Methods and compositions for preventing or treating food allergies

Abstract

Described herein are methods for preventing or treating food allergy or food intolerance by administering a peptide related to chaperonin 60.1.

Description

Technical Field

[0001] Field of the Invention Embodiments of the present invention relate to compositions and methods for the prevention or treatment of food allergy and food intolerance using peptides related to chaperonin 60.1 (Cpn60.1).

Background Art

[0002] Background of the Invention Food allergy is a damaging and potentially life-threatening condition. It is a medical condition in which exposure to food induces a harmful immune response. Food allergy is increasingly recognized as an increasing public health burden and is called the "second wave" of the allergy epidemic, following asthma. 1 Current evidence suggests that food allergy is common and affects up to 10% of infants in some countries 2 and that the prevalence has increased over the past few decades. These increases in prevalence preferentially affect industrial areas, but evidence of increases in prevalence in rapidly developing countries commensurate with increased economic growth is also now increasing. 3 In the United States, it is estimated that 32 million Americans, including 5.6 million children under 18 years old, have food allergies. 4,5 Along with the United States, Germany, Italy, and Norway have been reported to have the highest prevalence of food sensitivity, and about 22% of people in each country show antibodies to some type of food. 6 .

[0003] Given the dramatic increase in the prevalence of food allergies worldwide, effective prevention strategies are a public health priority. Several models have emerged around the etiology of food allergies, including the hygiene hypothesis, the dual allergen exposure hypothesis, and the vitamin D hypothesis. Other nutritional interventions, including the use of whey-based partially hydrolyzed infant formula in non-breastfed infants, also play an important role. In recent years, there has been a shift from long-term food allergen avoidance to early allergen introduction starting at four months of age. However, implementation of this strategy at the population level still poses significant logistic problems, including patient selection and development of suitable food forms for young infants. Treatment of food allergies has also seen major changes. On the one hand, emphasis has been placed on desensitization and tolerance induction by oral immunotherapy and epicutaneous immunotherapy. In addition, special hypoallergenic infant formulas for the treatment of infants with cow's milk allergy are undergoing reformulation to include the addition of lactose and probiotics to modulate the gut microbiota and early immune response. For several reasons, these strategies are insufficient, and strict allergen avoidance remains the main principle of prevention and treatment.

[0004] Therefore, there is a need for effective compositions and methods for the prevention or treatment of food allergies and food intolerances. SUMMARY OF THE INVENTION

[0005] Brief Summary of the Invention Embodiments of the present invention provide a method for treating or preventing the onset of food allergy or food intolerance to food-derived dietary antigens in a subject in need thereof by administering a peptide related to chaperonin 60.1 (Cpn60.1). In some embodiments, the Cpn60.1-related peptide is selected from DGSVVVNKVSELPAGHGLNVNTLSYGDLAAD (SEQ ID NO: 1) (PIN201104); DGSVVVNKVSELPAGH (SEQ ID NO: 2); GLNVNTLSYGDLAAD (SEQ ID NO: 3); SELPAGHGLNVNLTS (SEQ ID NO: 4); DGSVVVNKVS (SEQ ID NO: 5); ELPAGHGLNV (SEQ ID NO: 6); NTLSYGDLAAD (SEQ ID NO: 7); or a functionally equivalent fragment or variant thereof. In one embodiment, the Cpn60.1-related peptide is SEQ ID NO: 1.

[0006] In some embodiments, the method further comprises diagnosing a subject having or at risk of developing food allergy or food intolerance, or receiving the results of an assay diagnosing a subject having or at risk of developing food allergy or food intolerance, prior to administration of the Cpn60.1-related peptide.

[0007] In some embodiments, the Cpn60.1-related peptide is administered prior to the first exposure to a potential causative food allergen. In alternative embodiments, the Cpn60.1-related peptide is administered at the clinical signs of atopic symptoms.

[0008] In some embodiments, the Cpn60.1-related peptide is administered to a subject diagnosed with at least one food allergy or food intolerance to a dietary antigen. In some embodiments, the dietary antigen is derived from milk and its products; eggs and their products; meat and its products; fish, mollusks, and crustaceans, and their products; oils, fats, and their products; grains and their products; legumes, seeds, grains, nuts, and their products; vegetables and their products; fruits and their products; mushrooms and their products; sugars, sugar products, chocolate products, and confectioneries; and spices and herbs.

[0009] Other embodiments are also described and recited herein.

Brief Description of the Drawings

[0010] For illustrative purposes, certain embodiments of the invention are shown in the drawings described below. However, it should be understood that the invention is not limited to the exact arrangements, dimensions, and devices shown.

[0011]

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Mode for Carrying Out the Invention

[0012] Detailed Description of the Invention Here, the innovation of the subject matter will be described in relation to the drawings, and like reference numbers are used throughout to refer to like elements. In the following description, for purposes of explanation, numerous specific details are set forth in order to provide a thorough understanding of the present invention. It will be apparent, however, that the present invention may be practiced without these specific details. In other instances, well-known structures and devices are shown in block diagram form in order to facilitate the description of the present invention. Of course, specific aspects, modes, embodiments, variations and features of the present invention will be described below at various levels of detail in order to provide a substantial understanding of the present invention.

[0013] Definitions For the sake of simplicity, the meanings of some terms and phrases used in this specification, the examples, and the appended claims are provided below. Unless otherwise specified or implicitly suggested by the context, the following terms and phrases include the meanings provided below. Since the scope of the present invention is limited only by the claims, the definitions are provided to assist in the description of specific embodiments and are not intended to limit the invention of the claims. Unless otherwise defined, all technical and scientific terms used in this specification have the same meaning as commonly understood by one of ordinary skill in the technical field to which the present invention pertains. In the case of an obvious inconsistency between the use of a term in the art and its definition provided herein, the definition provided herein shall prevail.

[0014] As used in this specification and the appended claims, the singular forms "a", "an", and "the" include plural references unless the content clearly dictates otherwise. For example, a reference to "a cell" includes combinations of two or more cells and the like.

[0015] As used in this specification, the terms "approximately" or "about" with respect to a value or parameter generally encompass (and describe) embodiments that target that value or parameter, including numbers within a range of 5%, 10%, 15%, or 20% in either direction (greater than or less than), unless otherwise specified or not apparent from the context (except when such numbers are less than 0% of the possible values or exceed 100%). For example, a description referring to "about X" includes a description of "X".

[0016] As used herein, the term "or" means "and / or". The term "and / or" as used in phrases such as "A and / or B" herein is intended to include both A and B, A or B, A alone, and B alone. Similarly, the term "and / or" as used in phrases such as "A, B, and / or C" is intended to include each of the following embodiments: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A alone; B alone; and C alone.

[0017] As used herein, the term "comprising" means that other elements may be present in addition to the recited defined elements. The use of "comprising" indicates inclusion rather than limitation.

[0018] The term "consisting of" refers to the compositions, methods, and their respective components described herein and does not include any element not recited in the description of the embodiment.

[0019] As used herein, the term "consisting essentially of" refers to those elements necessary for a given embodiment. This term allows for the presence of additional elements that do not substantially affect the basic and novel or functional characteristics of that embodiment of the invention.

[0020] The terms "statistically significant" or "significant" refer to statistical significance and generally mean a difference of two times the standard deviation (2SD) or more.

[0021] As used herein, the term "subject" refers to mammals including, but not limited to, dogs, cats, horses, cows, pigs, sheep, goats, chickens, rodents, or primates. The subject can be a pet (e.g., dog, cat), an agricultural livestock animal (e.g., cow, horse, pig, chicken, etc.), a laboratory animal (e.g., mouse, rat, rabbit, etc.), but is not so limited. The subject includes human subjects. The human subject can be a pediatric, adult, or geriatric subject. The human subject can be of either sex.

[0022] As used herein, the terms "effective amount" and "therapeutically effective amount" include an amount sufficient to prevent or ameliorate the symptoms or medical conditions of diseases such as immune disorders including, for example, eosinophilic esophagitis, hemolytic anemia, thrombocytopenia, thyroiditis, pernicious anemia, Addison's disease, autoimmune diabetes, myasthenia gravis, rheumatoid arthritis, systemic lupus erythematosus, atherosclerosis, and autoimmune encephalitis, allergic conditions such as eczema, dermatitis, allergic rhinitis, allergic conjunctivitis, allergic airway diseases, eosinophilic syndrome, contact dermatitis, food allergy, etc., and respiratory diseases characterized by eosinophilic airway inflammation and airway hypersensitivity such as allergic asthma, intrinsic asthma, allergic bronchopulmonary aspergillosis, eosinophilic pneumonia, allergic bronchitis bronchiectasis, occupational asthma, reactive airway dysfunction syndrome, interstitial lung disease, eosinophilic syndrome, or pulmonary parasitosis. Of course, there are many methods known in the art for determining the effective amount for a given use. For example, pharmacological methods for dose determination can be used in a therapeutic context. In the context of therapeutic or prophylactic use, the amount of the composition administered to a subject depends on the type and severity of the disease, as well as individual characteristics such as general health, age, sex, weight, and drug tolerance. It also depends on the degree, severity, and type of the disease. One of ordinary skill in the art would be able to determine an appropriate dosage depending on these and other factors. The composition can also be administered in combination with one or more additional therapeutic compounds.

[0023] As used herein, the terms "treating" or "treatment", or "to treat", or "alleviating" or "to alleviate" refer to (1) therapeutic means for curing, slowing, reducing, and / or halting the progression of a diagnosed disease or infection, as well as (2) prophylactic or disease-preventing means for preventing or delaying the onset of a disease or infection.

[0024] As used herein, when used in connection with a disease, disorder, or medical condition, the terms "treat", "treatment", "treating", or "amelioration" refer to a therapeutic intervention for the condition, the goal of which is to reverse, alleviate, improve, inhibit, slow down, or stop the progression or severity of the symptoms or condition. The term "treatment" includes reducing or alleviating at least one adverse effect or symptom state. A treatment is generally "effective" if one or more symptoms or clinical markers are reduced. Alternatively, a treatment is "effective" if the progression of the condition is reduced or stopped. That is, "treatment" includes not only an improvement in symptoms or markers, but also halting, or at least slowing, the progression or worsening of symptoms that would be expected in the absence of treatment. Beneficial or desired clinical outcomes include, but are not limited to, alleviation of one or more symptoms, diminution of the extent of a deficit, stabilization (i.e., non-worsening) of an immunological disorder, delay or slowing of an immunological disorder, and extension of lifespan compared to what would be expected in the absence of treatment.

[0025] As used herein, the terms "short-term administration" or "acute administration" mean that a therapeutic agent or drug is administered as a single dose or daily dose over a period of 2, 3, 4, 5, 6, 7 or more days. "Short-term administration" can be administered prophylactically before potential exposure to one or more causative food allergens or after possible exposure to one or more causative food allergens, or therapeutically after the onset of symptoms following exposure to one or more causative food allergens.

[0026] As used herein, the term "long-term administration" means that a therapeutic agent or drug is administered over a period of at least 12 weeks. This includes that the therapeutic agent or drug is administered so as to be effective over a period of at least 12 weeks or more or over a period of at least 12 weeks. For example, when a sustained-release composition or a long-acting therapeutic agent or drug is used, it is included that the administration itself is not necessarily carried out for 12 weeks. Thus, the subject is treated over a period of at least 12 weeks. In many cases, long-term administration is at least 4, 5, 6, 7, 8, 9 months or more, or at least 1, 2, 3, 5, 7, or 10 years or more.

[0027] Administration of the compositions contemplated herein can be carried out in any convenient manner, including aerosol inhalation, injection, ingestion, transfusion, implantation, or transplantation. In a preferred embodiment, the composition is administered parenterally. As used herein, the phrases "parenteral administration" and "administered parenterally" refer to a mode of administration other than enteral administration and topical administration, usually by injection, and include, but are not limited to, intravascular, intravenous, intramuscular, intraarterial, intrathecal, intraarticular, intraorbital, intratumoral, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subepidermal, intra-articular, subcapsular, subdural, intraspinal, and intrasternal injections and infusions. In one embodiment, the compositions contemplated herein are administered to a subject by direct injection into a tumor, lymph node, or site of infection.

[0028] The terms "decrease," "reduced," "reduction," or "inhibit" are all used herein to mean a statistically significant amount of decrease. In some embodiments, "reduce," "reduction," "decrease," or "inhibit" typically means at least a 10% decrease compared to a baseline level (e.g., in the absence of any given treatment or drug administration), and can include, for example, at least about 10%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or more decrease. As used herein, "reduction" or "inhibition" does not include complete inhibition or reduction compared to a baseline level. "Complete inhibition" is 100% inhibition compared to a baseline level. The decrease can preferably be reduced to a level that is acceptable within the normal range for an individual without a given disorder.

[0029] The terms "increased", "increase", "enhance", or "activate" are all used herein to mean a statistically significant amount of increase. In some embodiments, the terms "increased", "increase", "enhance", or "activate" mean an increase of at least 10%, for example, at least about 20%, or at least about 30%, or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, at least about 90% increase, or up to 100% increase, or any increase from 10% to 100% compared to the baseline level, or at least about 2-fold, or at least about 3-fold, or at least about 4-fold, or at least about 5-fold or at least about 10-fold increase, or any increase from 2-fold to 10-fold or more compared to the baseline level. In the context of a marker or symptom, "increase" is a statistically significant increase in such level.

[0030] As used herein, the terms "protein" and "polypeptide" are used interchangeably herein to designate a series of amino acid residues joined to each other by peptide bonds between the alpha-amino and carboxy groups of adjacent residues. The terms "protein" and "polypeptide" refer to polymers of amino acids, including modified amino acids (e.g., phosphorylation, glycosylation, glycosylation, etc.) and amino acid analogs, regardless of their size or function. The terms "protein" and "polypeptide" are often used with respect to relatively large polypeptides, while the term "peptide" is often used with respect to small polypeptides, but the use of these terms in the art overlaps. The terms "protein" and "polypeptide" are used interchangeably herein when referring to gene products and fragments thereof. Thus, exemplary polypeptides or proteins include gene products, natural proteins, homologs, orthologs, paralogs, fragments, and other equivalents, variants, fragments, and analogs as described above.

[0031] In various embodiments described herein, it is further contemplated that variants (natural or otherwise), alleles, homologs, conservatively modified variants, and / or conservatively substituted variants of any of the specific polypeptides described are included. With respect to amino acid sequences, one of ordinary skill in the art will recognize that individual substitutions, deletions, or additions to a nucleic acid, peptide, polypeptide, or protein sequence that changes a single amino acid or a small percentage of amino acids in the encoded sequence are "conservatively modified variants" and that the modifications result in amino acid substitutions with chemically similar amino acids that retain the desired activity of the polypeptide. Such conservatively modified variants are in addition to, and do not exclude, polymorphic variants, interspecies homologs, and alleles that are consistent with the present disclosure.

[0032] In some embodiments, the polypeptides described herein (or nucleic acids encoding such polypeptides) may be a functional fragment of one of the amino acid sequences described herein. As used herein, "functional fragment" is a fragment or segment of a peptide that retains at least 50% of the activity of the wild-type reference polypeptide according to the assays described below. Functional fragments may include conservative substitutions of the sequences disclosed herein.

[0033] In some embodiments, the polypeptides described herein may be variants of the sequences described herein. In some embodiments, the variant is a conservative modification variant. Conservative substitution variants can be obtained, for example, by mutation of a native nucleotide sequence. As referred to herein, a "variant" is a polypeptide that is substantially homologous to a native or reference polypeptide but has an amino acid sequence that differs from the amino acid sequence of the native or reference polypeptide due to one or more deletions, insertions, or substitutions. A DNA sequence encoding a variant polypeptide includes sequences that contain one or more additions, deletions, or substitutions of nucleotides as compared to the native DNA sequence or reference DNA sequence, but encode a variant protein or a fragment thereof that retains activity. A variety of PCR-based site-directed mutagenesis approaches are known in the art and can be applied by those skilled in the art.

[0034] As used herein, the term "nucleic acid" or "nucleic acid sequence" refers to any molecule, preferably a macromolecule, incorporating units of ribonucleic acid, deoxyribonucleic acid, or analogs thereof. The nucleic acid may be either single-stranded or double-stranded. A single-stranded nucleic acid may be one nucleic acid strand of denatured double-stranded DNA. Alternatively, it may be a single-stranded nucleic acid not derived from any double-stranded DNA. In one aspect, the nucleic acid may be DNA. In another aspect, the nucleic acid may be RNA. Suitable DNA includes, for example, genomic DNA or cDNA. Suitable RNA includes, for example, mRNA.

[0035] In some embodiments of any of the aspects, the polypeptides, nucleic acids, or cells described herein can be engineered. As used herein, "engineered" refers to a form that has been manipulated by human hand. For example, a polypeptide is considered to be "engineered" when at least one aspect of the polypeptide, such as its sequence, has been manipulated by human hand such that it differs from forms that exist in nature. As is common practice and as will be understood by those of skill in the art, progeny of engineered cells are typically still referred to as "engineered" even though the actual manipulation was performed on a previous entity.

[0036] In some embodiments, a nucleic acid encoding a polypeptide described herein (e.g., an antibody or antibody reagent) consists of a vector. In some aspects of the embodiments described herein, a nucleic acid sequence encoding a given polypeptide described herein, or any module thereof, is operably linked to a vector. Vectors can include, but are not limited to, cloning vectors, expression vectors, plasmids, phages, transposons, cosmids, chromosomes, viruses, virions, and the like.

[0037] As used herein, the term "expression vector" refers to a vector that directs the expression of RNA or polypeptide from a sequence linked to a transcriptional regulatory sequence on the vector. The sequence to be expressed is often, but not necessarily, heterologous to the cell. The expression vector may contain additional elements; for example, the expression vector may have two replication systems and thus be maintained in two organisms, for example, human cells for expression and a prokaryotic host for cloning and amplification. The term "expression" means the cellular processes involved in the production of RNA and proteins, and, where appropriate, secreted proteins, including, but not limited to, transcription, transcriptional processing, translation and protein folding, modification and processing, where applicable. "Expression product" includes RNA transcribed from a gene and polypeptide obtained by translation of mRNA transcribed from a gene. The term "gene" means a nucleic acid sequence that is transcribed (DNA) into RNA in vitro or in vivo when operably linked to appropriate regulatory sequences. A gene may or may not include regions before and after the coding region, for example, 5' untranslated (5'UTR) sequences or "leader" sequences and 3'UTR sequences or "trailer" sequences, as well as intervening sequences (introns) between individual coding segments (exons).

[0038] As used herein, the terms “isolated” or “partially purified,” in the case of a nucleic acid or polypeptide, refer to a nucleic acid or polypeptide that is separated from at least one other component (e.g., a nucleic acid or polypeptide) that is present in the nucleic acid or polypeptide as it exists in its natural source and / or that is present in or secreted by a cell when expressed by the cell. Chemically synthesized nucleic acids or polypeptides, or nucleic acids or polypeptides synthesized using in vitro transcription / translation, are considered “isolated.” The terms “purified” or “substantially purified” refer to an isolated nucleic acid or polypeptide that is at least 95% by weight of the subject nucleic acid or polypeptide, including, for example, at least 96% by weight, at least 97% by weight, at least 98% by weight, at least 99% by weight, or more. In some embodiments, the antibodies, antigen-binding portions thereof, or chimeric antigen receptors (CARs) described herein are isolated. In some embodiments, the antibodies, antibody reagents, antigen-binding portions thereof, or CARs described herein are purified.

[0039] As used herein, “engineered” refers to a manner that has been manipulated by human hand. For example, an antibody, antibody reagent, antigen-binding portion thereof, CAR, or bispecific antibody is considered “engineered” when the sequence of the antibody, antibody reagent, antigen-binding portion thereof, CAR, or bispecific antibody has been manipulated by human hand such that, if it were to exist in nature, it would differ from the sequence of the antibody. As is common practice and as will be understood by those of skill in the art, progeny and copies of engineered polynucleotides and / or polypeptides typically are still referred to as “engineered,” even though the actual manipulation was performed on a previous entity.

[0040] Pharmaceutical composition The compositions and methods of the present invention can be utilized to treat an individual in need thereof. In certain embodiments, the individual is a mammal, such as a human, or a non-human mammal. When administered to an animal such as a human, the composition or compound is preferably administered as a pharmaceutical composition comprising, for example, a compound of the present invention and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are well known in the art and include, for example, aqueous solutions such as water or physiologically buffered saline, or oils such as glycols, glycerol, olive oil, or other solvents or vehicles such as injectable organic esters. In a preferred embodiment, when such a pharmaceutical composition is for human administration, particularly for invasive routes of administration (i.e., routes such as injection or implantation that avoid transport or diffusion through an epithelial barrier), the aqueous solution is pyrogen-free or substantially pyrogen-free. Excipients can be selected, for example, to provide for a delay in drug release or to selectively target one or more cells, tissues, or organs. The pharmaceutical composition can be in unit dosage forms such as tablets, capsules (including sprinkle capsules and gelatin capsules), granules, lyophilized agents for reconstitution, powders, solutions, syrups, suppositories, injectables, and the like. The composition can also be present in a transdermal delivery system, such as in a skin patch. The composition can also be present in a solution suitable for topical administration, such as a lotion, cream, or ointment.

[0041] A pharmaceutically acceptable carrier may contain, for example, a physiologically acceptable agent that acts to stabilize, increase the solubility, or increase the absorption of a compound such as a compound of the present invention. Such physiologically acceptable agents include, for example, carbohydrates such as glucose, sucrose, or dextran, antioxidants such as ascorbic acid or glutathione, chelating agents, low molecular weight proteins, or other stabilizers or excipients. The selection of a pharmaceutically acceptable carrier containing a physiologically acceptable agent depends, for example, on the route of administration of the composition. The preparation or pharmaceutical composition may be a self-emulsifying drug delivery system or a self-microemulsifying drug delivery system. The pharmaceutical composition (preparation) may also be a liposome or other polymeric matrix, which may, for example, incorporate therein a compound such as a compound of the present invention. For example, liposomes containing phospholipids or other lipids are carriers that are relatively simple to manufacture and administer, non-toxic, physiologically acceptable, and metabolizable.

[0042] As used herein, the term "pharmaceutically acceptable" refers to compounds, materials, compositions, and / or dosage forms that are suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response, and other problems and complications within the scope of sound medical judgment and commensurate with a reasonable benefit / risk ratio.

[0043] As used herein, the term "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition, or vehicle such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials that can function as pharmaceutically acceptable carriers include the following: (1) sugars (such as lactose, glucose, and sucrose), (2) starches (such as corn starch and potato starch), (3) cellulose and its derivatives (such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate), (4) powdered tragacanth, (5) malt, (6) gelatin, (7) talc, (8) excipients (such as cocoa butter and suppository waxes), (9) oils (such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil), (10) glycols (such as propylene glycol), (11) polyols (such as glycerin, sorbitol, mannitol, and polyethylene glycol), (12) esters (such as ethyl oleate and ethyl laurate), (13) agar, (14) buffering agents (such as magnesium hydroxide and aluminum hydroxide), (15) alginic acid, (16) pyrogen-free water, (17) isotonic saline, (18) Ringer's solution, (19) ethyl alcohol, (20) phosphate buffer solutions, and (21) other non-toxic compatible substances used in pharmaceutical formulations.

[0044] The pharmaceutical compositions (preparations) can be administered to a subject by any of a number of routes of administration, including, for example, oral administration (e.g., drenches, tablets, capsules (including sprinkle capsules and gelatin capsules), boluses, powders, granules, pastes for application to the tongue, such as in aqueous or non-aqueous solutions or suspensions); absorption through the oral mucosa (e.g., sublingual); subcutaneous administration; transdermal administration (e.g., as a patch applied to the skin); and topical administration (e.g., as a cream, ointment or spray applied to the skin). The compounds may also be formulated for inhalation. In certain embodiments, the compounds may simply be dissolved or suspended in sterile water. Details of suitable routes of administration and compositions suitable therefor can be found, for example, in U.S. Pat. Nos. 6,110,973, 5,763,493, 5,731,000, 5,541,231, 5,427,798, 5,358,970, and 4,172,896, and patents cited therein.

[0045] The formulations may be conveniently presented in unit dosage form and may be prepared by any method known in the art of pharmacy. The amount of active ingredient that can be combined with carrier materials to produce a single dosage form varies depending on the host treated, the particular mode of administration. The amount of active ingredient that can be combined with carrier materials to produce a single dosage form is generally the amount of compound that produces a therapeutic effect. Generally, out of one hundred percent, this amount will range from about 1 percent to about 99 percent of active ingredient, preferably from about 5 percent to about 70 percent, most preferably from about 10 percent to about 30 percent.

[0046] Methods of preparing these formulations or compositions include the step of bringing into association an active compound, such as a compound of the present invention, with the carriers and, optionally, one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association a compound of the present invention with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product.

[0047] The formulations of the present invention suitable for oral administration may be in the form of capsules (including sprinkle capsules and gelatin capsules), cachets, pills, tablets, troches (using flavored bases, usually sucrose and acacia or tragacanth), lyophilized agents, powders, granules, or solutions or suspensions in aqueous or non-aqueous liquids, or as water-in-oil or oil-in-water liquid emulsions, or as elixirs or syrups, or as medicinal drops (using inert bases such as gelatin and glycerin, or sucrose and acacia), and / or as mouthwashes, etc., each containing a predetermined amount of the compound of the present invention as an active ingredient. The composition or compound may also be administered as a bolus, lozenge, or paste.

[0048] To prepare solid dosage forms for oral administration (including capsules (including sprinkle capsules and gelatin capsules), tablets, pills, dragees, powders, granules, etc.), the active ingredient is mixed with one or more pharmaceutically acceptable carriers, such as sodium citrate or dicalcium phosphate, and / or any of the following: (1) fillers or extenders (such as starch, lactose, sucrose, glucose, mannitol, and / or silicic acid, etc.), (2) binders (such as carboxymethyl cellulose, alginic acid, gelatin, polyvinylpyrrolidone, sucrose and / or acacia, etc.), (3) humectants (such as glycerol, etc.), (4) disintegrants (agar, calcium carbonate, and potato or tapioca starch, alginic acid, certain silicates, sodium carbonate, etc.), (5) dissolution retardants (such as paraffin, etc.), (6) absorption promoters (such as quaternary ammonium compounds, etc.), (7) wetting agents (such as cetyl alcohol and glycerol monostearate, etc.), (8) absorbents (such as kaolin and bentonite clay, etc.), (9) lubricants (such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof, etc.), (10) complexing agents (such as modified cyclodextrin and unmodified cyclodextrin, etc.), and (11) coloring agents. In the case of capsules (including sprinkle capsules and gelatin capsules), tablets, and pills, the pharmaceutical composition may also contain a buffering agent. Similar types of solid compositions may also be used as fillers in soft and hard-filled gelatin capsules using excipients such as lactose or milk sugar, and high molecular weight polyethylene glycol, etc.

[0049] Tablets may be made by compression or molding, optionally with the use of one or more accessory ingredients. Compressed tablets can be prepared using binders (such as gelatin or hydroxypropylmethylcellulose), lubricants, inert diluents, preservatives, disintegrants (such as sodium starch glycolate or cross-linked sodium carboxymethylcellulose), surfactants or dispersing agents. Molded tablets may be made by molding a mixture of powdered compounds moistened with an inert liquid diluent in a suitable machine.

[0050] Tablets and other solid dosage forms of pharmaceutical compositions, such as, for example, dragees, capsules (including sprinkle capsules and gelatin capsules), pills, and granules, may optionally be scored or prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulation art. They may also be formulated to provide sustained or controlled release of the active ingredient therein, using, for example, various proportions of hydroxypropylmethylcellulose, other polymeric matrices, liposomes, and / or microspheres to provide the desired release profile. They may be sterilized, for example, by filtration through a bacteria retaining filter or by incorporating a sterilizing agent in the form of a sterile solid composition that can be dissolved in sterile water or some other sterile injectable medium immediately before use. These compositions may also optionally contain opacifying agents and may be compositions that release only the active ingredient or preferentially, optionally in a delayed manner, release the active ingredient in a particular part of the gastrointestinal tract. Examples of implant compositions that may be used include polymeric substances and waxes. The active ingredient may also, where appropriate, be in the form of microcapsules with one or more of the excipients described above.

[0051] Liquid dosage forms useful for oral administration include pharmaceutically acceptable emulsions, lyophilized powders for reconstitution, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active ingredient, liquid dosage forms may contain inert diluents commonly used in the art (e.g., water or other solvents), cyclodextrins and their derivatives, solubilizing and emulsifying agents (e.g., ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3 - butylene glycol, oils (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol, and sorbitan fatty acid esters, and mixtures thereof).

[0052] In addition to the inert diluent, the oral composition may also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening agents, flavoring agents, coloring agents, perfuming agents and preservatives.

[0053] In addition to the active compound, the suspension may contain suspending agents such as, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar and tragacanth, and mixtures thereof.

[0054] Dosage forms for topical or transdermal administration include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches, and inhalants. The active compound can be mixed under sterile conditions with a pharmaceutically acceptable carrier and any preservatives, buffers, or propellants as may be required.

[0055] Ointments, pastes, creams, and gels may contain excipients such as animal and vegetable fats, oils, waxes, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silicic acid, talc, and zinc oxide, or mixtures thereof, in addition to the active compound.

[0056] Powders and sprays may contain excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicate, and polyamide powder, or mixtures of these substances, in addition to the active compound. Sprays can additionally contain conventional propellants such as chlorofluorohydrocarbons, as well as volatile unsubstituted hydrocarbons such as butane and propane.

[0057] The transdermal patch has the additional advantage of providing controlled delivery of the compounds of the present invention into the body. Such dosage forms can be prepared by dissolving or dispersing the active compound in a suitable medium. Penetration enhancers can also be used to increase the flux of the compound across the skin. The rate of such flux can be controlled either by providing a rate-controlling membrane or by dispersing the compound in a polymeric matrix or gel.

[0058] As used herein, the terms "parenteral administration" and "administered parenterally" refer to a mode of administration other than enteral and topical administration, usually by injection, including, but not limited to, intravenous, intraocular (such as intravitreal), intramuscular, intraarterial, intrathecal, intraarticular, intracapsular, subarachnoid, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subepidermal, intraarticular, subcapsular, subdural, intraspinal and intrasternal injections and infusions. Pharmaceutical compositions suitable for parenteral administration consist of one or more active compounds in combination with one or more pharmaceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, or sterile powders that can be reconstituted into sterile injectable solutions or dispersions immediately before use, and these compositions can contain antioxidants, buffers, bacteriostats, solutes that render the formulation isotonic with the blood of the intended recipient, or suspending or thickening agents.

[0059] Examples of suitable aqueous and non-aqueous carriers that can be used in the pharmaceutical compositions of the present invention include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, etc.), and suitable mixtures thereof, vegetable oils such as olive oil, and organic esters for injection such as ethyl oleate. Examples of suitable aqueous and non-aqueous carriers that can be used in the pharmaceutical compositions of the present invention include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, etc.), and suitable mixtures thereof, vegetable oils such as olive oil, and organic esters for injection such as ethyl oleate. Appropriate fluidity can be maintained, for example, by the use of coating materials such as lecithin, by maintaining the required particle size in the case of dispersions, and by the use of surfactants. Appropriate fluidity can be maintained, for example, by the use of coating materials such as lecithin, by maintaining the required particle size in the case of dispersions, and by the use of surfactants.

[0060] These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents, and dispersing agents. Prevention of the action of microorganisms can be ensured by including various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenolsorbic acid, etc. Also, it may be desirable to include in the composition isotonic agents such as sugars, sodium chloride, etc. Furthermore, the prolonged absorption of injectable pharmaceutical forms can be brought about by the inclusion of agents that delay absorption such as aluminum monostearate and gelatin.

[0061] In some cases, it is desirable to delay the absorption of a drug from a subcutaneous or intramuscular injection in order to extend the effect of the drug. This can be achieved by the use of a liquid suspension of a crystalline or amorphous material with low water solubility. The absorption rate of the drug then depends on its dissolution rate, which in turn can depend on the crystal size and crystal form. Alternatively, the delay in absorption of the pharmaceutical form in parenteral administration is achieved by dissolving or suspending the drug in an oily vehicle.

[0062] Injectable depot forms are prepared by forming a microencapsulation matrix of the subject compound in a biodegradable polymer such as polylactide - polyglycolide. The drug release rate can be controlled according to the drug - to - polymer ratio and the nature of the specific polymer used. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations can also be prepared by entrapping the drug in liposomes or microemulsions that are compatible with body tissues.

[0063] For use in the methods of the present invention, the active compound can be given by itself or as a pharmaceutical composition containing from 0.1 to 99.5% (more preferably 0.5 to 90%) active ingredient, for example in combination with a pharmaceutically acceptable carrier.

[0064] The introduction method may also be provided by a rechargeable or biodegradable device. For the controlled delivery of drugs, including protein - based biopharmaceuticals, various sustained - release polymer devices have been developed and tested in vivo in recent years. A variety of biocompatible polymers (including hydrogels), both biodegradable and non - biodegradable, can be used to form implantable tablets for the sustained release of a compound at a specific target site.

[0065] The actual dosage level of the active ingredient in the pharmaceutical composition can be varied so as to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration without being toxic to the patient.

[0066] The selected dosage level will depend on a variety of factors including the activity of the specific compound or combination of compounds or its esters, salts, or amides used, the route of administration, the time of administration, the rate of excretion of the specific compound used, the duration of the treatment, other drugs, compounds, and / or substances used in combination with the specific compound, the age, sex, weight, condition, general health, and previous medical history of the patient being treated, as well as similar factors well known in the medical arts.

[0067] A physician or veterinarian having ordinary skill in the art can readily determine and prescribe the necessary therapeutically effective amount of the pharmaceutical composition. For example, a physician or veterinarian can initiate the dosage of the pharmaceutical composition or compound at a level lower than that required to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. "Therapeutically effective amount" means a concentration of the compound sufficient to elicit the desired therapeutic effect. It is generally understood that the effective amount of a compound will vary depending on the weight, sex, age, and medical history of the subject. Other factors that can affect the effective amount include the severity of the patient's condition, the disorder being treated, the stability of the compound, and, if desired, another type of therapeutic agent administered in combination with the compounds of the present invention, but are not limited thereto. Larger total doses can be delivered by multiple administrations of the agent. Methods for determining efficacy and dosage are known to those of ordinary skill in the art. See, for example, Isselbacher et al. (1996). 7 。

[0068] Generally, the appropriate daily dosage of the active compound used in the compositions and methods of the present invention is the amount of the compound that is the lowest dosage effective to produce a therapeutic effect. Such effective dosages generally depend on the factors described above.

[0069] Optionally, the effective daily dosage of the active compound may be administered as one, two, three, four, five, six or more partial doses administered separately at appropriate intervals throughout the day, optionally in unit dosage forms. In certain embodiments of the present invention, the active compound may be administered one or two or three times a day. In other embodiments, the active compound is administered once a day.

[0070] The patient being treated is any animal in need thereof and generally includes primates (particularly humans), as well as other mammals such as horses, cows, pigs, sheep, cats, and dogs, poultry, and pets.

[0071] In certain embodiments, the compounds of the invention may be used alone or administered in combination with another type of therapeutic agent.

[0072] The present disclosure includes the use of pharmaceutically acceptable salts of the compounds of the invention in the compositions and methods of the invention. In certain embodiments, the intended salts of the invention include, but are not limited to, alkyl, dialkyl, trialkyl, or tetraalkylammonium salts. In certain embodiments, the intended salts of the invention include, but are not limited to, L-arginine, benethamine, benzathine, betaine, calcium hydroxide, choline, deanol, diethanolamine, diethylamine, 2-(diethylamino)ethanol, ethanolamine, ethylenediamine, N-methylglucamine, hydrabamine, 1H-imidazole, lithium, L-lysine, magnesium, 4-(2-hydroxyethyl)morpholine, piperazine, potassium, 1-(2-hydroxyethyl)pyrrolidine, sodium, triethanolamine, tromethamine, and zinc salts. In certain embodiments, the intended salts of the invention include, but are not limited to, Na, Ca, K, Mg, Zn, or other metal salts.In certain embodiments, the intended salts of the invention include, but are not limited to, 1-hydroxy-2-naphthoic acid, 2,2-dichloroacetic acid, 2-hydroxyethanesulfonic acid, 2-oxoglutaric acid, 4-acetamidobenzoic acid, 4-aminosalicylic acid, acetic acid, adipic acid, l-ascorbic acid, l-aspartic acid, benzenesulfonic acid, benzoic acid, (+)-camphoric acid, (+)-camphor-10-sulfonic acid, capric acid (decanoic acid), caproic acid (hexanoic acid), caprylic acid (octanoic acid), carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecyl sulfate, ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, d-glucoheptonic acid, d-gluconic acid, d-glucuronic acid, glutamic acid, glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, hydrobromic acid, hydrochloric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, l-malic acid, malonic acid, mandelic acid, methanesulfonic acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, nitric acid, oleic acid, oxalic acid, palmitic acid, pamoic acid, phosphoric acid, propionic acid, l-pyroglutamic acid, salicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, l-tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroacetic acid, and undecylenate.

[0073] Pharmaceutically acceptable acid addition salts can also exist as various solvates with water, methanol, ethanol, dimethylformamide, and the like. Mixtures of such solvates can also be prepared. The source of such solvates may be inherent in the solvent of preparation or crystallization, derived from the solvent of crystallization, or may be exogenous to such solvents.

[0074] Wetting agents, emulsifying agents, and lubricants such as sodium lauryl sulfate and magnesium stearate, as well as colorants, release agents, coating agents, sweeteners, flavoring agents and perfuming agents, preservatives, and antioxidants may also be present in the composition.

[0075] Examples of pharmaceutically acceptable antioxidants include the following: (1) water-soluble antioxidants (such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, and the like), (2) oil-soluble antioxidants (such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol, and the like), and (3) metal chelating agents (such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like).

[0076] Unless otherwise defined herein, scientific and technical terms used in connection with this application shall have the meanings commonly understood by those of ordinary skill in the art to which this disclosure pertains. It is to be understood that the present invention is not limited to the specific methodologies, protocols, and reagents, etc. described herein and thus can vary. The terms used herein are for the purpose of describing particular embodiments only and are not intended to limit the scope of the invention, which is defined only by the claims. Definitions of common terms in immunology and molecular biology can be found in Merck Manual of Diagnostic and Therapy 8 , The Encyclopedia of Molecular Cell Biology and Molecular Medicine 9 , Molecular Biology and Biotechnology: a Comprehensive Desk Reference 10 , Immunology 11 , Janeway’s Immunobiology 12 , Lewin’s Genes XI 13 , Molecular Cloning: A Laboratory Manual 14 , Basic Methods in Molecular Biology 15 , Laboration Methods in Enzymology16 、Current Protocols in Molecular Biology (CPMB) 17 、Current Protocols in Protein Science (CPPS) 18 、and can be found in Current Protocols in Immunology (CPI). 19 。

[0077] In some embodiments of any of the aspects, the disclosure described herein is not directed to a process of cloning a human, a process of modifying the genetic identity of a human germ line, the use of a human embryo for industrial or commercial purposes, or a process of modifying the genetic identity of an animal that may cause pain to the animal without any substantial medical benefit to a human or animal, and animals resulting from such processes.

[0078] Other terms are defined herein within the description of the various aspects of the invention.

[0079] Food allergy An allergy is an immunodeficiency in which an individual is hypersensitized to react immunologically to a substance, typically harmless in itself, called an allergen. The main antibody involved in allergic reactions is immunoglobulin E (IgE). IgE plays an important role in type I hypersensitivity, which appears in various allergic diseases such as allergic asthma, most types of rhinitis, allergic rhinitis, food allergy, and certain types of chronic urticaria and atopic dermatitis. 20 IgE also plays an important role in reactions to allergens such as anaphylactic reactions to drugs, bee stings, and antigen preparations used in desensitization immunotherapy.

[0080] The constant region of IgE (Fc region) can bind to specific receptors on cells that can release histamine or other inflammatory mediators, cytokines, and / or proteases into the surrounding tissues. Histamine-releasing cells are mainly mast cells and basophils. The release of histamine is initiated when cell-bound IgE is cross-linked by contact with and allergens. Histamine, which is mainly stored in mast cells and basophils, is a prominent contributing factor in allergic diseases. Elevated plasma or tissue histamine levels have been observed during anaphylaxis and experimental allergic reactions of the skin, nose, and airways. Histamine released into the nose, eyes, and sinuses stimulates, for example, sneezing, runny nose, and itchy eyes; when released into the lungs, it causes constriction and swelling of the inner layer of the airways and secretion of thick mucus; in the skin, it causes rashes and hives; and in the digestive system, it causes stomach cramps and diarrhea. Typical allergens are derived from plant pollen (e.g., rye, mugwort, Japanese cedar, oak pollen), mold spores, drugs (e.g., penicillin, sulfonamides, salicylates, and local anesthetics), foods (e.g., nuts, seafood, eggs, peas, beans, peanuts, and other legumes, milk), insect products (e.g., bee sting venom, wasp sting venom, cockroach calyx, dust mites), and animal hair and dander.

[0081] Food allergy is an abnormal immune response to food. The signs and symptoms can range from mild to severe and may include itching, tongue swelling, vomiting, diarrhea, hives, difficulty breathing, or low blood pressure, and in severe cases, anaphylaxis. This typically occurs within minutes to hours of exposure. Sensitivity levels vary by country, but the most common food allergies are allergies to milk, eggs, peanuts, tree nuts, seafood, shellfish, soy, and wheat. One of the most common food allergies is sensitivity to peanuts, which are members of the legume family. Peanut allergy can be severe, but children with peanut allergy may outgrow it as they grow. Tree nuts, including cashew, Brazil nut, hazelnut, macadamia nut, pecan, pistachio, pine nut, coconut, and walnut, are also common allergens. Patients can be sensitive to one specific tree nut or many different tree nuts. Additionally, seeds, including sesame seeds and poppy seeds, contain oils that have proteins and may trigger an allergic reaction.

[0082] Diagnosis is usually based on the medical history, elimination diet, skin prick test, blood test for food-specific IgE antibodies, or oral food challenge test. In the skin prick test, a small board with protruding needles is used. The allergen is placed on the board or directly on the skin. Then, the board is placed on the skin, the skin is pricked, and the allergen is allowed to enter the body. If hives appear, the person is considered positive for the allergy. This test only functions for IgE antibodies. Allergy reactions caused by other antibodies cannot be detected by the skin prick test. The patch test is used to determine whether a particular substance causes allergic inflammation of the skin. This tests for delayed food reactions. Blood tests are another way to test for allergies, but they raise the same drawback, detecting only IgE allergens and not functioning for all possible allergens. Food challenges test for allergens other than those caused by IgE allergens. The allergen is given to the person in the form of a pill, allowing the person to directly ingest the allergen. The person is observed for signs and symptoms. The problem with food challenges is that they must be performed under careful hospital supervision because of the possibility of anaphylaxis. For tests that involve the subject's own reaction, the subject cannot be given many different tests in a short period of time. Additionally, these types of tests are expensive and invasive. Additional diagnostic tools for the evaluation of eosinophilic reactions or non-IgE antibody-mediated reactions include endoscopy, colonoscopy, and biopsy.

[0083] Chaperonin 60.1-related peptide Before the compositions and methods are described, it is to be understood that the invention is not limited to the specific compositions, methods, and experimental conditions described, and that such compositions, methods, and conditions may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting, as the scope of the invention is defined only by the appended claims.

[0084] Compositions useful in the method of the present invention include DGSVVVNKVSELPAGHGLNVNTLSYGDLAAD (SEQ ID NO: 1) (PIN201104), DGSVVVNKVSELPAGH (SEQ ID NO: 2), GLNVNTLSYGDLAAD (SEQ ID NO: 3), SELPAGHGLNVNLTS (SEQ ID NO: 4), DGSVVVNKVS (SEQ ID NO: 5), ELPAGHGLNV (SEQ ID NO: 6), NTLSYGDLAAD (SEQ ID NO: 7), or functionally equivalent fragments or variants thereof are included, but are not limited thereto.

[0085] Other compositions useful in the method of the present invention include U.S. Patent Application Publication No. 20040132163 21 and U.S. Patent Nos. 11,098,090 22 , 9,320,791 23 , 9,085,632 24 disclosed Cpn60.1-related peptides, but are not limited thereto.

[0086] In certain embodiments, the Cpn60.1-related peptide is composed of amino acid residues. As used herein, the term "amino acid residue" is used interchangeably with the terms "amino acid" or "aa" to refer to an amino acid that is part of a peptide or protein. In some such embodiments, the agonist or ligand of the invention is composed of amino acids having the standard structure NH2-C(H)(R)-COOH, where R represents the individual amino acid side chain. In certain embodiments, the agonist or ligand is composed of amino acid residues that are natural amino acids. In certain embodiments, the natural amino acids include one of the 20 standard amino acids found in natural peptides and proteins. In some such embodiments, the agonist or ligand is composed of at least one natural amino acid residue that is alanine ("A"), arginine ("R"), asparagine ("N"), aspartic acid ("D"), cysteine ("C"), glutamine ("Q"), glutamic acid ("E"), glycine ("G"), histidine ("H"), isoleucine ("I"), leucine ("L"), lysine ("K"), methionine ("M"), phenylalanine ("F"), proline ("P"), serine ("S"), threonine ("T"), tryptophan ("W"), tyrosine ("Y"), or valine ("V").

[0087] In other embodiments, the Cpn60.1-related peptide used in the methods of the invention is composed of at least one amino acid residue that is a non-natural amino acid or a synthetic amino acid. In some such embodiments, the non-natural amino or synthetic amino acids include, but are not limited to, chemically modified amino acids modified by methylation, amidation, acetylation, protecting groups, and / or substitution with other chemical groups that can alter the physiochemical properties of the peptide. In further embodiments, the non-natural amino or synthetic amino acids are chemically modified amino acids modified with one or more chemical entities (e.g., methyl group, acetic acid group, acetyl group, phosphate group, formyl moiety, isoprenoid group, sulfate group, polyethylene glycol moiety, lipid moiety, sugar chain, biotin moiety, etc.).

[0088] In a further aspect, the Cpn60.1-related peptide used in the method of the present invention is composed of at least one amino acid having the L configuration (chirality of L-amino acids). In an additional aspect, the Cpn60.1-related peptide used in the method of the present invention is composed of at least one amino acid having the D configuration (chirality of D-amino acids).

[0089] In certain aspects, the agonist or ligand of the present invention is about 50 aa in length, 49 aa in length, 48 aa in length, 47 aa in length, 46 aa in length, 45 aa in length, 44 aa in length, 43 aa in length, 42 aa in length, 41 aa in length, 40 aa in length, 39 aa in length, 39 aa in length, 38 aa in length, 37 aa in length, 36 aa in length, 35 aa in length, 34 aa in length, 33 aa in length, 32 aa in length, 31 aa in length, 30 aa in length, 29 aa in length, 28 aa in length, 27 aa in length, 26 aa in length, 25 aa in length, 24 aa in length, 23 aa in length, 22 aa in length, 21 aa in length, or 20 aa in length, 19 aa in length, 18 aa in length, 17 aa in length, 16 aa in length, 15 aa in length, 14 aa in length, 13 aa in length, 12 aa in length, 11 aa in length, 10 aa in length, 9 aa in length, 8 aa in length, 7 aa in length, 6 aa in length, or 5 aa in length.

[0090] In certain aspects, the composition of the present invention is administered to a patient by any suitable route known and / or employed by those skilled in the art. In some aspects, the composition of the present invention is administered orally (PO), intravenously (IV), intramuscularly (IM), intraarterially, intramedullary, intrathecal, subcutaneously (SQ), intraventricularly, transdermally, intradermally, rectally (PR), vaginally, intraperitoneally (IP), intragastrically (IG), topically (e.g., powders, ointments, creams, gels, lotions and / or drops), mucosally, intranasally, buccally, enterally, intravitreally, sublingually, by tracheal instillation, bronchial instillation and / or inhalation, oral spray, nasal spray, as an aerosol, and / or via a portal catheter.

[0091] In certain embodiments, the Cpn60.1-related peptides used in the methods and / or pharmaceutical compositions of the present invention may be administered intravenously, for example, by intravenous infusion. In further embodiments, the Cpn60.1-related peptides used in the methods and / or pharmaceutical compositions of the present invention may be administered by intramuscular injection. In yet additional embodiments, the Cpn60.1-related peptides used in the methods and / or pharmaceutical compositions of the present invention may be administered by intratumoral injection. In certain embodiments, the Cpn60.1-related peptides used in the methods and / or pharmaceutical compositions of the present invention may be administered by subcutaneous injection. In further embodiments, the Cpn60.1-related peptides used in the methods and / or pharmaceutical compositions of the present invention may be administered via a portal vein catheter. In yet additional embodiments, the present invention encompasses delivery of the Cpn60.1-related peptides used in the methods and / or pharmaceutical compositions of the present invention by any suitable route, considering the potential for advancement in the art of drug delivery.

[0092] In some embodiments, the Cpn60.1-related peptides used in the methods and / or pharmaceutical compositions of the present invention may be administered at dosage levels sufficient to deliver from about 0.001 mg / kg to 100 mg / kg, about 0.01 mg / kg to 50 mg / kg, about 0.1 mg / kg to 40 mg / kg, about 0.5 mg / kg to 30 mg / kg, about 0.01 mg / kg to 10 mg / kg, about 0.1 mg / kg to 10 mg / kg, or about 1 mg / kg to 25 mg / kg per day per patient weight to obtain a desired therapeutic effect. In certain embodiments, the desired dosage may be delivered more than 3 times a day, 3 times a day, 2 times a day, once a day, once every other day, once every 3 days, once a week, once every 2 weeks, once every 3 weeks, once every 4 weeks, once every 2 months, once every 6 months, or once every 12 months. In certain embodiments, the desired dosage may be delivered using multiple administrations (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or more administrations). In certain embodiments, the desired dosage may be delivered using one or more administrations during an initial period, followed by a period during which no dosage is administered.

[0093] In a further aspect, the Cpn60.1-related peptide used in the method of the present invention can be utilized for prophylactic use. In additional aspects, the prophylactic use includes systems and methods for preventing, inhibiting the progression of, and / or delaying the onset of food allergy in individuals who are susceptible to and / or exhibit symptoms of food allergy.

[0094] In certain aspects, the Cpn60.1-related peptide used in the method of the present invention is administered to target cells in vivo. In other aspects, the Cpn60.1-related peptide used in the method of the present invention is administered to target cells ex vivo. In additional aspects, the Cpn60.1-related peptide used in the method of the present invention is administered to target cells ex vivo, and then those target cells are reintroduced into an organism. In some such aspects, the target cells are cultured ex vivo into a plurality of progeny cells before being reintroduced into the organism. In further additional aspects, the organism is a human. In a further aspect, the organism is a human patient. In certain aspects, the target cells are originally derived from the organism into which they are reintroduced. In other aspects, the target cells are originally derived from a different organism into which they are reintroduced.

[0095] In certain aspects, the Cpn60.1-related peptide used in the method of the present invention and / or its pharmaceutical composition is used in combination therapy for treating food allergy or reducing its risk. In such aspects, the administration can be combined with one or more additional therapeutic agents. As used herein, phrases such as "combination therapy," "in combination with," "combined with," etc. refer to the simultaneous use of multiple agents or treatments to increase the response. In certain aspects, the Cpn60.1-related peptide used in the method of the present invention and / or its pharmaceutical composition is administered simultaneously with, before, or after one or more other desired treatments or medical procedures. In certain aspects, the Cpn60.1-related peptide used in the method of the present invention and / or its pharmaceutical composition is administered together in a single composition or separately in different compositions.

[0096] In certain embodiments, the particular combination of therapies employed in a combination regimen generally takes into account the suitability of the desired therapeutic agent and / or treatment, as well as the desired therapeutic effect to be achieved. In further embodiments, the therapies employed can achieve the desired effect for the same purpose (e.g., the Cpn60.1-related peptides used in the methods of the invention useful for the treatment, prevention, and / or delay of onset of food allergies can be administered concomitantly with another therapeutic agent that is similarly useful for the treatment, prevention, and / or delay of onset of food allergies), or can achieve different effects. In further embodiments, the combination of therapies employed can achieve the same or substantially similar desired effect for the same disease, condition, or disorder, can achieve the same or substantially similar desired effect for one or more different diseases, conditions, or disorders, can achieve different desired effects for the same disease, condition, or disorder, or can achieve different desired effects for one or more different diseases, conditions, or disorders.

[0097] In additional embodiments, the delivery of the Cpn60.1-related peptides used in the methods of the invention as pharmaceutical compositions is combined with one or more additional components that can improve the bioavailability of the Cpn60.1-related peptides used in the methods of the invention, reduce and / or modify their metabolism, inhibit their excretion, and / or modify their distribution in the body.

[0098] In certain embodiments, combination therapy can involve the administration of multiple Cpn60.1-related peptides according to the invention. In further embodiments, combination therapy can involve the administration of multiple Cpn60.1-related peptides that treat, prevent, ameliorate, achieve remission of, and / or reduce the risk of food allergies. In still further embodiments, combination therapy can be multiple Cpn60.1-related peptides that treat, prevent, ameliorate, achieve remission of, and / or reduce the risk of multiple food allergies.

[0099] In certain embodiments, the Cpn60.1-related peptide used in the methods of the invention is in the form of a pharmaceutical composition and is combined with at least one pharmaceutically acceptable excipient. As used herein, "pharmaceutical composition" refers to a formulation containing an active ingredient and optionally a pharmaceutically acceptable carrier, diluent, or excipient. The term "active ingredient" may interchangeably refer to "active agent" and means any agent capable of inducing the desired effect upon administration. Examples of active ingredients include, but are not limited to, compounds, drugs, therapeutic agents, small molecules, and the like.

[0100] In certain embodiments of the invention, the active ingredient is a Cpn60.1-related peptide disclosed herein. In certain embodiments, the active ingredient is PIN201104 or a derivative thereof. In certain embodiments, the active ingredient is PIN201360, PIN201361, PIN201362, PIN201116, PIN201105, or a derivative thereof. In further embodiments, the active ingredient is a peptide described in WO2009 / 106819 or a derivative thereof.

[0101] In certain embodiments, the pharmaceutical composition is useful for the manufacture of a medicament or pharmaceutical product. In further embodiments, the pharmaceutical composition is useful for one or more of the therapeutic uses disclosed herein, for example, in an individual suffering from an autoimmune disorder. In additional embodiments, the pharmaceutical composition is formulated for administration to a human patient.

[0102] In certain embodiments, the pharmaceutical composition is in a sterile injectable form (e.g., a form suitable for subcutaneous injection or intravenous infusion). In further additional embodiments, the pharmaceutical composition is in a liquid dosage form suitable for injection. In yet further embodiments, the pharmaceutical composition is in a powder (e.g., freeze-dried and / or sterilized) state, optionally under vacuum, which is reconstituted with an aqueous diluent (e.g., water, buffer, salt solution, etc.) prior to injection. In additional embodiments, the pharmaceutical composition is diluted and / or reconstituted in an aqueous diluent (e.g., water, sodium chloride solution, sodium acetate solution, benzyl alcohol solution, phosphate-buffered saline, etc.). In certain embodiments, the pharmaceutical composition is in a form that can be cooled and / or frozen. In further embodiments, the pharmaceutical composition is in a form that cannot be cooled and / or frozen. In certain embodiments, the pharmaceutical composition is a reconstituted solution and / or liquid dosage form that can be stored for a specific period (e.g., 2 hours, 12 hours, 24 hours, 2 days, 5 days, 7 days, 10 days, 2 weeks, 1 month, 2 months, or more) after reconstitution.

[0103] In certain embodiments, a method of preparing a pharmaceutical composition comprises combining an active ingredient (e.g., a Cpn60.1-related peptide used in the methods of the present invention) with one or more pharmaceutically acceptable excipients and then shaping and / or packaging the product into a desired single-dose unit or multiple-dose units. The pharmaceutical composition according to the present invention may be prepared, bulk packaged, packaged as a single unit dose, and / or packaged as multiple single unit doses. As used herein, "unit dose" refers to an individual amount of a pharmaceutical composition that contains a predetermined amount of an active ingredient. The amount of the active ingredient is generally equal to the dose administered to a subject and / or a convenient fraction of such a dose, such as, for example, half or one-third of such a dose. The relative amounts of the active ingredient, pharmaceutically acceptable excipients, and / or any additional components in the pharmaceutical composition according to the present invention may vary depending on the identity, size, and / or condition of the subject being treated and / or the route by which the composition is administered. In certain embodiments, for example, the composition may contain from about 0.1% to 100% (w / w) of the active ingredient.

[0104] In another aspect, the invention includes a kit useful for practicing the methods of the invention. The components included in the kit depend on a number of factors, including a particular use (e.g., a particular route of administration employed, or a particular disease, condition, or disorder being treated). In certain aspects, the invention provides a kit for administering a Cpn60.1-related peptide according to the invention for treating a disease, condition, or disorder disclosed herein. In some such aspects, the kit further includes instructions for administration. In certain aspects, the kit is for administering a Cpn60.1-related peptide for treating a patient having a food allergy. In certain aspects, the kit contains one or more Cpn60.1-related peptides. In certain aspects, the kit includes multiple unit doses of a pharmaceutical composition containing a Cpn60.1-related peptide. In additional aspects, a kit for use in accordance with the invention includes instructions (e.g., for administration, storage, etc.), buffers, and / or other reagents. In some such aspects, the kit includes (i) at least one Cpn60.1-related peptide, (ii) a syringe, needle, applicator, etc. for administering the at least one Cpn60.1-related peptide to a patient, and (iii) instructions for use. In further aspects, the kit includes a treatment schedule indicating when unit doses are to be administered. In yet additional aspects, a placebo dose in either a form similar to or separate from the dose of the pharmaceutical composition is included. In certain aspects, the kit includes one or more containers such that particular ones of the individual components or reagents can be separately housed. In certain aspects, the kit may include means for enclosing the individual containers in a relatively close confinement for commercial sale, e.g., a plastic box, which may enclose packaging materials such as instructions, styrofoam, etc.

[0105] The description of embodiments of the present disclosure is not intended to be exhaustive or to limit the disclosure to the exact forms disclosed. Specific embodiments and examples of the present disclosure are described herein for illustrative purposes, but various equivalent modifications are possible within the scope of the present disclosure, as will be recognized by those of ordinary skill in the relevant art. For example, while method steps or functions are presented in a given order, alternative embodiments may perform the functions in a different order or substantially simultaneously. The teachings of the present disclosure provided herein can be applied to other procedures or methods as needed. Combinations of the various embodiments described herein can provide further embodiments. Aspects of the present disclosure can be modified to adopt the compositions, functions, and concepts of the above references and applications, as needed, to provide still further embodiments of the present disclosure. Additionally, considering biological functional equivalency, some changes can be made to the protein structure without affecting the biological or chemical actions in terms of type or amount. These and other changes can be made to the present disclosure in view of the detailed description. All such modifications are intended to be included within the scope of the appended claims.

[0106] Any specific elements of any of the foregoing embodiments may be combined with or substituted for elements of other embodiments. Further, while the advantages associated with specific embodiments of the present disclosure have been described in the context of those embodiments, other embodiments may also exhibit such advantages, and not all embodiments necessarily exhibit such advantages in order to fall within the scope of the present disclosure.

[0107] The techniques described herein are further illustrated by the following examples, which should not be construed in any way as a further limitation. Methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, but the preferred methods and materials are described below.

Examples

[0108] The present invention will now be described generally. It is included only for the purpose of exemplifying specific aspects and embodiments of the present invention and is not intended to limit the present invention. It will be more readily understood by reference to the following examples.

[0109] Example 1 Effect of CPN60.1-related Peptide in an Animal Model of Ovalbumin-induced Food Allergy Chaperonin 60.1 (Cpn60.1)-related peptides such as PIN201104 ("1104") have been shown to be very effective in blocking inflammation associated with the late-phase reaction. See Figure 1. However, previous studies did not consider the effect of Cpn60.1-related peptides on the immediate hypersensitivity reaction characterized by acute anaphylaxis in which mast cells and basophils play important roles. For some indications such as food allergy, therapeutic efficacy in the immediate phase is important. This study evaluated whether 1104 has an effect on the immediate reaction in addition to its known effect on the late-phase reaction. A mouse model of food allergy was developed that showed a progressive severity of anaphylaxis induced by ovalbumin (OVA).

[0110] Allergens can induce hypersensitivity reactions mediated by IgE antibodies regardless of whether they are derived from food, air pollution, dust mites, pet dander, or mold. The result of an allergic reaction is a wheezing reaction characterized by airflow obstruction and bronchospasm. In an allergic reaction, T-helper cells induce the expression of many cytokines including TNF-α, IL-2, IL-4, IL-5, IL-6, IL-10, and IL-13 (1-4). Among these cytokines, IL-4 and IL-13 play important roles in the allergic reaction. IL-13 has a unique role in mucus production, while IL-4 has been found to be a major driver of IgE and IgG1 synthesis by B cells. Furthermore, IL-5 drives the eosinophilia in blood and tissues that is a characteristic feature of the allergic reaction 25,26 。

[0111] Ovalbumin (OVA) and house dust mite (HDM) are antigens widely used to induce allergic reactions. 27,28,29,30 For example, both OVA-induced and HDM-induced asthma models can induce airway inflammation, which can lead to airflow obstruction and airway remodeling (AR).

[0112] There are many chemokines related to allergic reactions, including CCL11, CCL17, and CCL22, all of which have been found to be upregulated in bronchoalveolar lavage tests of asthma patients. In mouse models, inhibition of CCL11 has been shown to reduce eosinophil recruitment and airway hyperresponsiveness. In fact, asthma patient samples showed higher levels of CCL11 in plasma than healthy volunteers. Chemokines CCL17 and CCL22 mainly bind to CCR4 expressed by Th2 cells and play a major role in T cell recruitment in both mice and humans. CCL17 and CCL22 have been well reported in many allergic reactions, including skin allergy and food allergy. Furthermore, Th1 / Th2 cytokines such as IL-4, IL-5, IL-13, and INF-γ have been reported to be upregulated. 31 。

[0113] Mouse model of food allergy In this study, a mouse model of food allergy was developed that showed a progressive severity of anaphylaxis induced by ovalbumin (OVA). The design protocol and evaluation items are shown in Figure 2 and summarized in Table 1.

[0114] (Table 1) Food allergy model TIFF2025521110000002.tif91161

[0115] The clinical scale used for anaphylaxis scoring is shown in Table 2.

[0116] (Table 2) Clinical scoring scale * TIFF2025521110000003.tif78145

[0117] Animal: Male BalbC mice (20 - 30 g upon arrival, Charles Rivers, UK) were used in this study.

[0118] Animal welfare: Upon arrival from the supplier, the animals were acclimatized for 7 days before the start of the experimental procedure. The mice were housed in cages of four based on their body weight at arrival (equal distribution of animal weight between cages by an animal technician) and placed on a 12 - hour light - dark cycle. The room temperature and humidity were maintained at 17 - 24°C and 40 - 70% respectively. Environmental enrichment was provided in all cages. The mice had free access to standard solid feed and water was available ad libitum from bottles.

[0119] As outlined below, animals were placed in cages of four and the welfare of all animals was checked daily. Throughout the study, the following guidelines were used to assess non - specific or unexpected adverse effects in animals subject to procedures or administration of test compounds: · Weight loss exceeding 20% of the highest measured individual body weight. · Marked piloerection associated with other signs of dehydration such as skin tenting. · Non - responsive to activity or elicitation. · Persistent curling of the body (immobile). · Distress - persistent vocalization. · Persistent and excessive ocular and nasal discharge. · Labored breathing. · Persistent tremors. · Persistent convulsions.

[0120] Animals showing two or more of any limited clinical signs in the category corresponding to the severity limit of the protocol were removed from the study and euthanized by the method of Schedule 1 (cervical dislocation). If an animal reached the limit of either or both of the first two signs, regardless of the presence or absence of other signs, the animal was removed from the test at the time of determination and euthanized by the method of Schedule 1. However, no animals were removed during this test.

[0121] OVA sensitization and challenge: Mice were intraperitoneally sensitized on day 0 with 100 μg of ovalbumin adsorbed to 1 mg of alum (Imjec alum - aluminum content 40 mg / mL) using 25 μL. Then the mice were challenged on days 14, 16, 18, 21, 23, 25, and 28 via gavage with 5 mg of OVA in 200 μL, or 200 μL of PBS (Groups 1 - 5).

[0122] Evaluation of body temperature and anaphylaxis: After each challenge, the animals were monitored for 60 minutes, and rectal temperatures were recorded at 0, 15, 30, and 60 minutes after gavage with PBS or OVA. Based on anaphylaxis symptoms, a score of 0 - 5 was assigned to each mouse. The clinical scale used for anaphylaxis scoring is provided in Table 2 above.

[0123] Treatment of animals with ‘1104 or vehicle: Fifteen minutes before the OVA challenge on days 14, 16, 18, 21, 23, and 25, the mice were weighed, placed in a hot box for 5 minutes, and then placed in a whole - body restraint device for intravenous administration (5 mL / kg) via the tail vein with either ‘1104 (80 μg / kg or 800 μg / kg) or vehicle.

[0124] Blood sample collection: One hour after the final OVA challenge on day 28 or 38, the final blood sample was collected via cardiac puncture and placed in a serum tube. Each serum sample was held at room temperature for 45 minutes to clot, then centrifuged (2000×g, 15 minutes at 4°C), and the supernatant obtained was extracted, aliquoted, and stored at - 80°C for analysis.

[0125] Tissue collection: Immediately after collecting the final blood sample, the animals were sacrificed by an overdose of pentobarbital. The abdomen was opened, the small intestine was detached from each animal, and flushed with PBS. The tissue was then divided into duodenum, jejunum, and ileum. Each section was opened along the long axis, coiled around a wooden stick to create a roll. Each tissue roll was then sectioned, and one section was placed in 10% formalin for 48 hours and then transferred to 70% ethanol for future histopathological examination. A second section was placed in a sterile Eppendorf, snap-frozen, and then stored at -80 °C for future biomarker analysis.

[0126] After collection of the small intestine, mesenteric lymph nodes were detached from each animal. After dissection, the lymph nodes were placed in a sterile Eppendorf, snap-frozen, and then stored at -80 °C for future analysis.

[0127] OVA-specific IgE and mMCP1 ELISA assays: Serum supernatants were evaluated for OVA-specific IgE and mMCP1 concentrations using ELISA kits (AssayGenie and Invitrogen, respectively) according to the manufacturer's instructions. Optical density was measured at 450 nM using a microplate reader (SpectraMax 340PC). The concentration of IgE was determined using SoftMax Pro v.6.4 (Molecular Devices). Data were reported as OVA-specific IgE (pg / mL) or mMCP1 concentration (pg / mL), mean ± S.E.M. (standard error of the mean).

[0128] Cytokine / chemokine assays: Cytokine / chemokine concentrations in serum samples (all groups) (see below for details of biomarkers to be evaluated) were measured using a magnetic multiplex assay according to the manufacturer's instructions. Levels were measured using a Magpix system (Luminex Corp.).

[0129] Optical density was measured at 450 nM using a microplate reader (SpectraMax 340PC). Mouse cytokine / chemokine magnetic multiplex panel (Biotechne): IL-4, IL-5, IL-13, and eotaxin. Data were reported as cytokine / chemokine (pg / mL), mean ± S.E.M. (standard error of the mean).

[0130] Data analysis: Deviations between groups were statistically analyzed by one-way analysis of variance (ANOVA). When there was a significant difference in the mean values between different levels of treatment, comparisons against the vehicle group were performed using the Dunnett's test. p < 0.05 was considered statistically significant.

[0131] Results Clinical scoring: This model provides a progressive model of food allergy. As shown in Figure 3, an increase in response was observed over time in all OVA-exposed mice. Clinical scoring of the animals was performed on days 21, 23, 25, and 28. As expected, mice challenged daily with 5 mg of OVA (intragastrically) from days 14 to 28 TIFF2025521110000004.tif4128 were PBS-treated animals TIFF2025521110000005.tif4128 showed a statistically significant progressive increase in anaphylaxis severity compared to. By day 28, some animals in the OVA / vehicle arm were scored 4 on the anaphylaxis symptom scale. 3 doses Treatment of animals with '1104 (80 μg / kg) of TIFF2025521110000006.tif4128, on day 28, the vehicle group exposed to OVA TIFF2025521110000007.tif4128 significantly reduced the severity of anaphylaxis compared to, but 6 doses Treatment with '1104 (80 μg / kg) of TIFF2025521110000008.tif4128, on days 23, 25, and 28, the vehicle group exposed to OVA Compared with TIFF2025521110000009.tif4128, the severity of anaphylaxis was significantly reduced. Treatment with three higher doses of '1104 (160 μg / kg) TIFF2025521110000010.tif4128, like the group administered six lower doses of 80 μg / kg, was the vehicle group exposed to OVA on days 25 and 28 Compared with TIFF2025521110000011.tif4128, the severity of anaphylaxis was significantly reduced. Interestingly, from the results on the last day, the hypothesis that the Cpn60.1-related peptide has short pharmacokinetics (PK) and long pharmacodynamics (PD) was confirmed. Because the serum PK of '1104 was about 15 minutes, while the last OVA challenge was 3 days after the last administration of '1104

[0132] Biometric measurements: Biomarker analysis was performed on day 28, and the most significant differences between the vehicle-treated group and the '1104-treated group were observed in this analysis. OVA-specific IgE, mMCP, body temperature, and clinical scores were recorded. As shown in Figure 4, prophylactic and therapeutic administration with '1104 (80 or 160 μg / kg, 3 or 6 doses) significantly reduced all four biometric measurements of food allergy on day 28, 3 days after the last administration of '1104: OVA-specific immunoglobulin E (IgE) (upper left panel), mouse mast cell protease (mMCP-1) (upper right panel), body temperature (lower left panel), and clinical scoring (lower right panel).

[0133] Major Th2 cytokines / chemokines: Serum levels of major Th2 cytokines / chemokines were also measured on day 28: IL-4, IL-5, IL-13, and eotaxin. As expected, mice challenged daily with 5 mg of OVA (intragastrically) from days 14 - 28 showed statistically increased levels of these major Th2 cytokines / chemokines compared to PBS-treated animals. See Figure 5: IL-4 (upper left panel), IL-5 (upper right panel), IL-13 (lower left panel), and eotaxin (lower right panel). Confirming the above biometric results, 6 doses of 80 μg / kg of ‘1104 and 3 doses of 160 μg / kg of ‘1104 significantly decreased the levels of major Th2 cytokines / chemokines in the ovalbumin-induced food allergy mouse model, as expected by the reduction of the severity of anaphylaxis in these animals by ‘1104.

[0134] Conclusion The results of this study using a mouse model of food allergy suggest that Cpn60.1-related peptides are effective in the prevention and treatment of food allergy and represent a novel non-allergen-specific therapy for food allergy. This study provides the first evidence that Cpn60.1-related peptides have an effect on immediate reactions in addition to their previously known effect on immediate reactions.

[0135] Example 2 Long-term efficacy of CPN60.1-related peptides in an animal model of ovalbumin-induced food allergy As shown in Example 1, Cpn60.1-related peptides have been shown to be effective in the prevention and treatment of food allergy. This study evaluated the efficacy of higher doses of ‘1104 (800 μg / kg) and the long-term efficacy of ‘1104 in the prevention of food allergy after a longer period had elapsed since the administration of ‘1104 in the above ovalbumin (OVA)-induced animal model of food allergy.

[0136] Materials and methods Test Design: The design protocol and endpoints are shown in Figure 6 and summarized in Table 3.

[0137] (Table 3) Test Design TIFF2025521110000012.tif121160

[0138] Animals: As described in Example 1.

[0139] Animal Welfare: As described in Example 1.

[0140] OVA Sensitization and Challenge: For animals from Groups 1 - 5, as described in Example 1. Animals from Groups 6 - 10 were rested for 10 days and then received an additional challenge on Day 38.

[0141] Evaluation of Body Temperature and Anaphylaxis: As described in Example 1.

[0142] Treatment of Animals with ‘1104 or Vehicle: Fifteen minutes before the OVA challenge on Days 14, 16, 18, 21, 23, and 25, mice were weighed, placed in a hot box for 5 minutes, and then placed in a whole - body restrainer for intravenous administration (5 mL / kg) via the tail vein of either ‘1104 (80 μg / kg or 800 μg / kg) or vehicle.

[0143] Blood Sample Collection: One hour after the final OVA challenge on Day 28 or Day 38, a final blood sample was collected via cardiac puncture and placed in a serum tube. Each serum sample was held at room temperature for 45 minutes to clot, then centrifuged (2000×g, 15 minutes at 4°C), and the supernatant obtained was extracted, aliquoted, and stored at - 80°C for analysis.

[0144] Tissue Collection: As described in Example 1.

[0145] OVA - specific IgE and mMCP1 ELISA Assays: As described in Example 1.

[0146] Cytokine / Chemokine Assay: Cytokine / chemokine concentrations in serum samples (all groups) (see below for details of biomarkers being evaluated) were measured using a magnetic multiplex assay according to the manufacturer's instructions. Levels were measured using a Magpix system (Luminex Corp.).

[0147] IL-9 and CCL-17 were measured using commercially available ELISA kits (Biotechne, UK) according to the manufacturer's instructions. Optical density was measured at 450 nM using a microplate reader (SpectraMax 340PC). Concentrations of IL-9 and CCL-17 were determined using SoftMax Pro v. 6.4 (Molecular Devices). Mouse Cytokine / Chemokine Magnetic Multiplex Panel (Biotechne): IL-4, IL-5, IL-10, IL-13, IL-17A, Eotaxin, CCL-22, IFN-gamma, TNF alpha. Data were reported as cytokine / chemokine (pg / mL), mean ± S.E.M. (standard error of the mean).

[0148] Data Analysis: As described in Example 1.

[0149] Results Clinical Scoring: This model provides a progressive model of food allergy. As shown in Figure 7, an increase in response was observed over time in all mice exposed to OVA. Clinical scoring of the animals was performed on days 21, 23, 25, and 28. As expected, treatment of animals with 6 doses TIFF2025521110000013.tif4128 of ‘1104 (80 μg / kg) significantly reduced the severity of anaphylaxis on days 23, 25, and 28 compared to the vehicle group exposed to OVA TIFF2025521110000014.tif4128. Treatment with higher doses of ‘1104 (800 μg / kg) TIFF2025521110000015.tif4128 showed a significantly reduced severity of anaphylaxis compared to the vehicle group exposed to OVA on days 23, 25, and 28. Interestingly, both doses of ‘1104 (80 and 800 μg / kg) showed long-term efficacy by significantly reducing the severity of anaphylaxis 13 days after the last ‘1104 administration, compared to the vehicle group exposed to OVA. From these results, it was reconfirmed that the Cpn60.1-related peptide has short pharmacokinetics (PK) and long pharmacodynamics (PD). TIFF2025521110000016.tif4128 showed a significantly reduced severity of anaphylaxis compared to the vehicle group exposed to OVA on days 23, 25, and 28. Interestingly, both doses of ‘1104 (80 and 800 μg / kg) showed long-term efficacy by significantly reducing the severity of anaphylaxis 13 days after the last ‘1104 administration, compared to the vehicle group exposed to OVA. From these results, it was reconfirmed that the Cpn60.1-related peptide has short pharmacokinetics (PK) and long pharmacodynamics (PD).

[0150] Biometric measurements: Biomarker analysis was performed on day 28 and again on day 38. Consistent with the results of Example 1, ‘1104 (80 and 800 μg / kg, 6 doses) significantly improved the symptom score and body temperature on day 28 compared to the vehicle group exposed to OVA (Figure 8, left panel). Interestingly, these beneficial effects were maintained on day 38 without further administration of ‘1104 (Figure 8, right panel).

[0151] OVA-specific IgE and mMCP1 levels on days 28 and 38: In a group of mice, serum samples obtained on day 28 from OVA-sensitized mice (day 0) challenged with OVA via orogastric gavage on days 14, 16, 18, 21, 23, and 25 with vehicle or ‘1104 (80 or 800 μg / kg, intravenous) were subjected to biomarker analysis. Samples were collected 1 hour after the final OVA challenge on day 28. Both doses of ‘1104 significantly reduced the clinical scores of OVA-specific IgE levels (Figure 9, upper left panel) and mMCP-1 levels (Figure 9, lower left panel) on day 28, 3 days after the last ‘1104 administration. These data confirmed the results of Example 1.

[0152] In another group of mice, serum samples obtained on day 38 from OVA-sensitized mice (day 0) challenged with OVA on days 14, 16, 18, 21, 23 and 25 via oral gavage with vehicle or ‘1104 (80 or 800 μg / kg, intravenous) were subjected to biomarker analysis. Animals from the groups sampled on day 38 also received an additional OVA challenge on day 38. Samples were taken 1 hour after the final OVA challenge on day 38. Again, both doses of ‘1104 decreased OVA-specific IgE (Figure 9, upper right panel) and mMCP-1 levels (Figure 9, lower right panel), but only the higher dose reached statistical significance. Interestingly, these data suggest that ‘1104 shows long-term effects in this model of food allergy, despite its very short half-life (about 10 - 15 minutes across species), and thus supports not only a therapeutic but also a prophylactic role for food allergy.

[0153] Major cytokines / chemokines: Serum levels of major Th2 cytokines / chemokines were measured on days 28 and 38. As expected, mice challenged daily with 5 mg of OVA (intragastrically) from days 14 - 28 showed statistically increased levels of these major cytokines / chemokines compared to PBS-treated animals. See Figures 10 - 14.

[0154] When the results of the above-mentioned biological measurements were examined, the ‘1104 at 6 dosages of 80 μg / kg and 800 μg / kg significantly decreased the levels of major cytokines / chemokines in the ovalbumin-induced food allergy mouse model, which was as expected by the reduction of the severity of anaphylaxis by the ‘1104 in these animals. The major cytokines / chemokines included IL-4 (Figure 10, upper panel), IL-5 (Figure 10, lower panel), IL-9 (Figure 12, upper panel), IL-10 (Figure 12, lower panel), IL-13 (Figure 11, upper panel), IL-17 (Figure 13, upper panel), CCL-17 (Figure 13, lower panel), CCL-22 (Figure 14, upper panel), IFN (Figure 14, lower panel), and eotaxin (Figure 11, lower panel). A significant decrease in the levels of major Th2 cytokines / chemokines was observed on day 28, which was 3 days after the final administration of the ‘1104 (left panel), and on day 28, which was 13 days after the final administration of the ‘1104 (right panel).

[0155] Interestingly, IL-9 has recently been reported to have an important role in the development of IgE-mediated food allergy 34 . For example, IL-9 has been reported to be an important cytokine that promotes mast cell proliferation and has been shown to be derived from peTh2 cells in food allergy 35 . Therefore, it is considered a potential target for the treatment of food allergy. The above results showing a significant decrease in IL-9 induced by the ‘1104 in the animal model of food allergy (Figure 12, upper panel) support the use of the Cpn60.1-related peptide for the prevention and treatment of food allergy.

[0156] Conclusion The results of this study using a mouse model of food allergy suggest that Cpn60.1-related peptides are effective in the prevention and treatment of food allergy. Cpn60.1-related peptides attenuated immediate anaphylactic allergic reactions in a model of ovalbumin-driven food allergy. This effect was consistent across both "clinical" symptoms (anaphylaxis score and body temperature) and related serum biomarkers (i.e., mMCP-1, OVA-IgE, IL-4, IL-5, IL-13, and eotaxin) after both prophylactic and therapeutic administrations. Despite a short plasma half-life (approximately 15 minutes), Cpn60.1-related peptides demonstrated a long pharmacodynamic effect (13 days) consistent with the effects observed in a model of delayed allergic reactions in this immediate allergic reaction model. Therefore, these studies support the use of Cpn60.1-related peptides as an alternative non-allergen-specific therapy for the treatment of food allergy.

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[0158] All patents and other publications, including references, issued patents, published patent applications, and co-pending patent applications, cited throughout this application are hereby expressly incorporated by reference herein for the purpose of describing and disclosing, for example, the methodologies described in such publications that may be used in connection with the techniques described herein. These publications are provided solely for the purpose of their disclosure prior to the filing date of this application. It should not be construed as an admission that the inventors are not entitled to antedate such disclosure by virtue of prior invention or otherwise. All statements as to the date or representation of the content of these documents are based on the information available to the applicant and do not constitute any admission as to the accuracy of the date or content of these documents.

[0159] The foregoing written description is considered to be sufficient to enable one of ordinary skill in the art to practice the present aspect and embodiments. The present aspect and embodiments are provided by way of example only as single illustrations of one aspect, and the scope is not limited by the provided examples because other functionally equivalent embodiments are within the scope of the present disclosure. In addition to what is shown and described herein, various modifications will become apparent to one of ordinary skill in the art from the foregoing description and are included in the appended claims. The advantages and objectives described herein are not necessarily encompassed by each embodiment. One of ordinary skill in the art will be able to recognize or confirm many equivalents to the specific embodiments described herein using only routine experimentation. Such equivalents are intended to be encompassed by the following claims.

[0160] sequence DGSVVVNKVSELPAGHGLNVNTLSYGDLAAD (SEQ ID NO: 1, '1104), DGSVVVNKVSELPAGH (SEQ ID NO: 2), GLNVNTLSYGDLAAD (SEQ ID NO: 3), SELPAGHGLNVNLTS (SEQ ID NO: 4), DGSVVVNKVS (SEQ ID NO: 5), ELPAGHGLNV (SEQ ID NO: 6), NTLSYGDLAAD (SEQ ID NO: 7)

Claims

1. A pharmaceutical composition for use in a method for treating or preventing the development of immunoglobulin E (IgE)-mediated food allergy to a food-derived dietary antigen in a subject in need thereof, the method comprising administering the pharmaceutical composition to the subject.

2. The pharmaceutical composition for use according to claim 1, wherein the administration reduces the level of antigen-specific immunoglobulin E (IgE) in the subject compared to an untreated control.

3. The pharmaceutical composition for use according to claim 1, wherein the administration reduces the level of mast cell protease in the subject.

4. The pharmaceutical composition for use according to claim 1, wherein the administration reduces the serum level of at least one Th2 cytokine or chemokine selected from interleukin-4 (IL-4), interleukin-5 (IL-5), interleukin-13 (IL-13), and eotaxin in the subject.

5. The pharmaceutical composition for use according to claim 1, wherein the pharmaceutical composition is used before the subject's first exposure to a potential causative food allergen or at the time of the appearance of clinical signs of atopic symptoms.

6. The pharmaceutical composition for use according to claim 1, further comprising, before administering the pharmaceutical composition, diagnosing the subject as having or being likely to develop an IgE-mediated food allergy.

7. A pharmaceutical composition for use according to claim 1, wherein the dietary antigen is derived from milk and its products; eggs and their products; meat and its products; fish, mollusks, and crustaceans, and their products; oils, fats, and their products; grains and their products; legumes, seeds, cereals, nuts, and their products; vegetables and their products; fruits and their products; mushrooms and their products; sugars, sugar products, chocolate products, and confectionery; and spices and herbs.

8. The pharmaceutical composition for use according to claim 1, wherein the pharmaceutical composition is administered in a route comprising 3 to 6 doses, each dose being between 80 μg / Kg and 800 μg / Kg per body weight of the subject.

9. The pharmaceutical composition for use according to claim 8, wherein the therapeutic effect lasts for at least 7 days after the final administration of the pharmaceutical composition.

10. The pharmaceutical composition for use according to claim 1, wherein the pharmaceutical composition is administered subcutaneously.

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