 12 mg / kg. In various embodiments, the TIMP-PPE is administered at a dose of about 0.001-10 mg / kg, about 0.005-12 mg / kg, about 0.01-12 mg / kg, about 0.05-12 mg / kg, about 0.1-12 mg / kg, about 0.5-10 mg / kg, about 1-8 mg / kg, about 1.5-10 mg / kg, about 2-12 mg / kg, about 2-10 mg / kg, about 3-10 mg / kg, about 4-10 mg / kg, about 4-12 mg / kg, or about 5-12 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, the TIMP-PPE is administered at a concentration of about 0.0005 mg / mL to about 50 mg / mL, hi various embodiments, the 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, the TIMP-PPE is administered via intravenous infusion lasting 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, one week apart. In various embodiments, TIMP-PPE is administered once a week, once every two weeks, once every three weeks, once every four weeks, once every two months, once every three months, once every six 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 initial TIMP-PPE administration. In various embodiments, the booster dose of TIMP-PPE is administered once a week, once every two weeks, once every three weeks, once every four weeks, once every two months, once every three months, once every six months, or once a year. In various embodiments, TIMP-PPE is administered in two doses, one week apart, followed by a booster dose of TIMP-PPE re-administered as a single dose once every three months.

[0012] In various embodiments, the booster dose of TIMP-PPE is re-administered at a dose of 0.001 mg / kg to 12 mg / kg. In various embodiments, the booster dose of TIMP-PPE is about 0.001 to 10 mg / kg, about 0.005 to 12 mg / kg, about 0.01 to 12 mg / kg, about 0.05 to 12 mg / kg, about 0.1 to 12 mg / kg, about 0.5 to 10 mg / kg, about 1 to 8 mg / kg, about 1.5 to 10 mg / kg, about 2 to 12 mg / kg, about 2 to 10 mg / kg, about 3 to 10 mg / kg, about 4 to 10 mg / kg, about 4 to 12 mg / kg, about 5 to 10 mg / kg, about 6 to 10 mg / kg, about 7 to 10 mg / kg, about 8 to 12 mg / kg, about 9 to 10 mg / kg, about 11 to 12 mg / kg, about 12 to 14 mg / kg, about 14 to 16 mg / kg, about 15 to 16 mg / kg, about 16 to 18 mg / kg, about 17 to 18 mg / kg, about 18 to 19 mg / kg, about 19 to 20 mg / kg, about 20 to 22 mg / kg, about 21 to 22 mg / kg, about 22 to 24 mg / kg, about 23 to 24 mg / kg, about 24 to 24 mg / kg, about 25 to 26 mg / kg, about 26 to 28 mg / kg, about 27 to 28 mg / kg, about 28 to 29 mg / kg, about 29 to 30 mg / kg, about 30 to 31 mg / kg, about 31 to 32 mg / kg, or about 5-12 mg / kg, or 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, 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 of about 0.0005 mg / mL to 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 TIMP-PPE booster dose is administered via intravenous infusion lasting about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 10, 12, 18, or 20 hours.

[0014] In various embodiments, the TIMP-PPE encapsulates peanut protein and / or antigenic fragments of peanut protein 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, or Ara h18. In various embodiments, the TIMP-PPE encapsulates peanut protein and / or antigenic fragments of peanut protein 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, Ara h18, or combinations thereof. In various embodiments, 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.

[0015] In various embodiments, 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 zeta potential. In various embodiments, the negative zeta potential of the TIMP-PPE particles is about -100 mV to about 0 mV. In various embodiments, the zeta potential of the particles is about -100 mV to about -25 mV, about -100 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 to about -35 mV, about -70 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 zeta potential is about -25mV, -30mV, -35mV, -40mV, -45mV, -50mV, -55mV, -60mV, -65mV, -70mV, -75mV, -80mV, -85mV, -90mV, -95mV, or -100mV.

[0016] In various embodiments, the size, or diameter, of the TIMP-PPE particles is from 0.05 μm to about 10 μm. In various embodiments, the diameter of the TIMP-PPE particles is from 0.1 μm to about 10 μm. In various embodiments, the diameter of the TIMP-PPE particles is from 0.1 μm to about 5 μm. In various embodiments, the diameter of the TIMP-PPE particles is from 0.1 μm to about 3 μm. In various embodiments, the diameter of the TIMP-PPE particles is from 0.3 μm to about 5 μm. In various embodiments, the diameter of the TIMP-PPE particles is from about 0.3 μm to about 3 μm. In various embodiments, the diameter of the TIMP-PPE particles is from about 0.3 μm to about 1 μm. In various embodiments, the diameter of the TIMP-PPE particles is from about 0.4 μm to about 1 μm. In various embodiments, TIMP-PPE particles have a diameter of about 100-10,000 nm, about 100-5,000 nm, about 100-3,000 nm, about 100-2,000 nm, about 300-5,000 nm, about 300-3,000 nm, about 300-1,000 nm, about 300-800 nm, about 400-800 nm, or about 200-700 nm. In various embodiments, TIMP-PPE particles have a diameter of about 50 nm, 100 nm, 200 nm, 300 nm, 400 nm, 500 nm, 600 nm, 700 nm, 800 nm, 900 nm, 1,000 nm, 1,100 nm, 1,200 nm, 1,300 nm, 1,400 nm, 1,500 nm, or 2,000 nm. In various embodiments, the negatively charged particles have a diameter of between 400 nm and 800 nm.

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

[0018] In various embodiments, the TIMP-PPE is administered alone or in combination with one or more additional therapeutic agents. In various embodiments, the present disclosure provides methods of treating peanut allergy or peanut allergy-related symptoms in a subject in need thereof, comprising administering to the subject a composition comprising a 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 IL-2 therapy, which induces Tregs. In various embodiments, the IL-2 therapy is low-dose IL-2, an IL-2 mutein 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 conjugate, or an IL-2 / CD25 fusion protein. In various embodiments, the additional therapeutic agent is an inhibitor of IgE, an inhibitor of basophil activation, an inhibitor of mast cell activation, an antihistamine, or a small molecule or biological therapeutic. In various embodiments, the additional therapeutic agent is an inhibitor of IgE, a competitor of IgE for allergen binding sites, an inhibitor of basophil activation, an inhibitor of mast cell activation, an antihistamine, a cytokine inhibitor, microbiome therapy, a small molecule, or a biological therapeutic agent. In various embodiments, the additional therapeutic agent inhibits IgE. In various embodiments, the additional therapeutic agent inhibits IgE antibodies. In various embodiments, the additional therapeutic agent inhibits basophil activation. In various embodiments, the additional therapeutic agent inhibits mast cell activation. In various embodiments, the additional therapeutic agent is a biological or small molecule. In various embodiments, the 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 nonsteroidal anti-inflammatory drug (NSAID).

[0020] In various embodiments, a therapeutic agent useful for treating peanut allergy is an inhibitor of IgE, a competitor of IgE for allergen binding sites, an inhibitor of basophil activation, an inhibitor of mast cell activation, an antihistamine, a cytokine inhibitor, a microbiome therapy, a small molecule, or a biological 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 biological or 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 nonsteroidal 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, levocetirizine, 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 mutein 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 conjugate, an IL-2 / CD25 fusion protein, a prebiotic, a probiotic, a histone deacetylase inhibitor, a short-chain fatty acid (e.g., acetate, butyrate, propionate, butyrate polymers), an inhibitor of IgE, a competitor of IgE for allergen binding sites, an inhibitor of basophil activation, an inhibitor of mast cell activation, a cytokine inhibitor, a microbiome therapy, a small molecule or biological therapeutic, or a nonsteroidal 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 nonsteroidal anti-inflammatory drug (NSAID). In various embodiments, the therapeutic agent useful for treating peanut allergy is a nonsteroidal 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 antileukotriene. 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, anti-IL-4Rα, anti-IL-13, or 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 administration of the TIMP-PPE. In various embodiments, the additional therapeutic agent is administered intravenously, subcutaneously, intramuscularly, intraperitoneally, intranasally, or orally.

[0028] In various embodiments, a therapeutic agent useful for treating peanut allergy is administered before, during, or after administration of the TIMP-PPE. In various embodiments, a 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 prior to, simultaneously with, or subsequent to administration of TIMP-PPE. In various embodiments, the therapeutic agent is administered 1, 2, 3, 4, 5, 6, or 7 days before administration of TIMP-PPE. In various embodiments, the therapeutic agent is administered 1, 2, 3, 4, 5, 6, or 7 weeks before 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 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 administration of TIMP-PPE. In various embodiments, the therapeutic agent is administered 1, 2, 3, 4, 5, 6, or 7 days after administration of TIMP-PPE. In various embodiments, the therapeutic agent is administered 1, 2, 3, 4, 5, 6, or 7 weeks after 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 after administration of the TIMP-PPE. In various embodiments, the therapeutic agent is administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 years after administration of the TIMP-PPE.

[0030] The present disclosure describes a method of treating peanut allergy in a subject, comprising administering to the subject a TIMP-PPE in combination with an anti-IgE antibody, wherein the TIMP-PPE is administered at a dose of about 0.001 to 12 mg / kg and 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, the TIMP-PPE is administered at a concentration of about 0.001-10 mg / kg, about 0.005-12 mg / kg, about 0.01-12 mg / kg, about 0.05-12 mg / kg, about 0.1-12 mg / kg, about 0.5-10 mg / kg, about 1-8 mg / kg, about 1.5-10 mg / kg, about 2-12 mg / kg, about 2-10 mg / kg, about 3-10 mg / kg, about 4-10 mg / kg, or about 4-12 mg / kg. g, or about 5-12 mg / kg, or 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, 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. 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 serum IgE levels.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 subject's weight. In various embodiments, the subject weighs between 30 and 40 kg, 40 and 50 kg, 50 and 60 kg, 60 and 70 kg, 70 and 80 kg, 80 and 90 kg, 90 and 125 kg, or 125 and 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 weekly, once every two weeks, once every three weeks, or once every four weeks. In various embodiments, the anti-IgE antibody is administered before, simultaneously with, or after administration of TIMP-PPE. In various embodiments, the anti-IgE antibody is administered 1 week, 2 weeks, 3 weeks, 4 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 administration of TIMP-PPE. In various embodiments, the anti-IgE antibody is administered 1 week, 2 weeks, 3 weeks, or 4 weeks after administration of TIMP-PPE.

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

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

[0034] The present disclosure describes a method of treating peanut allergy in a subject, comprising administering to the subject a TIMP-PPE in combination with an anti-IL-4Rα antibody, wherein the TIMP-PPE is administered at a dose of about 0.001 mg / kg to 12 mg / kg and 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, the TIMP-PPE is administered at a concentration of about 0.001-10 mg / kg, about 0.005-12 mg / kg, about 0.01-12 mg / kg, about 0.05-12 mg / kg, about 0.1-12 mg / kg, 0.5-10 mg / kg, about 1-8 mg / kg, about 1.5-10 mg / kg, about 2-12 mg / kg, about 2-10 mg / kg, about 3-10 mg / kg, about 4-10 mg / kg, or about 4-12 mg / kg. g, or about 5-12 mg / kg, or 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, 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. 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 serum IgE levels, which are 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 anti-IL-4Rα antibody dose level is determined based on the subject's body weight. In various embodiments, the subject's body weight 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 in 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, simultaneously with, or after administration of TIMP-PPE. In various embodiments, the anti-IL-4Rα antibody is administered 1 week, 2 weeks, 3 weeks, or 4 weeks before administration of TIMP-PPE. In various embodiments, the anti-IL-4Rα antibody is administered 1 week, 2 weeks, 3 weeks, or 4 weeks after administration of TIMP-PPE. In various embodiments, the anti-IL-4Rα antibody is administered at an initial dose level of 400 mg to 600 mg over two doses, followed by a maintenance dose level of 200 mg to 300 mg over subsequent doses.

[0035] In various embodiments, the antihistamine administered in combination with the TIMP-PPE is administered in a dose of 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 daily, once weekly, once every two weeks, once every three weeks, or once every four weeks. In various embodiments, the antihistamine is administered twice, three times, four times, five times, or six times daily. In various embodiments, the antihistamine is administered before, simultaneously with, or after administration of the TIMP-PPE. In various embodiments, the antihistamine is administered 1 week, 2 weeks, 3 weeks, 4 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 administration of the TIMP-PPE. In various embodiments, the antihistamine is administered 1 week, 2 weeks, 3 weeks, 4 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 after administration of the TIMP-PPE. In various embodiments, the antihistamine is administered 5, 10, 15, 30, 45, or 60 minutes before administration of the TIMP-PPE. In various embodiments, the antihistamine is administered 5, 10, 15, 30, 45, or 60 minutes after administration of the 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 administration of the 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 administration of the TIMP-PPE. In various embodiments, the antihistamine is administered 1, 2, 3, 4, 5, 6, or 7 days before administration of the TIMP-PPE. In various embodiments, the antihistamine is administered 1, 2, 3, 4, 5, 6, or 7 days after administration of the TIMP-PPE.

[0036] In various embodiments, the steroid administered in combination with the TIMP-PPE is administered at a dose of 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 daily, once weekly, 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 daily. In various embodiments, the steroid is administered before, simultaneously with, or after administration of TIMP-PPE. In various embodiments, the steroid is administered 1 week, 2 weeks, 3 weeks, 4 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 administration of TIMP-PPE. In various embodiments, the steroid is administered 1 week, 2 weeks, 3 weeks, 4 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 after administration of TIMP-PPE. In various embodiments, the steroid is administered 5, 10, 15, 30, 45, or 60 minutes before 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 prior to administration of the TIMP-PPE, hi various embodiments, the steroid is administered 1, 2, 3, 4, 5, 6, or 7 days prior to administration of the TIMP-PPE.

[0037] In various embodiments, the corticosteroid administered in combination with a TIMP-PPE is administered in a dose of 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 daily, once weekly, once every two weeks, once every three weeks, or once every four weeks. In various embodiments, the corticosteroid is administered twice, three times, four times, five times, or six times daily. In various embodiments, the corticosteroid is administered before, simultaneously with, or after administration of the TIMP-PPE. In various embodiments, the corticosteroid is administered 1 week, 2 weeks, 3 weeks, 4 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 administration of the TIMP-PPE. In various embodiments, the corticosteroid is administered 1 week, 2 weeks, 3 weeks, 4 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 after administration of the TIMP-PPE. In various embodiments, the corticosteroid is administered 5, 10, 15, 30, 45, or 60 minutes before administration of the TIMP-PPE. In various embodiments, the corticosteroid is administered 5, 10, 15, 30, 45, or 60 minutes after administration of the 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 administration of the 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 administration of the TIMP-PPE. In various embodiments, the corticosteroid is administered 1, 2, 3, 4, 5, 6, or 7 days before administration of the TIMP-PPE. In various embodiments, the corticosteroid is administered 1, 2, 3, 4, 5, 6, or 7 days after administration of the TIMP-PPE.

[0038] In various embodiments, the NSAID administered in combination with the 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, 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 twice, three times, four times, five times, or six times a day. In various embodiments, the NSAID is administered before, simultaneously with, or after administration of TIMP-PPE. In various embodiments, the NSAID is administered 1 week, 2 weeks, 3 weeks, 4 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 administration of TIMP-PPE. In various embodiments, the NSAID is administered 1 week, 2 weeks, 3 weeks, 4 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 after administration of TIMP-PPE. In various embodiments, the NSAID is administered 5, 10, 15, 30, 45, or 60 minutes before administration of the TIMP-PPE. In various embodiments, the NSAID is administered 5, 10, 15, 30, 45, or 60 minutes after administration of the 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 administration of the 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 administration of the TIMP-PPE. In various embodiments, the NSAID is administered 1, 2, 3, 4, 5, 6, or 7 days before administration of the TIMP-PPE. In various embodiments, the NSAID is administered 1, 2, 3, 4, 5, 6, or 7 days after administration of the TIMP-PPE.

[0039] In various embodiments, the leukotriene modifier administered in combination with TIMP-PPE is administered in a dose of about 0.05 mg to 2000 mg. In various embodiments, the leukotriene modifier 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 modifier is administered in a single dose or multiple doses. In various embodiments, the leukotriene modifier is administered once daily, once weekly, once every two weeks, once every three weeks, or once every four weeks. In various embodiments, the leukotriene modifier is administered twice, three times, four times, five times, or six times daily. In various embodiments, the leukotriene modifier is administered before, simultaneously with, or after administration of TIMP-PPE. In various embodiments, the leukotriene modifier is administered 1 week, 2 weeks, 3 weeks, 4 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 administration of TIMP-PPE. In various embodiments, the leukotriene modifier is administered 1 week, 2 weeks, 3 weeks, 4 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 after administration of TIMP-PPE. In various embodiments, the leukotriene modifier is administered 5, 10, 15, 30, 45, or 60 minutes before administration of TIMP-PPE. In various embodiments, the leukotriene modifier is administered 5, 10, 15, 30, 45, or 60 minutes after administration of TIMP-PPE. In various embodiments, the leukotriene modifying agent 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 prior to administration of the TIMP-PPE.In various embodiments, the leukotriene modifier 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 administration of TIMP-PPE. In various embodiments, the leukotriene modifier is administered 1, 2, 3, 4, 5, 6, or 7 days before administration of TIMP-PPE. In various embodiments, the leukotriene modifier is administered 1, 2, 3, 4, 5, 6, or 7 days after administration of TIMP-PPE.

[0040] When TIMP-PPE is administered with an additional therapeutic agent described herein, it is contemplated that the TIMP-PPE may be administered in a single dose or multiple doses, hi various embodiments, the TIMP-PPE is administered in two doses spaced one week apart.

[0041] When TIMP-PPE is administered in combination with a therapeutic agent useful for treating peanut allergy, as described herein, it is contemplated that the TIMP-PPE may be administered in a single dose or multiple doses. In various embodiments, the TIMP-PPE is administered in two doses spaced one week apart. In various embodiments, the TIMP-PPE is administered in two doses spaced one week apart, followed by a booster dose of TIMP-PPE administered as a single dose once every three months.

[0042] In various embodiments, administering a 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, which in various embodiments are selected from the group consisting of skin reactions, hives, skin redness, skin swelling, itching, throat tightness, 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, which in various embodiments 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 disorders such as diarrhea, stomach cramps, nausea, or vomiting, low 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 reduces the duration and severity of an 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 reduces the duration and severity of an allergic immune response after 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 a TIMP-PPE in alleviating one or more symptoms of peanut allergy and / or reducing the duration and severity of an allergic immune response to peanut proteins is determined by assaying one or more biological samples from the 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), stool, tissue biopsy, and / or bone marrow biopsy. In various embodiments, the efficacy of a TIMP-PPE in alleviating one or more symptoms of peanut allergy and / or reducing the duration and severity of an allergic immune response to peanut proteins is determined by a double-blind, placebo-controlled food challenge (DBPCFC). In various embodiments, the efficacy of a TIMP-PPE in alleviating one or more symptoms of peanut allergy and / or reducing the duration and severity of an allergic immune response to peanut proteins is determined by a skin prick test (SPT).

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

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

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

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

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

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

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

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

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

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

[0057] It is understood that each feature or embodiment, or combination, described herein is a non-limiting, illustrative example of any of the aspects of the invention, and as such is meant to be combinable with any other feature, embodiment, or combination described herein. For example, when features are described in language such as "one embodiment," "some embodiments," "particular embodiments," "further embodiments," "particular exemplary embodiments," and / or "another embodiment," each of these types of embodiments is a non-limiting example of the feature that is intended to be combined with any other feature or combination of features described herein, and it is not necessary to list all possible combinations. Such features or combinations of features apply to any of the aspects of the invention. When example values ​​that fall within a range are disclosed, any of these examples are contemplated as possible endpoints of the range, and any and all numerical values ​​between such endpoints are contemplated, with any and all combinations of upper and lower endpoints envisioned.

[0058] The headings herein are for the convenience of the reader and are not intended to be limiting. Additional aspects, embodiments, and variations of the invention will be apparent from the detailed description and / or drawings and / or claims. [Brief explanation of the drawings]

[0059] [Figure 1A] 1 is a timeline of events for subjects participating in Part A of the human trial of CNP-201 particles. [Figure 1B] 1 is a timeline of events for subjects participating in Part B of the human trial of CNP-201 particles. [Figure 1C] 1 is a timeline of events for subjects participating in a low starting dose study of CNP-201 particles. [Figure 2] Summary of results for patients receiving low-dose CNP-201 particles for peanut allergy: Patients received approximately one dose of 1 mg or 25 mg of CNP-201 intravenously. Patients receiving low-dose CNP-201 had increased BAT thresholds, decreased peanut-specific IgE-to-IgG ratios, induction of antigen-specific Tregs, and / or decreased pathogenic peanut-specific T cell subsets (Th2a, TGF-β, B cell plasmablasts). [Figure 3] In a basophil sensitivity test with crude peanut extract: peanut-allergic patients who received one 25 mg intravenous dose of CNP-201 showed an increase in the EC50 (the concentration of peanut allergen) required to increase the expression of activation markers on the surface of basophils after stimulation with peanut allergen. [Figure 4] Shift from IgE to IgG phenotype at CNP-201 doses up to 25 mg: Peanut-allergic patients receiving intravenous doses of CNP-201 up to 25 mg showed an increase in peanut-specific IgG and a decrease in the peanut-specific IgE / IgG ratio at 15 and 38 days post-dose compared to placebo. [Figure 5] Reduction of peanut allergy-associated immune cell subsets: Peanut-allergic patients who received one 25 mg intravenous dose of CNP-201 showed a reduction in the allergy-promoting subsets T helper 2A (Th2A), T follicular helper cells (TFH), terminally differentiated effector memory cells (TEMRA), and B cell plasmablasts. [Figure 6]Lower pathogenic activated peanut-specific CD4+ T cells were observed after a single 25 mg dose of CNP-201. There was a decrease in the peanut-specific activated T cell subsets CD4+CD25+, CD4+CD69+, and CD4+PD-1+ in low-dose CNP-201-treated patients compared with placebo. [Figure 7] Induction of antigen-specific regulatory T cells (Tregs): Peanut-allergic patients who received one 25 mg intravenous dose of CNP-201 showed an increase in peanut-specific Tregs compared to placebo. [Figure 8] 1 is a timeline of events for subjects participating in a 1 mg starting dose study of CNP-201 particles. DETAILED DESCRIPTION OF THE INVENTION

[0060] The present disclosure provides methodologies for monitoring the induction and maintenance of immune tolerance in a subject after undergoing immunotherapy.

[0061] definition Unless otherwise stated, the following terms used in this Application, including the specification and claims, have the definitions given below.

[0062] As used in this specification and the appended claims, the indefinite articles "a" and "an" and the definite article "the" include plural and singular referents unless the context clearly dictates otherwise.

[0063] The term "about" or "approximately" refers to a tolerance for a particular value as determined by one 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 number in a series of two or more numbers, it is understood that the term "about" or "approximately" applies to every single number in the series.

[0064] As used herein, "particle" refers to any non-tissue-derived composition and may be a sphere or sphere-like entity, a bead, or a liposome. The terms "particle," "immunomodulating particle," "carrier particle," and "bead" may be used interchangeably depending on the context. Additionally, the term "particle" may be used to encompass beads and spheres.

[0065] As used herein, "negatively charged particles" refers to particles that have been modified to have a net surface charge that is less than zero.

[0066] "Carboxylated particles" or "carboxylated beads" or "carboxylated spheres" include any particles modified to contain carboxyl groups on their surface. In some embodiments, the addition of carboxyl groups enhances phagocytic / monocyte uptake of the particles from the circulation, for example, through interaction with scavenger receptors such as MARCO. Carboxylation of particles can be achieved using any compound that adds carboxyl groups.

[0067] As used herein, "TIMP-PPE" refers to a negatively charged tolerizing immunomodulating particle (TIMP) comprising 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, Ara h14, Ara h15, Ara h16, Ara h17, and Ara h18.

[0068] As used herein, the term "Th cells" or "helper T cells" refers to CD4 + Refers to cells. CD4 + T cells assist other leukocytes in immunological processes, including the maturation of B cells into plasma cells and memory B cells, and the activation of cytotoxic T cells and macrophages. T cells are activated when peptide antigens are presented to them by MHC class II molecules expressed on the surface of antigen-presenting cells (APCs).

[0069] As used herein, the term "Th1 cells" refers to a subset of Th cells that produce proinflammatory mediators. Th1 cells secrete cytokines to promote immune responses and play a role in host defense against pathogens, in part by mediating the recruitment of neutrophils and macrophages to infected tissues. Th1 cells secrete cytokines, including IFN-gamma, IL-2, IL-10, and TNF alpha / beta, to orchestrate defense against intracellular pathogens such as viruses and some bacteria.

[0070] As used herein, the term "Th2 cells" refers to a subset of Th cells that mediate the activation and maintenance of antibody-mediated immune responses to 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 involved in antibody production, eosinophil activation, and inhibition of several macrophage functions, thus providing a phagocyte-independent protective response.

[0071] "Polypeptide" and "protein" refer to polymers composed of amino acid residues, related naturally occurring structural variants, and synthetic, non-naturally occurring analogs thereof, linked via peptide bonds or peptide bond isosteres. Synthetic polypeptides can be synthesized, for example, using an automated polypeptide synthesizer. The terms "polypeptide" and "protein" are not limited to a minimum length of the product. The term "protein" typically refers to a large polypeptide. The term "peptide" typically refers to a short polypeptide. Thus, peptides, oligopeptides, dimers, multimers, and the like, are included within the definition. Both full-length proteins and fragments thereof are encompassed by the definition. The terms "polypeptide" and "protein" also include post-expression modifications of polypeptides or proteins, e.g., glycosylation, acetylation, phosphorylation, and the like. Furthermore, for purposes of this disclosure, "polypeptide" can include "modifications" to the native sequence, such as deletions, additions, substitutions (which may be conservative in nature or may include substitutions with any of the 20 amino acids commonly found in human proteins, or any other naturally occurring, non-naturally occurring, or atypical amino acids), and chemical modifications (e.g., addition of or substitution with peptidomimetics). These modifications may be deliberate, through site-directed mutagenesis or through chemical modification of amino acids to remove or attach chemical moieties, or may be accidental, such as through mutations that occur via the host cell producing the protein or through errors due to PCR amplification prior to host cell transfection.

[0072] As used herein, "antigenic moiety" or "antigen" refers to any moiety, e.g., peptide, that is recognized by a host's immune system. Examples of antigenic moieties include, but are not limited to, autoantigens, allergens, enzymes, and / or bacterial or viral proteins, peptides, drugs, or components.

[0073] "Pharmaceutically acceptable carrier" refers to any of the standard pharmaceutical carriers, buffers, and the like, such as phosphate-buffered saline, 5% aqueous dextrose, 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, glidants, sweeteners, flavoring agents, and coloring agents. Suitable pharmaceutical carriers, excipients, and diluents are described in Remington's Pharmaceutical Sciences, 19th Ed. (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, intramuscular, intravenous, or intraperitoneal injection, or topical, transdermal, or transmucosal administration) or via inhalation.

[0074] "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 undesired biological effects or interacting in a deleterious manner with any of the components of the composition in which it is contained, or with any components present on or in the individual's body.

[0075] As used herein, the term "subject" encompasses mammals and non-mammals. Examples of mammals include, but are not limited to, any member of the mammalian class: humans, non-human primates such as chimpanzees, and other ape and monkey species; livestock such as cows, horses, sheep, goats, and pigs; domestic animals such as rabbits, dogs, and cats; and laboratory animals including rodents such as rats, mice, and guinea pigs. Examples of non-mammals include, but are not limited to, birds, fish, and the like. The term does not denote a particular age or sex.

[0076] The term "epitope" refers to a portion of any molecule capable of being recognized and bound by a selective binding agent at one or more of its antigen-binding regions. Epitopes typically consist of chemically active surface groupings of molecules, such as amino acids or carbohydrate side chains, and have specific three-dimensional structural and charge characteristics. As used herein, an epitope can be continuous or discontinuous. An epitope can also be a mimetic (mimotope) in that it contains a three-dimensional structure identical to the epitope used to generate antibodies but that includes none or only some of the amino acid residues found in the target used to stimulate the antibody immune response. As used herein, a mimotope is not considered a distinct antigen from the epitope bound by the selective binding agent; the selective binding agent recognizes the same three-dimensional structure of the epitope and mimotope.

[0077] The term "therapeutically effective amount" is used herein to indicate an amount of an antigen-specific composition of the present disclosure that is effective in ameliorating or alleviating the symptoms or signs of the disease being treated.

[0078] The terms "treat," "treated," "treating," and "treatment," as used with respect to methods herein, refer to eliminating, reducing, inhibiting, or ameliorating, either temporarily or permanently, either partially or completely, the clinical symptoms, signs, or progression of an event, disease, or condition. Such treating need not be absolute to be useful.

[0079] As used herein, the term "symptom" refers to any physical or observable manifestation of a disorder, whether or not it is generally characteristic of that disorder. The term "symptom" can refer to all such manifestations or any subset thereof.

[0080] As used herein, a "booster dose" refers to the re-administration of a TIMP after an initial or initial administration. The administration or re-administration of a booster dose enhances the tolerogenic immune response induced by the initial administration. The booster dose may be greater than, equal to, or less than the initial dose of TIMP particles administered to the subject. Subjects treated with tolerization therapy are monitored to ensure maintenance of immune tolerance. The decision to administer a booster dose is based on observing a change, weakening, or loss of immune tolerance. Methods for monitoring the immune tolerance status of subjects treated with tolerization therapy have been previously described (see International Patent Publication No. WO 2022 / 221622, incorporated herein by reference). The administration of a booster dose of TIMP-PPE may be with the same combination therapy as described herein, a different combination therapy, or no combination therapy, which may be administered with the initial TIMP-PPE dose.

[0081] particle The size and charge of particles are important for tolerance induction. Particles vary in size and charge based on the antigen encapsulated within the particle; however, generally, the particles described herein are effective for inducing tolerance when they are between about 100 nanometers and about 1500 nanometers and have a charge of 0 to about -100 mV. In various embodiments, the particles are between 400 and 800 nanometers in diameter and have a charge of between about -25 mV and -70 mV. In various embodiments, the particles are between 400 and 800 nanometers in diameter and have a charge of between about -30 mV and -60 mV. In various embodiments, the particles are between 400 and 800 nanometers in diameter and have a charge of between about -30 mV and -80 mV. The average particle size and charge may be slightly altered during the lyophilization process; therefore, both the post-synthesis average and the post-lyophilization average 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. The terms "post-lyophilization size" and "post-lyophilization charge" refer to the size and charge of the particles after lyophilization.

[0082] In some embodiments, the particles are non-metallic. In these embodiments, the particles may be formed from a polymer. In a preferred embodiment, the particles are biodegradable in an individual. In this embodiment, the particles can be provided to an individual over multiple doses without accumulation of the particles in the individual. Examples of suitable particles include polystyrene particles, PLGA particles, PLURONICS stabilized polypropylene sulfide particles, and diamond particles.

[0083] Preferably, the particle surface is composed of a material that minimizes nonspecific or undesirable biological interactions. Interactions between the particle surface and the interstitium may be a factor that plays a role in lymphatic uptake. The particle surface may be coated with a material that prevents or reduces nonspecific interactions. Steric stabilization by coating particles with a hydrophilic layer, such as poly(ethylene glycol) (PEG) and its copolymers, e.g., PLURONICS® (comprising copolymers of poly(ethylene glycol)-bl-poly(propylene glycol)-bl-poly(ethylene glycol)), may reduce nonspecific interactions with interstitial proteins, as demonstrated by improved lymphatic uptake after subcutaneous injection. All of these facts demonstrate the relevance of particle physical properties in terms of lymphatic uptake. Biodegradable polymers may be used to fabricate all or part of the polymer and / or particle and / or layer. Biodegradable polymers may degrade, for example, as a result of functional groups reacting with water in solution. As used herein, the term "degradation" refers to becoming soluble either by a reduction in molecular weight or by converting hydrophobic groups to hydrophilic groups. Polymers containing ester groups, such as polylactide and polyglycolide, are generally subject to spontaneous hydrolysis.

[0084] The particles disclosed herein may also contain additional components. For example, the carrier may have a contrast agent incorporated or conjugated thereto. An example of a carrier nanosphere with a contrast agent currently on the market is Kodak X-sight nanospheres. Inorganic quantum-confined luminescent nanocrystals, known as quantum dots (QDs), have emerged as ideal donors for FRET applications: their high quantum yield and tunable size-dependent Stokes shift allow them to emit light of different sizes from blue to infrared when excited by a single ultraviolet wavelength. (Bruchez, et al., Science, 1998, 281, 2013; Niemeyer, CM Angew. Chem. Int. Ed. 2003, 42, 5796; Waggoner, A. Methods Enzymol. 1995, 246, 362; Brus, LEJ Chem. Phys. 1993, 79, 5566). Quantum dots, including hybrid organic / inorganic quantum dots based on a class of polymers known as dendrimers, can be used in biological labeling, imaging, and optical biosensing systems (Lemon, et al., J. Am. Chem. Soc. 2000, 122, 12886). Unlike traditional synthesis of inorganic quantum dots, synthesis of these hybrid quantum dot nanoparticles does not require high temperatures or highly toxic, unstable reagents (Etienne, et al., Appl. Phys. Lett. 87, 181913, 2005).

[0085] Particles can be formed from a wide range of materials. Particles are preferably composed of materials suitable for biological use. For example, 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, carrier particles can be composed of polyesters of linear or branched, substituted or unsubstituted, saturated or unsaturated, linear or crosslinked, alkanyl, haloalkyl, thioalkyl, aminoalkyl, aryl, aralkyl, alkenyl, aralkenyl, heteroaryl, or alkoxyhydroxy acids, or linear or branched, substituted or unsubstituted, saturated or unsaturated, linear or crosslinked, alkanyl, haloalkyl, thioalkyl, aminoalkyl, aryl, aralkyl, alkenyl, aralkenyl, heteroaryl, or alkoxydicarboxylic acids. Additionally, carrier particles can be quantum dots or can be 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 linkages (e.g., copolymers of glycolic acid and sebacic acid) can also be used. For example, the carrier particles can comprise materials including polyglycolic acid polymers (PGA), polylactic acid polymers (PLA), polysebacic acid polymers (PSA), poly(lactic-co-glycolic) acid copolymers (PLGA or PLG, the terms are interchangeable), poly(lactic-co-sebacic) acid copolymers (PLSA), poly(glycol-co-sebacic) acid copolymers (PGSA), polypropylene sulfide polymers, poly(caprolactone), 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 urethanes, as well as copolymers thereof with linear or branched, substituted or unsubstituted alkanyl, haloalkyl, thioalkyl, aminoalkyl, alkenyl, or aromatic hydroxy or dicarboxylic acids.In addition, biologically important amino acids with reactive side 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 aforementioned materials to provide reactive groups and conjugate moieties for conjugation to antigenic peptides and proteins. Suitable biodegradable materials for the present invention include diamond, PLA, PGA, polypropylene sulfide, and PLGA polymers. Biocompatible but non-biodegradable materials can also be used for the carrier particles of the present invention. For example, non-biodegradable polymers of acrylate, ethylene-vinyl acetate, acyl-substituted cellulose acetate, non-degradable urethane, styrene, vinyl chloride, vinyl fluoride, vinyl imidazole, chlorosulfonated olefins, ethylene oxide, vinyl alcohol, TEFLON® (DuPont, Wilmington, Del.), and nylon can be used.

[0086] In certain embodiments, the particles are copolymers having a molar ratio of about 80:20 to about 100:0. Suitable copolymer ratios for the 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 polypropylene sulfide particles, polyglycolic acid particles (PGA), polylactic acid particles (PLA), or poly(lactic-co-glycolic acid) particles. In certain embodiments, the particles have a copolymer ratio of 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 have a copolymer ratio of about 50:50 polylactic acid:polyglycolic acid.

[0087] It is contemplated that the particles may further comprise a surfactant. The surfactant may be anionic, cationic, or nonionic. Poloxamer and poloxamine family surfactants 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 include, but are not limited to, vitamin E TPGS (D-α-tocopheryl 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 a double emulsion method, the two surfactants can include a hydrophobic surfactant for the first emulsion and a hydrophobic surfactant for the second emulsion.

[0088] In certain embodiments, polypeptide antigens are encapsulated in particles by a single emulsion process. In further embodiments, the polypeptide antigen is more hydrophobic. Sometimes, the double emulsion process results in the formation of large particles, which can lead to leakage of hydrophilic active ingredients and low entrapment efficiency. Coalescence and Ostwald ripening are two mechanisms that can destabilize double emulsion droplets, while diffusion of hydrophilic active ingredients through the organic phase is the primary mechanism responsible for low levels of entrapped active ingredients. In some embodiments, it can be beneficial to reduce the nanoparticle size. One strategy to achieve this is to apply a second, strong shear rate. The leakage effect can be reduced by using high polymer concentrations and high polymer molecular weights, which are accompanied by increased viscosity of the internal aqueous phase and increased surfactant molecular weight. In certain embodiments, particles encapsulating antigens are produced by nanoprecipitation, coprecipitation, inert gas condensation, sputtering, microemulsion, sol-gel processing, layer-by-layer techniques, or ionic gelation. Several methods for producing nanoparticles have been described in the literature and are incorporated herein by reference (Sanchez, Mejia, and Orozco 2020; Zielinska et al. 2020).

[0089] antigen An antigen refers to a discrete portion of a molecule, such as a polypeptide or peptide sequence, a 3D structural formation of a polypeptide or peptide, a polysaccharide or polynucleotide that can be recognized by host immune cells. Antigen specificity refers to the ability of a subject's host cells to recognize and generate an immune response to the antigen alone or to a molecule closely similar to the antigen, as well as to an epitope or mimotope.

[0090] "Anergy," "tolerance," or "antigen-specific tolerance" refers to the insensitivity of T cells to T cell receptor-mediated stimulation. Such insensitivity is generally antigen-specific and persists after exposure to antigenic peptides has ceased. For example, anergy in T cells 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 CD3-mediated signal) in the absence of a second signal (costimulatory signal). Under these conditions, re-exposure of the cells to the same antigen (even if re-exposure occurs in the presence of costimulatory molecules) results in a failure of cytokine production and subsequent failure to proliferate. Thus, the failure of cytokine production prevents proliferation. However, anergy T cells can proliferate when cultured with cytokines (e.g., IL-2).

[0091] The tolerization therapy described herein is intended to be antigen-specific. For example, the TIMP administered as a tolerization therapy encapsulates one or more antigens associated with the tolerization therapy and the associated disease or condition being treated. The TIMP used in the tolerization therapy is intended to include one or more peanut antigens. The one or more peanut antigens can be derived from peanut protein extracts or peptides derived from known peanut proteins.

[0092] More than 15 peanut allergens, Ara h1 through 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, are officially recognized by the WHO / IUIS Allergen Nomenclature Sub-Committee (www.allergen.org). Peanut allergens can be classified into different groups based on their architecture (e.g., trimer, monomer, cupin, albumin, prolamin, profilin, oleosin, defensin, vincilin, and nonspecific lipid transfer protein (nsLTP)) based on Ara h1, h2, h3, h5, h6, and h8, 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 those derived from Arachis hypogaea Ara h1, Ara h2, Ara h3, Ara h5, Ara h6, Ara h7, Ara h8, and Ara h18.For example, UNIPROT database number E5G076, which indicates the Ara h1 polypeptide sequence (SEQ ID NO: 1), UNIPROT database number A0A445BYI5 for the Ara h2 polypeptide (SEQ ID NO: 2), UNIPROT database number E5G077 for the Ara h3 polypeptide (SEQ ID NO: 3) (see also UNIPROT database numbers O82580 (SEQ ID NO: 4) and Q9SQH7 (SEQ ID NO: 5) for Ara h3 isoallergens 1 and 2 (formerly Ara h4), respectively), UNIPROT database number L7QH52 for the Ara h5 polypeptide (SEQ ID NO: 6), UNIPROT database number A5Z1R0 for the Ara h6 polypeptide (SEQ ID NO: 7), UNIPROT database number B4XID4 for the Ara h7 polypeptide (SEQ ID NO: 8), UNIPROT database number Q6VT83 for the Ara h8 polypeptide sequence (SEQ ID NO: 9), and UNIPROT database number L7QH52 for the Ara h5 polypeptide (SEQ ID NO: 10). h9, isoallergens 1 and 2, UNIPROT database numbers B6CEX8 and B6CG41, respectively (SEQ ID NOs: 10 and 11), Ara h10, isoallergens 1 and 2, UNIPROT database numbers Q647G5 and Q647G4, respectively (SEQ ID NOs: 12 and 13), Ara h11, isoallergens 1 and 2, UNIPROT database numbers Q45W87 and Q45W86, respectively (SEQ ID NOs: 14 and 15), Ara h12 UNIPROT database number B3EWP3 (SEQ ID NO: 16), Ara h13, isoallergens 1 and 2, UNIPROT database numbers B3EWP4 and C0HJZ1, respectively (SEQ ID NOs: 17 and 18), Ara See h14, isoallergens 1, 2, and 3, UNIPROT database numbers Q9AXI1, Q9AXI0, and Q6J1J8, respectively (SEQ ID NOs: 19-21); Ara h15, UNIPROT database number Q647G3 (SEQ ID NO: 22); Ara h16, UNIPROT database number A0A509ZX51 (SEQ ID NO: 23); Ara h17, UNIPROT A database number 0A510A9S3 (SEQ ID NO: 24); and Ara h18, UNIPROT database number A0A444XS96 (SEQ ID NO: 25).

[0093] In various embodiments, the allergenic peanut protein comprises one or more 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. In various embodiments, the peptide derived from a peanut protein comprises an allergenic epitope from one or more 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 proteins.

[0094] In certain embodiments, one, two, three or more antigens or antigenic peptides are used in TIMP.In certain embodiments, one or more peanut antigens are encapsulated in TIMP by covalent binding to the inner surface of the particle (see, for example, US Patent Publication No. 2019 / 0282707, which is incorporated herein by reference).In certain embodiments, two or more peanut protein sequences, for example, from Ara h1, Ara h2, Ara h3, Ara h5, Ara h6, Ara h7, and / or Ara h8, are contemplated to be combined in a fusion protein and encapsulated in the TIMP described herein. In certain embodiments, sequences of two or more peanut proteins, e.g., from 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, are combined in a fusion protein and are contemplated to be encapsulated within the TIMPs described herein. Methods for producing TIMPs with combined epitopes are described in U.S. Patent Publication No. 2019 / 0365656, which is incorporated herein by reference.

[0095] How to use Peanut allergy is the most common food allergy in the United States, diagnosed in up to 0.6% of adults and up to 0.8% of children. In addition, peanut allergy is the most common cause of fatal food-related anaphylaxis due to its high prevalence in the population and the widespread use of peanut-derived products as fillers in many packaged foods. As a result, peanut allergy poses a significant health threat in the United States.

[0096] The four major peanut allergenic proteins are Ara h1, Ara h2, Ara h3, and Ara h6. Ara h1 and Ara h2 are recognized by more than 95% of patients with peanut allergy.

[0097] Peanut avoidance through dietary changes has long been the most effective treatment for peanut allergy. However, severe cases of peanut allergy can also be caused by foods processed near peanuts and / or breathing in close proximity to peanut products. If complete avoidance of both peanut product intake and areas contaminated with peanut products is not achieved, severe allergic reactions can occur, requiring patients to receive medications such as epinephrine, antihistamines, and / or oral steroids.

[0098] Therapies currently approved for the treatment of peanut allergy (such as PALFORZIA™) and those in clinical trials are designed only to protect against accidental peanut exposure by desensitizing subjects. Treatment regimens with PALFORZIA™ also involve fairly extensive daily dose escalation and maintenance schedules, which can present subject compliance challenges and still require subjects to maintain a peanut-free diet.

[0099] A therapeutic approach that renders T cells tolerant to peanut proteins could potentially cure peanut allergy, thus eliminating the burden associated with strict avoidance of peanut products.

[0100] Provided herein are methods for treating peanut allergy in a subject, comprising administering TIMP-PPE to the subject, wherein the TIMP-PPE is administered at a dose level determined based on the subject's body weight. It is also contemplated that the TIMP-PPE may be administered at a fixed dose regardless of the subject's body weight. Provided herein are methods for treating peanut allergy in a subject, comprising administering TIMP-PPE to the subject, wherein the TIMP-PPE is administered at a dose of 0.1 to 12 mg / kg. Provided herein are methods for treating peanut allergy in a subject, comprising administering TIMP-PPE to the subject, wherein the TIMP-PPE is administered at a dose of 0.001 to 12 mg / kg. In various embodiments, contemplated are methods for treating peanut allergy in a subject, comprising administering TIMP-PPE to the subject, wherein the TIMP-PPE is administered at a dose of 0.001 to 12 mg / kg or at a fixed dose of 0.1 mg to 800 mg based on the subject's body weight. Also provided herein is a method for reducing an allergic immune response to a peanut antigen in a subject suffering from PA, comprising administering TIMP-PPE to the subject, wherein the TIMP-PPE is administered at a dose of 0.001 to 12 mg / kg.Further contemplated herein is a method for reducing an allergic immune response to a peanut antigen in a subject suffering from PA, comprising administering TIMP-PPE to the subject, wherein the TIMP-PPE is administered at a dose of 0.001 to 12 mg / kg or at a fixed dose of 0.1 mg to 800 mg based on the subject's body weight.

[0101] It is also contemplated that the TIMP-PPE is administered at a dose of about 0.001 to 10 mg / kg, about 0.005 to 12 mg / kg, about 0.01 to 12 mg / kg, about 0.05 to 12 mg / kg, about 0.1 to 12 mg / kg, about 0.5 to 10 mg / kg, about 1 to 8 mg / kg, about 1.5 to 10 mg / kg, about 2 to 12 mg / kg, about 2 to 10 mg / kg, about 3 to 10 mg / kg, about 4 to 10 mg / kg, about 4 to 12 mg / kg, or about 5 to 12 mg / kg. Optionally, 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, 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, the TIMP-PPE is administered in 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, the TIMP-PPE is administered at a concentration of about 0.0005 mg / mL to 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.

[0102] 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 four weeks, once every two months, once every three months, once every six months, or once a year. In certain embodiments, TIMP-PPE is administered in two doses, one week apart.

[0103] In various embodiments, the TIMP-PPE is administered intravenously, subcutaneously, intramuscularly, intraperitoneally, intranasally, or orally. It is contemplated that when the TIMP-PPE is given intravenously, it may be via an intravenous infusion lasting about 1, 2, 3, 4, 5, 6, 7, 8, 10, 12, 18, or 20 hours, or an intravenous infusion lasting about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 10, 12, 18, or 20 hours.

[0104] Also provided herein are methods for enhancing the duration or efficacy of tolerance induced to a peanut allergen by TIMP-PPE treatment, comprising administering a booster dose of TIMP-PPE. In various embodiments, the booster dose of TIMP-PPE is administered as a single dose or multiple doses after the initial or initial TIMP-PPE administration. In various embodiments, the booster dose of TIMP-PPE is administered once a week, once every two weeks, once every three weeks, once every four weeks, once every two months, once every three months, once every six months, or once a year. In various embodiments, the booster dose of TIMP-PPE is administered as a single dose every three months.

[0105] It is further contemplated that TIMP-PPEs may be administered alone or in combination with one or more additional therapeutic agents, including, but not limited to, inhibitors of IgE, inhibitors of basophil activation, inhibitors of mast cell activation, antihistamines, nonsteroidal anti-inflammatory drugs (NSAIDs), or small molecule or biological therapeutic agents.

[0106] In various embodiments, TIMP-PPE is administered alone or in combination with one or more therapeutic agents useful in treating peanut allergy. Exemplary therapeutic agents include, but are not limited to, inhibitors of IgE, inhibitors of basophil activation, inhibitors of mast cell activation, antihistamines, nonsteroidal 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 therapy, steroids, corticosteroids, leukotriene modifiers, or small molecule or biological therapeutic agents. Exemplary additional therapeutic agents or therapeutic agents useful in treating peanut allergy also include those that increase the number, frequency, or activity of Tregs.

[0107] In various embodiments, the biologic is an antibody. In various embodiments, the antibody is an anti-IgE, anti-IL-4Rα, anti-IL-13, or anti-IL-33 antibody. In various embodiments, the anti-IgE antibody is omalizumab (XOLAIR®). Exemplary anti-IL-4Rα antibodies include dupilumab (DUPIXENT®), and anti-IL-33 antibodies include etokinumab.

[0108] 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, levocetirizine, 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.

[0109] In various embodiments, the additional therapeutic agent is a nonsteroidal anti-inflammatory drug (NSAID). In various embodiments, the therapeutic agent useful for treating peanut allergy is a nonsteroidal 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.

[0110] 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 antileukotriene. 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.

[0111] In various embodiments, the additional therapeutic agent is administered before, during, or after administration of the TIMP-PPE. In various embodiments, the therapeutic agent useful in treating peanut allergy is administered before, during, or after administration of the TIMP-PPE.

[0112] In various embodiments, the additional therapeutic agent is administered intravenously, subcutaneously, intramuscularly, intraperitoneally, intranasally, via inhalation, or orally. In various embodiments, the therapeutic agent useful in treating peanut allergy is administered intravenously, subcutaneously, intramuscularly, intraperitoneally, intranasally, via inhalation, or orally.

[0113] The present disclosure provides methods of treating peanut allergy in a subject, comprising administering to the subject a TIMP-PPE in combination with an anti-IgE antibody, wherein the TIMP-PPE is administered at a dose of about 0.001 mg / kg to 12 mg / kg and 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®), kiruizumab, or ligelizumab.

[0114] 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 dosage level of the anti-IgE antibody is determined based on the serum IgE level, which may be 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 dosage level of the anti-IgE antibody is determined based on the subject's body weight. In various embodiments, the subject weighs between 30 and 40 kg, 40 and 50 kg, 50 and 60 kg, 60 and 70 kg, 70 and 80 kg, 80 and 90 kg, 90 and 125 kg, or 125 and 150 kg.

[0115] 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, simultaneously with, or subsequently / after administration of TIMP-PPE. In various embodiments, the anti-IgE antibody is administered one week, two weeks, three weeks, or four weeks before administration of TIMP-PPE. In various embodiments, the anti-IgE antibody is administered one week, two weeks, three weeks, or four weeks after administration of TIMP-PPE.

[0116] The combination or simultaneous administration of two therapeutic agents does not require that the agents be administered at the same time or by the same route, so long as there is an overlap in the time periods during which the agents are exerting their therapeutic effect. Simultaneous or sequential administration is contemplated, as is administration on different days or weeks. Therapeutic agents are further contemplated to refer to agents administered in separate formulations, administered simultaneously or in combination, and given simultaneously within 30 minutes of each other. Prior administration refers to administration of a therapeutic agent one week prior to treatment with TIMP-PPE, up to 30 minutes prior to administration of TIMP-PPE. Subsequent administration is intended to describe administration from 30 minutes after TIMP-PPE treatment up to one week after TIMP-PPE administration.

[0117] The present disclosure provides a method for treating peanut allergy comprising administering to a subject TIMP-PPE in combination with an anti-IgE antibody, wherein the subject is administered the anti-IgE antibody once weekly for two weeks or once weekly for four weeks prior to administration of TIMP-PPE, wherein the TIMP-PPE is administered in two doses, one week apart, at a dosage level of 0.001 mg / kg to 12 mg / kg, and the anti-IgE antibody is administered at a dosage level of about 50 mg to 500 mg.

[0118] Also provided is a method of treating peanut allergy in a subject, comprising administering to the subject TIMP-PPE in combination with an anti-IL-4Rα antibody, wherein the TIMP-PPE is administered at a dose of about 0.001 mg / kg to 12 mg / kg and 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®).

[0119] 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 may be 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 subject's body weight. In various embodiments, the subject weighs between 30 and 40 kg, 40 and 50 kg, 50 and 60 kg, 60 and 70 kg, 70 and 80 kg, 80 and 90 kg, 90 and 125 kg, or 125 and 150 kg.

[0120] In various embodiments, the anti-IL-4Rα antibody is administered in a single dose or multiple doses. In various embodiments, the anti-IL-4Rα antibody is administered once weekly, once every two weeks, once every three weeks, or once every four weeks. In various embodiments, the anti-IL-4Rα antibody is administered before, simultaneously with, or after administration of TIMP-PPE. In various embodiments, the anti-IL-4Rα antibody is administered 1 week, 2 weeks, 3 weeks, or 4 weeks before administration of TIMP-PPE. In various embodiments, the anti-IL-4Rα antibody is administered 1 week, 2 weeks, 3 weeks, or 4 weeks after administration of TIMP-PPE. In various embodiments, the anti-IL-4Rα antibody is administered at an initial dose level of 400 mg to 600 mg over two doses, followed by a maintenance dose level of 200 mg to 300 mg over subsequent doses.

[0121] In various embodiments, the antihistamine administered in combination with the TIMP-PPE is administered in a dose of 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 daily, once weekly, once every two weeks, once every three weeks, or once every four weeks. In various embodiments, the antihistamine is administered twice, three times, four times, five times, or six times daily. In various embodiments, the antihistamine is administered before, simultaneously with, or after administration of the TIMP-PPE. In various embodiments, the antihistamine is administered 1 week, 2 weeks, 3 weeks, 4 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 administration of the TIMP-PPE. In various embodiments, the antihistamine is administered 1 week, 2 weeks, 3 weeks, 4 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 after administration of the TIMP-PPE. In various embodiments, the antihistamine is administered 5, 10, 15, 30, 45, or 60 minutes before administration of the TIMP-PPE. In various embodiments, the antihistamine is administered 5, 10, 15, 30, 45, or 60 minutes after administration of the 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 administration of the 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 administration of the TIMP-PPE.In various embodiments, the antihistamine is administered 1, 2, 3, 4, 5, 6, or 7 days before administration of the TIMP-PPE. In various embodiments, the antihistamine is administered 1, 2, 3, 4, 5, 6, or 7 days after administration of the 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, levocetirizine, doxylamine, ebastine, embramin, epinephrine, fexofenadine, loratadine, and olopatadine.

[0122] In various embodiments, the steroid administered in combination with the TIMP-PPE is administered at a dose of 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 daily, once weekly, 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 daily. In various embodiments, the steroid is administered before, simultaneously with, or after administration of TIMP-PPE. In various embodiments, the steroid is administered 1 week, 2 weeks, 3 weeks, 4 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 administration of TIMP-PPE. In various embodiments, the steroid is administered 1 week, 2 weeks, 3 weeks, 4 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 after administration of TIMP-PPE. In various embodiments, the steroid is administered 5, 10, 15, 30, 45, or 60 minutes before administration of TIMP-PPE. In various embodiments, the steroid is administered 5, 10, 15, 30, 45, or 60 minutes after administration of the 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 administration of the 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 administration of the TIMP-PPE.In various embodiments, the steroid is administered 1, 2, 3, 4, 5, 6, or 7 days before administration of the TIMP-PPE. In various embodiments, the steroid is administered 1, 2, 3, 4, 5, 6, or 7 days after administration of the TIMP-PPE. In various embodiments, the steroid is selected from the group consisting of beclomethasone, ciclesonide, fluticasone furoate, mometasone, budenoside, fluticasone, triamcinolone, and loteprednol.

[0123] 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 in a dose of 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 daily, once weekly, once every two weeks, once every three weeks, or once every four weeks. In various embodiments, the corticosteroid is administered twice, three times, four times, five times, or six times daily. In various embodiments, the corticosteroid is administered before, simultaneously with, or after administration of the TIMP-PPE. In various embodiments, the corticosteroid is administered 1 week, 2 weeks, 3 weeks, 4 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 administration of the TIMP-PPE. In various embodiments, the corticosteroid is administered 1 week, 2 weeks, 3 weeks, 4 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 after administration of the TIMP-PPE. In various embodiments, the corticosteroid is administered 5, 10, 15, 30, 45, or 60 minutes before administration of the TIMP-PPE. In various embodiments, the corticosteroid is administered 5, 10, 15, 30, 45, or 60 minutes after administration of the 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 administration of the 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 administration of the TIMP-PPE. In various embodiments, the corticosteroid is administered 1, 2, 3, 4, 5, 6, or 7 days before administration of the TIMP-PPE. In various embodiments, the 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.

[0124] In various embodiments, the NSAID administered in combination with the 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 twice, three times, four times, five times, or six times a day. In various embodiments, the NSAID is administered before, simultaneously with, or after administration of TIMP-PPE. In various embodiments, the NSAID is administered 1 week, 2 weeks, 3 weeks, 4 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 administration of TIMP-PPE. In various embodiments, the NSAID is administered 1 week, 2 weeks, 3 weeks, 4 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 after administration of TIMP-PPE. In various embodiments, the NSAID is administered 5, 10, 15, 30, 45, or 60 minutes before administration of the TIMP-PPE. In various embodiments, the NSAID is administered 5, 10, 15, 30, 45, or 60 minutes after administration of the 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 administration of the 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 administration of the TIMP-PPE.In various embodiments, the NSAID is administered 1, 2, 3, 4, 5, 6, or 7 days before administration of the TIMP-PPE. In various embodiments, the NSAID is administered 1, 2, 3, 4, 5, 6, or 7 days after administration of the 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, felbinac, piroxicam, mefenamic acid, meclofenamate sodium, meloxicam, nabumetone, oxaprozin, piroxicam, celecoxib, etodolac, etoricoxib, lumiracoxib, rofecoxib, and valdecoxib.

[0125] In various embodiments, the leukotriene modifier administered in combination with TIMP-PPE is administered in a dose of about 0.05 mg to 2000 mg. In various embodiments, the leukotriene modifier 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 modifier is administered in a single dose or multiple doses. In various embodiments, the leukotriene modifier is administered once daily, once weekly, once every two weeks, once every three weeks, or once every four weeks. In various embodiments, the leukotriene modifier is administered twice, three times, four times, five times, or six times daily. In various embodiments, the leukotriene modifier is administered before, simultaneously with, or after administration of TIMP-PPE. In various embodiments, the leukotriene modifier is administered 1 week, 2 weeks, 3 weeks, 4 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 administration of TIMP-PPE. In various embodiments, the leukotriene modifier is administered 1 week, 2 weeks, 3 weeks, 4 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 after administration of TIMP-PPE. In various embodiments, the leukotriene modifier is administered 5, 10, 15, 30, 45, or 60 minutes before administration of TIMP-PPE. In various embodiments, the leukotriene modifier is administered 5, 10, 15, 30, 45, or 60 minutes after administration of TIMP-PPE. In various embodiments, the leukotriene modifying agent 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 prior to administration of the TIMP-PPE.In various embodiments, the leukotriene modifier 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 administration of the TIMP-PPE. In various embodiments, the leukotriene modifier is administered 1, 2, 3, 4, 5, 6, or 7 days before administration of the TIMP-PPE. In various embodiments, the leukotriene modifier is administered 1, 2, 3, 4, 5, 6, or 7 days after administration of the 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.

[0126] Administration of TIMP-PPE, alone or in combination with one or more additional therapeutic agents, to a subject in need thereof is provided to alleviate one or more symptoms of peanut allergy. Peanut allergy symptoms include skin reactions, hives, redness of the skin, swelling of the skin, itching, tightness in the throat, difficulty breathing, shortness of breath, and anaphylaxis. Additional symptoms include digestive disorders, such as diarrhea, stomach cramps, nausea, or vomiting, and decreased blood pressure.

[0127] Administration of TIMP-PPE, alone or in combination with one or more therapeutic agents useful in treating peanut allergy, to a subject in need thereof provides relief from one or more symptoms of peanut allergy, including skin reactions, hives, skin redness, skin swelling, itching, throat tightness, difficulty breathing, shortness of breath, digestive disorders such as diarrhea, stomach cramps, nausea, or vomiting, low blood pressure, and anaphylaxis.

[0128] It is also contemplated that administering TIMP-PPE, alone or in combination with one or more additional therapeutic agents, to a subject in need thereof reduces the duration and severity of allergic immune responses to peanut protein or after 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, including T helper type 2 (Th2) cytokine production (e.g., IL-4, IL-5, IL-9, and IL-13), B cell class switching leading to IgE antibody production, and / or upregulation of the IgE:IgG ratio.

[0129] It is also contemplated that administering TIMP-PPE, alone or in combination with one or more therapeutic agents useful in treating peanut allergy, to a subject in need thereof will reduce the duration and severity of an allergic immune response to or after exposure to peanut proteins. The allergic immune response contemplated herein includes T cell-dependent mechanisms, including T helper type 2 (Th2) cytokine production (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 upregulation of the IgE:IgG ratio.

[0130] Screening Methods It is contemplated that the induction of immune tolerance and its maintenance will be monitored in subjects suffering from peanut allergy who are being treated with, or about to be treated with, an antigen-specific tolerizing therapy consisting of TIMPs encapsulating peanut antigens as described herein.

[0131] Methods for screening for cell types, cytokines, or other measures of tolerance from subjects receiving the tolerizing therapies described herein are known in the art. Methods for assessing tolerance are performed using techniques such as flow cytometry, mass cytometry (CyTOF), ELISA, ELISPOT, in vivo / ex vivo cell stimulation assays (including, but not limited to, cell proliferation assays, basophil activation test (BAT), macrophage stimulation assays), measuring autoantibodies or measuring Ig serotypes, e.g., by ImmunoCap assay.

[0132] In various embodiments, the subject's immune tolerance state is determined from assaying one or more biological samples from the subject. The biological samples include whole blood, peripheral blood, peripheral blood mononuclear cells (PBMCs), serum, plasma, urine, cerebrospinal fluid (CSF), stool, tissue biopsy, and / or bone marrow biopsy. In various embodiments, assaying the biological sample includes analyzing the level and / or presence or absence of cell surface proteins, extracellular proteins, intracellular proteins, nucleic acids, metabolites, and / or combinations thereof.

[0133] The cells assayed from the biological sample include immune cells, non-immune cells, and / or combinations thereof. Immune cells include innate immune cells, adaptive immune cells, and / or combinations thereof. The innate immune cells assayed from the biological sample include antigen-presenting cells (APCs). Exemplary innate immune cells assayed from the biological sample include monocytes, macrophages, neutrophils, granulocytes, dendritic cells, mast cells, eosinophils, basophils, and / or combinations thereof. The adaptive immune cells assayed from the biological sample 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.

[0134] In certain embodiments, the cells assayed from the biological sample are epithelial cells, stromal cells, endothelial cells, fibroblasts, pericytes, adipocytes, mesenchymal stem cells, hematopoietic stem cells, hematopoietic progenitor cells, hepatic sinusoidal endothelial cells (LSECs), and / or Kupffer cells.

[0135] One aspect of a subject's immune tolerance state and immune signature is determined by analyzing one or more proteins from one or more biological samples from the subject. In various embodiments, the proteins are cytokines and / or chemokines. In various embodiments, the proteins are cell signaling proteins. In various embodiments, the cytokines and chemokines include 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-18, IL-20, IL-21, IL-22, IL-23, IL-24, IL-25, IL-26, IL-27, IL-28, IL-29, IL-30, IL-31, IL-32, IL-33, IL-34, IL-35, IL-36, IL-37, IL-38, IL-39, IL-40, IL-41, IL-42, IL-43, IL-44, IL-45, IL-46, IL-47, IL-48, IL-49, IL-50, IL-51, IL-52, IL-53, IL-54, IL-55, IL-56, IL-57, IL-58, IL-59, IL-60, IL-61, IL-62, IL-63, IL-64, IL-65, IL-66, IL-67, IL-68, IL-69, IL-70, IL-71, IL-72, IL-73 , 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-3 3, IL-35, IL-36, CCL1, CCL2, CCL3, CCL4, CCL5, CCL6, CCL7, CCL8, CCL9, CCL10, CCL11, CCL12, CC 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.

[0136] In various embodiments, the protein is a protease, hi various embodiments, the protease is an aspartic acid protease, a cysteine ​​protease, a metalloprotease, a serine protease, and / or a threonine protease. In various embodiments, the polypeptide 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 apoptosis-related protein is 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 assaying proteins from biological samples have been described in the literature, including enzyme-linked immunosorbent assay (ELISA), Western blot, 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 immunoglobulins are antigen-specific. Several methods for the detection of immunoglobulins from biological samples have been described in the literature, including ELISA and ImmunoCap.

[0137] Lists of human metabolites that can be assayed from biological samples can be found in the literature, including (Psychogios et al., 2011), (Wishart et al., HMDB: the Human Metabolome Database. Nucleic Acids Res. 2007 Jan;35(Database issue):D521-6, 2007), and the Human Metabalome Database (HMDB), which are incorporated herein by reference.

[0138] One aspect of a subject's immune tolerance state and immune signature is determined 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, CD48, CD49, CD50, CD51, CD52, CD53, CD54, CD55, CD56, CD57, CD58, CD59, CD60, CD61, CD62, CD63, CD64, CD65, CD66, CD67, CD68, CD69, CD70, CD71, CD72, CD73, CD74, CD75, CD76, CD77, CD78, CD79, CD80, CD81, CD82, CD83, CD84, CD85, CD86, CD87, CD88, CD89, CD90, CD91, CD92, CD93, CD94, CD95, CD96, CD97, CD98, CD99, CD100, CD101, CD102, CD103, CD104, CD105, CD106, CD107, CD108, CD110, CD111, CD112, CD113, CD114, CD115, CD116, CD117, D45RO, CD48, CD52, CD55, CD56, CD58, CD61, CD66b, CD69, CD70, CD72, CD79, CD68, CD84, CD86, CD93, CD94, CD95, CRACC, BLAME, BCMA, CD103, CD107, CD11 2, 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, KIR 2DL3, KIR2DL5A, KIR2DL5B, KIR3DL1, KIR3DL2, KIR3DL3, KIR3DL4, KIR2DS1, KIR2DS2, KIR2DS3, KIR2DS4, KIR2DS5, DAP12, KIR3DS, N Kp44, 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, L IGHT, 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, BA FF, 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, IG SF4A, 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 Integrins include 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 assaying cell surface protein expression have been described in the literature, including flow cytometry and mass cytometry (CyTOF).

[0139]

[0013] One aspect of a subject's immune tolerance state and immune signature is determined by analyzing one or more metabolites from a biological sample. In various embodiments, the metabolite is a pro-inflammatory metabolite. In various embodiments, the metabolite is an anti-inflammatory metabolite. In various embodiments, examples of pro-inflammatory metabolites include acids, lipids, sugars, amino acids, lactate, trimethylamine N-oxide, O-acetylcreatine, L-carnitine, choline, succinate, glutamine, fatty acids, cholesterol, 3-hydroxybutyrate, 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 6-semialdehyde, picolinic acid, anthranilic acid, 3-hydroxyanthranilic acid, glutaryl co-A, NAD+, quinolinic acid, arginine, butyrate, and adenosine. Lists of human metabolites that can be assayed from biological samples can be found in literature, including (Psychogios et al., 2011 PLoS One 6(2):e16957), (Wishart et al., HMDB: the Human Metabolome Database. Nucleic Acids Res. 2007 Jan;35(Database issue):D521-6,2007), and the Human Metabolome Database (HMDB), each of which is incorporated herein by reference.

[0140] In certain embodiments, the tolerance state of a subject is determined by analyzing nucleic acids from a biological sample. In various embodiments, the nucleic acid is 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 tolerance state of a subject is determined by assaying gene expression from a biological sample. In various embodiments, the tolerance state 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 tolerance state is determined by assaying gene expression associated with immune suppression. In various embodiments, the tolerance state is determined by assaying gene expression associated with immune activation. In various embodiments, the tolerance state is determined by assaying gene expression associated with regulatory function. In various embodiments, nucleic acid analysis is used to generate immune tolerance signatures. Several methodologies 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.

[0141] The biological sample is optionally assayed after in vivo and / or ex vivo stimulation with one or more stimuli, such as an antigen, an allergen, and one or more activating agents. 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, THO cells, or other antigen-specific T cells. B cells include effector B cells, memory B cells, plasma cells, and regulatory B (Breg) cells. In certain embodiments, T cells or B cells are identified based on the expression of proteins listed in Table 1. [Table 1]

[0142] In various embodiments, the subject's immune tolerance state is determined by obtaining one or more samples, such as whole blood, from the subject before the first TIMP-PPE dose (day 1), 14 days after 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 of the isolated cells from one or more samples are collected from the subject and analyzed using methods described below.

[0143] In various embodiments, the subject's immune tolerance state is determined by obtaining one or more samples, such as whole blood, from the subject before the first TIMP-PPE dose (day 1), 14 days after the second dose (day 15), 60 days, 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 of the isolated cells from one or more samples are collected from the subject and analyzed using methods described below.

[0144] In various embodiments, the subject's immune tolerance state determined prior to administration of TIMP-PPE serves as a baseline. In various embodiments, the subject's baseline is determined from assay of one or more biological samples 1, 2, 3, 4, 5, 6, or 7 days prior to administration of TIMP-PPE. In various embodiments, the subject's baseline is determined from assay of one or more biological samples 1, 2, 3, or 4 weeks prior to administration of TIMP-PPE. In various embodiments, the subject's baseline is determined from assay of one or more biological samples 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months prior to administration of TIMP-PPE.

[0145] In various embodiments, the subject's immune tolerance status is determined after administration of TIMP-PPE. In various embodiments, the subject's immune tolerance status is determined from assays of one or more biological samples 1, 2, 3, 4, 5, 6, or 7 days after administration of TIMP-PPE. In various embodiments, the immune tolerance is determined from assays of one or more biological samples 1, 2, 3, or 4 weeks after administration of TIMP-PPE. In various embodiments, the subject's immune tolerance is determined from assays of one or more biological samples 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months after administration of TIMP-PPE. In various embodiments, the subject's immune tolerance status determined after administration of TIMP-PPE is compared to a baseline. In various embodiments, the subject's immune tolerance status determined after administration of TIMP-PPE is compared to healthy subjects or subjects administered a placebo.

[0146] In various embodiments, the subject's immune tolerance state is determined after administration of TIMP-PPE, whether it be an initial or priming dose of TIMP-PPE or a booster dose of TIMP-PPE.

[0147] The following analyses are intended to follow the immune status and tolerance induction of subjects undergoing TIMP-PPE tolerization therapy.

[0148] The percentage of peanut-specific Th2a+ cells (Th2a+ cells / total peanut-specific T cells) stimulated ex vivo with purified antigenic peanut protein can be measured, for example, by flow cytometry. Th2a+ cells are defined as CRTH2+ / CD161+ / CD154+ / CD27-. Total peanut-reactive cells are defined as CRTH2- / CD161+ / CD154+ / CD27-. In various embodiments, administration of TIMP-PPE in a subject maintains or increases peanut-specific Th2a+ cells by about 1% to 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%) compared to one or more baseline measurements taken from a placebo-administered and / or treated subject. %, or about 100%, including all values ​​and ranges therebetween), 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 therebetween).

[0149] The percentage 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+. Non-activated peanut-specific T cells are defined as CD154+. In various embodiments, administration of TIMP-PPE in a subject maintains or increases activated peanut-specific T cells by about 1% to 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%) compared to one or more baseline measurements taken from a placebo-administered and / or treated subject. , or about 100%, including all values ​​and ranges therebetween), 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 therebetween).

[0150] T regulatory cell population (CD4+ / CD25+ / FoxP3+ / Helios+ / IL-10+) or (CD4+CD45RA 低 CD4+CD137+CD25+CD127 低The frequency of peanut-specific T regulatory cells (peanut-specific T regulatory cells / peanut-specific CD4+ effector memory cells) is determined by flow cytometry. Multicolor flow analysis is performed to provide the percentage of peanut-specific T regulatory cells (peanut-specific T regulatory cells / peanut-specific CD4+ effector memory cells). In various embodiments, administration of TIMP-PPE in a subject increases Treg cells by about 1% to 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%) compared to one or more baseline measurements taken from placebo-administered and / or treated subjects. %, including all values ​​and ranges therebetween), 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 therebetween).

[0151] The ratio of cellular IL-5 to IFN-γ in the PBMC culture supernatant is measured, for example, as detected by Luminex 200.

[0152] The following indicators of immune tolerance status can be determined from assays of basophils isolated from one or more blood samples collected from a subject and stimulated ex vivo with purified antigenic peanut protein: the percentage of activated CD203+ / CD63+ basophils after ex vivo stimulation with purified antigenic peanut protein using the Basophil Activation Test (BAT) (Santos and Lack 2016 Clin Transl Allergy. 6:10), and the effective concentration at 50% of maximum basophil activation (EC50) after ex vivo stimulation with purified antigenic peanut protein, measured using the Basophil Activation Test, where activated basophils are CD203+ / CD63+ / -. Analysis is performed to provide the effective concentration at 50% of maximum basophil activation (EC50).

[0153] The following indicators of immune tolerance status can be determined from assays of serum isolated from one or more blood samples obtained from a subject: ratio of peanut-specific IgE to IgG as measured by ImmunoCap assay. In various embodiments, administration of TIMP-PPE in a subject increases the peanut-specific IgE:IgG ratio by about 1% to 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%) compared to one or more baseline measurements taken from a placebo-administered and / or treated subject. , or about 100%, including all values ​​and ranges therebetween), 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 therebetween).

[0154] Change in peanut-specific IgE as measured by ImmunoCap assay. In various embodiments, administration of TIMP-PPE in a subject maintains or increases the level of peanut-specific IgE by about 1% to 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%) compared to one or more baseline measurements taken from a placebo-administered and / or treated subject. , about 95%, or about 100%, including all values ​​and ranges therebetween), 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 therebetween).

[0155] The following indicators of immune tolerance can be determined from assays of basophils isolated from one or more blood samples collected from a subject and stimulated ex vivo with purified antigenic peanut protein: the percentage of activated CD203+ / CD63+ basophils after ex vivo stimulation with purified antigenic peanut protein using the Basophil Activation Test (BAT) (Santos and Lack 2016 Clin Transl Allergy. 6:10), and the effective concentration at 50% of maximum basophil activation (EC50) after ex vivo stimulation with purified antigenic peanut protein, measured using the Basophil Activation Test, in which activated basophils are CD203+ / CD63+ / -. Analysis is performed to provide the effective concentration at 50% of maximum basophil activation (EC50). In various embodiments, administration of TIMP-PPE reduces basophil activation. In various embodiments, administration of a TIMP-PPE in a subject increases the EC50 of maximal basophil activation by about 1% to 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%, overall) compared to one or more baseline measurements taken from a placebo-administered and / or treated subject. an increase of 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).

[0156] In certain embodiments, the efficacy of TIMP-PPE in alleviating one or more symptoms of peanut allergy, increasing tolerance to peanut protein, and / or reducing the duration and severity of the allergic immune response to peanut protein is determined by double-blind placebo-controlled food challenge (DBPCFC), the change in cumulative tolerated dose of peanut protein administered during DBPCFC, and / or skin prick testing (SPT). Procedures for performing DBPCFC and SPT have been previously described (Sampson et al., J Allergy Clin Immunol. 2012;130(6):1260-1274; Heinzerling et al. The skin prick test - European standards. Clin Transl Allergy. 2013;3(1):3).

[0157] The effectiveness of TIMP-PPE in alleviating one or more symptoms of peanut allergy and / or reducing the duration and severity of an allergic immune response to peanut proteins is determined from assays of one or more biological samples from the subject. Biological samples include whole blood, peripheral blood, peripheral blood mononuclear cells (PBMCs), serum, plasma, urine, cerebrospinal fluid (CSF), stool, tissue biopsy, and / or bone marrow biopsy. In various embodiments, assaying the biological sample includes analyzing the level and / or presence or absence of cell surface proteins, extracellular proteins, intracellular proteins, nucleic acids, metabolites, and / or combinations thereof.

[0158] A subject's immune tolerance signature is generated using one or more of the following parameters assayed from one or more biological samples obtained from the subject and stimulated in vivo and / or ex vivo: a. The proportion of effector T cells in the total T cell population; b. The percentage of Treg cells in the total T cell population; c. the proportion of effector B cells in the total B cell population; d. specific IgG, IgA, IgM, and / or IgE levels; e. levels of inflammatory cytokines and chemokines; f. levels of anti-inflammatory cytokines and chemokines; g. levels of inflammatory metabolites, and h. Levels of anti-inflammatory metabolites.

[0159] An immune tolerance signature indicates maintenance of immune tolerance if 1, 2, 3, 4, 5, 6, 7, or 8 of the parameters listed in (a)-(h) above indicate maintenance of immune tolerance. In various embodiments, an immune tolerance signature indicates maintenance of immune tolerance if at least 2 / 8 of the parameters listed in (a)-(h) above indicate maintenance of immune tolerance. In various embodiments, a determination is made that the subject does not require treatment with a TIMP if 1, 2, 3, 4, 5, 6, 7, or 8 of the parameters listed in (a)-(h) above indicate maintenance of immune tolerance. In various embodiments, a determination is made that the subject does not require treatment with a TIMP if at least 3 / 8 of the parameters listed in (a)-(h) above indicate maintenance of immune tolerance.

[0160] A subject's immune tolerance signature generated using one or more parameters described herein indicates weakened and / or absent immune tolerance before or after treatment with TIMP-PPE if:

[0161] a. The percentage of effector T cells in the total T cell population is between 5% and 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 therebetween); and / or

[0162] b. The percentage of Treg cells in the total T cell population is 1-3%; and / or

[0163] c. The percentage of effector B cells in the total B cell population is between 5% and 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 therebetween); and / or

[0164] d. The levels of IgG, IgA, IgM, and / or IgE increase by about 5% to 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%, all values) compared to one or more baseline measurements taken from a healthy subject and / or a subject under treatment. and / or is increased by 10-95%, 15-90%, 20-85%, 25-75%, 30-70%, 35-65%, 40-60%, 45-55%, or 50%, or by 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); and / or

[0165] e. The level of inflammatory cytokines / chemokines is increased by about 5% to 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%, all values ​​and all combinations thereof) compared to one or more baseline measurements taken from a healthy subject and / or a subject under treatment. and / or is increased by 10-95%, 15-90%, 20-85%, 25-75%, 30-70%, 35-65%, 40-60%, 45-55%, or 50%, or by 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);

[0166] f. The levels of anti-inflammatory cytokines and chemokines are increased by about 5% to 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%, all values ​​and all values) compared to one or more baseline measurements taken from a healthy subject and / or a subject under treatment. and / or is reduced by 10-95%, 15-90%, 20-85%, 25-75%, 30-70%, 35-65%, 40-60%, 45-55%, or 50%, or by 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); and / or

[0167] g. The level of an inflammatory metabolite is increased by about 5% to 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%, all values ​​and values ​​therein) compared to one or more baseline measurements taken from a healthy subject and / or a subject under treatment. and / or is increased by 10-95%, 15-90%, 20-85%, 25-75%, 30-70%, 35-65%, 40-60%, 45-55%, or 50%, or by 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);

[0168] h. The level of an anti-inflammatory metabolite is increased by about 5% to 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%, all values ​​and any combination thereof) compared to one or more baseline measurements taken from a healthy subject and / or a subject under treatment. The antibody is reduced by 10-95%, 15-90%, 20-85%, 25-75%, 30-70%, 35-65%, 40-60%, 45-55%, or 50%, or by 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).

[0169] It is assumed that the percentage of Th2a+ cells on day 1 pre-dose is expected to be >15% in peanut-allergic subjects. Treatment with TIMP-PPE is expected to reduce the percentage of Th2a+ cells to <15% 14 days after the second dose, indicating the induction of immune tolerance. An increase in the percentage of Th2a+ cells to >15% at any of the subsequent time points (e.g., days 90, 180, 270, and 360 post-dose) would indicate weakening of immune tolerance and justify re-administration of TIMP-PPE to restore immune tolerance. In combination, the results of the above analyses can be used to determine the immune tolerance signature and whether the subject has maintained immune tolerance. If such analyses indicate weakening and / or loss of immune tolerance, TIMP-PPE can be re-administered to the subject to restore immune tolerance.

[0170] Pharmaceutical preparations Pharmaceutical compositions of the present disclosure containing the TIMP-PPE described herein as an active ingredient may contain pharmaceutically acceptable carriers or additives depending on the route of administration. Examples of such carriers or additives include water, pharmaceutically acceptable organic solvents, collagen, polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymers, sodium carboxymethylcellulose, sodium polyacrylate, sodium alginate, water-soluble dextran, sodium carboxymethyl starch, pectin, methylcellulose, ethylcellulose, xanthan gum, gum arabic, casein, gelatin, agar, diglycerin, glycerin, propylene glycol, polyethylene glycol, petrolatum, paraffin, stearyl alcohol, stearic acid, human serum albumin (HSA), mannitol, sorbitol, lactose, and pharmaceutically acceptable surfactants. The additives used may be selected from the above or combinations thereof as needed depending on the dosage form of the present disclosure, but are not limited to these.

[0171] The formulation of a pharmaceutical composition will vary according to the route of administration (e.g., solution, emulsion) selected. A suitable composition 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 or alcoholic / aqueous solutions, emulsions, or suspensions, including saline and buffered media. Parenteral vehicles can include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's, or fixed oils. Intravenous vehicles can include various additives, preservatives, or fluid, nutrient, or electrolyte replenishers.

[0172] Various aqueous carriers, e.g., sterile phosphate-buffered saline, bacteriostatic water, water, buffered water, 0.4% saline, 0.3% glycine, etc., may contain other proteins for enhanced stability, such as albumin, lipoproteins, globulins, etc., that have been subjected to mild chemical modifications, etc.

[0173] Therapeutic formulations of the inhibitor are prepared for storage by mixing the inhibitor of the desired purity, in the form of a lyophilized formulation or aqueous solution, with any physiologically acceptable carrier, excipient, or stabilizer (Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)). Acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations employed and include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzylammonium chloride; hexamethonium chloride; benzalkonium chloride; benzethonium chloride; phenol, butyl, or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; serum albumin, gelatin, etc. hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, alginate, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions such as sodium; metal complexes (e.g., Zn-protein complexes); and / or non-ionic surfactants such as TWEEN™, PLURONICS™, or polyethylene glycol (PEG).

[0174] Formulations to be used for in vivo administration must be sterile, which is readily accomplished by filtration through sterile filtration membranes.

[0175] Aqueous suspensions may contain the active compound mixed with excipients suitable for the manufacture of aqueous suspensions. Such excipients include suspending agents, such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, tragacanth gum, and acacia gum; dispersing or wetting agents may be naturally occurring phosphatides, such as lecithin, or condensation products of alkylene oxides with fatty acids, such as polyoxyethylene stearate, or condensation products of ethylene oxide with long-chain aliphatic alcohols, such as heptadecaethyl-enoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitols, such as sorbitol polyoxyethylene monostearate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, such as polyethylene sorbitan monostearate. Aqueous suspensions may also contain one or more preservatives, such as ethyl or n-propyl p-hydroxybenzoates.

[0176] The TIMP-PPEs described herein can be lyophilized for storage and reconstituted in a suitable carrier prior to use.

[0177] 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 dicalcium phosphate, and / or a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, b) binders such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, c) humectants such as glycerol, d) disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) solution retardants such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) humectants such as cetyl alcohol and glycerol monostearate, h) absorbents such as kaolin and bentonite clay, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium laurate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also comprise buffering agents.

[0178] kit In an additional aspect, the present disclosure includes kits containing one or more compounds or compositions packaged in a manner that facilitates their use to practice the methods of the present disclosure. In one embodiment, such kits include a compound or composition described herein (e.g., a composition comprising a TIMP alone or in combination with a second agent) packaged in a container such as a sealed bottle or vessel, with a label affixed to the container or included in the package that describes the use of the compound or composition in practicing the method. Preferably, the compound or composition is packaged in a unit dosage form. The kit may further include a suitable device for administering the composition according to a specific route of administration or for performing a screening assay. Preferably, the kit includes a label that describes the use of the inhibitor composition.

[0179] Additional aspects and details of the present disclosure will be apparent from the following examples, which are intended to be illustrative rather than limiting.

[0180] Example 1 - Phase I / II Study of TIMP-PPE in Peanut Allergy This example describes a Phase 1b / 2a, randomized, double-blind, placebo-controlled, two-part study to evaluate the safety, tolerability, pharmacodynamics, and efficacy of TIMP-PPE (CNP-201) in subjects aged 16 to 55 years with peanut allergy.

[0181] CNP-201 consists of PLGA nanoparticles encapsulating purified peanut extract. The CNP-201 particles have an average diameter of 400-800 nm and a negative zeta potential of -32 mV to -50 mV. CNP-201 particles are supplied as a lyophilized formulation. CNP-201 particles are reconstituted in sterile water for injection and diluted in sterile saline (0.9% sodium chloride) prior to administration.

[0182] The test includes Part A and Part B.

[0183] Part A: Part A is a randomized, double-blind, placebo-controlled study of the safety and tolerability of CNP-201 at ascending dose levels. Part A will enroll three cohorts to receive CNP-201 or placebo at multiple ascending dose levels. Part B will continue as a randomized, double-blind, placebo-controlled, repeat-dose study using the safe and tolerated dose levels of CNP-201 determined from Part A.

[0184] Subjects who meet all inclusion criteria but no exclusion criteria after the 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 allergy to peanuts. The SPT and DBPCFC will be conducted by study physicians or staff trained to manage clinical emergencies present on-site, with ready access to emergency medications and equipment, and in close proximity to a hospital emergency department for rapid delivery of emergency treatment if needed. Subjects who continue to meet all inclusion criteria and no exclusion criteria after completing both days of DBPCFC will be eligible to enroll in the study.

[0185] All subjects who continue to meet all I / E criteria after DBPCFC will receive subcutaneous injections of omalizumab (XOLAIR®). The dose of omalizumab (XOLAIR®) will be determined by the subject's weight and serum IgE at initial screening according to the protocol-specified product label. Subjects will be dosed either every 2 weeks or every 4 weeks according to the product label.

[0186] Subjects who continue to meet the inclusion / exclusion criteria will be randomized in a 2:1 ratio (Part A) or a 1:1 ratio (Part B) on Day 1 to receive either CNP-201 or placebo (0.9% sodium chloride USP) by intravenous (IV) infusion. Subjects will receive CNP-201 or placebo on Days 1 and 8.

[0187] Subjects with peanut allergy eligible for enrollment in this study will be defined based on the following inclusion criteria: 1. Men and non-pregnant women aged 16-55. 2. Subjects with a body mass index (BMI) of ≥ 18 and ≤ 32 and a weight of > 30 kg and ≤ 150 kg at screening. 3. Subjects with serum IgE ≥ 30 IU / mL and ≤ 1500 IU / mL at screening. 4. Subjects with a physician-diagnosed peanut allergy or a documented history of peanut allergy. 5. Subject with a documented history of non-severe anaphylaxis (grade ≤ 3) to peanut, including mild wheezing or dyspnea without hypoxia. 6. Subjects with peanut-specific IgE >5kU / L as measured by ImmunoCAP at screening. 7. Subject self-reported to be on a peanut-free diet with no suspected peanut exposure, including any peanut food challenge, for at least 14 days prior to screening and agrees to continue limiting peanut exposure during the study, excluding Study DBPCFC. 8. Subjects with a positive skin prick test (SPT) to peanut with a change in wheal diameter of >3 mm compared to the negative control (50% glycerin) at screening. 9. Subjects willing and able to provide Institutional Review Board (IRB) approved written informed consent. 10. Subjects with a positive peanut DBPCFC at screening (baseline DBPCFC) at a challenge dose of peanut protein of ≥ 10 mg and ≤ 300 mg. 11. Subjects with ≥ 15% peanut-specific Th2a+ T cells (peanut-specific Th2a+ cells / total peanut-specific T cells) after ex vivo stimulation of PBMCs at screening.

[0188] After completing both screening visits, including both sets of baseline DBPCFC (peanut and placebo challenges, administered on two separate days), subjects who meet all inclusion criteria and no exclusion criteria will be enrolled in one of three dose-escalation cohorts. Subjects will be randomized 2:1 to receive either CNP-201 or placebo (0.9% sodium chloride injection) as a 200 mL intravenous infusion on Days 1 and 8.

[0189] The dose levels for the three cohorts are as follows: Cohort 1: 250 mg, Cohort 2: 450 mg, Cohort 3: 650 mg. Dosing of subjects within a dose cohort will be separated by at least 48 hours.

[0190] Subjects who meet all inclusion criteria but no exclusion criteria after the 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 is treated, the peanut and placebo challenges will be administered at least 48 hours apart. The procedural schedule for this study is shown in Figure 1C.

[0191] After completion of the second set of DBPCFC and 2-hour observation period, subjects who continue to meet all inclusion criteria and do not meet any exclusion criteria will receive their first subcutaneous injection of omalizumab. The dose of omalizumab (XOLAIR®) will be determined by the subject's weight and serum IgE at screening, according to the product label and as described in Table 2. Subjects will be dosed either every 2 weeks (days -29, -15, and -1) or every 4 weeks (days -29 and -1), according to the product label. [Table 2]

[0192] Any subject who experiences a severe hypersensitivity reaction to omalizumab will be provided appropriate treatment, withdrawn from the study, and then replaced with an additional subject at the same dose level.

[0193] Subjects return to the clinic on Day 1 for a final assessment of eligibility and collection of laboratory samples. Subjects who continue to meet all inclusion criteria and do not meet any exclusion criteria will be randomized to an open dose cohort at that time. Subjects will receive CNP-201 or placebo on Days 1 and 8. CNP-201 or placebo will be administered by intravenous infusion over approximately 3-4 hours using a graded infusion rate. Subjects will undergo medical observation in the clinic for acute adverse events (AEs), including infusion reactions (IRs), for 4 hours after the infusion. Antihistamine / epinephrine will be immediately available to treat allergic reactions, if any occur.

[0194] Subjects return for office visits 2 days after each infusion (Days 3 and 10) for collection of safety laboratory values, medication review, and assessment of AEs, and are followed daily via telephone visits between infusions (Days 33-36) to assess and document any AEs and medication changes. In the post-dose period, subjects return to the clinic on Day 15 for collection of safety laboratory values, PD measurements, and assessment of AEs and medication changes.

[0195] After all subjects in a dose cohort have completed the day 15 office visit (7 days after the second dose), a Data Monitoring Committee (DMC) will convene to review all available safety data and determine whether progression to the next escalating dose cohort is acceptable, whether cohort expansion is justified (a minimum of 3 additional subjects randomized 2:1 to receive CNP-201 or placebo (0.9% sodium chloride USP)), or whether any other clinical recommendations should be made.

[0196] Subjects return to the clinic on Day 60 for collection of immune safety laboratory values, PD measurements, and a second SPT, followed by DBPCFC (completed on Day 61). Subjects return to the clinic at the end of the Day 90 study visit for collection of safety laboratory values, PD measurements, and final assessment of AEs and medication changes. Once all subjects have completed the Day 15 visit, a recommendation will be made regarding continuation into Part B at the safe and tolerated dose of CNP-201 identified in Part A.

[0197] Part B: Subjects in Part B will be randomized 1:1 to receive CNP-201 or placebo (0.9% sodium chloride, USP) as determined in Part A for safety and tolerability levels. Subjects who meet all inclusion criteria but not exclusion criteria after the 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 is treated, the peanut and placebo challenges will be administered at least 48 hours apart. After completion of the DBPCFC and the second set of 2-hour observation periods, subjects who continue to meet all inclusion criteria but not exclusion criteria will receive their first subcutaneous injection of omalizumab.

[0198] The dose of omalizumab (XOLAIR®) will be determined by the subject's weight and serum IgE at screening according to the product labeling specified in the protocol and is listed in Table 2. Subjects will be dosed either every 2 weeks or every 4 weeks according to the product labeling. Any subject who experiences a severe hypersensitivity reaction to omalizumab will be provided appropriate treatment and discontinued from the study. These subjects will be replaced in Part B.

[0199] Subjects will return to the clinic on Day 1 for final assessment of eligibility and collection of laboratory samples. Subjects who continue to meet all inclusion criteria and do not meet any exclusion criteria will be randomized to receive either CNP-201 or placebo.

[0200] Subjects will receive CNP-201 or placebo on days 1 and 8. The study product will be administered by IV infusion over approximately 3-4 hours using a gradual infusion rate. Subjects will undergo medical observation in the clinic for acute AEs for 4 hours after the infusion. Antihistamine / epinephrine will be immediately available to treat allergic reactions if they occur. Subjects will be followed through daily telephone visits between each infusion (days 2-7 and 9-14) to assess and document any AEs and medication changes.

[0201] In the post-dose period, subjects return to the clinic on Day 15 for collection of safety laboratory values, PD measurements, and assessment of AEs and medication changes. Subjects also return to the clinic on Day 60 for immune safety laboratory values, PD measurements, and a second SPT, followed by DBPCFC (completed on Day 61). Subjects return to the clinic at the end of the study visit on Day 90 for collection of safety laboratory values, PD measurements, and final assessment of AEs and medication changes.

[0202] Emerging safety and tolerability data will be monitored in Part B. The Medical Monitor will be notified of any serious adverse events (SAEs) and any >Grade 2 adverse events (CTCAE v. 5.0) within 24 hours of becoming aware of such events. The Medical Monitor may then convene an ad hoc DMC meeting to evaluate the safety and tolerability data, determine whether continued dosing remains acceptable, and make recommendations, including but not limited to, continuing dosing and discontinuing or suspending dosing in subjects. The DMC may recommend discontinuing or suspending the study at any time during the study if, in their opinion, continued dosing poses a safety risk to the subject.

[0203] In both Part A and Part B of the study, subjects will receive CNP-201 via intravenous infusion lasting approximately 3 to 4 hours according to 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 remainder of the infusion.

[0204] Study duration: 2 doses, 7 days apart (Parts A and B). The total duration of the study for an individual subject is approximately 134 days, with 14 days for screening, 30 days for omalizumab administration, 60 days for IP administration, and a 30-day follow-up period.

[0205] Primary endpoints (Part A and Part B) included: frequency of adverse events (AEs) and serious adverse events (SAEs), MedDRA 23.0 (CTCAE v.5.0); laboratory safety assessments (hematology, serum chemistry, coagulation panel, urinalysis); physical examination including vital signs (blood pressure, heart rate, 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 peanut-specific Th2a expression after ex vivo stimulation of PBMCs with placebo versus CNP-201 at baseline (pre-dose Day 1) and Day 15. + T cells (peanut-specific Th2a + and the change in the percentage of activated peanut-specific T cells vs. total peanut-specific T cells after ex vivo stimulation of PBMCs between placebo and CNP-201 at baseline (day 1 pre-dose) and day 15.

[0206] Secondary endpoints (Part A and Part B) include: change in the ratio of IL-5 to IFN-γ after ex vivo stimulation of PBMCs between placebo and CNP-201 at baseline (day 1 pre-dose) and day 15.

[0207] Exploratory endpoints (Part A and Part B) include: change in the percentage of peanut-specific T regulatory cells (peanut-specific T regulatory cells / peanut-specific CD4+ effector memory cells) after ex vivo stimulation of PBMCs between placebo and CNP-201 at baseline (pre-dose day 1) and day 15; effective concentration (EC) at 50% of maximum basophil activation as measured by basophil activation test (CD203c+ / CD63+ / - basophil activation) between placebo and CNP-201 at baseline (pre-DBPCFC day -30) and day 60. 50 ); change in the ratio of peanut-specific IgE to IgG as measured by ImmunoCap assay between placebo and CNP-201 at baseline (day -30 before DBPCFC) and day 60; and change in cumulative tolerated dose (CTD) of peanut protein (mg) administered during DBPCFC between placebo and CNP-201 at baseline (days -30 to -29) and days 60 to 61. The cellular markers used to assess the exploratory endpoints are listed in Table 3 below: [Table 3]

[0208] Example 1A - Phase I / II Study of TIMP-PPE in Peanut Allergy An alternative protocol is provided for conducting a Phase 1b / 2a randomized, double-blind, placebo-controlled, two-part study to evaluate the safety, tolerability, pharmacodynamics, and efficacy of TIMP-PPE (CNP-201) in subjects aged 16 to 35 years with peanut allergy.

[0209] CNP-201 consists of PLGA nanoparticles encapsulating purified peanut extract with an average diameter of 400-800 nm and a negative zeta potential of -30 mV to -60 mV. CNP-201 particles are supplied as a lyophilized formulation. CNP-201 particles are reconstituted in sterile water for injection and diluted in sterile saline (0.9% sodium chloride) prior to administration.

[0210] The study includes Part A and Part B. Part A: Part A is a randomized, double-blind, placebo-controlled study of the safety and tolerability of CNP-201 at ascending dose levels. Part A will enroll three cohorts to receive CNP-201 or placebo at multiple ascending dose levels. Part B will continue as a randomized, double-blind, placebo-controlled, repeat-dose study using the safe and tolerated dose levels of CNP-201 determined from Part A.

[0211] Subjects who meet all inclusion criteria (except IgG and IgE results, which may not be available at Visit 2) after the initial screening assessment but do not meet any exclusion criteria 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 allergy to peanuts. The SPT and DBPCFC will be conducted by study physicians or staff trained to manage clinical emergencies present on-site, with ready access to emergency medications and equipment, and in close proximity to a hospital emergency department for rapid delivery of emergency care if needed. Subjects who continue to meet all inclusion criteria after completing both days of DBPCFC and who do not meet any exclusion criteria will be eligible to enroll in the study.

[0212] All subjects who continue to meet all I / E criteria after DBPCFC will receive subcutaneous injections of omalizumab (XOLAIR). The dose of omalizumab (XOLAIR®) will be determined by the subject's weight and serum IgE at initial screening according to the product labeling specified in the protocol. Subjects will be dosed either every 2 weeks or every 4 weeks according to the product labeling.

[0213] Subjects who continue to meet the inclusion criteria and do not meet the exclusion criteria will be randomized in a 2:1 ratio (Part A) or a 1:1 ratio (Part B) on Day 1 to receive either CNP-201 or placebo (0.9% sodium chloride USP) by intravenous (IV) infusion. Subjects will receive CNP-201 or placebo on Days 1 and 8. Subjects will remain in the clinic on Days 1 and 8 from the time of admission (before administration of CNP-201 or placebo) until their final procedure, which will be performed 4 hours after the dose on that day, unless an infusion reaction, anaphylaxis, or other adverse event requires an extended monitoring period. If safety parameters are acceptable to the investigator, subjects will be discharged.

[0214] Subjects with peanut allergy eligible for enrollment in this study will be defined based on the following inclusion criteria: 1. Men and non-pregnant women aged 16-35. 2. Subjects with a body mass index (BMI) of ≥ 18 and ≤ 32 and a weight of > 30 kg and ≤ 150 kg at screening. Subjects outside this range may be included at the discretion of the investigator. 3. Subjects with 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 documented history of peanut allergy. 5. Subject with a documented history of non-severe anaphylaxis (grade ≤ 3) to peanut, including mild wheezing or dyspnea without hypoxia. 6. Subjects with peanut-specific IgE >2kU / L as measured by ImmunoCAP at screening, and / or a positive skin prick test (SPT) to peanut with a change in wheal diameter of >3mm compared to the negative control (50% glycerin) at screening. 7. Subject self-reported to be on a peanut-free diet with no suspected peanut exposure, including any peanut food challenge, for at least 14 days prior to screening and agrees to continue limiting peanut exposure during the study, excluding Study DBPCFC. 8. Female subjects and male subjects and their female spouses / partners willing to practice highly effective methods of contraception, which may include, but are not limited to, abstinence, sexual intercourse only with persons of the same sex, monogamous relationships with a vasectomized partner, vasectomy, hysterectomy, bilateral tubal ligation, approved hormonal methods, intrauterine device (IUD), or the use of spermicides in combination with barrier methods (e.g., condoms, diaphragms) beginning at screening and continuing throughout the study through Day 90 (EOS / ET). 9. Female subjects agree to abstain from breastfeeding throughout the study, starting at initial screening and continuing through Day 90 (EOS / ET). 10. Female subjects who agree not to donate eggs starting at initial screening and continuing throughout the study until Day 90 (EOS / ET). 11. Subjects willing and able to provide Institutional Review Board (IRB) approved written informed consent. 12. Subject willing to perform and comply with all study procedures, including attending study visits as scheduled and completing two DBPCFCs. 13. Male subjects agree not to donate sperm starting at screening and throughout the study until Day 90 (EOS / ET).

[0215] Subjects must have a positive peanut DBPCFC at screening at challenge doses of ≥ 10 mg and ≤ 300 mg peanut protein to be included in statistical analyses for exploratory endpoints. Subjects who tolerate > 444 mg peanut (cumulative tolerated dose) will be followed for safety and evaluated separately.

[0216] After completing both screening visits, including both sets of baseline DBPCFC (peanut and placebo challenges, administered on two separate days), subjects who meet all inclusion criteria and none of the exclusion criteria will be enrolled in one of three dose-escalation cohorts. Subjects will be randomized in a 2:1 ratio to receive either CNP-201 or placebo (0.9% sodium chloride injection) as a 200 mL intravenous infusion on days 1 and 8. The dose levels for the three cohorts are as follows: Cohort 1: 250 mg, Cohort 2: 450 mg, and Cohort 3: 650 mg. Dosing of subjects within a dose cohort will be separated by at least 48 hours.

[0217] Subjects who meet all inclusion criteria but no exclusion criteria after the 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 is treated, the peanut and placebo challenges will be administered at least 48 hours apart.

[0218] After completion of the second DBPCFC challenge and the 2-hour observation period, subjects who continue to meet all inclusion criteria and do not meet any exclusion criteria will proceed to their first subcutaneous injection of XOLAIR®.

[0219] The dose and frequency of administration of XOLAIR® (every 2 weeks or every 4 weeks) will be determined by the subject's serum IgE and body weight at screening measured at XOLAIR® dose 1 as described in Table 2.

[0220] Subjects return to the clinic on Day 1 for a final assessment of eligibility and collection of laboratory samples. Subjects who continue to meet all inclusion criteria and do not meet any exclusion criteria will be randomized to an open dose cohort at that time. Subjects will receive CNP-201 or placebo on Days 1 and 8. CNP-201 or placebo will be administered by intravenous infusion over approximately 3-4 hours using a graded infusion rate. Subjects will undergo medical observation in the clinic for acute adverse events (AEs), including infusion reactions (IRs), for 4 hours after the infusion. Antihistamine / epinephrine will be immediately available to treat allergic reactions, if any occur.

[0221] If two of the same Grade 3 adverse events (AEs) considered likely to be related to CNP-201 or one ≥ Grade 4 (CTCAE v.5.0 or CoFAR v.1 for allergy-related AEs) occur, dosing will be suspended and the DMC will convene to review all available safety data obtained to date. Any Grade 4 or Grade 5 AEs considered to be serious AEs (SAEs) have been reported to the Agency. After reviewing all available safety data, the DMC will make recommendations, including but not limited to, discontinuing dosing, de-escalating dosing, continuing dosing, expanding the current cohort, adding cohorts in Part A, or determining dose-limiting toxicities. The DMC may also convene ad hoc to address emerging safety concerns during dosing of any subject in Part A, based on ongoing daily monitoring of safety data by the sponsor medical monitor. The DMC will evaluate available safety data, including but not limited to AEs, physical examination, vital signs, 12-lead ECG, and available laboratory results.

[0222] Subjects return for an office visit 2 days after each infusion (Days 3 and 10) for collection of safety laboratory values, medication review, and assessment of AEs, and are followed daily via telephone visits after each infusion (Days 4-7 and 11-14) to assess and document any AEs and medication changes. In the post-dose period, subjects return to the clinic 7 days after administration of Dose 2 of CNP-201 or placebo for collection of safety laboratory values, PD measurements, and assessment of AEs and medication changes.

[0223] After all subjects in a dose cohort have completed the Day 15 office visit (7 days after the second dose), the DMC will convene to review all available safety data and determine whether progression to the next escalating dose cohort is permitted, whether cohort expansion is justified (a minimum of 3 additional subjects randomized 2:1 to receive CNP-201 or placebo), or whether any other clinical recommendations (e.g., expansion to Part B) should be made.

[0224] Subjects will return to the clinic on Day 60 for immune safety laboratory data collection, PD measurements, and a second SPT, followed by a post-dose DBPCFC. They will also return to the clinic at the end of the study visit on Day 90 for safety laboratory data collection, PD measurements, and a final assessment of AEs and medication changes. Emerging safety and tolerability data will be continuously monitored. The medical monitor will be notified of any serious adverse events (SAEs) likely to be related to the study product (CTCAE v.5.0 or CoFAR v.1) and any Grade 2 adverse events within 24 hours of becoming aware of such events. The medical monitor will then evaluate the safety and tolerability data to determine whether continued administration remains acceptable and may convene an ad hoc DMC meeting to make recommendations, including but not limited to, continuing administration and discontinuing or suspending administration in subjects. The DMC may recommend discontinuing or suspending the study at any time during the study if, in their opinion, continued administration poses an unacceptable safety risk to the subject. Once all subjects have completed the Day 15 visit, a recommendation will be made regarding continuation into Part B at the safe and tolerated dose of CNP-201 identified in Part A.

[0225] Part B: Subjects in Part B are randomized in a 1:1 ratio to receive CNP-201 or placebo as determined in Part A for safety and tolerability levels. Subjects in Parts A and B undergo the same evaluations; the only difference between the two parts is dose escalation in Part A.

[0226] Subjects who meet all inclusion criteria but no exclusion criteria after the 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 is treated, the peanut and placebo challenges will be administered at least 48 hours apart.

[0227] After completion of the second DBPCFC challenge and the 2-hour observation period, subjects who continue to meet all inclusion criteria and no exclusion criteria will proceed to their first subcutaneous injection of XOLAIR®. The dose and administration frequency (every 2 weeks or every 4 weeks) of XOLAIR® will be determined by the subject's serum IgE and body weight at screening measured at XOLAIR® dose 1 as described in Table 2.

[0228] Subjects will return to the clinic on Day 1 for final assessment of eligibility and collection of laboratory samples. Subjects who continue to meet all inclusion criteria and do not meet any exclusion criteria will be randomized to receive either CNP-201 or placebo.

[0229] Subjects will receive CNP-201 or placebo on days 1 and 8. CNP-201 or placebo will be administered by intravenous infusion over approximately 3-4 hours using a graded infusion rate. Subjects will undergo medical observation in the clinic for acute adverse events (AEs), including infusion reactions (IRs), for 4 hours after the infusion. Antihistamine / epinephrine will be immediately available to treat allergic reactions, if any occur.

[0230] Subjects return for an office visit 2 days after each infusion (Days 3 and 10) for collection of safety laboratory values, medication review, and assessment of AEs, and are followed daily via telephone visits after each infusion (Days 4-7 and 11-14) to assess and document any AEs and medication changes. In the post-dose period, subjects return to the clinic 7 days after administration of Dose 2 of CNP-201 or placebo for collection of safety laboratory values, PD measurements, and assessment of AEs and medication changes.

[0231] Subjects will return to the clinic on Day 60 for immune safety laboratory data collection, PD measurements, and a second SPT, followed by a post-dose DBPCFC. They will also return to the clinic at the end of the study visit on Day 90 for safety laboratory data collection, PD measurements, and a final assessment of AEs and medication changes. Emerging safety and tolerability data will be continuously monitored. The medical monitor will be notified of any serious adverse events (SAEs) likely to be related to the study product (CTCAE v.5.0 or CoFAR v.1) and any Grade 2 adverse events within 24 hours of becoming aware of such events. The medical monitor will then evaluate the safety and tolerability data to determine whether continued administration remains acceptable and may convene an ad hoc DMC meeting to make recommendations, including but not limited to, continuing administration and discontinuing or suspending administration in subjects. The DMC may recommend discontinuing or suspending the study at any time during the study if, in their opinion, continued administration poses an unacceptable safety risk to the subject.

[0232] In both Part A and Part B of the study, subjects will receive CNP-201 via intravenous infusion lasting approximately 3 to 4 hours according to 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 remainder of the infusion.

[0233] In both Part A and Part B, subjects randomized to placebo will receive 0.9% sodium chloride for injection (normal saline [NS]) administered as a 200 mL intravenous infusion on Days 1 and 8 according to the following stepped infusion rates: 20 mL / hour for the first 15 minutes, 40 mL / hour for the next 15 minutes, and 80 mL / hour for the remainder of the infusion.

[0234] Study duration: 2 doses, 7 days apart (Parts A and B). The total duration of the study for an individual subject is approximately 134 days, with 14 days of screening, approximately 30 days of XOLAIR® administration, 60 days of study product administration, and approximately 30 days of post-dose follow-up.

[0235] Primary endpoints (Part A and Part B) included: frequency of adverse events (AEs) and serious adverse events (SAEs), MedDRA 23.0 (CTCAE v.5.0 or CoFAR V.1 for allergy-related AEs); laboratory safety assessments (hematology, serum chemistry, coagulation panel, and urinalysis); physical examination including vital signs (blood pressure, heart rate, and 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).

[0236] Exploratory endpoints (Part A and Part B) include: change in the percentage 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 (pre-dose Day 1) and Day 15; change in the effective concentration at 50% of maximal basophil activation (EC50) as measured by a basophil activation test (CD203c+ / CD63+ / - basophil activation) between placebo and CNP-201 at baseline (screening, pre-DBPCFC Visit 2) and Visit 11; change in peanut-specific IgE as measured by ImmunoCap assay .... Change in the ratio of peanut-specific IgE to IgG as measured by ImmunoCap assay between placebo and CNP-201 at Question 11; Change in cumulative tolerated dose (CTD) of peanut protein (mg) administered during DBPCFC between placebo and CNP-201 at baseline (screening DBPCFC, Visits 2 and 3) and post-dose (post-DBPCFC administration, Visits 11 and 12); Change in the percentage 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 (pre-dose on Day 1) and Day 15.

[0237] Example 2 - Phase I / II study of TIMP-PPE without IgE inhibitors in peanut allergy 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 administration of an IgE inhibitor in subjects aged 16 to 55 years with peanut allergy.

[0238] CNP-201 consists of PLGA nanoparticles encapsulating purified peanut extract with an average diameter of 400-800 nm and a negative zeta potential of -30 mV to -60 mV. CNP-201 particles are supplied as a lyophilized formulation. CNP-201 particles are reconstituted in sterile water for injection and diluted in sterile saline (0.9% sodium chloride) prior to administration.

[0239] The study is a randomized, double-blind, placebo-controlled study of the safety and tolerability of CNP-201 at ascending dose levels. The study will enroll three cohorts to receive CNP-201 or placebo at multiple ascending dose levels.

[0240] Subjects who meet all inclusion criteria and none of the exclusion criteria will be enrolled in the study. Subjects who continue to meet the inclusion / exclusion criteria will be randomized in a 2:1 ratio on Day 1 to receive either CNP-201 or placebo (0.9% sodium chloride USP) via intravenous (IV) infusion. Subjects will receive CNP-201 or placebo on Days 1 and 8.

[0241] Subjects may receive a premedication with an antihistamine (e.g., 10 mg IV cetirizine) and a corticosteroid (e.g., 125 mg IV methylprednisolone) 30 minutes before each infusion of CNP-201 or placebo. Subjects will maintain their current SoC throughout the study, except for a 12-hour washout period (for beta-agonists, theophylline, and cromolyn only) and a 7-day washout period for the antihistamine prior to the skin prick test (SPT). Subjects will resume their SoC regimen after the SPT and maintain SoC throughout the study administration / duration.

[0242] Subjects with peanut allergy eligible for enrollment in this study will be defined based on the following inclusion criteria: 1. Men and non-pregnant women aged 16-55. 2. Subjects with a physician-diagnosed peanut allergy or a documented history of peanut allergy. 3. Subjects weighing ≥ 31.25 kg at screening. Subjects outside this range may be included at the discretion of the investigator. 4. Subject with a documented history of non-severe anaphylaxis (grade ≤ 3) to peanut, including mild wheezing or dyspnea without hypoxia. 5. Subjects with peanut-specific IgE >5kU / L as measured by ImmunoCAP at screening. 6. Subject self-reports having been on a peanut-free diet with no suspected peanut exposure, including any peanut food challenge, for at least 14 days prior to screening and agrees to continue limiting peanut exposure during the study. 7. Subjects with a positive skin prick test (SPT) to peanut with a change in wheal diameter of >3 mm compared to the negative control (50% glycerin) at screening. 8. Female subjects and male subjects and their female spouses / partners willing to practice highly effective methods of contraception, which may include, but are not limited to, abstinence, sexual intercourse only with persons of the same sex, monogamous relationship with a vasectomized partner, vasectomy, hysterectomy, bilateral tubal ligation, approved hormonal methods, intrauterine device (IUD), or the use of spermicides in combination with barrier methods (e.g., condoms, diaphragms) beginning at screening and continuing throughout the study through Day 38 (EOS / ET). 9. Female subjects agree to abstain from breastfeeding starting at initial screening and throughout the study until Day 38 (EOS / ET). 10. Female subjects who agree not to donate eggs starting at initial screening and continuing throughout the study until Day 38 (EOS / ET). 11. Subjects willing and able to provide Institutional Review Board (IRB) approved written informed consent. 12. Subject is willing to perform and comply with all study procedures. 13. Male subjects agree not to donate sperm starting at screening and throughout the study until Day 38 (EOS / ET).

[0243] After completing the screening visit, subjects who meet all inclusion criteria and no exclusion criteria will be enrolled in one of three dose escalation cohorts. Subjects will be randomized in a 2:1 ratio to receive either CNP-201 or placebo (0.9% sodium chloride injection) as a 200mL intravenous infusion on days 1 and 8. The dose levels for the three cohorts are as follows: Cohort 1: 250mg, Cohort 2: 450mg, and Cohort 3: 650mg. Dosing of subjects within a dose cohort will be separated by at least 48 hours.

[0244] Subjects who meet all inclusion criteria and none of the exclusion criteria at screening will be enrolled in the study. Subjects will return to the clinic on Day 1 for a final assessment of eligibility and collection of laboratory samples. Subjects who continue to meet all inclusion criteria and none of the exclusion criteria will be randomized to the next open dose cohort at that time. Subjects may receive a premedication with an antihistamine (e.g., 10 mg IV cetirizine) and a corticosteroid (e.g., 125 mg IV methylprednisolone) 30 minutes before each infusion of CNP-201 or placebo on Days 1 and 8. CNP-201 or placebo will be administered by intravenous infusion over approximately 3-4 hours using a graded infusion rate. Subjects will be medically monitored in the clinic for acute adverse events (AEs), including infusion reactions (IRs), for 4 hours after the infusion. Antihistamine / epinephrine, including intramuscular (IM) and intravenous (IV) epinephrine, will be readily available to treat allergic reactions, if they occur.

[0245] Subjects will be followed daily through telephone visits after each infusion (Days 2-7 and Days 9-14) to assess and document any AEs and medication changes. During the post-dose period, subjects will return to the clinic 7 days after administration of Dose 2 of CNP-201 or placebo (Day 15) for collection of safety laboratory values, PD measurements, and assessment of AEs and medication changes.

[0246] After all subjects in a dose cohort have completed their day 15 office visit (7 days after the second dose), a DMC will convene to review all available safety data and determine whether progression to the next escalating dose cohort is permissible.

[0247] Subjects return to the clinic at the end of study visit on Day 38 for collection of safety laboratory values, PD measurements, and a final assessment of AEs and medication changes and a second SPT.

[0248] Emerging safety and tolerability data will be continuously monitored. The Medical Monitor will be notified of any serious adverse events (SAEs) likely to be related to the study product (CTCAE v.5.0 or CoFAR v.3.0) and any Grade 2 or greater adverse events within 24 hours of becoming aware of such events. The Medical Monitor may convene ad hoc DMC meetings to evaluate safety and tolerability data, determine whether continued dosing remains acceptable, and make recommendations, including but not limited to, continuing dosing and discontinuing or suspending dosing in subjects. The DMC may recommend discontinuing or suspending the study at any point during the study if, in their opinion, continued dosing poses an unacceptable safety risk to the subject.

[0249] Subjects will receive CNP-201 via intravenous infusion lasting approximately 3-4 hours according to the following step-wise infusion rate: 20 mL / hour for the first 15 minutes, 40 mL / hour for the next 15 minutes, and 80 mL / hour for the remainder of the infusion.

[0250] Study duration: 2 doses, 7 days apart. The total duration of the study for each subject is approximately 45 days, with 7 days for screening, 8 days for test product administration, and 30 days for post-dose evaluation.

[0251] Primary endpoints included: frequency 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 examination including vital signs (blood pressure, heart rate, and 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).

[0252] Exploratory endpoints include: change in the ratio of peanut-specific IgE to IgG as measured by ImmunoCap assay between placebo and CNP-201 at baseline and day 38; change in peanut-specific IgE as measured by ImmunoCap assay between placebo and CNP-201 at baseline and day 38.

[0253] Preferred embodiments of the present disclosure are described herein. Variations of those preferred embodiments may become apparent to those skilled in the art upon reading the foregoing description. The inventors expect those skilled in the art to adopt such variations as appropriate, and the inventors intend for the present disclosure to be practiced otherwise than as specifically described herein. Accordingly, this disclosure includes all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Also, any combination of the above-described elements in all possible variations thereof is encompassed by the present disclosure unless otherwise indicated herein or otherwise clearly contradicted by context.

[0254] Example 3 - Phase I / II Study of Low-Dose TIMP-PPE in Peanut Allergy 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) in subjects aged 16 to 55 years with peanut allergy.

[0255] CNP-201 consists of PLGA nanoparticles encapsulating purified peanut extract with an average diameter of 400-800 nm and a negative zeta potential of -30 mV to -60 mV. CNP-201 particles are supplied as a lyophilized formulation. CNP-201 particles are reconstituted in sterile water for injection and diluted in sterile saline (0.9% sodium chloride) prior to administration.

[0256] The study is a randomized, double-blind, placebo-controlled study of the safety and tolerability of CNP-201 at ascending dose levels. The study will enroll four cohorts to receive CNP-201 or placebo at multiple ascending dose levels.

[0257] Subjects who meet all inclusion criteria and none of the exclusion criteria will be enrolled in the study. Subjects who continue to meet the inclusion / exclusion criteria will be randomized in a 2:1 ratio on Day 1 to receive either CNP-201 or placebo (0.9% sodium chloride USP) via intravenous (IV) infusion. Subjects will receive CNP-201 or placebo on Days 1 and 8.

[0258] Subjects may receive premedication with an NSAID (e.g., 325 mg oral acetylsalicylic acid) daily for two days and 60 minutes before each infusion of CNP-201 or placebo. Subjects may receive premedication and / or postmedication with a leukotriene modifier (e.g., 10 mg oral montelukast) 12 hours, 45 minutes, and 12 hours before each infusion of CNP-201 or placebo. Subjects may receive premedication with an antihistamine (e.g., 50 mg IV diphenhydramine) and a corticosteroid (e.g., 125 mg IV methylprednisolone) 30 minutes before each infusion of CNP-201 or placebo.

[0259] Subjects will maintain their current SoC throughout the study, except for a 12-hour washout period (for beta-agonists, theophylline, and cromolyn only) and a 7-day washout period for antihistamines prior to the skin prick test (SPT). Subjects will resume their SoC regimen after the SPT and maintain SoC throughout the administration / duration of the study.

[0260] Subjects with peanut allergy eligible for enrollment in this study will be defined based on the following inclusion criteria: 1. Men and non-pregnant women aged 16-55. 2. Subjects with a physician-diagnosed peanut allergy or a documented history of peanut allergy. 3. Subjects weighing ≥ 31.25 kg at screening. Subjects outside this range may be included at the discretion of the investigator. 4. Subject with a documented history of non-severe anaphylaxis to peanut (grade a§3), including mild wheezing or dyspnea without hypoxia. 5. Subjects with peanut-specific IgE ≥ 5 kU / L as measured by ImmunoCAP at screening, unless they have previously undergone OIT for peanut allergy. Subjects who have previously undergone OIT for peanut allergy and do not have peanut-specific IgE ≥ 5 kU / L as measured by ImmunoCap at screening may be included at the discretion of the investigator, or subjects with a positive SPT to peanut with a change in wheal diameter of ≥ 3 mm compared to the negative control (50% glycerin) at screening. Subjects who have previously undergone OIT for peanut allergy and have a positive skin prick test (SPT) to peanut with a change in wheal diameter of ≥ 3 mm at screening may be included at the discretion of the investigator. 6. Subject self-reports having been on a peanut-free diet with no suspected peanut exposure, including any peanut food challenge, for at least 14 days prior to screening and agrees to continue limiting peanut exposure during the study. 7. Subjects with a positive skin prick test (SPT) to peanut with a change in wheal diameter of >3 mm compared to the negative control (50% glycerin) at screening. 8. Female subjects and male subjects and their female spouses / partners willing to practice highly effective methods of contraception, which may include, but are not limited to, abstinence, sexual intercourse only with persons of the same sex, monogamous relationship with a vasectomized partner, vasectomy, hysterectomy, bilateral tubal ligation, approved hormonal methods, intrauterine device (IUD), or the use of spermicides in combination with barrier methods (e.g., condoms, diaphragms) beginning at screening and continuing throughout the study through Day 38 (EOS / ET). 9. Female subjects agree to abstain from breastfeeding starting at initial screening and throughout the study until Day 38 (EOS / ET). 10. Female subjects who agree not to donate eggs starting at initial screening and continuing throughout the study until Day 38 (EOS / ET). 11. Subjects willing and able to provide Institutional Review Board (IRB) approved written informed consent. 12. Subject is willing to perform and comply with all study procedures. 13. Male subjects agree not to donate sperm starting at screening and throughout the study until Day 38 (EOS / ET).

[0261] After completing the screening visit, subjects who meet all inclusion criteria and no exclusion criteria will be enrolled in one of several dosing cohorts. Subjects will be randomized in a 2:1 ratio to receive either CNP-201 or placebo (0.9% sodium chloride injection) as a 200mL intravenous infusion on days 1 and 8. The dose level of the first cohort is 25mg. Additional dose levels of the remaining three cohorts may be 0.1mg, 0.25mg, 0.5mg, 1mg, 2mg, 2.5mg, 5mg, 10mg, or may be increased to 650mg. Dosing of subjects within a dose cohort will be separated by at least 48 hours.

[0262] Subjects who meet all inclusion criteria and no exclusion criteria at screening will be enrolled in the study. Subjects will return to the clinic on Day 1 for a final assessment of eligibility and collection of laboratory samples. Subjects who continue to meet all inclusion criteria and no exclusion criteria will be randomized to the currently open dose cohort. Subjects may receive a premedication with an NSAID (e.g., 325 mg oral acetylsalicylic acid) 2 days daily and 60 minutes before each infusion of CNP-201 or placebo on Days 1 and 8. Subjects may receive a premedication and / or postmedication with a leukotriene modifier (e.g., 10 mg oral montelukast) 12 hours, 45 minutes, and 12 hours after each infusion of CNP-201 and placebo on Days 1 and 8. Subjects may receive premedication 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 days 1 and 8.

[0263] CNP-201 or placebo will be administered by intravenous infusion over approximately 3-4 hours using a gradual infusion rate. Subjects will undergo medical observation in the clinic for acute adverse events (AEs), including infusion reactions (IRs), for 4 hours after infusion. Antihistamines / epinephrine, including intramuscular (IM) and intravenous (IV) epinephrine, will be readily available to treat allergic reactions should they occur.

[0264] Subjects will be followed daily through telephone visits after each infusion (Days 2-7 and Days 9-14) to assess and document any AEs and medication changes. During the post-dose period, subjects will return to the clinic 7 days after administration of Dose 2 of CNP-201 or placebo (Day 15) for collection of safety laboratory values, PD measurements, and assessment of AEs and medication changes.

[0265] After all subjects in a dose cohort have completed their day 15 office visit (7 days after the second dose), a DMC will convene to review all available safety data and determine whether progression to the next escalating dose cohort is permissible.

[0266] Subjects return to the clinic at the end of the study visit on Day 38 for collection of safety laboratory values ​​and PD measurements, assessment of AEs and medication changes, and a second SPT. On Day 60, subjects return to the clinic for safety laboratory values, PD measurements, and undergo DBPCFC. On Days 90 and 120, subjects optionally return to the clinic for collection of safety laboratory values ​​and PD measurements. Subjects optionally return on Day 180 for a second DBPCFC consisting of peanut and placebo (oat) challenges, safety laboratory values, PD measurements, and a final assessment of AEs and medication changes.

[0267] Emerging safety and tolerability data will be continuously monitored. The Medical Monitor will be notified of any serious adverse events (SAEs) likely to be related to the study product (CTCAE v.5.0 or CoFAR v.3.0) and any Grade 2 or greater adverse events within 24 hours of becoming aware of such events. The Medical Monitor may convene ad hoc DMC meetings to evaluate safety and tolerability data, determine whether continued dosing remains acceptable, and make recommendations, including but not limited to, continuing dosing and discontinuing or suspending dosing in subjects. The DMC may recommend discontinuing or suspending the study at any point during the study if, in their opinion, continued dosing poses an unacceptable safety risk to the subject.

[0268] Subjects will receive CNP-201 via intravenous infusion lasting approximately 3 to 4 hours according to the following stepwise 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 remainder of the infusion.

[0269] Study duration: 2 doses, 7 days apart. The total duration of the study for an individual subject is approximately 67 days, with 7 days for screening, 8 days for test product administration, and 52 days for post-dose assessments. An additional 120-day post-dose assessment is optional.

[0270] Primary objectives include: safety and tolerability of CNP-201;

[0271] Exploratory objectives include: change in response to DBPCFC between patients treated with CNP-201 or placebo; change in the ratio of peanut-specific IgE to IgG between patients treated with CNP-201 or placebo; change in peanut-specific IgE between patients treated with CNP-201 or placebo; change in the proportion of peanut-specific Th2a+ T cells between patients treated with CNP-201 or placebo; change in response to a basophil activation test between patients treated with CNP-201 or placebo; change in the proportion of peanut-specific T regulatory cells between patients treated with CNP-201 or placebo.

[0272] Primary endpoints included: frequency 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 examination including vital signs (blood pressure, heart rate, and 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).

[0273] Exploratory endpoints include: difference in proportion of subjects passing DBPCFC (2000 mg dose level, not reaching induction dose at or before 4043 mg cumulative) between placebo and CNP-201 at days 60 and 180; change in cumulative tolerated dose (CTD) of peanut protein (mg) administered during DBPCFC between placebo and CNP-201 at days 60 and 180; change in peanut-specific IgE to IgG ratio as measured by ImmunoCap assay between placebo and CNP-201 at baseline and day 38; change in peanut-specific IgE to IgG ratio as measured by ImmunoCap assay between placebo and CNP-201 at baseline and day 38. Change in gE, change in the percentage 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, change in the effective concentration at 50% of maximum basophil activation (EC50) as measured by basophil activation test (CD203c+ / CD63+ / - basophil activation) between placebo and CNP-201 at baseline and day 60 and day 180, change in the percentage 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.

[0274] Example 4 - Interim Results from a Phase I / II Study of Low-Dose TIMP-PPE in Peanut Allergy This example describes the results of a Phase 1b / 2a randomized, double-blind, placebo-controlled study demonstrating that administration of low doses of CNP-201 is effective in treating peanut allergy.

[0275] Subjects received a single intravenous dose of either 1 mg or 25 mg of CNP-201 on day 1. Figure 2 summarizes the results for patients with peanut allergy who received low-dose CNP-201 particles. Three of four patients who received low-dose CNP-201 had increased BAT thresholds, with prolonged duration in two patients, by day 60. Peanut-allergic patients who received the maximum 25 mg dose of CNP-201 demonstrated a decrease in the peanut-specific IgE-to-IgG ratio on days 15 and 38. Peanut-allergic patients who received a single 25 mg dose of CNP-201 demonstrated induction of antigen-specific Tregs and a decrease in pathogenic peanut-specific T cell subsets (Th2a, TfH, B cell plasmablasts).

[0276] For the BAT test, blood samples were collected before and after low-dose CNP-201 treatment. PBMCs isolated from the blood were stimulated with increasing concentrations of peanut allergen. Basophil degranulation was measured by flow cytometry based on the expression of CD203 and CD63 positive markers on basophils. The activation threshold, i.e., cutoff, for a positive response was 50% CD203. + CD63 + In Figure 3, basophil activation was detected before treatment with 25 mg of CNP-201. Peanut-allergic subjects who received one 25 mg intravenous dose of CNP-201 showed an increase in the EC50 (the concentration of peanut allergen) required to increase the 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 demonstrates non-responsiveness to peanut allergen.

[0277] Peanut-specific IgE and IgG levels were determined from patient serum. Peanut-allergic patients receiving up to 25 mg intravenous doses of CNP-201 showed an increase in peanut-specific IgG, but peanut-specific IgE levels remained unchanged (Figure 4). Compared to placebo, a decrease in the peanut-specific IgE / IgG ratio was detected on days 15 and 38 after administration (Figure 4).

[0278] PBMCs obtained from the blood of peanut-allergic patients who received a single dose of 25 mg of CNP-201 were stimulated with whole peanut extract and CD40 ligand. Subpopulations of peanut-specific activated T cells were further characterized by the expression of lineage-specific markers. Figure 5 shows the reduction in peanut allergy-related immune cell subsets up to 100 days after administration of a single 25 mg dose of CNP-201. Peanut-allergic patients who received a single 25 mg intravenous dose of CNP-201 showed a reduction in the allergy-promoting subsets T helper 2A (Th2A), T follicular helper cells (TFH), terminally differentiated effector memory cells (TEMRA), and B cell plasmablasts. Figure 6 shows the lower pathogenic activation of peanut-specific CD4A after administration of 25 mg of CNP-201. + In low-dose CNP-201-treated patients, there was a decrease in the peanut-specific activated T cell subsets CD4+CD25+, CD4+CD69+, and CD4+PD-1+ compared to placebo. Peanut-allergic patients who received one 25 mg intravenous dose of CNP-201 had a decrease in peanut-specific Tregs (CD4 + CD137 + CD25 + CD127 lo ) showed an increase (Figure 7).

[0279] Example 5: Phase I / II Study of 1 mg Dose TIMP-PPE in Peanut Allergy 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) in subjects aged 16 to 55 years with peanut allergy.

[0280] CNP-201 consists of PLGA nanoparticles encapsulating purified peanut extract with an average diameter of 400-800 nm and a negative zeta potential of -30 mV to -80 mV. CNP-201 particles are supplied as a lyophilized formulation. CNP-201 particles are reconstituted in sterile water for injection and diluted in sterile saline (0.9% sodium chloride) prior to administration.

[0281] The study is a randomized, double-blind, placebo-controlled study of the safety, pharmacodynamics, and tolerability of multiple ascending dose levels of CNP-201 (titration phase), with the goal of identifying a safe and tolerated dose level that will be further evaluated in a larger number of subjects (titration phase). The study will enroll four cohorts to receive CNP-201 or placebo at multiple ascending dose levels.

[0282] Subjects will maintain their current SoC throughout the study, except for a 12-hour washout period (for beta-agonists, theophylline, and cromolyn only) and a 7-day washout period for antihistamines prior to the skin prick test (SPT). Subjects will resume their SoC regimen after the SPT and maintain SoC throughout the administration / duration of the study.

[0283] Subjects with peanut allergy eligible for enrollment in this study will be defined based on the following inclusion criteria: 1. Men and non-pregnant women aged 16-55. 2. Subjects with a physician-diagnosed peanut allergy or a documented history of peanut allergy. 3. Subjects weighing ≥ 31.25 kg at screening. Subjects outside this range may be included at the discretion of the investigator. 4. Subject with a documented history of non-severe anaphylaxis (grade ≤ 3) to peanut, including mild wheezing or dyspnea without hypoxia. 5. Subjects with peanut-specific IgE ≥ 5 kU / L as measured by ImmunoCAP at screening, unless they have previously undergone OIT for peanut allergy. Subjects who have previously undergone OIT for peanut allergy and do not have peanut-specific IgE ≥ 5 kU / L as measured by ImmunoCap at screening may be included at the discretion of the investigator. 6. Subjects with a positive SPT to peanut with a change in wheal diameter of ≥ 5 mm compared to the negative control (50% glycerin) at screening. Subjects who have previously undergone OIT for peanut allergy and have a positive skin prick test (SPT) to peanut with a change in wheal diameter of ≥ 5 mm at screening may be included at the discretion of the investigator. 7. Subject self-reports having been on a peanut-free diet with no suspected peanut exposure, including any peanut food challenge, for at least 14 days prior to screening and agrees to continue limiting peanut exposure during the study. 8. Female subjects and male subjects and their female spouses / partners willing to practice highly effective methods of contraception, which may include, but are not limited to, abstinence, sexual intercourse only with persons of the same sex, monogamous relationship with a vasectomized partner, vasectomy, hysterectomy, bilateral tubal ligation, approved hormonal methods, intrauterine device (IUD), or the use of spermicides in combination with barrier methods (e.g., condoms, diaphragms) beginning at screening and continuing throughout the study through Day 38 (EOS / ET). 9.Female subjects who agree to abstain from breastfeeding continuously through Day 60 starting at initial screening. 10.Female subjects who agree not to continue donating eggs through day 60 starting at the initial screening. 11. Subjects willing and able to provide Institutional Review Board (IRB) approved written informed consent. 12. Subject is willing to perform and comply with all study procedures. 13. Male subjects who agree not to donate sperm starting at screening and continuing through day 60.

[0284] Subjects who meet all inclusion criteria and no exclusion criteria after completing the screening visit will be enrolled in one of several dosing cohorts. During the titration phase, subjects will receive CNP-201 as an intravenous infusion on days 1 and 8. The dose level for the first cohort will be 1 mg. Additional dose levels for the remaining three cohorts may be 0.1 mg, 0.25 mg, 0.5 mg, 1 mg, 2 mg, 2.5 mg, 5 mg, 10 mg, or may be increased up to 650 mg.

[0285] Subjects who meet all inclusion criteria and no exclusion criteria at screening will be enrolled in the study. CNP-201 or placebo will be administered by intravenous infusion using a stepwise infusion rate. CNP-201 will be administered using the following stepwise 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, and 20 mL / hour for the remainder of the infusion.

[0286] Unless infusion reactions, anaphylaxis, or other adverse events require an extended monitoring period, subjects will remain in the clinic from the time of admission (before administration of TP) until the final procedure, which will be performed 4 hours after the dose on the same day, on Days 1 and 8. If safety parameters are acceptable to the investigator, subjects will be discharged.

[0287] Subjects will be followed daily through telephone visits after each infusion (Days 2-7 and 9-14) to assess and document any AEs and medication changes.

[0288] Seven days after the second dose of CNP-201 (Day 15), subjects must return to the clinic for collection of safety laboratory values, evaluation of AEs and medication changes, and collection of PD assessments. Subjects continue to be followed for safety and tolerability during the post-dose period.

[0289] On Day 60 (end of study), subjects will return to the clinic for collection of immune safety laboratory values, PD measurements, and will undergo a double-blind, placebo-controlled food challenge (DBPCFC) consisting of a peanut and placebo (oat) challenge. The DBPCFC will be conducted by study physicians or staff trained to manage clinical emergencies present on-site, with ready access to emergency medications and equipment, and close proximity to a hospital emergency department for rapid delivery of emergency treatment if needed.

[0290] Subjects then return to the clinic at the end of the 60 day study visit for collection of safety laboratory values, PD measurements, and final assessment of AEs and medication changes.

[0291] The total duration of the study for an individual subject is approximately 240 days, with a maximum of 180 days for screening, 8 days for CNP-201 or placebo administration, and 52 days for post-dose assessments (Figure 8).

[0292] After all subjects in a dose cohort have completed their day 15 office visit (7 days after the second dose), the DMC will convene to review all available safety data. At this point, the DMC will determine whether it is acceptable to proceed to the next dose cohort and make a recommendation regarding the dose level for that cohort.

[0293] If a safe and effective dose is identified in the titration phase, the study will proceed to the expansion phase. Subjects in the titration phase will be randomized 1:1 to receive CNP-201 at the safe and tolerated dose level identified in the expansion phase or placebo. Subjects in the titration and expansion phases will undergo the same evaluations.

[0294] Primary objectives include: safety and tolerability of CNP-201;

[0295] Exploratory objectives include: change in response to DBPCFC between patients treated with CNP-201 or placebo; change in the ratio of peanut-specific IgE to IgG between patients treated with CNP-201 or placebo; change in peanut-specific IgE between patients treated with CNP-201 or placebo; change in the proportion of peanut-specific Th2a+ T cells between patients treated with CNP-201 or placebo; change in response to a basophil activation test between patients treated with CNP-201 or placebo; change in the proportion of peanut-specific T regulatory cells between patients treated with CNP-201 or placebo.

[0296] Primary endpoints included: frequency 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 examination including vital signs (blood pressure, heart rate, and 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).

[0297] Exploratory endpoints include: difference in proportion of subjects passing DBPCFC (2000 mg dose level, not reaching induction dose at or before 4043 mg cumulative) between placebo and CNP-201 at day 60; change in cumulative tolerated dose (CTD) of peanut protein (mg) administered during DBPCFC between placebo and CNP-201 at day 60; change in peanut-specific IgE to IgG ratio as measured by ImmunoCap assay between placebo and CNP-201 at baseline and day 60; change in peanut-specific IgE as measured by ImmunoCap assay between placebo and CNP-201 at baseline and day 60; Change in the percentage 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; change in the effective concentration at 50% of maximum basophil activation (EC50) as measured by basophil activation test (CD203c+ / CD63+ / - basophil activation) between placebo and CNP-201 at baseline and day 15 and day 60; change in the percentage 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.

[0298] Preferred embodiments of the present disclosure are described herein. Variations of those preferred embodiments may become apparent to those skilled in the art upon reading the foregoing description. The inventors expect those skilled in the art to adopt such variations as appropriate, and the inventors intend for the present disclosure to be practiced otherwise than as specifically described herein. Accordingly, this disclosure includes all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Also, any combination of the above-described elements in all possible variations thereof is encompassed by the present disclosure unless otherwise indicated herein or otherwise clearly contradicted by context.

Claims

1. 1. A method of treating peanut allergy in a subject, comprising administering to the subject tolerizing immune modifying particles encapsulating peanut proteins (TIMP-PPE), alone or in combination with one or more therapeutic agents, wherein the TIMP-PPE is administered at a dosage level of about 0.001 mg / kg to 12 mg / kg.

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

3. The method of claim 1 or 2, wherein the TIMP-PPE particles have a negative zeta potential.

4. 10. A method according to any one of the preceding claims, wherein the particles have a negative zeta potential of between -30mV and -100mV.

5. 10. The method of any one of the preceding claims, wherein the TIMP-PPE is administered at a concentration of about 0.0005 mg / mL to 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.

6. 10. The method of any one of the preceding claims, wherein 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. 10. The method of any one of the preceding claims, wherein the TIMP-PPE is administered at a dosage level of about 0.1 mg to 800 mg.

8. 10. The method of 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. 10. The method of any one of the preceding claims, wherein the TIMP-PPE is administered in a single dose or in multiple doses.

10. 10. The method of any one of the preceding claims, wherein TIMP-PPE is administered once per week, once every two weeks, once every three weeks, once every four weeks, once every two months, once every three months, once every six months, or once per year.

11. 10. The method of any one of the preceding claims, wherein the TIMP-PPE is administered in two doses, one week apart.

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

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

14. 10. The method of any one of the preceding claims, wherein TIMP-PPE is administered in two doses spaced one week apart, followed by a booster dose of TIMP-PPE re-administered as a single dose once every three months.

15. The therapeutic agent administered in combination with TIMP-PPE may be 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 mutein 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, or an IL-2 / CD25 fusion protein.

15. The method of any one of claims 1 to 14, wherein the active ingredient is a synthetic protein, a prebiotic, a probiotic, a histone deacetylase inhibitor, a short chain fatty acid (e.g., acetate, butyrate, propionate, butyrate polymers), an inhibitor of IgE, a competitor of IgE for allergen binding sites, an inhibitor of basophil activation, an inhibitor of mast cell activation, a cytokine inhibitor, a microbiome therapy, a small molecule or biological therapeutic agent, or a nonsteroidal anti-inflammatory drug (NSAID).

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

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

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

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

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

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

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

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

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

26. 20. The method of claim 19, wherein the one or more 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 disorders such as diarrhea, stomach cramps, nausea, or vomiting, drop in blood pressure, and anaphylaxis.

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

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

30. 30. The method of claim 29, wherein 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), stool, tissue biopsy, and / or bone marrow biopsy.

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

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

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

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

35. 35. The method of claim 34, wherein basophil activation is assayed from ex vivo stimulation of basophils with peanut protein in the Basophil Activation Test (BAT).

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

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

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

39. 39. The method of claim 38, wherein the Th2 cytokine is selected from the group consisting of IL-4, IL-5, IL-9, and IL-13.

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

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

42. 41. The method of claim 40, wherein tolerance to peanut protein is determined by skin prick testing (SPT).

43. 10. The method of 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. 10. The method of 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. 10. The method of any one of the preceding claims, wherein the subject is administered an antihistamine, a corticosteroid, an NSAID, and / or a leukotriene modifier in combination with TIMP-PPE.

46. 46. ​​The method of claim 45, wherein the antihistamine is diphenhydramine.

47. 47. The method of claim 45 or 46, wherein 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.

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

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

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

51. 46. ​​The method of claim 45, wherein the corticosteroid is methylprednisone.

52. 52. The method of any one of claims 45-51, wherein 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.

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

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

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

56. 46. ​​The method of claim 45, wherein the NSAID is acetylsalicylic acid.

57. 57. The method of any one of claims 45 and 56, wherein the NSAID 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.

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

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

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

61. 46. ​​The method of claim 45, wherein the leukotriene modifier is montelukast.

62. 62. The method of claim 45 or 61, wherein the leukotriene modifying agent 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. 63. The method of any one of claims 45 or 61-62, wherein the leukotriene modifying agent is administered orally.

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

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

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