The present invention provides fusion proteins comprising an allergen sequence linked via an IgG hinge region to another polypeptide sequence capable of specifically binding to a native IgG inhibitory receptor containing an immune receptor tyrosine based inhibitory motif (ITIM). They are designed to cross-link an Fc receptor for IgE (e.g., Fc.epsilon.R1) and an IgG inhibitory receptor (e.g., Fc.gamma.RIIb), thereby inhibiting the IgE-driven mediators released from mast cells and basophils. In addition, the present invention provides nucleic acid molecules encoding the fusion proteins, vectors and host cells for producing the fusion proteins, pharmaceutical compositions comprising the fusion proteins, and methods for ameliorating or reducing the risk of IgE-medicated allergic diseases.

Description of the Invention

BRIEF SUMMARY OF THE INVENTION

The present invention provides fusion proteins that comprise an allergen sequence linked via an IgG hinge region to another polypeptide sequence capable of specifically binding to a native IgG inhibitory receptor containing an immune receptor tyrosine based inhibitory motif (ITIM). They are designed to cross-link an Fc receptor for IgE (e.g., Fc.epsilon.R1) and an IgG inhibitory receptor (e.g., Fc.gamma.RIIb), thereby inhibiting the IgE-driven mediators released from mast cells and basophils (e.g., during the presentation of allergens). In addition, the present invention provides nucleic acid molecules encoding the fusion proteins, vectors and host cells for producing the fusion proteins, pharmaceutical compositions comprising the fusion proteins, and methods for ameliorating or reducing the risk of IgE-medicated allergic diseases.

In one aspect, the present invention provides a fusion protein comprising (i) a first polypeptide sequence capable of specifically binding to a native IgE molecule, (ii) a second polypeptide sequence capable of specifically binding to a native IgG inhibitory receptor comprising an immune receptor tyrosine based inhibitory motif (ITIM), and (iii) an IgG hinge region, wherein the first polypeptide sequence comprises an allergen sequence, and wherein the first polypeptide sequence and the second polypeptide sequence is functionally connected via the IgG hinge region.

In certain embodiments, the allergen sequence within the fusion protein is native cat allergen Fel d1 or a portion thereof. For example, the allergen sequence may comprise both a portion of native cat allergen Fel d1 chain 1 as set forth in SEQ ID NO:18 and a portion of native cat allergen Fel d1 chain 2 as set forth in SEQ ID NO:19.

In certain embodiments, the allergen sequence within the fusion protein is native mite allergen protein Der p1 as set forth in SEQ ID NO:4 or a portion thereof.

In certain embodiments, the allergen sequence within the fusion protein is native peanut allergen Ara has set forth in SEQ ID NO:10 or a portion thereof.

In certain embodiments, the IgG hinge region within the fusion protein is human IgG1 hinge region as set forth in SEQ ID NO:20.

In certain embodiments, the second polypeptide sequence is capable of specifically binding to a low-affinity IgG receptor Fc.gamma.RIIb.

In certain embodiments, the second polypeptide within the fusion comprises the CH2 and CH3 portion of an IgG immunoglobulin heavy chain constant region. For example, the second polypeptide may comprise the CH2 and CH3 portion of human IgG1 immunoglobulin heavy chain constant region as set forth in SEQ ID NO:21.

In certain embodiments, the fusion protein is the protein referred to as "kitcin," which has the amino acid sequence as set forth in SEQ ID NO:17.

In another aspect, the present invention provides nucleic acid molecules comprising nucleotide sequences encoding the fusion proteins described herein.

In certain embodiments, the nucleic acid molecule comprises a nucleotide sequence that encodes the kitcin fusion protein having the amino acid sequence as set forth in SEQ ID NO:17. For example, in certain embodiments, the nucleic acid molecule comprises the nucleotide sequence as set forth in SEQ ID NO:16.

In certain embodiments, the present invention provides vectors comprising and capable of expressing the nucleotide sequence encoding the fusion protein described herein.

In certain embodiments, the vector comprises and is capable of the nucleotide sequence encoding the katcin fusion protein as set forth in SEQ ID NO:17. For example, in certain embodiments, the vector comprises the nucleotide sequence as set forth in SEQ ID NO:16.

In another aspect, the present invention provides host cells transformed with the vectors described herein.

In another aspect, the present invention provides pharmaceutical compositions comprising a fusion protein described herein and a pharmaceutically acceptable ingredient.

In another aspect, the present invention provides methods for ameliorating or reducing the risk of an IgE-mediated allergic disease, comprising administering to a patient in need thereof an effective amount of a fusion protein described herein.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides fusion proteins capable of cross-linking an Fc receptor for an IgE (e.g., Fc.epsilon.R1) and an IgG inhibitory receptor that contains an IMIT (e.g., Fc.gamma.RIIb), thereby inhibiting the IgE-driven mediators released from mast cells and basophils. In addition, the present invention provides nucleic acid molecules encoding the fusion proteins, vectors and host cells for producing the fusion proteins, pharmaceutical compositions comprising the fusion proteins, and methods for ameliorating or reducing the risk of IgE-mediated allergic diseases or other disorders.

In one aspect, the present invention provides a fusion protein comprising (i) a first polypeptide sequence capable of specifically binding to a native IgE molecule, (ii) a second polypeptide sequence capable of specifically binding to a native IgG inhibitory receptor comprising an immune receptor tyrosine based inhibitory motif (ITIM), and (iii) an IgG hinge region, wherein the first polypeptide sequence comprises an allergen sequence, and wherein the first polypeptide sequence and the second polypeptide sequence is functionally connected via the IgG hinge region. Because by specifically binding to the native IgE molecule, the first polypeptide is indirectly bound to an IgE receptor for the native IgE molecule (e.g., Fc.epsilon.RI receptor) via the native IgE molecule, the fusion protein is capable of cross-linking the IgE receptor with the IgG inhibitory receptor.

The first polypeptide sequence of the fusion protein comprises an allergen sequence. The allergen sequence may be a naturally occurring allergen sequence, a portion of a naturally occurring allergen sequence, or a variant of a naturally occurring allergen sequence, that is capable of specifically binding to an allergen-specific IgE molecule.

In certain embodiments, the first polypeptide sequence comprises an allergen sequence with an at least 80%, 85%, 90%, 95%, or 99% sequence identify with a naturally occurring allergen sequence or a portion thereof that is capable of binding to an allergen-specific IgE molecule.

"Sequence identity" is defined as the percentage of amino acid residues in one sequence that are identical with the amino acid residues in another reference polypeptide sequence after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, and not considering any conservative substitutions as part of the sequence identity. The percentage sequence identity values are generated by the NCBI BLAST2.0 software as defined by Altschul et al., (1997), "Gapped BLAST and PSI-BLAST: a new generation of protein database search programs", Nucleic Acids Res., 25:3389-3402 with the parameters set to default values.

The sequences of a large number of allergens are known in the art. Exemplary allergens include mite allergen Der p1 (see, SEQ ID NOS:3 and 4 for its nucleotide and amino acid sequences), cat allergen Fel d1 (see, SEQ ID NOS:5 and 6 for the nucleotide and amino acid sequences of its chain 1, and SEQ ID NOS:7 and 8 for the nucleotide and amino acid sequences of its chain 2), and peanut allergen Ara h (see, SEQ ID NOS:9 and 10 for its nucleotide and amino acid sequences). Numerous other allergens and their sequences are available from the SWISS-PROT database, some of which are listed in Table 1 of U.S. Patent Application Publication No. US 2003/0082190 (which is incorporated herein by reference).

In certain embodiments, the first polypeptide sequence of the fusion protein of the present invention comprises native cat allergen Fel d1 or a portion thereof. For example, in certain embodiments, the first polypeptide sequence comprises both a portion of native cat allergen Fel d1 chain 1 as set forth in SEQ ID NO:18 and a portion of native cat allergen Fed d1 chain 2 as set forth in SEQ ID NO:19.

In certain embodiments, the first polypeptide sequence of the fusion protein of the present invention comprises a sequence having an at least 80%, 85%, 90%, 95%, or 99% sequence identity with native cat allergen Fel d1 or a portion thereof and capable of specifically binding to a cat allergen Fel d1-specific IgE antibody.

In certain embodiments, the first polypeptide sequence of the fusion protein of the present invention comprises both a sequence having an at least 80%, 85%, 90%, 95%, or 99% sequence identity with native cat allergen Fel d1 chain 1 and another sequence having an at least 80%, 85%, 90%, 95%, or 99% sequence identity with native cat allergen Fel d1 chain 2, wherein the first polypeptide is capable of specifically binding to a cat allergen Fel d1-specific IgE antibody.

In certain embodiments, the first polypeptide sequence of the fusion protein of the present invention comprises native mite allergen protein Der p1 as set forth in SEQ ID NO:4 or a portion thereof.

In certain embodiments, the first polypeptide sequence of the fusion protein of the present invention comprises a sequence having an at least 80%, 85%, 90%, 95%, or 99% sequence identity with native mite allergen protein Der p1 set forth in SEQ ID NO:4 or a portion thereof and capable of specifically binding to a mite allergen protein Der p1-specific IgE antibody.

In certain embodiments, the first polypeptide sequence of the fusion protein of the present invention comprises native peanut allergen Ara h as set forth in SEQ ID NO:10 or a portion thereof.

In certain embodiments, the first polypeptide sequence of the fusion protein of the present invention comprises a sequence having an at least 80%, 85%, 90%, 95%, or 99% sequence identity with native peanut allergen Ara h as set forth in SEQ ID NO:10 or a portion thereof and capable of specifically binding to a peanut allergen Ara h-specific IgE antibody.

The second polypeptide sequence in the fusion protein of the present invention is capable of specifically binding to a native IgG inhibitory receptor comprising an ITIM, such as Fc.gamma.RIIb receptor and gp49b1.

In certain embodiments, the second polypeptide sequence comprises the CH2 and CH3 portion of human IgG immunoglobulin heavy chain constant region. In certain embodiments, the second polypeptide sequence comprises the CH2 and CH3 portion of human IgG.sub.1 immunoglobulin heavy chain constant region as set forth in SEQ ID NO:21. In certain other embodiments, the second polypeptide sequence comprises the CH2 and CH3 portion of human IgG.sub.2, IgG.sub.3, or IgG.sub.4 immunoglobulin heavy chain constant region.

In certain embodiments, the second polypeptide sequence comprises a sequence having an at least 80%, 85%, 90%, 95%, or 99% sequence identity with the CH2 and and CH3 portion of human IgG immunoglobulin heavy chain constant region. In certain embodiments, the second polypeptide sequence comprises having an at least 80%, 85%, 90%, 95%, or 99% sequence identity with the CH2 and CH3 portion of human IgG.sub.1 immunoglobulin heavy chain constant region as set forth in SEQ ID NO:21 and capable of specifically binding to a native IgG inhibitory receptor containing an ITIM (e.g., Fc.epsilon.RIIb). In certain other embodiments, the second polypeptide sequence comprises a sequence having an at least 80%, 85%, 90%, 95%, or 99% sequence identity with the CH2 and CH3 portion of human IgG.sub.2, IgG.sub.3, or IgG.sub.4 immunoglobulin heavy chain constant region and capable of specifically binding to a native IgG inhibitory receptor containing an ITIM (e.g., Fc.epsilon.RIIb).

The first and second polypeptides of the fusion protein of the present invention are functionally connected via an IgG hinge region.

The term "IgG hinge region" refers to the hinge region of a native IgG immunoglobulin heavy chain constant region, a portion of the hinge region of a native IgG immunoglobulin heavy chain constant region that consists of at least 10 consecutive amino acid residues (e.g., 10, 11, 12, 13, or 14 amino acid residues), or a sequence that has an at least 80% (e.g., 80%, 85%, 90%, 95%, or 99%) sequence identity with the hinge region of a native IgG immunoglobulin heavy chain constant region or a portion thereof that consists of at least 10 consecutive amino acid residues (e.g., 10, 11, 12, 13, or 14 amino acid residues).

The first polypeptide of the fusion protein of the present invention is "functionally connected" to the second polypeptide if in the resulting fusion protein, the first polypeptide is still capable of specifically binding to a native IgE molecule, and the second polypeptide is still capable of specifically binding to a native IgG inhibitory receptor comprising an ITIM.

In certain embodiments, the first and second polypeptides of the fusion protein of the present invention are functionally connected via the hinge region of an IgG.sub.1 immunoglobulin. For example, the first and second polypeptides may be functionally connected via the 15 amino acid hinge region of human IgG.sub.1 immunoglobulin as set forth in SEQ ID NO:20.

In certain embodiments, the first and second polypeptides of the fusion protein of the present invention are functionally connected via a portion of the hinge region of an IgG.sub.1 immunoglobulin (e.g., human IgG.sub.1 immunoglobulin) that consist of at least 10, 11, 12, 13, or 14 amino acid residues.

In certain embodiments, the first and second polypeptides of the fusion protein of the present invention are functionally connected via a sequence having an at least 80%, 85%, 90%, 95%, 99% of the hinge region of an IgG.sub.1 immunoglobulin (e.g., human IgG.sub.1 immunoglobulin) or a portion thereof that consists of at least 10 consecutive amino acid residues (e.g., 10, 11, 12, 13, or 14 amino acid residues).

In certain embodiments, the first and second polypeptides of the fusion protein of the present invention are functionally connected via the hinge region of an IgG.sub.2, IgG.sub.3, or IgG.sub.4 immunoglobulin. For example, the first and second polypeptides may be functionally connected via the hinge region of human IgG.sub.2, IgG.sub.3, or IgG.sub.4 immunoglobulin as set forth in SEQ ID NO:22, 23, or 24.

In certain embodiments, the first and second polypeptides of the fusion protein of the present invention are functionally connected via a portion of the hinge region of an IgG.sub.2, IgG.sub.3, or IgG.sub.4 immunoglobulin (e.g., human IgG.sub.2, IgG.sub.3, or IgG.sub.4 immunoglobulin) that consist of at least 10, 11, 12, 13, or 14 amino acid residues.

In certain embodiments, the first and second polypeptides of the fusion protein of the present invention are functionally connected via a sequence having an at least 80%, 85%, 90%, 95%, 99% of the hinge region of an IgG.sub.2, IgG.sub.3, or IgG.sub.4 immunoglobulin (e.g., human IgG.sub.2, IgG.sub.3, or IgG.sub.4 immunoglobulin) or a portion thereof that consists of at least 10 consecutive amino acid residues (e.g., 10, 11, 12, 13, or 14 amino acid residues).

In certain embodiments, the first and second polypeptides of the fusion protein of the present invention may be linked with an IgG hinge region so that the first polypeptide is located N-terminus to the second polypeptide. In certain other embodiments, the first polypeptide may be linked to the second polypeptide via an IgG hinge region so that it is C-terminus to the second polypeptide.

In certain embodiments, the fusion proteins of the present invention is capable of (1) indirect binding of a native high-affinity Fc.epsilon.R1 receptor via directly binding between a native IgE molecule to which the native high-affinity Fc.epsilon.R1 receptor binds and the first polypeptide sequence in the fusion protein, and (2) direct binding of a native low-affinity Fc.gamma.RIIb receptor via the second polypeptide sequence in the fusion protein.

In certain embodiments, the fusion protein of the present invention comprises an allergen sequence and the CH2 and CH3 portion of human IgG.sub.1 heavy chain constant region linked by the hinge region of human IgG.sub.1 heavy chain constant region (see, FIG. 1, see Original Patent). The cDNA sequence encoding the hinge-CH2--CH3 portion of the human IgG.sub.1 heavy chain constant region and the amino acid sequence of this portion of the human IgG.sub.1 heavy chain constant region are set forth in SEQ ID NOS:1 and 2, respectively.

In certain embodiments, the fusion protein of the present invention comprises mite allergen protein (or a portion thereof) and the CH2 and CH3 portion of human IgG.sub.1 heavy chain constant region linked by the hinge region of human IgG.sub.1 heavy chain constant region (see, FIG. 2, see Original Patent).

In certain embodiments, the fusion protein of the present invention comprises cat allergen Fel d1 protein (or a portion thereof) and the CH2 and CH3 portion of human IgG.sub.1 heavy chain constant region linked by the hinge region of human IgG.sub.1 heavy chain constant region (see, FIG. 3, see Original Patent).

In certain embodiments, the fusion protein of the present invention comprises peanut allergen Ara protein (or a portion thereof) and the CH2 and CH3 portion of human IgG.sub.1 heavy chain constant region linked by the hinge region of human IgG.sub.1 heavy chain constant region (see, FIG. 4, see Original Patent).

The fusion proteins of the present invention are capable of specific binding of a native IgE molecule via its first polypeptide and specific binding of a native IgG inhibitory receptor comprising an ITIM via its second polypeptide. Such specific binding may be tested using any known assays, such as competitive binding assays including RIAs and ELISAs. Protein-protein complexes (e.g., complexes formed between the fusion protein and a native IgE molecule, between the fusion protein and a native IgG inhibitory receptor comprising an ITIM, and among the fusion protein, a native IgE molecule, and a native IgG inhibitory receptor comprising an ITIM) can be identified and isolated using various methods such as filtration, centrifugation, and flow cytometry. The assays may be performed using a purified protein of interest (e.g., a first or second polypeptide sequence of a fusion protein of the present invention, a fusion protein of the present invention, a native IgE molecule, or a native IgG inhibitory receptor comprising an ITIM), intact cells expressing a protein of interest, or cell lysate containing a protein of interest. The protein of interest may be immobilized or labeled prior to or during the assays.

The fusion proteins of the present invention may be prepared using standard techniques of recombinant DNA technology.

In a related aspect, the present invention provides a nucleic acid molecule comprising a nucleotide sequence encoding the fusion proteins described herein. In certain embodiments, the nucleic acid molecule comprises a nucleotide sequence encoding a fusion protein that comprises an allergen sequence capable of specifically binding to a native IgE molecule functionally linked to a second polypeptide sequence capable of specifically binding to a native IgG inhibitory receptor comprising an ITIM (e.g., Fc.epsilon.RIIb) via an IgG hinge region.

In certain embodiments, the nucleic acid molecule comprises a nucleotide sequence encoding a fusion protein that comprises a native allergen polypeptide sequence or a portion thereof linked to the CH2--CH3 portion of human IgG.sub.1 heavy chain constant region via the hinge region of human IgG.sub.1 heavy chain constant region.

In certain embodiments, the nucleic acid molecule comprises a nucleotide sequence encoding the katcin fusion protein that comprises a portion of cat allergen Fel d1 and the hinge-CH2--CH3 portion of human IgG1 as set forth in SEQ ID NO:17.

In certain embodiments, the nucleic acid molecule comprises a nucleotide sequence encoding the katcin fusion protein and has a nucleotide sequence as set forth in SEQ ID NO:16.

In certain embodiments, the nucleic acid molecule comprises a nucleotide sequence encoding a fusion protein of the present invention and is capable of specifically binding to SEQ ID NO:16 or its full length complement under stringent hybridization and wash conditions.

"Stringent hybridization and wash conditions" are defined as hybridization in 50% formamide, 6.times.SSC (0.75 M NaCl, 0.075 M sodium citrate), 50 mM sodium phosphate (pH 6.8), 0.1% sodium pyrophosphate, 5.times. Denhardt's solution, sonicated salmon sperm DNA (100 .mu.g/ml), 0.5% SDS, and 10% dextran sulfate at 42.degree. C. for at least 12 hours, with washes in 2.times.SSC (sodium chloride/sodium citrate) containing 0.1% SDS at 42.degree. C. for 30 minutes twice and 0.2.times.SSC containing 0.1% SDS at 42.degree. C. for 30 minutes.

The nucleic acid molecules encoding the fusion proteins of the present invention may be obtained via standard recombinant DNA techniques, chemical synthesis, or the combination thereof. For example, the coding sequences of various portions of the fusion proteins (e.g., first and second polypeptide sequences, and IgG hinge regions) may be obtained from natural sources or synthesized. Such sequences may be appropriately linked to produce the nucleic acid molecules encoding the fusion proteins of the present invention using various commercially available cloning and/expression vectors (e.g., those available from companies such as Invitrogen, San Diego, Calif.).

In certain embodiments, the present invention provides vectors comprising and capable of expressing the nucleotide sequences encoding the fusion proteins described herein. In certain embodiments, the vector comprises a nucleic acid sequence encoding the katcin fusion protein as set forth in SEQ ID NO:17. In certain embodiments, the vector comprises a nucleic acid sequence encoding the katcin fusion protein and has a nucleotide sequence as set forth in SEQ ID NO:16.

The vectors of the present invention include those useful for recombinant production in E. coli, S. cerevisiae strains of yeast, a baculovirus expression system for production in insect cells, fungal cells, avian cells, mammalian cells such as Chinese Hamster Ovary cells, and plant cells.

In one aspect, the present invention provides host cells transformed with the vectors described herein. Host cells useful for transformation with the vectors of the present invention include prokaryotic and eukaryotic host cells such as bacterial cells (e.g., E. coli cells), yeast cells (e.g., S. cerevisiae cells), insect cells, fungal cells, insect cells, mammalian cells, avian cells, and cells of higher plants.

Construction of expression systems suitable for desired hosts are known in the art. For recombinant production of the fusion protein of the present invention, the DNA encoding the fusion protein is suitably ligated into the expression vector of choice and then used to transform the compatible host. The transformed host cells are then cultured and maintained under conditions appropriate for expression of the foreign gene. The fusion protein of the present invention thus produced is recovered from the culture, either by lysing the cells or from the culture medium as appropriate and known in the art. Such fusion protein may be further purified if needed using standard techniques known in the art.

Alternatively, the fusion proteins of the present invention may be produced by chemical synthesis. Such methods are well known in the art and employ either solid or solution phase synthesis methods. Information about chemical synthesis of proteins may be found, for example, in Stewart and Young, Solid Phase Peptide Synthesis, 2nd Ed., Pierce Chemical Co., Rockford, Ill., 1984; Barany and Merrifield, The Peptide: Analysis Synthesis, Biology, editors E. Gross and J. Meienhofer, Vol. 2, Academic Press, New York, 1980; and Bodansky, Principles of Peptide Synthesis, Springer-Verlag, Berlin, 1984.

In one aspect, the present invention provides pharmaceutical compositions that comprise the fusion proteins of the present invention and pharmaceutically acceptable ingredients, such as physiologically acceptable excipients, additives, carriers or diluents. Suitable physiological acceptable ingredients are described in Remington's Pharmaceutical Sciences, 18.sup.th Edition, Mack Publishing Co., Easton, Pa., 1990. Additional exemplary physiological acceptable ingredients include coloring, stabilizing agents, osmotic agents, and antibacterial agents.

The fusion proteins of the present invention may be used as vaccines, and thus also referred to as "allergen vaccine proteins." Accordingly, in one aspect, the present invention provides methods for ameliorating or reducing the risk of an IgE-mediated allergic disease, comprising administering to a patient in need thereof an effective amount of the fusion proteins described herein. In certain embodiments, the present invention provides a method for ameliorating or reducing the risk of cat allergy, comprises administering to a patient in need thereof an effective amount of the katcin fusion protein with the amino acid sequence as set forth in SEQ ID NO:17.

A disease is "ameliorated" if the symptoms of the disease are alleviated, the extent of the disease is diminished, the progression of the disease is delayed or slowed, the disease state is ameliorated or palliated, and/or partial or total remission occurs.

The risk of a disease is "reduced" if the likelihood of a patient to have the disease is reduced, or the onset of the disease in a patient is delayed.

A "patient in need" refers to a patient already with an allergic disease or is prone to have an allergic disease. The patient may be a human or a non-human mammalian subject.

An "effective amount" is an amount sufficient to effect beneficial or desired therapeutic (including preventive) results. An effective amount can be administered in one or more administrations.

The IgE-mediated allergic diseases that the fusion proteins of the present invention are useful in ameliorating or reducing the risk of the diseases include, but are not limited to, allergic asthma, allergic rhinitis, hay fever, food allergy, such as those caused by peanut or other nuts, shellfish, milk, fish, soy, wheat, and egg, atopic dermatitis, pet allergy (e.g., cat allergy and dog allergy), eczema, drug allergy, chronic urticaria, ear infections associated with allergy, angioedema, allergy caused by pollen, mold, dust mite droppings, insect stings, or cockroaches or other insects, and anaphylactic shock.

The fusion proteins or pharmaceutical compositions of the present invention may be administered by any means that enables the fusion proteins to reach the targeted cells. These methods include, but are not limited to, oral, topical, transdermal, subcutaneous, intravenous, intramuscular, intra-arterial, intranasal, intrapulmonary, and intraparenteral modes of administration. The fusion proteins may be administered singularly or in combination with other compounds useful for anti-IgE therapy or allergen immunotherapy.

In certain embodiments, the fusion proteins or pharmaceutical compositions of the present invention are administered by inhalation. In certain other embodiments, the fusion proteins or pharmaceutical compositions of the present invention are administered by injection.

For parenteral administration, the fusion proteins of the invention can be, for example, formulated as a solution, suspension, emulsion or lyophilized powder in association with a physiologically acceptable parenteral vehicle such as water, saline, Ringer's solution, dextrose solution, and 5% human serum albumin. Liposomes and nonaqueous vehicles such as fixed oils may also be used. For example, a parenteral composition suitable for administration by injection is prepared by dissolving 1.5% by weight of active ingredient in 0.9% sodium chloride solution. The compositions of the present invention may be administered as a single dose or in multiple doses.

The compositions of the present invention may be provided in the form of an oral liquid, tablet, or capsule, nasal spray, aerosol, suspension, solution, emulsion, and/or eye drop. The appropriate dosage can be extrapolated from the dosages that indicate efficacy in vitro or in animal studies. The dosage administered varies depending upon factors such as: pharmacodynamic characteristics; its mode and route of administration; age, health, and weight of the recipient; nature and extent of symptoms; type of concurrent treatment; and frequency of treatment. Usually, the dosage of the fusion protein can be about 0.01 to 100 mg/kg of body weight, such as about 0.1 to 100, 0.05 to 50, or 1.0 to 10 mg/kg of body weight. The fusion proteins or pharmaceutical compositions may be administered to an individual per day in divided doses one or more times per day to obtain desired results.
 

Claim 1 of 2 Claims

1. A fusion protein comprising (i) a first polypeptide sequence capable of specifically binding to a native IgE molecule, (ii) a second polypeptide sequence capable of specifically binding to a native IgG inhibitory receptor comprising an immune receptor tyrosine based inhibitory motif (ITIM), and (iii) an IgG hinge region, wherein the first polypeptide sequence comprises an allergen sequence, the first polypeptide sequence and the second polypeptide sequence is functionally connected via the IgG hinge region, and the fusion protein comprises the amino acid sequence as set forth in SEQ ID NO:17.

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