Anti-inflammatory agents

Abstract

To provide therapies that provide for prevention or treatment of inflammatory responses associated with or caused by aberrant activation of the immune system.SOLUTION: The invention relates to anti-inflammatory peptides, to pharmaceutical compositions comprising the anti-inflammatory peptides, and to uses thereof for treatment of inflammation including, but not limited to inflammation associated with immune activation.SELECTED DRAWING: None

Description

[Technical Field]

[0001] Related Applications This application claims priority from Australian provisional application AU2019901968, the entire contents of which are incorporated herein by reference.

[0002] FIELD OF THE INVENTION The present invention relates to anti-inflammatory peptides, pharmaceutical compositions containing same, and their use for the treatment of inflammation, including but not limited to inflammation associated with an immune response. [Background technology]

[0003] The reference to prior art herein is not an admission or suggestion that this prior art forms part of the common general knowledge in any jurisdiction, or that this prior art could reasonably be expected to be understood, considered relevant, and / or combined with other prior art by a person skilled in the art.

[0004] Inflammatory processes can be associated with or caused by aberrant activation of the immune system, resulting in a self-perpetuating cycle of inflammation and immune activation and subsequent immune responses that can be very debilitating.

[0005] These processes can result in acute and chronic inflammation, including that observed in cutaneous, musculoskeletal, intestinal or pulmonary disorders or diseases.

[0006] Of particular concern are processes associated with or caused by abnormal immune cell responses, including abnormal activation of macrophages or dendritic cells.One particular example is psoriasis, a particularly problematic and incurable condition characterized by epidermal hyperplasia and excessive inflammatory infiltration of the skin, estimated to affect 1-4% of the Western population.

[0007] Other inflammatory diseases and disorders may be idiopathic with suspected immune involvement, examples of skin diseases include contact and atopic dermatitis, polymorphous light eruption, and vitiligo.

[0008] Some of the current treatments have significant immunosuppressive side effects or other adverse events.

[0009] There is a need for therapies that provide prevention or treatment of inflammatory responses.

[0010] There is a need for therapies that provide prevention or treatment of inflammatory responses associated with or caused by aberrant activation of the immune system. Summary of the Invention

[0011] The present invention seeks to address one or more of the above limitations or needs. Accordingly, in a first aspect, the present invention provides an anti-inflammatory peptide for preventing or treating inflammation in an individual in need thereof, the peptide comprising: the amino acid sequence TVLTVV (SEQ ID NO: 1) or the amino acid sequence LLLLLTVLTVV (SEQ ID NO: 2) or a functional variant thereof The present invention provides a peptide comprising, consisting of, or consisting essentially of (wherein the peptide consists of less than 21 amino acid residues, preferably 6 to 20 amino acid residues).

[0012] In a preferred embodiment, the present invention provides a peptide consisting of the amino acid sequence of SEQ ID NO:1.

[0013] In a further preferred embodiment, the present invention provides a peptide consisting of the amino acid sequence of SEQ ID NO:2.

[0014] In another embodiment of the invention, the anti-inflammatory peptide comprises, consists of, or consists essentially of the amino acid sequence of SEQ ID NO: 30, or a functional variant or fragment thereof. More preferably, the peptide consists of the amino acid sequence of SEQ ID NO: 30.

[0015] In a further aspect of the invention, - a peptide comprising or consisting of the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 2; and - one or more further amino acids flanking the sequences of SEQ ID NO: 1 and 2, with the proviso that the one or more further amino acids do not define the sequence MTPGTQSPFF at the N-terminal position adjacent to the sequence of SEQ ID NO: 1 or 2, or the sequence TGSGHASSTP at the C-terminal position adjacent to the sequence of SEQ ID NO: 1 or 2. The present invention provides a polypeptide comprising the above, which comprises at least 20 amino acid residues, preferably 20 to 1000 residues.

[0016] In another aspect of the present invention, - a peptide as defined herein, preferably a peptide comprising the sequence of any one of SEQ ID NO: 1, 2 (preferably SEQ ID NO: 3 or 30), or a functional variant or fragment thereof; - In some cases, Modifications to enhance the solubilization of the peptide in biological fluids, tissues or formulations, a moiety for facilitating the entry of the peptide into cells, and Modifications or moieties to allow detection of the peptide one or more modifications selected from Anti-inflammatory peptides are provided, comprising:

[0017] In an embodiment of this aspect of the invention, the peptide preferably comprises an amino acid sequence that is at least 90%, at least 95% or at least 98% identical to the sequence of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3 or SEQ ID NO:30.

[0018] The present invention provides peptides consisting of any one of SEQ ID NOS: 1-65, or functional variants or fragments thereof. Functional variants or fragments generally exhibit at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 70%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% amino acid identity and retain their function in treating or preventing inflammatory responses as described herein, including in the Examples.

[0019] In some embodiments, the modification to enhance peptide solubilization is in the form of an amino acid residue (or series thereof), polymer, or other functional moiety that enhances peptide solubilization. In certain embodiments, the modification to enhance peptide solubilization comprises polyethylene glycol (PEG) of various lengths. For example, the PEG may be at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 1 PEG may contain 4, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100 or more ethylene glycol units. Alternatively, the PEG may be free of ethylene glycol units.

[0020] In a further embodiment, modifications to enhance solubilization of the peptide include one or more charged amino acid residues, preferably one or more cationic amino acid residues, more preferably one or more lysine (K), arginine (R), or histidine (H).

[0021] It will be appreciated that modifications to enhance peptide solubilization can be located in the N-terminal or C-terminal regions of the peptide. Preferably, the modifications are located in the N-terminal region of the peptide.

[0022] One or more modifications to improve the solubilization of the peptide can be used, for example, a combination of a polymer and a cationic amino acid residue.

[0023] In either embodiment, the moiety for detecting the peptide is a label known in the art to facilitate detection of the peptide in vivo or in vitro. The label can be a fluorescent label such as GFP, YFP, BFP, CFP, Y-FAST, AF594, mCherry, or dsRed. The label can be biotin. Alternatively, the label can be a metal label such as (Gd)Dota.

[0024] In yet another embodiment, the peptides described herein may contain synthetic isomers of amino acid residues. In a particular embodiment, the synthetic amino acid isomer is photoleucine.

[0025] In some embodiments, the peptides described herein may contain a cell-penetrating peptide (CPP) conjugated to the N-terminal or C-terminal region to facilitate or assist the peptides herein in entering cells. Non-limiting examples of cell-penetrating peptides are well known in the art and are described herein. Non-limiting examples include Tat and Tat-based peptides.

[0026] In a particularly preferred embodiment of the invention, the anti-inflammatory peptide comprises, consists of, or consists essentially of the peptide defined in SEQ ID NO: 12: KKKK-PEG-MTPGTQSPFFLLLLLTVLTVV. More preferably, the peptide is as defined in SEQ ID NO: 12.

[0027] In certain embodiments, the peptides of the present invention may be further conjugated to anti-inflammatory peptides known in the art.

[0028] In some embodiments of the present invention, the peptides described herein include a linker comprising glycine or serine repeats of various lengths. The peptides of the present invention and the linker can be directly or indirectly linked. Preferably, the linker is located at the N-terminus of the peptide, although it is understood that the linker can also be located at the C-terminus of the peptide. In some embodiments, the linker comprises the sequence GGG, and the peptides of the present invention comprise or consist of KKKK-GGG-MTPGTQSPFFLLLLLTVLTVV (SEQ ID NO: 22) or a functional variant thereof, including KKKK-GGG-TPGTQSPFFLLLLLTVLTVV (SEQ ID NO: 23).

[0029] The anti-inflammatory peptide may comprise, consist of, or consist essentially of a peptide set forth in any of Tables A-E herein.

[0030] In one embodiment, there is provided a pharmaceutical composition comprising any one of the peptides described herein (including any one of the peptides of SEQ ID NOs: 1-66) or a functional variant thereof, and a pharmaceutically acceptable carrier, diluent or excipient, or a pharmaceutically acceptable salt thereof.

[0031] In certain embodiments, the pharmaceutical composition comprises a peptide comprising or consisting of the sequence of SEQ ID NO: 1, 2, 3, 12 or 30 and a pharmaceutically acceptable carrier, diluent or excipient, or a pharmaceutically acceptable salt thereof.

[0032] In yet another embodiment, the composition comprising a pharmaceutically effective amount of a peptide or any one of the peptides described herein or a functional variant thereof, or a pharmaceutically acceptable salt thereof, does not contain granulocyte-macrophage colony-stimulating factor (GM-CSF).

[0033] The present invention also provides compositions comprising a peptide described herein and an additional active agent, which can be an anti-inflammatory agent, an antisteroidal anti-inflammatory drug (NSAID), a disease-modifying antirheumatic drug (DMARD), or other active agent useful in treating an inflammatory disorder in need of treatment (e.g., an agent for treating psoriasis, dermatitis, rheumatoid arthritis, as the case may be).

[0034] In certain embodiments, the anti-inflammatory agent is a corticosteroid. Non-limiting examples of corticosteroids suitable for use in the present invention include aristocort, decadron, mometasone, cotrone, triamcinolone, cortisone, prednisone, methylprednisolone, and betamethasone dipropionate.

[0035] Alternatively, the additional anti-inflammatory agent is an antigen-binding protein for inhibiting the activity of a pro-inflammatory molecule. Preferably, the antigen-binding protein is an antibody or an antigen-binding fragment thereof. Typically, the antigen-binding protein is an antibody, for example, a monoclonal antibody.

[0036] In certain embodiments, antigen binding proteins (e.g., monoclonal antibodies) for inhibiting the activity of inflammatory molecules bind to anti-inflammatory molecules, thereby inhibiting the activity of the molecules in promoting an inflammatory response. The antigen binding proteins may bind to receptors for inflammatory molecules, thereby inhibiting the activity of the inflammatory molecules. Non-limiting examples of antigen binding proteins or monoclonal antibodies suitable for use in accordance with the present invention include those capable of inhibiting the activity of inflammatory molecules including interleukin-17 (IL-17), interleukin-12 (IL-12), interleukin-6 (IL-6), interleukin-22 (IL-22), interleukin-23 (IL-23), interleukin-36 (IL-36), and tumor necrosis factor alpha (TNFα). Examples of such antigen binding proteins or monoclonal antibodies include brodalumab, ixekizumab or secukinumab (to inhibit IL-17), tocilizumab or siltuximab (to inhibit IL-6), guselkumab or tildrakizumab or mirikizumab or brazikumab or risankizumab or ustekinumab (to inhibit IL-23), fezakinumab (to inhibit IL-22), etanercept, infliximab, adalimumab, certolizumab, certolizumab pegol or golimumab (to inhibit the activity of TNFα).

[0037] In embodiments where the additional active agent is a DMARD, the DMARD can be thalidomide, lenalidomide, methotrexate.

[0038] In still further embodiments, the additional active agent may be an agent known to be useful in the treatment of inflammatory disorders, non-limiting examples of which include calcipotriol, anakinra, inflammasome inhibitors, baricitinib, abrocitinib, upadacitinib (JAK inhibitors), tofacitinib, ruxolitinib, antihistamines, topical tacrolimus (calcineurin inhibitors), and crisaborole (phosphodiesterase 4 (PDE4) inhibitors).

[0039] Compositions comprising the peptides described herein and an additional anti-inflammatory agent can provide greater anti-inflammatory effects than the peptides alone. For example, the effects of the peptides described herein and a corticosteroid are synergistic compared to the effects of each component alone. Thus, the present invention also encompasses synergistic compositions comprising the peptides described herein and an additional anti-inflammatory agent, wherein the additional anti-inflammatory agent is preferably a corticosteroid, a DMARD, or an antigen-binding protein for binding to and inhibiting the activity of an inflammatory molecule.

[0040] In any embodiment, a composition comprising a peptide described herein and an additional anti-inflammatory agent may be formulated for administration in the form of a tablet, syrup or solution, inhalant or nasal spray, injection (including formulations for intradermal or subcutaneous injection, mesotherapy / microneedling), patch, or as a cream, ointment, spray, lotion, or gel.

[0041] In another embodiment, there is provided a nucleic acid encoding a peptide as defined herein, or a fragment thereof, comprising a DNA or RNA sequence, including one having an open reading frame, that encodes a therapeutic polypeptide and that is capable of being expressed under appropriate conditions as one of the therapeutic peptides of the invention. Alternatively, the invention provides a vector or plasmid comprising the nucleic acid or a fragment thereof.

[0042] In a further aspect of the invention, there is provided a method of preventing or treating inflammation or an inflammatory disorder in an individual, comprising administering to an individual in need thereof a pharmaceutically effective amount of a peptide or composition as described herein, thereby preventing or treating inflammation in said individual.

[0043] The inflammation may be in or on a tissue selected from skin or mucosal tissue, musculoskeletal tissue, lung tissue, or intestinal tissue. In certain preferred embodiments, the method of the invention is for treating inflammation in dermal tissue.

[0044] The inflammation can be a symptom of a disease or condition selected from the group consisting of an inflammatory skin disease or disorder, a musculoskeletal disease or disorder, a pulmonary disease or disorder, or an intestinal disease or disorder.

[0045] Inflammatory disorders and related conditions that may be treated or prevented by use of the peptides or compositions described herein include rheumatoid arthritis, bronchitis, contact dermatitis, atopic dermatitis, psoriasis, seborrheic dermatitis, eczema, allergic dermatitis, polymorphous light eruption, inflammatory skin diseases, folliculitis, alopecia, vitiligo, poison ivy, insect bites, inflammatory acne, and the like. inflammation), irritation induced by external factors (including but not limited to chemicals, trauma, pollutants (e.g., cigarette smoke) and sun exposure), secondary conditions resulting from inflammation (including but not limited to xeroderma, hyperkeratosis, pruritus, post-inflammatory hyperpigmentation, scarring, pulmonary inflammatory conditions (including influenza), lupus, rheumatoid arthritis, multiple sclerosis, sarcoidosis and scleroderma), hypersensitivity pneumonitis, wound healing, and inflammatory bowel disease (including colitis and Crohn's disease), or inflammation resulting from transplantation (graft versus host disease, GVHD).

[0046] In a preferred embodiment, when the inflammatory disorder and related condition is an inflammatory skin disorder, the disorder may be contact dermatitis, atopic dermatitis, allergic dermatitis, or psoriasis. In such an embodiment, administration of a peptide or composition described herein minimizes one or more symptoms selected from the group consisting of erythema, skin thickness, scaling, and inflammatory edema.

[0047] In another embodiment, the inflammation to be treated is associated with TNF, IL-6 or IL-17 or IL-12 / 23 production. Thus, administration of the peptides or compositions described herein may minimize the release of inflammatory cytokines, preferably TNF, IL-6, IL-17 and IL-12 / 23.

[0048] In another embodiment, administration of a peptide or composition described herein minimizes activation of lymphocytes, neutrophils or mast cells or antigen presenting cells, preferably macrophages or dendritic cells, at the site of inflammation.

[0049] In another embodiment, there is provided a method of modulating an immune response in an individual, comprising administering to an individual in need thereof a pharmaceutically effective amount of a peptide or composition described herein, thereby modulating the immune response in the individual. Modulation of the immune response is understood to be an effect observed in any of the examples provided herein or as would be understood by one of skill in the art in response to a peptide or composition of the invention.

[0050] In embodiments, the immune response is an adaptive immune response or an innate immune response. When the immune response is an innate immune response, the innate immune response is associated with the activation of antigen-presenting cells, preferably macrophages or dendritic cells.

[0051] In any embodiment, the methods of the present invention may further comprise the administration of an additional active agent, including an anti-inflammatory agent, an antisteroidal anti-inflammatory drug (NSAID), a disease-modifying antirheumatic drug (DMARD), or other active agent useful in treating the inflammatory disorder in need of treatment (e.g., an agent for treating psoriasis, dermatitis, or in some cases rheumatoid arthritis), or the administration of an additional treatment (e.g., the use of phototherapy).

[0052] In certain embodiments, the method includes administering an additional anti-inflammatory agent in the form of a corticosteroid. Non-limiting examples of corticosteroids suitable for use in the methods of the invention include aristocort, decadron, mometasone, cotrone, triamcinolone, cortisone, prednisone, methylprednisolone, and betamethasone dipropionate.

[0053] Alternatively, the method may include administering an anti-inflammatory agent that is an antigen-binding protein to inhibit the activity of inflammatory molecules. Preferably, the antigen-binding protein is an antibody or an antigen-binding fragment thereof. Typically, the antigen-binding protein is an antibody, for example, a monoclonal antibody. The antigen-binding protein (e.g., monoclonal antibody) may be an antigen-binding protein or a monoclonal antibody that has the ability to inhibit the activity of inflammatory molecules, including interleukin-17 (IL-17), interleukin-12 (IL-12), interleukin-6 (IL-6), interleukin-22 (IL-22), interleukin-23 (IL-23), interleukin-36 (IL-36), and tumor necrosis factor alpha (TNFα). Examples of such antigen binding proteins or monoclonal antibodies include brodalumab, ixekizumab or secukinumab (to inhibit IL-17), tocilizumab or siltuximab (to inhibit IL-6), guselkumab or tildrakizumab or mirikizumab or brazikumab or risankizumab or ustekinumab (to inhibit IL-23), fezakinumab (to inhibit IL-22), etanercept, infliximab, adalimumab, certolizumab, certolizumab pegol or golimumab (to inhibit the activity of TNFα).

[0054] In embodiments where the additional active agent is a DMARD, the DMARD can be thalidomide, lenalidomide, methotrexate.

[0055] In still further embodiments, the additional active agent may be an agent known to be useful in the treatment of inflammatory disorders, non-limiting examples of which include calcipotriol, anakinra, inflammasome inhibitors, baricitinib, abrocitinib, upadacitinib (JAK inhibitors), tofacitinib, ruxolitinib, antihistamines, topical tacrolimus (calcineurin inhibitors), and crisaborole (phosphodiesterase 4 (PDE4) inhibitors).

[0056] In another embodiment, - prevention or treatment of inflammation, preferably inflammation associated with abnormal immune activation, possibly contact dermatitis, atopic dermatitis, allergic dermatitis or psoriasis; - modulation of the immune response, preferably the innate immune response; - TCRαβ + Decreased levels of T cells; - Decreased levels of TCRγδ+ T cells; - regulatory cells, preferably regulatory CD4 + an increase in T cells; or - inhibiting the activation of neutrophils, mast cells or antigen-presenting cells, preferably macrophages or dendritic cells, at the site of inflammation There is provided the use of a peptide as described herein in the manufacture of a medicament for

[0057] In a further embodiment, the medicament may further comprise an anti-inflammatory agent, preferably a corticosteroid, for preventing or treating an inflammatory response. In yet another embodiment, the medicament may further comprise an antigen binding protein for binding to an inflammatory molecule, preferably selected from the group consisting of interleukin 17 (IL-17), interleukin 12 (IL-12), interleukin 6 (IL-6), interleukin 23 (IL-23), interleukin 36 (IL-36), and tumor necrosis factor (TNF).

[0058] In yet another embodiment, - prevention or treatment of inflammation, preferably inflammation associated with abnormal immune activation, more preferably contact dermatitis, atopic dermatitis, allergic dermatitis or psoriasis; - modulation of the immune response, preferably the innate immune response; - TCRαβ + Decreased levels of T cells; - Decreased levels of TCRγδ+ T cells; - regulatory cells, preferably regulatory CD4 + an increase in T cells; or - inhibiting the activation of neutrophils, mast cells or antigen-presenting cells, preferably macrophages or dendritic cells, at the site of inflammation For, - use of a peptide as described herein in the manufacture of a first medicament; and - the use of corticosteroids for the prevention or treatment of inflammatory reactions in the manufacture of a second medicine; or - use of an antigen-binding site for inhibiting the activity of a pro-inflammatory molecule, preferably selected from the group consisting of interleukin 6 (IL-6), interleukin 17 (IL-17), interleukin 12 (IL-12), interleukin 23 (IL-23), interleukin 36 (IL-36) and tumor necrosis factor (TNF), in the manufacture of a second medicament. is provided.

[0059] In another embodiment, there is provided a pharmaceutically effective amount of a peptide or composition described herein for use in the prevention or treatment of inflammation, preferably inflammation associated with aberrant immune activation.

[0060] In any embodiment, the peptides or compositions described herein are suitable for administration intradermally, intraarticularly, intramuscularly, intraperitoneally, intravenously, subcutaneously, intranasally, epidurally, orally, sublingually, intracerebrally, intralymphatic, intratracheally, intravaginally or transdermally, rectally, by inhalation, or topically, particularly to the ear, nose, eye or skin. In preferred embodiments, the peptides or compositions described herein are administered topically, intranasally, or by inhalation.

[0061] As used herein, unless the context requires otherwise, the term "comprise", and variations of that term such as "comprising", "comprises" and "comprised", are not intended to exclude additional additives, ingredients, integers or steps.

[0062] Further aspects of the present invention and further implementations of the aspects described in the preceding paragraphs will become apparent from the following description, given by way of example with reference to the accompanying drawings, in which: [Brief explanation of the drawings]

[0063] [Figure 1] Peptide administration accelerates skin tumor growth. Mice were subcutaneously injected with a mixture containing 5 nmol of peptide RP23 (SEQ ID NO: 12) (solid line), PBS, or liposome control (dotted line) 21 and 7 days before UV13-1 tumor cell implantation. When peptide RP23 was included in the vaccine formulation, tumors grew faster (A), and mice succumbed to tumors more quickly (B). Differences in survival rates were statistically analyzed using the log-rank test.

[0064] [Figure 2] Peptide administration reduces inflammation and inflammatory infiltration of bone marrow in a mouse model of contact dermatitis. (A) Experimental outline of DNFB-induced contact dermatitis. (B) Ear thickness of mice treated with peptide RP23 (SEQ ID NO: 12), replicated three times. (C) Inflammatory infiltrates in the ears were characterized by flow cytometry 72 hours after challenge with DNFB (D15), showing changes in the monocyte / macrophage population (CD11b+). Data are presented as mean ± SEM. Statistical significance was determined by multiple t-test (B) and ANOVA with Dunnett's multiple comparison test (C) (*p<0.05, **p<0.01, ***p<0.001).

[0065] [Figure 3] Peptide effects depend on the peptide sequence and dose. (A) PBS, scrambled peptide, or peptide MUC1SP (MTPGTQSPFFLLLLLTVLTVV; SEQ ID NO: 3) was subcutaneously injected into C57BL / 6 mice (n=5) 7 days before sensitization with DNFB in the abdominal region. Ear inflammation after ear challenge with DNFB was determined, indicating that the effect was specific to the sequence of SEQ ID NO: 3. (B) Comparison of the immunosuppressive effects of peptides RP23 (SEQ ID NO: 12) and MUC1SP (SEQ ID NO: 3). (C) SEQ ID NO: 12 suppressed immunity in a dose-dependent manner. (D) Addition of GM-CSF to SEQ ID NO: 12 injection abolished immunosuppression. Data are means ± SEM.

[0066] [Figure 4] Peptide administration reduces local disease severity in a mouse model of psoriasis. (A) Experimental overview. (B) Clinical scores of mice receiving PBS or peptide RP23 (SEQ ID NO: 12) injections compared with daily (days 0–4) topical application of glucocorticoids to the back and ears. (C) Measurement of local disease at the site of peptide RP23 injection. Redness was measured using an unbiased erythema meter, and skin thickness was measured using manual calipers. (D) Spleen weight on day 5 as a measure of systemic immunosuppression. Ear thickness measurements showed that peptide RP23 did not suppress disease at distal sites. Data are expressed as mean ± SEM. Statistical significance was determined by ANOVA with Dunnett's multiple comparison test (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001).

[0067] [Figure 5] Peptide administration attenuates proinflammatory DC and macrophage activation. (A) Splenocytes from C57BL / 6 mice were stimulated with LPS ± peptide RP23 (SEQ ID NO: 12), and the expression of activation markers on DCs was assessed by flow cytometry. (B) Mouse bone marrow-derived macrophages were incubated with peptide RP23 for 30 minutes and then stimulated with LPS overnight. The positive control was stimulated with LPS alone. The negative control was unstimulated. Inflammatory cytokine production was assessed by ELISA. (C) CD14+ cells were isolated from human blood and differentiated into macrophages by culturing with human serum. Cells were incubated with peptide RP23 for 30 minutes and then stimulated overnight with LPS (left panel) or imiquimod (right panel). Inflammatory cytokine (IL-6 or IL-12 / 23) production was assessed by ELISA. Representative of six donors. Data are presented as mean ± SEM. Statistical significance was determined by ANOVA with Dunnett's multiple comparison test (*p<0.05, **p<0.01, ***p<0.001).

[0068] [Figure 6] Peptide administration is not cytostatic or cytotoxic to mouse or human macrophages / monocytes. After overnight incubation, the cytostatic / cytotoxic effects of peptide RP23 (SEQ ID NO: 12) were assessed by (A) WST-1 assay in RAW and THP-1 cell lines (data are means of duplicates) and (B) LDH assay in primary mouse BMDM or human MDM (data are means ± SEM).

[0069] [Figure 7] Peptide administration reduces spontaneous myeloid cell activation in human skin. (A) Peptide RP23 (SEQ ID NO: 12) was injected intradermally into healthy human skin grafts. After overnight culture, myeloid cell activation was assessed. (B) Effect of peptide RP23 on spontaneous upregulation of activation markers on myeloid cell subsets in skin. Data are representative of six donors.

[0070] [Figure 8] Effect of peptide variants on inflammatory responses. Mice received subcutaneous injections (10 nmol) of PBS or one of the following peptides in the abdominal region: (A) peptide RP23 (SEQ ID NO: 12), peptide AF594-RP23 (SEQ ID NO: 21), peptide K4-G3-MUC1SP (SEQ ID NO: 22), or MUC1SP C fragment (SEQ ID NO: 2). Five days later, mice were challenged with DNFB at the distal site of the ear. Ear thickness was measured over 72 hours.

[0071] [Figure 9]Effect of additional peptide variants lacking an N-terminal methionine on inflammatory responses in a contact dermatitis model. Mice received a single subcutaneous abdominal injection of PBS, RP23 (SEQ ID NO: 12) (10 nmol), or one of the peptide variants shown in the table (equimolar amounts of RP23), including SEQ ID NOs: 32 and 35 described herein. Where indicated, photoleucine replaced leucine, and these molecules were used in photocrosslinking studies. Seven days later, the abdominal skin was sensitized with a contact irritant (DNFB), and five days later, mice were challenged with DNFB at a distal site in the ear. (A) Specific increase in ear thickness over 72 hours. Data are means + / - SEM (n = 6). Statistical significance was determined by ANOVA with Dunnett's multiple comparison test relative to the PBS-only control; **p < 0.01. (B) Specific increase in ear thickness 24 hours after challenge. Data are means + / - SEM (n = 6). Statistical significance was determined by t-test against relevant controls as indicated (*p<0.05, **p<0.01, ns=not significant).

[0072] [Figure 10]Therapeutic synergy between RP23 and topical steroids in an imiquimod-induced psoriasis mouse model. Psoriasis was induced in C57BL / 6 female mice by daily (days 0–4) application of Aldara cream (imiquimod 5%, 57.5 mg) to the backs. (A) Clinical scores (PASI; modified psoriasis area and severity index) were compared between mice receiving PBS or RP23 injections on day -1 and topical application of glucocorticoid (0.05% betamethasone dipropionate, "BD," 30 mg) to the back on day 0 (A) and mice receiving PBS or RP23 injections on day 1 and topical application of glucocorticoid on day 1 (B) and mice receiving PBS or RP23 injections on day 1 and topical application of glucocorticoid on day 1 (B). Data are mean ± SEM (n = 5). Statistical significance was determined relative to the PBS-only control by ANOVA with Dunnett's multiple comparison test (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001).

[0073] [Figure 11] RP23 treatment reduces TCRαβ+ T cells in the back skin and increases regulatory CD4+ T cells in the lymph nodes of mice in an imiquimod-induced psoriasis model. C57BL / 6 female mice received PBS or RP23 injections on day -1, followed by daily application of Aldara cream (imiquimod 5%, 57.5 mg) to the back (days 0–4) to induce psoriasis. On day 15, skin and inguinal lymph nodes were analyzed by flow cytometry to quantify (A) T cell subsets as a percentage of total leukocytes (CD45+) in psoriatic skin on the back, and (B) the number of CD3+CD4+ TCRαβ T cells in the skin-draining lymph nodes (inguinal) using transcription factor staining to identify cells with a regulatory phenotype (FoxP3+). Data are means + / - SEM (n = 4–5). Statistical significance was determined by multiple t-test (*p<0.05).

[0074] [Figure 12]RP23 formulated for topical application can penetrate human skin. RP23AF594 in a stock emulsion was applied topically to full-thickness human skin explants and cultured for 24 hours. Frozen sections were analyzed by fluorescence microscopy. RP23AF594 (associated with DAPI+ cells indicated by arrows) was detectable both within the epidermis (E) and beneath the basement membrane (line) in the dermis (D).

[0075] [Figure 13] RP23 treatment reduces inflammation in a mouse model of atopic dermatitis. C57BL / 6 female mice were sensitized (day 0) and challenged (days 7, 10, 14, 17, 19, 21, 23, 25, and 27) on the ears and shaved backs with topical application of the irritant oxazolone or vehicle alone. Mice were injected with RP23 on day -1 or both days -7 and -1 compared with injections of PBS alone (days -7 and -1) or topical application of steroid (betamethasone dipropionate 0.01%, 30 mg) to the back at each oxazolone application. Ear thickness was measured with manual calipers. Data are mean ± SEM (n = 5).

[0076] [Figure 14] RP23 interacts with innate immune cells in the skin and lungs. AF594-RP23+ cells were identified by flow cytometry 24 hours after administration to C57BL / 6 mice. (A) RP23 was injected subcutaneously into the lower back. RP23+ leukocytes (CD45+) in the back skin were primarily neutrophils (CD11b+ Ly6G+) or CD11b+ myeloid cell subsets. (B) RP23 was delivered to the lungs by intranasal instillation. The percentage distribution of total RP23+ leukocytes among immune cell subsets in the lungs is shown. Data are means + / - SEM (n = 4).

[0077] [Figure 15]Mucosal delivery of RP23 to the lungs reduces the activation of antigen-presenting cells in the lungs and bronchoalveolar spaces. (A) C57BL / 6 mice (n = 5) received either PBS or RP23 by intranasal instillation, or (B) C57BL / 6 mice (n = 4) received either PBS, scrambled RP23 peptide, or RP23 by intranasal instillation. 24 hours later, Pam2Cys-SK4-PEG(OH) was administered intranasally to induce an inflammatory state. After 24 hours, cells from bronchoalveolar lavage and perfused lungs were collected. Median MHCII fluorescence intensity on CD11c+ dendritic cells was determined by flow cytometry. Data are means + / - SEM. Statistical significance was determined relative to the PBS control by (A) t-test (***p < 0.001) and (B) ANOVA with Dunnett's multiple comparison test (*p < 0.05). DETAILED DESCRIPTION OF THE INVENTION

[0078] Detailed Description of the Embodiments The present inventors have herein identified a group of peptides useful for the suppression, prevention, or treatment of inflammation. In particular, the present inventors have established that the peptides characterized herein have the ability to suppress, prevent, or treat inappropriate inflammatory responses. In embodiments, the peptides can be used to suppress inflammatory responses associated with immune system activation. In another embodiment, the peptides described herein can be used to suppress inflammatory responses in the absence of concomitant immune system activation.

[0079] The peptides described herein have been shown to be useful in many commonly used models for evaluating the activation of inflammatory response, including dermatitis and psoriasis.These models are well characterized in the art, including acute and / or chronic inflammatory episodes.Therefore, the inventors recognize that the peptides defined are applicable to a wide range of diseases or conditions that involve inflammation, of which the disclosed ones are only a few examples.

[0080] The peptides described herein have particular utility in suppressing erythema, a key feature of the inflammatory process. In one aspect, this is achieved by suppressing monocyte / macrophage or other phagocyte infiltration into the affected area. In particular, the peptides have the ability to reduce the secretion of inflammatory chemokines or cytokines and the expression of surface activation markers of proinflammatory immune cells. Non-limiting examples include CD86, MHCII, and levels of IL-6, IL-12 / IL-23, TNF-α, and MCP-1. The inventors have also surprisingly discovered that the effect of suppressing the inflammatory response is in the absence of any cytostatic or cytotoxic effects on cells. The peptides described herein surprisingly can reduce the activation of antigen-presenting cells, reduce spontaneous activation of dermal dendritic cells, and suppress TCRαβ expression. + It can reduce the levels of T cells and regulatory CD4 + These data support the idea that inflammatory conditions mediated by TNF-α and other proinflammatory cytokines or regulatory CD4 T cells can be a factor in the proliferation of T cells. + This points to a potential role for the peptide in inhibiting inflammation in a wide range of inflammatory conditions, including those that are suppressed by the induction of regulatory cells such as T cells.

[0081] The data obtained by the present inventors suggest that the peptides of the present invention suppress proinflammatory responses by innate immune cells involved in inflammatory stimulation under various stimuli, including autoinflammation, tissue injury, cancer, or infectious diseases. Using models characterized by Th1-type inflammation or IL-17 / Th17-driven, the present inventors show that the peptides of the present invention can attenuate (reduce) stimulation of Th1 / Th17-type responses and / or promote regulatory responses (FoxP3+ CD4+ T cells). Thus, these data indicate that the peptides of the present invention may be suitable for the treatment of inflammatory disorders, including indications such as psoriasis, dermatitis, rheumatoid arthritis, IBD, and transplant rejection / tolerance (graft-versus-host disease, GVHD).

[0082] Thus, the peptides featured herein are 1. Erythema; 2. Monocyte / macrophage infiltration; 3. Increased levels of cytokines and / or chemokines. 4. Activation of cell receptors / markers on inflammatory cells. 5. Activation of dendritic cells, and / or 6. Lymphocyte activation The compounds have utility in suppressing, treating or preventing inflammatory responses characterized by one or more of the following:

[0083] The inventors have also established that the described peptides have utility in reducing local inflammation in a manner that avoids unnecessary immune system suppression. This is important because most clinically prescribed immunosuppressants cause systemic immunosuppression. Thus, the peptides described herein offer previously unrecognized advantages.

[0084] Inflammatory response The peptides described herein have utility in treating or suppressing acute or chronic inflammatory responses, with or without an accompanying immune response. Alternatively, the peptides described herein can suppress inappropriate acute inflammatory responses associated with an immune response, which may be an innate response, an adaptive immune response, or both.

[0085] The term suppression of an inappropriate inflammatory response or treatment of an inappropriate immune response refers to the alleviation of one or more symptoms of inflammation, including erythema (redness), edema (swelling), pain, and itching, which are characteristic of an inflammatory condition. This may also be understood as a reduction in one or more pro-inflammatory cytokines and / or chemokines, such as TNF-α, IL-6, CXCL1, CXCL2, IL-1β, prostaglandin E2, and IL-23 / IL-17, as well as other chemokines / cytokines known in the art.

[0086] In embodiments, the peptides described herein have utility in treating or suppressing acute inflammatory responses, whether or not there is an associated immune response, hi embodiments, the peptides can suppress acute inflammatory responses associated with the innate immune response.

[0087] Acute inflammation is understood as a short-term process that occurs in response to tissue injury and typically manifests within minutes or hours. It is characterized by five cardinal symptoms: pain, redness, immobility (loss of function), swelling, and heat. The acute inflammatory response involves the recruitment of blood leukocytes, the activation of tissue-resident cells (mast cells, dendritic cells, fibroblasts, macrophages), and the production of a series of mediators, including IL-1 and TNFα. The outcome can include the eventual resolution of the inflammatory process, the induction of events leading to cellular regeneration or wound healing, or the progression of an inflammatory response that can be characterized as chronic inflammation.

[0088] In embodiments, the peptides described herein have utility in treating or suppressing chronic inflammatory responses, with or without an associated immune response, hi embodiments, the peptides can suppress chronic inflammatory responses associated with an adaptive immune response.

[0089] Chronic inflammation is understood to be a slow, prolonged inflammation that persists for months to years. It can be caused by a number of different factors, including (1) failure to eliminate acute inflammation-causing agents such as infectious organisms, including Mycobacterium tuberculosis, protozoa, fungi, and other parasites that can resist host defenses and persist in tissues for long periods; (2) exposure to low levels of specific irritants or foreign substances that cannot be eliminated by enzymatic degradation or phagocytosis; (3) autoimmune disorders in which the immune system is sensitized to normal components of the body; (4) recurrent episodes of acute inflammation; or (5) inflammatory and biochemical triggers that cause oxidative stress and mitochondrial dysfunction.

[0090] It is understood that chronic inflammatory processes can be diagnosed by one of many tests known in the art, including (1) serum protein electrophoresis (SPE), which demonstrates hypoalbuminemia and a polyclonal increase in all gamma globulins (polyclonal gammopathy); (2) measurement of serum high-sensitivity C-reactive protein (hsCRP) and fibrinogen (normal serum levels of hsCRP are less than 0.55 mg / L in men and less than 1.0 mg / L in women, and normal levels of fibrinogen are 200-300 mg / dL); and detection of proinflammatory cytokines such as tumor necrosis factor alpha (TNF-α), interleukin 1 beta (IL-1β), interleukin 6 (IL-6), interleukin 8 (IL-8), interleukin 17 (IL-17), and interleukin 12 / 23 (IL-12 / 23).

[0091] Adaptive and innate immune responses The peptides described herein have utility in suppressing inflammatory processes associated with the adaptive or innate immune response.

[0092] Adaptive or acquired immunity includes humoral (B lymphocyte) and cell-mediated (T lymphocyte) immune responses. Humoral immunity involves B cells and their products, including antibodies. Cell-mediated immunity involves the activation of phagocytes, antigen-specific CD4+ and CD8+ T lymphocytes, and the release of various cytokines in response to antigens. Cell-mediated immunity activates antigen-specific cytotoxic T cells, which can induce apoptosis in cells displaying epitopes of foreign antigens on their surface, such as virus-infected cells or cells harboring intracellular bacteria. Cell-mediated immune responses, particularly CD4+ helper responses, also involve the activation of B cells. Cell-mediated immune responses also involve macrophages and innate lymphocytes (e.g., natural killer cells) for the destruction of pathogens by recognition and secretion of cytotoxic granules (in the case of natural killer cells) and phagocytosis (in the case of macrophages), as well as the stimulation of cells to secrete cytokines. Cell-mediated immunity is primarily T cell-driven, utilizing helper (CD4+) and cytotoxic (CD8+) subtypes, and is involved in responses to viral infections, transplant rejection, chronic inflammation, and tumor immunity. Activated T cells can take several days to mount a cell-mediated attack after exposure to a novel antigen, whereas memory T cells respond rapidly if primed by previous exposure. The peptides described herein have utility in treating chronic inflammatory processes associated with the adaptive immune response, as known in the art and as described herein.

[0093] In contrast, the innate immune system is composed of barriers to infection (epithelia of the skin, gastrointestinal tract, respiratory tract, and urogenital tract), antimicrobial peptides and proteins, humoral components (i.e., complement and opsonins), and cellular components (i.e., mast cells, neutrophils, monocytes / macrophages, dendritic cells, and innate lymphoid cells). Innate immunity serves as the first line of host defense and plays an essential role in preventing infection while tolerating normal host microbiota. The innate immune system directs the subsequent development of adaptive immune responses.

[0094] It is understood that the peptides described herein have utility in suppressing inappropriate inflammatory reactions associated with the innate immune response within such barrier layers of the body, including the skin, gastrointestinal tract, joints and respiratory tract.

[0095] Those skilled in the art will also appreciate that an inappropriate inflammatory response can involve activation of both the adaptive and innate immune responses.

[0096] GM-CSF In embodiments, a peptide pharmaceutical composition comprising a pharmaceutically effective amount of a peptide described herein or a pharmaceutically acceptable salt thereof does not include granulocyte-macrophage colony-stimulating factor (GM-CSF).

[0097] GM-CSF is a monomeric glycoprotein secreted by macrophages, T cells, mast cells, natural killer cells, endothelial cells, and fibroblasts, where it functions as a cytokine. It is known to have proinflammatory roles, primarily due to its role as a growth and differentiation factor for granulocytes, macrophages, and monocytes, as well as dendritic cell populations. Therefore, agents capable of blocking GM-CSF or its receptor have been used as anti-inflammatory therapies.

[0098] Therefore, it is contemplated that the peptides described herein having utility in suppressing, treating, or preventing inappropriate inflammatory responses will not be used in combination with growth factors, cytokines, chemokines, etc. that promote the inflammatory process, as this may negate the therapeutic effect of any of the peptides described herein. This is illustrated in the Examples herein, where GM-CSF abrogates the suppressive effect of the peptides of the invention (Figure 3).

[0099] Determination of such growth factors, cytokines, chemokines, etc. that can promote inflammation is well within the knowledge of one of ordinary skill in the art and can include GM-CSF, TNF-α, IL-6, IL-12, IL-17, IL-1β, MCP-1, MIP-1, IFN-γ, IL-18, IL-23, CCL-2, CCL-3, CCL-4 and CCL-5.

[0100] Conditions suitable for treatment with defined peptides The present invention provides a method for treating or preventing an inappropriate immune response in an individual in need thereof, comprising administering to the individual in need thereof a pharmaceutical composition comprising a therapeutically effective amount of a peptide as described herein, or a pharmaceutically acceptable salt thereof, thereby treating the inappropriate immune response in the individual. Thus, the peptides of the present invention find utility in the treatment of a wide range of inflammatory disorders.

[0101] The terms "individual" or "patient" or "subject," as used herein, refer to a mammal or human being undergoing treatment for an inflammatory condition and / or an inflammatory condition with visible symptoms. Preferably, the individual is a human.

[0102] "Therapeutically effective amount" is used herein to refer to the amount of a formulation of a composition comprising peptide that shows anti-inflammatory effect when applied to the affected area or affected region of tissue.One application of the formulation of the present invention may be sufficient, or the formulation may be repeatedly applied over a period of time, such as several times a day for several days or weeks.The amount of active ingredient varies depending on the condition to be treated, the stage of the condition, the age and type of host, and the type and concentration of the formulation to be applied.The appropriate amount in a given case will be readily apparent to those skilled in the art or can be determined by routine experimentation.

[0103] The term "treatment" or "treating" a subject includes the application or administration of a compound of the invention to a subject (or the application or administration of a compound of the invention to a cell or tissue from a subject) for the purpose of delaying, slowing, stabilizing, curing, healing, alleviating, relieving, altering, remedying, making less worsening, ameliorating, or ameliorating a disease or condition, a symptom of a disease or condition, or a risk of (or susceptibility to) a disease or condition, or affecting a disease or condition, a symptom of a disease or condition, or a risk of (or susceptibility to) a disease or condition. The term "treating" refers to indicators of success in treating or ameliorating an injury, pathology, or condition, including objective or subjective parameters, such as abatement; remission; reduction in the rate of deterioration; reduction in the severity of the disease; stabilization, diminishing of symptoms, or making the injury, pathology, or condition more tolerable to the subject; slowing the rate of degeneration or decline; making the final point of degeneration less debilitating; or improving the physical or mental health of the subject.

[0104] As used herein, the terms "preventing," "prevent" or "prevention" include stopping or preventing the occurrence of at least one symptom of an inflammatory condition by administering a peptide or composition of the invention. The term also encompasses treatment of a subject in remission to prevent or hinder recurrence.

[0105] Inflammatory disorders and associated conditions that may be treated or prevented by use of the peptides described herein include, but are not limited to, rheumatoid arthritis, bronchitis, contact dermatitis, atopic dermatitis, psoriasis, seborrheic dermatitis, eczema, allergic dermatitis, dermatitis herpetiformis, polymorphous light eruption, inflammatory skin diseases, folliculitis, alopecia, vitiligo, poison ivy, insect stings, inflammatory acne, transplant rejection / tolerance (graft versus host disease, GVHD), irritation induced by external factors (including, but not limited to, chemicals, trauma, pollutants (e.g., cigarette smoke), and sun exposure), secondary conditions resulting from inflammation (including, but not limited to, xeroderma, hyperkeratosis, pruritus, post-inflammatory hyperpigmentation, scarring, etc.).

[0106] Inflammatory disorders and related conditions that may be treated or prevented by use of the peptides described herein also include those that result in excessive pneumonia (including when caused by viral or microbial infections), lupus, rheumatoid arthritis, multiple sclerosis, sarcoidosis and scleroderma, hypersensitivity pneumonitis.

[0107] Inflammatory disorders and related conditions that may be treated or prevented by use of the peptides described herein also include those that may be present during wound healing and inflammatory bowel disease (including ulcerative colitis and Crohn's disease).

[0108] Preferably, inflammatory disorders and related conditions that may be treated or prevented using the methods of the invention include contact dermatitis, atopic dermatitis, psoriasis, viral and microbial infections, wound and inflammatory bowel disease (including Crohn's disease and ulcerative colitis), musculoskeletal inflammatory conditions (including, for example, rheumatoid arthritis), and transplant rejection / tolerance (graft versus host disease, GVHD).

[0109] Psoriasis, dermatitis and other inflammatory skin disorders Those skilled in the art will be familiar with how to identify individuals in need of prevention or treatment of inflammatory skin diseases, including psoriasis and dermatitis, and how to determine the effectiveness of treatment for such conditions.

[0110] As used herein, dermatitis may refer to contact dermatitis, atopic dermatitis, or dermatitis herpetiformis.

[0111] Atopic dermatitis may be a type of eczema selected from the group consisting of endogenous eczema, flexural eczema, infantile eczema, and may also be known as "venié prurigo," "neurodermatitis," or "diathesic prurigo."

[0112] When the condition is atopic dermatitis and the peptides administered are administered topically, the areas of skin that come into contact with the peptides of the invention can include broken and unbroken skin.

[0113] Psoriasis is an immune-mediated inflammatory skin disease characterized by thickened skin, red patches, and dry scales. Psoriasis can be caused by many factors, including injury, trauma, infection, and medication. The disease is clinically assessed using the Psoriasis Activity and Severity Index (PASI) scale, which ranks the severity of erythema (redness), induration (thickness), and scaling (scaling). Test parameters include in vivo clinical evaluation of the skin, histopathological evaluation of skin sections, and optional cytokine analysis of the skin and / or internal immune organs. The disease has specific distinct but overlapping clinical phenotypes, including chronic plaque lesions, rashes, and pustular lesions. There are several types of psoriasis, including pustular psoriasis, guttate psoriasis, inverse psoriasis, and erythrodermic psoriasis. Those skilled in the art will understand that the peptides described herein are suitable for treating or preventing all types of psoriasis.

[0114] Histologically, the disease is characterized by acanthosis with hyperkeratosis, immune cell infiltration in the dermis and epidermis, parakeratosis, and angiogenesis. IMQ also induces histopathological changes associated with human disease, including epidermal changes due to keratinocyte hyperproliferation and altered differentiation, increased IL-23, immune cell infiltration (T cells, DCs, and neutrophils), and altered angiogenesis.

[0115] The imiquimod-induced psoriasis model has many important markers of human disease, including histopathology of lesions and strong activation of the immune system, making it particularly promising for clinical application. Imiquimod (IMQ) is a ligand for TLRs, such as TLR-7 (Toll-like receptor), on immune cells (including macrophages, monocytes, and plasmacytoid dendritic cells), and therefore contributes to strong activation of the immune system. A hallmark of this disease in humans is the involvement of the IL-23 / IL-17 cytokine axis. This imiquimod-induced psoriasis model represents human plaque-type psoriasis, in which the IL-23 / IL-17 cytokine axis plays a crucial role.

[0116] Current treatment options for psoriasis include coal tar preparations, corticosteroids, triamcinolone (acetonide, Trianex), clobetasol (Temovate), synthetic forms of vitamin D such as calcipotriene and calcitriol (Vectical), retinoids such as tazarotene (Tazorac, Abazi), calcineurin inhibitors such as tacrolimus (Protopic) and pimecrolimus (Elidel), medications such as methotrexate, acitretin, cyclosporine and calcipotriol, and ultraviolet light therapy.

[0117] It is therefore envisioned that the peptides described herein may be administered to a patient in need thereof in combination with the above-mentioned therapies or therapies known in the art for the treatment of psoriasis.

[0118] Successful response to treatment can be determined by assessing the remission of one or more symptoms of psoriasis, using the Psoriasis Activity Severity Index (PASI) scale, which is outlined above and ranks the severity of erythema (redness), induration (thickness) and scaling (scales), or by other methods known in the art.For example, a subject who shows a positive response may have one or more symptoms of erythema, induration or scaling reduced.This positive response to treatment can be determined by clinical evaluation of skin, histopathological evaluation of skin sections, and optional cytokine analysis in skin and / or internal immune organs.Other means for assessing positive response are within the knowledge of those skilled in the art.

[0119] inflammatory bowel disease Inflammatory bowel disease (IBD) is a group of chronic inflammatory disorders of the gastrointestinal tract. It can manifest as ulcerative colitis, which causes persistent inflammation and ulcers in the mucosa of the large intestine and rectum, or as Crohn's disease, which is characterized by inflammation of the mucosa of the gastrointestinal tract (particularly the small intestine and large intestine) that spreads to affected tissues such as the mouth, esophagus, stomach, and anus.

[0120] Crohn's disease is a condition characterized by chronic inflammation in the mucous membranes of the digestive system. There may be small patches of inflammation, or it may be quite widespread along the intestine, or there may be several patches in different locations. Typical symptoms include frequent urgency to go to the bathroom, frequent inability to void, recurrent diarrhea, abdominal pain and cramps (usually worse after eating), extreme fatigue (malaise), and / or weight loss. Colonoscopy is used for diagnosis through the sampling of a small tissue sample (biopsy) for examination under a microscope.

[0121] Ulcerative colitis is a chronic inflammatory disease that usually occurs in the rectum (the part of the large intestine that is just inside the anus) and the lower part of the colon, but can affect the entire large intestine (colon).When the colon becomes inflamed, and this inflammation becomes severe, the mucous membrane of the colon can break down and form ulcers.The diagnosis of ulcerative colitis can be made by blood tests to check inflammation, anemia and protein level, stool samples to check infection, X-rays to help evaluate the degree of the condition, sigmoidoscopy to check the degree of inflammation in the rectum and the lower part of the colon, and / or colonoscopy to examine the inside of the entire colon.

[0122] Viral infections The peptides and compositions described herein have utility in treating or preventing viral infections that cause inflammation, including those of the respiratory tract. Such viral infections associated with an inflammatory response include the common cold, influenza, cough and bronchitis, chickenpox, HIV / AIDS, meningitis, pneumonia, herpes, rotavirus, severe acute respiratory syndrome (SARS), severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and Middle East respiratory syndrome (MERS).

[0123] Those skilled in the art will be familiar with how to identify individuals who need treatment for viral infections and how to determine the effectiveness of treatment for a given viral infection.For example, individuals with viral infections may have one or more of the following symptoms: fever, muscle pain, cough, sore throat, and headache.Therefore, a positive response to treatment with the peptides or compositions described herein includes the alleviation of one or more of these symptoms.For example, an individual who has a positive response to treatment with the peptides or compositions described herein may have a decrease in fever, a reduction in muscle pain, a decrease in cough, a reduction in sore throat, or a reduction in headache, or symptoms may completely disappear.

[0124] Influenza (commonly referred to as "the flu") is an infectious disease caused by an RNA virus (influenza virus) of the Orthomyxoviridae family that affects birds and mammals. The most common symptoms of the disease are chills, fever, sore throat, muscle aches, severe headache, cough, weakness / fatigue, and a general feeling of unease.

[0125] Influenza infection involves a multilayered series of events. The first is viral infection of the airway and alveolar epithelium and its replication in these cells, during which strategies to limit viral entry or replication can prevent or attenuate the severity of infection. The second is the innate immune response followed by an adaptive immune response to the virus, which is important for viral clearance but can also cause significant damage to the alveolar epithelium and endothelium. The third is the development of long-term immunity to the infecting virus strain, accompanied by resolution of infiltration and regeneration of damaged lung tissue, during which susceptibility to secondary bacterial infections increases.

[0126] Influenza viruses comprise three of the five genera in the family Orthomyxoviridae. Influenza A and B viruses co-circulate during seasonal epidemics and can cause severe influenza infections. Influenza C virus infections are less common but can be severe and cause localized epidemics.

[0127] Influenza A viruses are subdivided into different serotypes or subtypes based on the antibody response to these viruses. Influenza A viruses are divided into subtypes based on two proteins on the surface of the virus: hemagglutinin (H) and neuraminidase (N). There are 18 different hemagglutinin subtypes and 11 different neuraminidase subtypes (H1 to H18 and N1 to N11, respectively). The subtypes identified in humans are H1N1, H1N2, H2N2, H3N2, H5N1, H7N2, H7N3, H7N7, H9N2, and H10N7.

[0128] In either embodiment, the influenza infection for which treatment, prevention or suppression of an inflammatory response is required is an infection caused by a virus selected from the group consisting of influenza A, B or C.

[0129] dermatitis Dermatitis, also known as eczema, is a group of diseases that cause inflammation of the skin, including atopic dermatitis, allergic contact dermatitis, irritant contact dermatitis, and stasis dermatitis. Although the symptoms of each type of dermatitis vary, certain signs are common to all dermatitis, including redness, swelling, itching, and sometimes oozing and scarring.

[0130] Atopic dermatitis is a chronic inflammatory condition that presents as a red, itchy rash, most commonly on the insides of the elbows, behind the knees, and in areas where the skin bends, such as the front of the neck. Scratching can cause the rash to leak fluid and form scabs. People with atopic dermatitis may improve and then experience flare-ups.

[0131] Contact dermatitis is an acute or chronic inflammatory skin condition that can be classified as irritant or allergic. It develops on areas of the body that come into contact with substances that irritate the skin or cause an allergic reaction, such as poison ivy, soap, and essential oils. The red rash can be sore, stinging, or itchy. It may blister.

[0132] Dinitrofluorobenzene (DNFB) is a known irritant that causes contact dermatitis hypersensitivity. It acts as a hapten, inducing a cell-mediated response, leading to the activation of keratinocytes, resulting in the production of chemical mediators such as TNF-α, IL-1β, and prostaglandin E2, and the migration and maturation of skin dendritic cells. These cytokines then activate vascular endothelial cells to express adhesion molecules (i.e., ICAM-1 and P / E selectin) that induce T cell migration from the blood into tissues.

[0133] Haptens such as DNFB also activate mast cells and keratinocytes to produce neutrophil-recruiting chemokines, such as CXCL1 and CXCL2, leading to neutrophil recruitment and innate immune responses. The activation of dendritic cells (i.e., antigen-presenting cells) then leads to the induction of cell-mediated immunity and the infiltration of T cells, resulting in the production of IFN-γ and IL-17. Treg cells control contact dermatitis by attenuating leukocyte influx and degrading ATP. B cells and their products, particularly IgM antibodies, are also involved, particularly in the induction phase.

[0134] Treatments for dermatitis include hydrocortisone creams, gels, or ointments, calcineurin inhibitors, antihistamines to relieve severe itching, and monoclonal antibody therapy, including those with antibodies that inhibit the activity of IL4 / IL-13, IL-33, and TSLP. It is contemplated that the peptides described herein can be administered to a patient in need thereof in combination with the above therapies or therapies known in the art for treating dermatitis.

[0135] Rheumatoid arthritis Rheumatoid arthritis is an autoimmune disease in which a person's immune system attacks their own tissues, resulting in joint pain and swelling. Rheumatoid arthritis is a chronic disease characterized by symptoms of joint inflammation and pain. These symptoms and signs occur during periods known as flare-ups or exacerbations. Other periods are known as remissions, during which symptoms disappear completely. While rheumatoid arthritis symptoms can affect several organs in the body, the primary symptoms of rheumatoid arthritis include joint pain, swelling, stiffness, and decreased joint function and deformity.

[0136] Medications used to treat rheumatoid arthritis fall into two main groups. The first group reduces the activity of the immune system, which attacks and damages healthy joints. These drugs are called disease-modifying antirheumatic drugs (DMARDs) and include methotrexate, sulfasalazine, leflunomide, prednisone, and hydroxychloroquine. The second group treats pain and / or inflammation and includes agents like paracetamol and anti-inflammatory drugs like ibuprofen or celecoxib.

[0137] The peptides of the present invention can be used alone or in combination with known agents for the treatment of RA.

[0138] Peptides and appropriate modifications The present invention establishes the utility of a broad class of peptides in the suppression, treatment or prevention of inappropriate inflammatory responses, which may or may not be accompanied by an immune response. In a first embodiment, the peptides comprise: - the amino acid sequence of TVLTVV (SEQ ID NO: 1) or the amino acid sequence of LLLLLTVLTVV (SEQ ID NO: 2) or a functional variant thereof wherein the peptide consists of less than 21 amino acid residues, preferably 6 to 20 amino acid residues, preferably 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18 or 19 residues.

[0139] As described herein, the inventors believe that peptides having the amino acid sequence TVLTVV or the amino acid sequence LLLLLTVLTVV have anti-inflammatory effects on symptoms of inflammation and immunosuppressive responses as manifested by modulating macrophage and dendritic cell activation, cell surface marker expression, and changes in pro-inflammatory cytokine profiles. In some embodiments, TVLTVV and LLLLLTVLTVV may each be referred to as an "anti-inflammatory domain" or "AID." According to this embodiment of the invention, the peptide may consist of the sequence TVLTVV or LLLLLTVLTVV, or the peptide may comprise TVLTVV or LLLLLTVLTVV, and may further comprise additional amino acid residues, provided that the length of the peptide, measured in amino acids, is less than 21 amino acids.

[0140] As further described herein, the additional amino acids (i.e., additions to TVLTVV or LLLLLTVLTVV) can be residues such as lysine, arginine, or histidine. These can be provided in the form of polylysine, polyarginine, or polyhistidine. These residues can be located at the N-terminus of TVLTVV or LLLLLTVLTVV (or, alternatively, at the C-terminus of TVLTVV or LLLLLTVLTVV). These residues can function to help facilitate solubilization of the peptide in biological fluids or tissues or biologically compatible formulations. Examples of such peptides are shown in Table A.

[0141] [Table 1]

[0142] In a second embodiment, the peptide comprises: - a peptide comprising the sequence SEQ ID NO: 1 or SEQ ID NO: 2; - one or more further amino acids adjacent to the sequence SEQ ID NO: 1 or SEQ ID NO: 2, with the proviso that said one or more further amino acids do not define the sequence MTPGTQSPFF in the N-terminal position adjacent to the sequence SEQ ID NO: 1 or SEQ ID NO: 2, or the sequence TGSGHASSTP in the C-terminal position adjacent to the sequence SEQ ID NO: 1 or SEQ ID NO: 2. wherein the peptide contains at least 12 amino acid residues, preferably 20 to 100 residues.

[0143] In accordance with this embodiment, the peptides generally do not consist of the sequences TVLTVV and LLLLLTVLTVV. Rather, the peptides comprise TVLTVV or LLLLLTVLTVV and further comprise additional amino acid residues other than those defining the sequence MTPGTQSPFF at the N-terminal position adjacent to SEQ ID NO: 1 or 2 or the sequence TGSGHASSTP at the C-terminal position adjacent to SEQ ID NO: 1 or 2.

[0144] Thus, the minimum length of such peptides may be 12 amino acid residues, although the peptides may be longer and may have a length of at least 13, at least 14, at least 15 amino acid residues, or at least 20, 30, 40, 50, or up to 1000 residues. The additional amino acids (i.e., additions to TVLTVV or LLLLLTVLTVV) may be cationic amino acid residues such as lysine, arginine, or histidine, positioned as above.

[0145] Optionally, the peptide may also include a "solubilization domain," or a modification that enhances the solubilization of the peptide in biological fluids, tissues, and the like. Such improved solubilization can result in higher loading of the peptide in biological fluids, tissues, or cells, thereby improving the anti-inflammatory and / or immunosuppressive effects of the peptide. The inventors have established this effect by utilizing a polycationic amino acid sequence. As further described herein, the solubilization may also include polyethylene glycol (PEG) of various lengths.

[0146] Additionally, the peptide may also include a label as described herein (eg, biotin) or a molecule in the form of a label attached to the peptide to aid in its detection.

[0147] Examples of peptides containing modifications to improve solubility or moieties to allow detection of the peptide are shown in Table B.

[0148] [Table 2]

[0149] In a further embodiment, the peptide comprises: - an anti-inflammatory peptide comprising or consisting of the sequence of any one of SEQ ID NOs: 1, 2, preferably SEQ ID NOs: 3 or 30, or a functional variant or fragment thereof; - optionally modifications to facilitate solubilization of the peptide in biological fluids, tissues or formulations; and / or fluorescent labeling It includes:

[0150] In accordance with this embodiment, the peptide generally does not consist of TVLTVV or LLLLLTVLTVV. Rather, the peptide comprises TVLTVV or LLLLLTVLTVV, and further comprises additional amino acid residues to provide the peptide with a length of generally greater than 20 amino acids. The further additional amino acids may comprise the sequence MTPGTQSPFF and / or TGSGHASSTP (or alternatively may comprise the sequence TPGTQSPFF).

[0151] Thus, in one example, the peptide comprises or consists of the sequence of SEQ ID NO: 3 (MTPGTQSPFFLLLLLTVLTVV) or comprises or consists of the sequence of SEQ ID NO: 30 (TPGTQSPFFLLLLLTVLTVV), including functional variants and fragments thereof.

[0152] The peptides can be up to 1000 residues in length or longer. In this embodiment, the additional amino acids (i.e., additions to TVLTVV or LLLLLTVLTVV) can be cationic amino acid residues such as lysine, arginine, or histidine. These amino acids can form or be included in solubilizing domains, as described above, to facilitate solubilization of the peptide in biological fluids, tissues, or formulations. As further described herein, solubilizing domains can include polyethylene glycol (PEG) of various lengths. The peptides can also include molecules in the form of fluorescent labels attached to the peptide to aid in its detection. Examples of such peptides are shown in Table C.

[0153] [Table 3]

[0154] In an embodiment, the peptides of the present invention may include a linker containing glycine repeats or serine repeats of various lengths. The peptides of the present invention and the linker may be directly or indirectly linked to either the N- or C-terminal region of the peptide. Preferably, the linker is located at the N-terminus of the peptide. In an embodiment, the linker includes the sequence GGG, and the peptides of the present invention comprise or consist of the functional variant KKKKGGGMTPGTQSPFFLLLLLTVLTVV (SEQ ID NO: 22) or KKKKGGGTPGTQSPFFLLLLLTVLTVV (SEQ ID NO: 23).

[0155] As used herein, the term "peptide" refers to a compound composed of a single chain of D- or L-amino acids, or a mixture of D- and L-amino acids joined by peptide bonds. Generally, peptides contain at least two amino acid residues and are less than about 50 amino acids in length.

[0156] As used herein, the term "protein" refers to a compound composed of a linear arrangement of amino acids linked by peptide bonds, but which, in contrast to peptides, has a well-defined tertiary structure. Proteins, in contrast to peptides, generally consist of chains of 50 or more amino acids.

[0157] As used herein, "polypeptide" refers to a polymer of at least two amino acid residues that contains one or more peptide bonds. "Polypeptide" encompasses peptides and proteins, whether or not the polypeptide has a clearly defined conformation.

[0158] In embodiments, it is understood that the anti-inflammatory peptides of the present invention may be provided in the form of "functional variants," "biologically active fragments," or "analogs" of the TVLTVV or LLLLLTVLTVV sequence. Variants, fragments, or analogs are compounds that are capable of reproducing some or all of the anti-inflammatory and / or immunosuppressive effects exemplified for the peptides TVLTVV or LLLLLTVLTVV, as shown in the Examples herein. Functional variants or analogs of the peptides of the present invention will generally exhibit at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 70%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% amino acid identity with all or a portion of the amino acid sequence of the peptide. Methods for assaying the ability of biologically active fragments and analogs to treat the inflammatory conditions described herein in the DNFB and imiquimod models are known by those of skill in the art and include those described herein.

[0159] In one embodiment, functional variants of SEQ ID NO: 1 that form anti-inflammatory peptides may be as set forth in Table D.

[0160] [Table 4]

[0161] In one embodiment, functional variants of SEQ ID NO:2 that form anti-inflammatory peptides can be as set forth in Table E.

[0162] [Table 5]

[0163] In another embodiment, peptides according to the invention may include retro-inverso peptides of the peptides described herein, in which the D-amino acids represent the conformational mirror images of the naturally occurring L-amino acids.

[0164] Peptides of the present invention include polymeric forms of amino acids of any length, including naturally occurring amino acids, coded and non-coded amino acids, chemically or biochemically modified, derivatized or designer amino acids, amino acid analogs, peptidomimetics, and depsipeptides, as well as polypeptides with modified cyclic, bicyclic, depsicyclic, or depsibicyclic peptide backbones. They include single-chain proteins and multimers. They also include composite proteins, fusion proteins (including, but not limited to, glutathione S-transferase (GST) fusion proteins, fusion proteins with heterologous amino acid sequences, fusion proteins with heterologous and homologous leader sequences), fusion proteins with or without an N-terminal methionine residue, pegylated proteins, and immunologically or His-tagged proteins. Polypeptides of the present invention also include variants of naturally occurring proteins (such variants are homologous or substantially similar to the naturally occurring proteins) and corresponding homologs from different species. Variants of a polypeptide sequence include insertions, additions, deletions or substitutions compared to the reference polypeptide. Polypeptides of the present invention also include peptide aptamers.

[0165] Protein engineering can be used to improve or alter the properties of the therapeutic peptides of the present invention. Using recombinant DNA techniques known to those skilled in the art, novel mutant proteins or "muteins" containing single or multiple amino acid substitutions, deletions, additions, or fusion proteins can be created. Such modified polypeptides can exhibit desirable properties, such as enhanced activity or improved stability.

[0166] For example, the peptides herein may contain one or more positively charged amino acids. The positively charged group includes at least one positively charged amino acid, such as arginine (R), lysine (K), or histidine (H). Examples of such a group include KKKK-TVLTVV (SEQ ID NO: 4), KKKK-LLLLLTVLTVV (SEQ ID NO: 5), or KKKK-MTPGTQSPFFLLLLLTVLTVV (SEQ ID NO: 6), or functional variants thereof.

[0167] In an embodiment, the peptide of the present invention may include a solubilizing agent. The peptide of the present invention and the solubilizing agent may be linked directly or indirectly. An example of a solubilizing agent is polyethylene (PEG), and in this embodiment, the peptide of the present invention may be PEGylated. Examples of such peptides include PEG-TVLTVV (SEQ ID NO: 7), PEG-LLLLLTVLTVV (SEQ ID NO: 8), PEG-MTPGTQSPFFLLLLLTVLTVV (SEQ ID NO: 9), KKKK-PEG-TVLTVV (SEQ ID NO: 10), KKKK-PEG-LLLLLTVLTVV (SEQ ID NO: 11), KKKK-PEG-MTPGTQSPFFLLLLLTVLTVV (SEQ ID NO: 12), or functional variants thereof.

[0168] In embodiments, PEG is at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53 , 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100 or more ethylene glycol units. Alternatively, the PEG may be free of ethylene glycol units.

[0169] In an embodiment, the peptide of the present invention may contain a fluorescent tag or label. The fluorescent tag or label may be directly or indirectly linked to the peptide of the present invention and may be located at the N-terminus of the peptide. In an embodiment, the fluorescent tag or label is GFP, YFP, BFP, CFP, Y-FAST, AF594, mCherry, or dsRed. Examples of such peptides include AF594-TVLTVV (SEQ ID NO: 13) or AF594-LLLLLTVLTVV (SEQ ID NO: 14), AF594-MTPGTQSPFFLLLLLTVLTVV (SEQ ID NO: 15), AF594-PEG-TVLTVV (SEQ ID NO: 16), AF594-PEG-LLLLLTVLTVV (SEQ ID NO: 17), AF594-PEG-MTPGTQSPFFLLLLLTVLTVV (SEQ ID NO: 18), AF594-KKKK-PEG-TVLTVV (SEQ ID NO: 19), AF594-KKKK-PEG-LLLLTVLTVV (SEQ ID NO: 20), AF594-KKKK-PEG-MTPGTQSPFFLLLLLTVLTVV (SEQ ID NO: 21), or functional variants thereof.

[0170] In an embodiment, the peptides of the present invention may include a linker comprising glycine repeats or serine repeats of various lengths. The peptides of the present invention and the linker may be linked directly or indirectly. Preferably, the linker is located at the N-terminus of the peptide. In an embodiment, the linker comprises the sequence GGG, and the peptides of the present invention comprise or consist of KKKKGGGMTPGTQSPFFLLLLLTVLTVV (SEQ ID NO: 22) or a functional variant thereof (e.g., TPGTQSPFFLLLLLTVLTVV with the same modifications and linker, i.e., SEQ ID NO: 23: KKKKGGGPGTQSPFFLLLLLTVLTVV).

[0171] Furthermore, the protein may be purified in higher yields and exhibit greater solubility than the corresponding native polypeptide, at least under certain purification and storage conditions. It is known in the art that for many proteins, including the extracellular domain of a membrane-bound protein or the mature form of a secreted protein, one or more amino acids may be deleted from the N- or C-terminus without substantial loss of biological function.

[0172] However, even if the deletion of one or more amino acids from the N-terminus of a protein results in the alteration or loss of one or more biological functions of the protein, other biological activities may still be retained.Thus, the ability of a truncated protein to induce and / or bind to antibodies that recognize the full or mature form of a protein is generally retained when less than half of the residues of the full or mature protein are removed from the N-terminus.Whether a particular polypeptide that lacks the N-terminal residues of a full protein retains its necessary activity can be determined by conventional methods known in the art.

[0173] Thus, the invention further provides polypeptides in which one or more residues have been deleted from the amino terminus of the amino acid sequence of the molecule. Similarly, many examples of biologically functional C-terminal deletion mutants are known.

[0174] However, even if the deletion of one or more amino acids from the C-terminus of a protein results in the alteration or loss of one or more biological functions of the protein, other biological activities may still be retained.Thus, the ability of a truncated protein to induce and / or bind to antibodies that recognize the complete or mature form of a protein is generally retained when less than half of the residues of the complete or mature protein are removed from the C-terminus.Whether a particular polypeptide that lacks the C-terminal residues of a complete protein retains such biological activity can be determined by conventional methods known in the art.

[0175] In addition to the truncated forms of the proteins described above, those skilled in the art will also recognize that some amino acid sequences of the therapeutic polypeptides of the invention can be altered without significantly affecting the structure or function of the protein. When such differences in sequence are contemplated, those skilled in the art will understand how to determine which domains are important regions for retaining protein function.

[0176] Such variants include deletions, insertions, inversions, repeats, and type substitutions, which are selected according to general rules known in the art so as to have little effect on activity.For example, guidelines for making phenotypically silent amino acid substitutions are provided in Bowie et al., (1990) Science 247:1306-1310, and the authors point out that there are two main approaches to studying tolerance to amino acid sequence changes.The first method relies on the evolutionary process, in which mutations are either accepted or rejected by natural selection.

[0177] The second approach uses genetic engineering to introduce amino acid changes at specific positions in cloned genes, followed by selection or screening to identify sequences that maintain function. These studies have demonstrated that proteins are surprisingly tolerant of amino acid substitutions. It is known that certain amino acid changes are more likely to be tolerated at specific positions in proteins. For example, most buried amino acid residues require nonpolar side chains, whereas surface side chain features are generally poorly conserved.

[0178] Typically found as conservative substitutions are the replacement of the aliphatic amino acids Ala (A), Val (V), Leu (L), and Ile (I) with one another; the exchange of hydroxyl residues Ser (S) and Thr (T), the exchange of acidic residues Asp (D) and Glu (E), the exchange between amide residues Asn (N) and Gln (Q), the exchange of basic residues Lys (K) and Arg (R), and the exchange between aromatic residues Phe (F) and Tyr (Y). Thus, a polypeptide fragment, derivative, or analog can be one in which (i) one or more amino acid residues are substituted with a conserved or non-conserved amino acid residue; such substituted amino acid residues may or may not be those encoded by the genetic code; (ii) one or more amino acid residues include a substituent group; (iii) the mature polypeptide is fused to another compound, such as a compound that increases the half-life of the polypeptide (e.g., polyethylene glycol); or (iv) additional amino acids are fused to the above forms of the polypeptide, such as an IgG Fc fusion region peptide, a leader or secretory sequence, a sequence used to purify the above forms of the polypeptide, or a proprotein sequence. Such fragments, derivatives, and analogs are deemed to be within the scope of one skilled in the art from the teachings herein.

[0179] Thus, the therapeutic polypeptides of the present invention may contain one or more amino acid substitutions, deletions, or additions, either due to natural mutation or human manipulation. As indicated, these changes may be minor in nature, such as conservative amino acid substitutions, that do not significantly affect protein folding or activity. For example, one or more amino acid substitutions, deletions, or additions do not affect the ability of the peptides described herein to inhibit, treat, or prevent inappropriate inflammatory responses.

[0180] Conservative amino acid substitutions include aromatic substitutions Phe, Trp, and Tyr; hydrophobic substitutions Leu, Iso, and Val; polar substitutions Glu and Asp; basic substitutions Arg, Lys, and His; acidic substitutions Asp and Glu; and small amino acid substitutions Ala, Ser, Thr, Met, and Gly.

[0181] Amino acids essential for the function of the therapeutic polypeptides of the invention can be identified by methods known in the art, such as site-directed mutagenesis or alanine scanning mutagenesis. In the latter procedure, single alanine mutations are introduced. The resulting mutant molecules are then tested for biological activity, such as receptor binding, or in vitro or in vitro proliferative activity.

[0182] Furthermore, substitution of charged amino acids with other charged or neutral amino acids can produce proteins with highly desirable improved properties, such as reduced aggregation, which not only reduces activity but can also be problematic when preparing pharmaceutical formulations, for example, because aggregates can be immunogenic.

[0183] Amino acid substitutions can also alter the selectivity of ligand binding to cell surface receptors. Sites important for ligand-receptor binding can also be determined by structural analyses such as crystallization, nuclear magnetic resonance, or photoaffinity labeling.

[0184] The present invention also provides nucleic acids or fragments thereof encoding proteins defined herein, including DNA or RNA sequences, including those having an open reading frame that encodes a therapeutic polypeptide and that can be expressed as one of the therapeutic polypeptides of the present invention under appropriate conditions. The term "nucleic acid" also encompasses DNA, cDNA, mRNA, splice variants, antisense RNA, RNAi, DNA containing one or more single nucleotide polymorphisms (SNPs), and vectors containing the subject nucleic acid sequence. Also encompassed by this term are nucleic acids that are homologous or substantially similar or identical to a nucleic acid encoding a therapeutic protein. Thus, the subject invention provides genes encoding the subject proteins, as well as homologs thereof.

[0185] The polynucleotide or nucleic acid of the present invention refers to a polymeric form of nucleotides of any length. Polynucleotides can contain deoxyribonucleotides, ribonucleotides, and / or their analogs or derivatives. For example, nucleic acids can be naturally occurring DNA or RNA, or synthetic analogs as known in the art. Polynucleotides of the present invention also encompass genomic DNA, genes, gene fragments, exons, introns, control sequences, or regulatory elements, such as promoters, enhancers, start and stop regions, other control regions, expression regulators, and expression controls; DNA, including one or more single nucleotide polymorphisms (SNPs), allelic variants, isolated DNA of any sequence, and cDNA; mRNA, tRNA, rRNA, ribozymes, splice variants, antisense RNA, antisense conjugates, RNAi, and isolated RNA of any sequence; recombinant polynucleotides, heterologous polynucleotides, branched polynucleotides, labeled polynucleotides, hybrid DNA / RNA, polynucleotide constructs, vectors containing the nucleic acid of interest, nucleic acid probes, primers, and primer pairs.

[0186] The polynucleotides of the present invention encompass modified nucleic acid molecules with alterations in the backbone, sugar, or heterocyclic base, such as methylated nucleic acid molecules, peptide nucleic acids, and nucleic acid molecule analogs, which may be suitable, for example, as probes if they exhibit superior stability and / or binding affinity under assay conditions. They also encompass single-stranded, double-stranded, and triple-helical molecules, either DNA, RNA, or hybrid DNA / RNA, which may encode full-length genes or biologically active fragments thereof.

[0187] The polynucleotides of the present invention include single nucleotide polymorphisms. Single nucleotide polymorphisms (SNPs) occur frequently in the genomes of eukaryotic organisms. A nucleotide sequence determined from an individual of a species may differ from other allelic forms present in the population. The present invention encompasses such SNPs. The subject polynucleotides include those encoding variants of the polypeptides described herein. Thus, in some embodiments, the subject polynucleotides encode variant polypeptides containing insertions, deletions, or substitutions compared to the polypeptides described herein. Conservative amino acid substitutions include serine / threonine, valine / leucine / isoleucine, asparagine / glutamine, glutamic acid / aspartic acid, etc.

[0188] In some embodiments, the proteins or functional derivatives described herein may be modified by the covalent attachment of any type of molecule to the composition to enhance its suitability for administration, i.e., so that the covalent attachment does not interfere with the activity of the composition. For example, but not limited to, derivatives include compositions modified by glycosylation, lipidation, acetylation, pegylation, phosphorylation, amidation, derivatization with known protecting / blocking groups, proteolytic cleavage, conjugation to cellular ligands or other proteins, among others. Any of a number of chemical modifications can be performed by known techniques, including, but not limited to, specific chemical cleavage, acetylation, formylation, metabolic synthesis of tunicamycin, and the like. Additionally, derivatives may contain one or more non-classical amino acids.

[0189] In still other embodiments, the proteins or functional derivatives described herein may be modified to attach effector moieties such as chemical linkers, detectable moieties such as fluorescent dyes, enzymes, substrates, bioluminescent materials, radioactive materials and chemiluminescent moieties, or functional moieties such as streptavidin, avidin, biotin, cytotoxins, cytotoxic agents and radioactive materials.

[0190] In another embodiment, the peptides or functional derivatives described herein can be modified to attach metal ion labels such as gadoteric acid, a macrocyclic gadolinium-based MRI contrast agent (GBCA) containing the organic acid DOTA as a chelating agent and gadolinium (Gd), used in the form of its meglumine salt (gadoterate meglumine). Such labels are also referred to herein as (Gd)Dota.

[0191] In yet another embodiment, the peptides described herein may contain synthetic isomers of photoreactive amino acid residues for the observation and characterization of protein-protein interactions (PPIs). A non-limiting example is photoleucine, a synthetic derivative of the amino acid leucine that is used as a natural analog and is characterized by its photoreactivity. Those skilled in the art will understand that when a protein containing this amino acid is irradiated with ultraviolet light while interacting with another protein, the complex formed between the two proteins will remain attached and can be isolated.

[0192] In certain embodiments, any of the peptides described herein may contain a cell-penetrating peptide (CPP) attached to the N- or C-terminal region to facilitate or assist the peptide's entry into cells. CPPs are typically amphipathic and cationic peptides, approximately 7-30 amino acid residues in length, that facilitate the rapid translocation of molecules across cell membranes. Non-limiting examples of cell-penetrating peptides include Tat and Tat-based peptides, although those skilled in the art will appreciate that numerous CPPs exist that can be used in accordance with the present invention. Examples of such CPPs are described in the literature, e.g., Habault and Poyet, (2019) Molecules, 24: 927; Borerlli et al., (2018) Molecules, 23: 295 (incorporated herein by reference).

[0193] In certain embodiments, any of the peptides described herein can be lipidated at the N- or C-terminal region, for example, to facilitate or assist entry of the peptides of the invention into cells.

[0194] Nucleic acids encoding proteins of the subject invention can be cDNA or genomic DNA or fragments thereof. The term "gene" is intended to mean the open reading frame encoding the specific proteins and polypeptides of the subject invention and introns, as well as adjacent 5' and 3' untranslated nucleotide sequences involved in regulating expression outside the translated region but potentially extending further in either direction, up to about 20 kb. The gene can be introduced into an appropriate vector for extrachromosomal maintenance or for integration into the host genome. The subject polynucleotide is generally isolated and obtained in substantial purity, other than as an intact chromosome. Typically, the DNA is obtained substantially free of other nucleic acid sequences not including the sequence of the subject gene or fragments thereof, generally at least about 50%, usually at least about 90%, pure, and typically "recombinant," i.e., flanked by one or more nucleotides not normally associated on the naturally occurring chromosome.

[0195] The present invention provides a plasmid, i.e., a small, independently replicating fragment of extrachromosomal cytoplasmic DNA that can be transferred from one organism to another, containing the therapeutic polynucleotide of the present invention. The plasmid can be integrated into the host genome or remain independent. Artificially constructed plasmids are commonly used as cloning vectors. The present invention also provides a vector, i.e., a plasmid, that can be used to transfer DNA sequences from one organism to another.

[0196] Expression vectors can be used to express the therapeutic gene products of the present invention and typically contain restriction sites to provide for the insertion of nucleic acid sequences encoding heterologous proteins or RNA molecules.

[0197] Expression vectors can be used to introduce genes into cells.Such vectors generally have convenient restriction sites located near promoter sequences to allow the insertion of nucleic acid sequences.A transcription cassette can be prepared, comprising a transcription initiation region, a target gene or its fragment, and a transcription termination region.Transcription cassettes can be introduced into various vectors, such as plasmids; retroviruses, such as lentiviruses; adenoviruses; adeno-associated viruses; and the like, and the vectors can be maintained in cells transiently or stably, usually for at least about one day, more usually for at least about several days to several weeks.

[0198] anti-inflammatory agents In an aspect of the present invention, a composition for preventing or treating inflammatory reactions is provided, comprising the peptides described herein and additional anti-inflammatory agents.Those skilled in the art can easily identify different types of anti-inflammatory agents that can be used in combination with the peptides described herein.Non-limiting examples of such anti-inflammatory agents include non-selective nonsteroidal anti-inflammatory drugs (NSAIDs), selective NSAIDs, corticosteroids, antibodies that exhibit anti-inflammatory properties (for example, by inhibiting the activity of inflammatory molecules), and other anti-inflammatory agents known to those skilled in the art.

[0199] Examples of non-selective NSAIDs include diclofenac, ibuprofen, indomethacin, ketoprofen, ketorolac, mefenamic acid, naproxen, piroxicam, and sulindac. Examples of selective NSAIDs include celecoxib, etoricoxib, meloxicam, and paracoxib.

[0200] In a preferred embodiment, a composition for the prevention or treatment of an inflammatory response is provided, comprising a peptide described herein and a corticosteroid.

[0201] Corticosteroids are synthetic drugs that mimic cortisol and function by reducing inflammation and suppressing immune system activity. They are used to treat a variety of inflammatory diseases and conditions, including psoriasis, asthma, inflammatory conditions, chronic obstructive pulmonary disease (COPD), hay fever, hives, eczema, vasculitis (inflammation of blood vessels) and myositis (inflammation of muscles), arthritis, inflammatory bowel disease (including Crohn's disease), rheumatoid arthritis, lupus, Sjögren's syndrome, and gout. Examples of corticosteroid drugs used herein include aristocort, decadron, mometasone, cotrone, triamcinolone, cortisone, prednisone, methylprednisolone, and betamethasone dipropionate.

[0202] Those skilled in the art will appreciate that the corticosteroids used in the present invention may be in one of the following dosage forms: Tablets, syrups and liquids - for example, prednisolone; Inhalants and nasal sprays - for example, beclomethasone and fluticasone; Injectables (intramuscular, intravenous, intraarticular) - for example, methylprednisolone; Ophthalmology - for example, eye drops; and Creams, lotions and gels - for example, hydrocortisone skin cream.

[0203] Corticosteroids described herein or known in the art are suitable for administration in dosages thereof.

[0204] In embodiments, the use of corticosteroids in the present invention serves to treat or prevent inflammatory responses. Thus, in this embodiment, the peptides described herein can be used in combination with corticosteroids to effectively treat or prevent inflammatory responses. In embodiments, this effect can be greater than the effect of the peptide alone, and in some embodiments, can be synergistic.

[0205] The corticosteroid can be administered simultaneously with the peptide of the present invention, including when provided in the same or different formulations. Alternatively, the corticosteroid and peptide can be administered at different times and in different dosage forms (formulations). When provided in different formulations, the peptide and corticosteroid can be administered by the same or different routes of administration.

[0206] Monoclonal antibodies The present invention also provides a composition for preventing or treating an inflammatory response, comprising a peptide and an antigen-binding protein described herein. Preferably, the antigen-binding protein is an antibody or an antigen-binding fragment thereof. Typically, the antigen-binding protein is an antibody, e.g., a monoclonal antibody.

[0207] In certain embodiments, antigen binding proteins (e.g., monoclonal antibodies) for inhibiting the activity of inflammatory molecules bind to anti-inflammatory molecules, thereby inhibiting the activity of the molecules in promoting an inflammatory response. The antigen binding proteins may bind to receptors for inflammatory molecules, thereby inhibiting the activity of the inflammatory molecules. Non-limiting examples of antigen binding proteins or monoclonal antibodies suitable for use in accordance with the present invention include those capable of inhibiting the activity of inflammatory molecules including interleukin-17 (IL-17), interleukin-12 (IL-12), interleukin-6 (IL-6), interleukin-22 (IL-22), interleukin-23 (IL-23), interleukin-36 (IL-36), and tumor necrosis factor alpha (TNFα). Examples of such antigen binding proteins or monoclonal antibodies include brodalumab, ixekizumab or secukinumab (to inhibit IL-17), tocilizumab or siltuximab (to inhibit IL-6), guselkumab or tildrakizumab or mirikizumab or brazikumab or risankizumab or ustekinumab (to inhibit IL-23), fezakinumab (to inhibit IL-22), etanercept, infliximab, adalimumab, certolizumab, certolizumab pegol or golimumab (to inhibit the activity of TNFα).

[0208] Methods for generating antibodies are known in the art and / or described in Harlow and Lane (editors) Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, (1988). Generally, in such methods, an immunogenic fragment or epitope thereof, or a cell expressing and presenting it (i.e., immunogen), optionally formulated with an appropriate or desired carrier, adjuvant, or pharmaceutically acceptable excipient, is administered to a non-human animal, such as a mouse, chicken, rat, rabbit, guinea pig, dog, horse, cow, goat, or pig. The immunogen may be administered intranasally, intramuscularly, subcutaneously, intravenously, intradermally, intraperitoneally, or by other known routes.

[0209] The production of polyclonal antibodies can be monitored by sampling the blood of immunized animals at various times after immunization.If necessary to achieve the desired antibody titer, one or more additional immunizations can be performed.The boosting and titering process is repeated until an appropriate titer is achieved.Once the desired level of immunogenicity is achieved, the immunized animals are bled, and the serum is isolated and stored, and / or the animals are used to generate monoclonal antibodies (mAbs).

[0210] Monoclonal antibodies are one example of an antibody form contemplated by the present invention. The term "monoclonal antibody" or "mAb" refers to a homogeneous antibody population capable of binding to the same antigen, e.g., the same epitope within an antigen. The term is not intended to be limited with regard to the source of the antibody or its method of production.

[0211] Antigen-binding proteins suitable for use in accordance with the present invention may be synthetic antibodies, for example, the antibody may be a chimeric antibody, a humanized antibody, a human antibody synhumanized antibody, a primatized antibody, or a de-immunized antibody.

[0212] Antigen-binding proteins or antibodies, such as monoclonal antibodies, (i) single-chain Fv fragment (scFv); (ii) dimeric scFv (di-scFv); (iii) one of (i) or (ii) linked to the antibody constant region Fc, or heavy chain constant domain (CH)2 and / or CH3; or (iv) one of (i) or (ii) conjugated to a protein that binds to immune effector cells; It may be in the form of:

[0213] Furthermore, as described herein, an antigen-binding site or antibody may comprise: (i) bispecific antibodies; (ii) trispecific antibodies; (iii) tetraspecific antibodies; (iv) Fab; (v) F(ab')2; (vi) Fv; (vii) one of (i) to (iv) linked to the antibody constant region Fc, or the heavy chain constant domain (CH)2 and / or CH3; or (viii) one of (i) to (iv) bound to a protein that binds to immune effector cells; It may be in the form of:

[0214] Any one of a number of known techniques can be used for preparation of mAbs, for example, procedures exemplified in US Pat. No. 4,196,265 or Harlow and Lane (1988), supra.

[0215] For example, a suitable animal is immunized with an immunogen under conditions sufficient to stimulate antibody-producing cells. Exemplary animals include rodents such as rabbits, mice, and rats. For example, mice genetically engineered to express human antibodies that do not express mouse antibodies can also be used to generate antibodies of the present invention (e.g., as described in WO2002 / 066630).

[0216] Following immunization, somatic cells with the potential to produce antibodies, particularly B lymphocytes (B cells), are selected for use in mAb generation protocols. These cells can be obtained from biopsies of the spleen, tonsils, or lymph nodes, or from a peripheral blood sample. The B cells from the immunized animal are then fused with cells of an immortal myeloma cell, generally derived from the same species as the animal immunized with the immunogen.

[0217] The hybrids are amplified by culturing in a selective medium containing drugs that block de novo synthesis of nucleotides in tissue culture media. Exemplary drugs are aminopterin, methotrexate, and azaserine.

[0218] The amplified hybridomas are subjected to functional selection for antibody specificity and / or titer, for example, by flow cytometry and / or immunohistochemistry and / or immunoassays (e.g., radioimmunoassays, enzyme immunoassays, cytotoxicity tests, plaque assays, dot immunoassays, and the like).

[0219] Alternatively, ABL-MYC technology (NeoClone, Madison, WI 53713, USA) is used to generate MAb-secreting cell lines (e.g., as described in Largaespada et al, J. Immunol. Methods. 197: 85-95, 1996).

[0220] Antibodies can also be generated or isolated by screening display libraries, such as phage display libraries as described in U.S. Patent No. 6,300,064 and / or U.S. Patent No. 5,885,793. For example, the present inventors have isolated fully human antibodies from phage display libraries.

[0221] Suitable dosages of the antigen-binding proteins or antibodies described herein will vary depending on the particular antigen-binding site, the condition being treated, and / or the subject being treated. It is within the ability of a skilled physician to determine appropriate dosages, for example, by starting with a suboptimal dosage and gradually modifying the dosage to determine an optimal or useful dosage. Alternatively, data from cell culture assays or animal studies can be used to determine appropriate therapeutic / prophylactic dosages, where an appropriate dose is determined to be the ED of the active compound with little or no toxicity. 50The circulating concentration range includes the IC50 / IC50 / IC60 / IC70 / IC80 / IC9 ... 50 A dose can be formulated in animal models to achieve a circulating plasma concentration range that includes the compound (i.e., the concentration or amount of compound that achieves a half-maximal inhibition of symptoms). Such information can be used to more accurately determine useful doses in humans. Plasma concentrations can be measured, for example, by high performance liquid chromatography.

[0222] Pharmaceutical Compositions and Routes of Administration Provided herein are pharmaceutical compositions comprising a pharmaceutically effective amount of the peptides described herein. In additional embodiments, pharmaceutical compositions comprising pharmaceutically acceptable salts are provided.

[0223] The term "pharmaceutically acceptable salt" also refers to a salt of the composition of the present invention having an acidic functional group, such as a carboxylic acid functional group, and a base. Pharmaceutically acceptable salts include, but are not limited to, sulfate, citrate, acetate, oxalate, chloride, bromide, iodide, nitrate, bisulfate, phosphate, acid phosphate, isonicotine, lactate, salicylate, acid citrate, tartaric acid, oleate, tannic acid, pantothenic acid, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucaronate, saccharate, Formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, camphorsulfonate, pamoate, phenylacetate, trifluoroacetate, acrylate, chlorobenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, methylbenzoate, o-acetoxybenzoate, naphthalene-2-benzoate, isobutyrate, phenyl Mention may be made of butyrate, α-hydroxybutyrate, butyne-1,4-dicarboxylate, hexyne-1,4-dicarboxylate, caprate, caprylate, cinnamate, glycolate, heptanoate, hippurate, malate, hydroxymaleate, malonate, mandelate, mesylate, nicotinate, phthalate, terephthalate, propiolate, propionate, phenylpropionate, sebacate, suberate, p-brornobenzenesulfonate, chlorobenzenesulfonate, ethylsulfonate, 2-hydroxyethylsulfonate, methylsulfonate, naphthiene-1-sulfonate, naphthalene-2-sulfonate, naphthiene-1,5-sulfonate, xylenesulfonate and tartrate salts.

[0224] Furthermore, the peptides or functional variants described herein can be administered to a subject as a component of a composition comprising a pharmaceutically acceptable carrier or vehicle. Such compositions may contain a suitable amount of a pharmaceutically acceptable excipient to provide a dosage form for proper administration.

[0225] The pharmaceutical excipient may be a liquid such as water or an oil, including those of petroleum, animal, vegetable, or synthetic origin, such as peanut oil, soybean oil, mineral oil, and sesame oil. The pharmaceutical excipient may be, for example, saline, acacia gum, gelatin, starch paste, talc, keratin, colloidal silica, and urea. Additionally, auxiliary substances, stabilizers, thickeners, lubricants, and coloring agents may be used.

[0226] In one embodiment, pharmaceutically acceptable excipients are sterile when administered to a subject. When the agents described herein are administered intravenously, water is a useful excipient. Physiological saline and aqueous solutions of dextrose and glycerol can also be used as liquid excipients, particularly for injection solutions. Suitable pharmaceutical excipients also include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol, etc. The agents described herein can also contain minor amounts of wetting or emulsifying agents, or pH buffering agents, if desired.

[0227] In one embodiment, the peptides or functional variants described herein can be in the form of a solution, suspension, emulsion, drops, tablets, pills, pellets, capsules, liquid-containing capsules, powders, sustained-release formulations, suppositories, emulsions, aerosols, sprays, suspensions, nanoparticles or microneedles, or any other suitable dosage form. In one embodiment, the composition is in the form of a capsule. Other examples of suitable pharmaceutical excipients are described in Remington's Pharmaceutical Sciences 1447-1676 (Alfonso R. Gennaro eds., 19th ed. 1995), which is incorporated herein by reference.

[0228] Optionally, the peptides or functional variants described herein also include a solubilizing agent, and the agent can be delivered using an appropriate vehicle or delivery device, as known in the art.

[0229] The peptides outlined herein can be co-delivered in a single delivery vehicle or device. The composition for administration may also include a local anesthetic, such as lignocaine, to reduce pain at the injection site.

[0230] Formulations comprising the peptides or functional variants described herein, or the corticosteroids or monoclonal antibodies described herein, may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art. Such methods generally include the step of bringing the therapeutic agent into association with the carrier, which constitutes one or more accessory ingredients. Typically, the formulations are prepared by uniformly and intimately associating the therapeutic agent with liquid carriers, finely divided solid carriers, or both, and then, if necessary, shaping the product into the desired formulation (e.g., wet or dry granulation, powder blending, etc., followed by tableting using conventional methods known in the art).

[0231] In one embodiment, the peptides or functional variants described herein, or the corticosteroids or monoclonal antibodies described herein, are formulated according to conventional procedures into compositions suitable for the administration modes described herein. In one aspect, the pharmaceutical compositions are formulated for administration to the respiratory tract, skin, or gastrointestinal tract. Thus, pharmaceutical compositions comprising the peptides described herein for administration to the respiratory tract can be formulated as inhalable substances as is common in the art and described herein. In another embodiment, pharmaceutical compositions comprising the peptides described herein for administration to the gastrointestinal tract can be formulated with an enteric coating as is common in the art and described herein. In yet another embodiment, pharmaceutical compositions comprising the peptides described herein for administration to the skin include a carrier that allows the composition to be absorbed into the dermal layer of an individual in need thereof.

[0232] In embodiments, the pharmaceutical composition can be administered in a single dose or multiple doses. The pharmaceutical composition can be administered 1 to 3 times per 24 hours, or daily for a period of 7 days or more. The frequency and timing of administration can be as described in the Examples.

[0233] Routes of administration include, for example, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, oral, sublingual, intracerebral, intralymphatic, intratracheal, intraarticular, intravaginal, transdermal, rectal, by inhalation, or topically, particularly to the ear, nose, eye, or skin. In some embodiments, administration is oral or by parenteral injection. The mode of administration is left to the discretion of the practitioner and will depend in part on the site of the pathology. In most cases, administration will result in the release of the agents described herein into the bloodstream.

[0234] The peptides disclosed herein can be administered in a total amount of about 0.2 to about 2400 mg per day. The dose can be divided into two or three doses over the course of the day, or can be administered once daily. Specific total daily amounts of therapeutic agent contemplated by the disclosed methods include about 0.2 mg, about 0.5 mg, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg, about 95 mg, about 100 mg, about 105 mg, about 110 mg, about 115 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, about 190 mg, about 210 mg, about 220 mg, about 230 mg, about 240 mg, about 250 mg, about 260 mg, about 270 mg, about 280 mg, about 290 mg, about 300 mg, about 310 mg, about 320 mg, about 330 mg, about 340 mg, about 350 mg, about 360 mg, about 370 mg, about 380 mg, about 390 mg, about 400 mg, about 410 mg, about 420 mg, about 430 mg, about 440 mg, about 450 mg, about 460 mg, about 470 mg, about 480 mg, about 490 mg, about 500 mg, about 510 mg, about 520 mg, about 530 mg Examples include 20 mg, about 125 mg, about 130 mg, about 135 mg, about 140 mg, about 145 mg, about 150 mg, about 200 mg, about 250 mg, about 267 mg, about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 534 mg, about 550 mg, about 600 mg, about 650 mg, about 700 mg, about 750 mg, about 800 mg, about 850 mg, about 900 mg, about 950 mg, about 1000 mg or more.

[0235] In some embodiments, the patient or individual is a mammal, such as a human, mouse, rat, guinea pig, dog, cat, horse, cow, pig, rabbit, sheep, or a non-human primate such as a monkey, chimpanzee, or baboon. In other embodiments, the subject and / or animal is a non-mammal, such as a zebrafish. In some embodiments, the subject and / or animal may comprise fluorescently tagged cells (e.g., with GFP). In some embodiments, the subject and / or animal is a transgenic animal comprising fluorescent cells.

[0236] In some embodiments, the patient or individual is a human. In some embodiments, the human is a pediatric human. In other embodiments, the human is an adult human. In other embodiments, the human is an elderly human. In other embodiments, the human may be referred to as a patient.

[0237] In certain embodiments, the human is aged in the range of about 0 to about 6 months, about 6 to about 12 months, about 6 to about 18 months, about 18 to about 36 months, about 1 to about 5 years, about 5 to about 10 years, about 10 to about 15 years, about 15 to about 20 years, about 20 to about 25 years, about 25 to about 30 years, about 30 to about 35 years, about 35 to about 40 years, about 40 to about 45 years, about 45 to about 50 years, about 50 to about 55 years, about 55 to about 60 years, about 60 to about 65 years, about 65 to about 70 years, about 70 to about 75 years, about 75 to about 80 years, about 80 to about 85 years, about 85 to about 90 years, about 90 to about 95 years, or about 95 to about 100 years.

[0238] In other embodiments, the subject is a non-human animal, and thus the invention relates to veterinary uses. In a specific embodiment, the non-human animal is a domestic pet. In another specific embodiment, the non-human animal is a livestock animal.

[0239] Intestinal application In embodiments, there is provided a method of treating or preventing an inappropriate inflammatory response in the gastrointestinal tract of an individual, the method comprising administering to an individual in need thereof a pharmaceutical composition comprising a pharmaceutically effective amount of a peptide as described herein, or a pharmaceutically acceptable salt thereof, thereby treating or preventing an inappropriate inflammatory response in the gastrointestinal tract of the individual.

[0240] A positive response to treatment can be prevention or attenuation of inflammatory bowel symptom exacerbations, for example, reduction in inflammatory erythema or levels of cytokine / chemokine expression, ulcer formation, intestinal discomfort and pain.

[0241] The compositions provided herein can be formulated by various methods that are clear to those skilled in the art of pharmaceutical formulations.The above-mentioned various release characteristics can be achieved by various different methods.Suitable formulations include, for example, tablets, capsules, press-coated formulations and other easy-to-administer formulations.

[0242] Suitable pharmaceutical preparations may contain, for example, about 0.1% to about 99.9%, preferably about 1% to about 60%, of the active ingredient. Pharmaceutical preparations for combination therapy for enteral or parenteral administration are, for example, in unit dosage forms such as sugar-coated tablets, tablets, capsules, or suppositories, or ampoules. Unless otherwise specified, these are prepared in a manner known per se, for example, by conventional mixing, granulating, sugar-coating, dissolving, or lyophilizing processes. It will be understood that the unit content of the combination partner contained in each individual dose of each dosage form does not necessarily constitute an effective amount, since the required effective amount can be reached by administering multiple dosage units.

[0243] Respiratory applications In embodiments, methods are provided for treating or preventing an inappropriate inflammatory response in the airways of an individual, comprising administering to an individual in need thereof a pharmaceutical composition comprising a pharmaceutically effective amount of a peptide as described herein, or a pharmaceutically acceptable salt thereof, thereby treating or preventing an inappropriate inflammatory response in the airways of the individual.

[0244] The term "respiration" refers to the process of taking in oxygen and expelling carbon dioxide through the body's systems, including the nose, throat, larynx, trachea, bronchi, and lungs.

[0245] The term "respiratory disease" or "respiratory condition" refers to any one of several diseases that involve inflammation and affect components of the respiratory system, including the upper respiratory tract (including the nasal passages, pharynx, and larynx) and the lower respiratory tract (including the trachea, bronchi, and lungs).

[0246] Symptoms of respiratory illness may include cough, excessive sputum production, shortness of breath, or chest tightness with audible wheezing. Exercise capacity may be significantly limited. The impact of each of these conditions can also be measured by the number of days missed from work / school, sleep disturbances, need for bronchodilators, and need for glucocorticoids, including oral glucocorticoids.

[0247] The presence, improvement, treatment, or prevention of respiratory disease can be determined by clinical or biochemical methods of a subject or a biopsy from the subject. For example, the parameters measured can be the presence or degree of pulmonary function, signs and symptoms of obstruction, exercise tolerance, nighttime awakenings, days missed from school or work, bronchodilator use, inhaled corticosteroid (ICS) dose, oral (glucocorticoid) GC use, need for other medications, need for medical treatment, or hospitalization.

[0248] A positive response to treatment can also be prevention or reduction of worsening respiratory symptoms, such as a decrease in inflammatory erythema or levels of cytokine / chemokine expression, a decrease in cough or sputum production, or a decrease in chest tightness.

[0249] The peptides described herein may be in compositions formulated for administration to the upper respiratory tract (URT), lower respiratory tract (LRT), or both the URT and LRT. The peptides described herein may be formulated for intranasal administration, including as a dry powder, spray, mist, or aerosol. This may be particularly preferred for the treatment of respiratory infections. Suitable formulations for administration in which the carrier is a liquid, such as a nasal spray or nasal drops, include aqueous or oily solutions of the active ingredient. Alternatively, the composition may be a dry powder administered to the upper respiratory tract, including excipients such as mannitol, trehalose, lactose, etc. Liquid formulations or dry powders can be designed for delivery to the upper and lower respiratory tracts.

[0250] Other ingredients, such as preservatives, colorants, lubricants, or viscous mineral or vegetable oils, flavorings, natural or synthetic plant extracts, such as aromatic oils, and humectants and thickeners, such as glycerol, known in the art, can also be included to provide the formulation with additional viscosity, moisturizing properties, a pleasant texture, and odor.For nasal administration of the solutions or suspensions according to the present invention, various devices for producing droplets, droplets, and sprays are available in the art.For example, the compounds or compositions described herein can be administered intranasally using a simple dropper (or pipette) comprising a glass, plastic, or metal dispensing tube, the contents of which are expelled drop by drop by air pressure provided by a manual pump (e.g., a soft rubber bulb) attached to one end.

[0251] Because tear secretions from the eye drain from the orbit into the nasal cavity, if desired, a suitable pharmaceutically acceptable ophthalmic solution can be readily provided by one of skill in the art as a carrier for the peptides or compositions described herein to be delivered, and can be administered in the form of eye drops into the orbit of the eye to provide both ophthalmic and intranasal administration.

[0252] In one embodiment, a pre-measured unit dose dispenser, including a dropper or spray device containing a solution or suspension for delivery as droplets or a spray, is prepared to contain one or more doses of the drug to be administered. The invention also includes kits containing one or more unit dehydrated doses of the compound, along with any necessary salts and / or buffers, preservatives, colorants, etc., ready to be prepared into a solution or suspension by the addition of an appropriate amount of water. The water can be sterile or non-sterile, although sterile water is generally preferred.

[0253] Skin application In embodiments, methods are provided for treating or preventing an inappropriate inflammatory response in the dermis layer or skin (including the epidermis, hypodermis / subcutaneous tissue) of an individual, comprising administering to an individual in need thereof a pharmaceutical composition comprising a pharmaceutically effective amount of a peptide, a peptide as described herein, or a pharmaceutically acceptable salt thereof, thereby treating or preventing an inappropriate inflammatory response in the dermis layer or skin of the individual.

[0254] A positive response to treatment can be prevention or attenuation of worsening of skin symptoms, such as a decrease in inflammatory erythema or levels of cytokine / chemokine expression, swelling, rash, scaling, and / or fluid leakage from the affected area.

[0255] The pharmaceutical composition suitable for treating inappropriate inflammatory reactions in the skin can be formulated with one or more excipients.In addition, the pharmaceutical composition can also be administered together with a steroid cream, a moisturizer, a coal tar, a cream or ointment, or a retinoid cream.

[0256] Other substances, such as biologically active agents, pharmaceutical excipients and cosmetic agents, may be included in the topical compositions of the present invention.

[0257] Biologically active agents can include, but are not limited to, flavanoid / flavone compounds, including, but not limited to, tanetin, 3,7,3'-trimethoxyquercetagetin, apigenin and its derivatives. When present, flavanoid / flavone compounds are present at a concentration of between about 0.001% and about 0.5%, preferably between about 0.005% and 0.2%, by weight of the topical composition.

[0258] Additional biologically active agents include, but are not limited to, sunscreens, anti-wrinkle / anti-aging agents, anti-fungals, antibiotics, anti-acne and anti-psoriasis agents, depigmenting agents, and such agents may be utilized so long as they are physically and chemically compatible with the other components of the topical composition.

[0259] The composition of the present invention may contain additional skin active agents. The active agents may be, but are not limited to, vitamin compounds. Skin lightening agents (such as kojic acid, ascorbic acid, and derivatives such as ascorbyl palmitate); antioxidants such as tocopherol and esters; nicotinamide, metal chelators, retinoids and derivatives, moisturizers, hydroxy acids such as salicylic acid, sunscreens such as octyl methoxycinnamate, oxybenzone, and avonzone, sunblocks such as titanium dioxide and zinc oxide, and skin protectants. Mixtures of the above skin active agents may be used.

[0260] Sunscreens that can be used in the compositions of the present invention can include, but are not limited to, organic or inorganic sunscreens such as octyl methoxycinnamate and other cinnamic acid compounds, titanium dioxide, and zinc oxide.

[0261] Antifungal agents include, but are not limited to, miconazole, econazole, ketoconazole, itraconazole, fluconazole, bifoconazole, terconazole, butoconazole, tioconazole, oxiconazole, sulconazole, saperconazole, clotrimazole, undecylenic acid, haloprogin, butenafine, tolnaftate, nystatin, ciclopirox olamine, terbinafine, amorolfine, naftifine, elubiol, griseofulvin, and pharmaceutically acceptable salts thereof.

[0262] Antibiotics (or antiseptics) include, but are not limited to, mupirocin, neomycin sulfate, bacitracin, polymyxin B, l-ofloxacin, tetracyclines (chlortetracycline hydrochloride, oxytetracycline hydrochloride, and tetracycline hydrochloride), clindamycin phosphate, gentamicin sulfate, benzalkonium chloride, benzethonium chloride, hexylresorcinol, methylbenzethonium chloride, phenol, quaternary ammonium compounds, triclocarbon, triclosan, tea tree oil, benzoyl peroxide, and pharmaceutically acceptable salts thereof.

[0263] Anti-acne ingredients include, but are not limited to, agents that normalize epidermal differentiation (e.g., retinoids), keratolytic agents (e.g., salicylic acid and alpha-hydroxy acids), benzoyl peroxide, antibiotics, and sebum-regulating compounds or plant extracts.

[0264] Antipsoriatic agents include, but are not limited to, corticosteroids (e.g., betamethasone dipropionate, betamethasone valerate, clobetasol propionate, diflorasone diacetate, halobetasol propionate, amcinonide, desoximetasone, fluocinonide, fluocinolone acetonide, halcinonide, triamcinolone acetate, hydrocortisone, hydrocortisone valerate, hydrocortisone butyrate, aclometasone dipropionate, flurandrenolide, mometasone furoate, methylprednisolone acetate), vitamin D and analogs thereof (e.g., calcipotriene, retinoids (e.g., tazarotene), and anthraline).

[0265] Cosmetic agents that can be used in the compositions of the present invention include, but are not limited to, emollients, vitamins and antioxidants (e.g., vitamin E), and herbal extracts (e.g., aloe vera) to prevent potential skin irritation. Cosmetic agents can also include moisturizers, antioxidants / preservatives, botanical extracts, fragrances, fragrances, and surfactants. Examples of moisturizers include glycerol, sorbitol, propylene glycol, ethylene glycol, 1,3-butylene glycol, polypropylene glycol, xylitol, maltitol, lactitol, oat protein, allantoin, acetamine MEA, and hyaluronic acid. These may be used alone or in combination.

[0266] Cosmetic agents can also include substances that mask the symptoms of inflammatory disorders and related conditions; such substances include, but are not limited to, pigments, dyes, and other additives (e.g., silica, talc, zinc oxide, titanium oxide, clay powder). Pharmaceutical excipients include, but are not limited to, pH-modifying agents, organic solvents (e.g., propylene glycol, glycerol, etc.), cetyl alcohol, kaolin, talc, zinc oxide, titanium oxide, corn starch, sodium gluconate, oils (e.g., mineral oil), ceteareth-20, ceteth-2, surfactants and emulsifiers, thickeners (or binders), fragrances, antioxidants, preservatives, and water.

[0267] Binder or thickener can be used in the composition of the present invention to give the composition durability and physical stability.Binder or thickener suitable for use in the composition of the present invention includes cellulose derivatives such as carboxymethylcellulose, methylcellulose, hydroxyethylcellulose and alkali metal salts of sodium carboxymethylhydroxyethylcellulose, alkali metal alginates such as sodium alginate, propylene glycol alginate, gums such as carrageenan, xanthan gum, tragacanth gum, karaya gum and gum arabic, and synthetic binders such as polyvinyl alcohol, sodium polyacrylate and polyvinylpyrrolidone.Thickeners such as natural gums and synthetic polymers, as well as coloring agents and fragrances, are also generally included in this composition.

[0268] Examples of preservatives that can be used in the compositions of the present invention include, but are not limited to, salicylic acid, chlorhexidine hydrochloride, phenoxyethanol, sodium benzoate, methyl parahydroxybenzoate, ethyl parahydroxybenzoate, propyl parahydroxybenzoate, and butyl parahydroxybenzoate.

[0269] Examples of flavors and fragrances that can be used in the compositions of the present invention include menthol, anethole, carvone, eugenol, limonene, ocimene, n-decyl alcohol, citronellol, α-terpineol, methyl salicylate, methyl acetate, citronellyl acetate, cineole, linalool, ethyl linalool, vanillin, thymol, spearmint oil, peppermint oil, lemon oil, orange oil, sage oil, rosemary oil, cinnamon oil, pimento oil, cinnamon leaf oil, perilla oil, wintergreen oil, clove oil, and eucalyptus oil.

[0270] The compositions of the present invention can be prepared in many forms for topical application to a patient. For example, the compositions can be applied as gels, creams, ointments, shampoos, scalp conditioners, liquids, spray-on preparations, paint / brush-on preparations, aerosols, powders, or adhesive bandages. Additionally, the compositions can be impregnated into bandages, hydrocolloids or other wound dressings, therapeutic patches, or cloth wipe products such as baby wipes or facial wipes.

[0271] The compositions of the present invention may be in the form of emulsions, such as creams and lotions. Such compositions may have two or more phases and may contain surfactants that allow for the production of multiple emulsions.

[0272] Examples of surfactants that can be used in the compositions of the present invention include sodium alkyl sulfates, such as sodium lauryl sulfate and sodium myristyl sulfate, sodium N-acylsarcosinates, such as sodium N-lauroylsarcosinate and sodium N-myristoylsarcosinate, sodium dodecylbenzenesulfonate, hydrogenated coconut fatty acid monoglyceride sodium sulfate, sodium lauryl sulfoacetate, and N-acylglutamates, such as N-palmitoylglutamate, N-methylacyltaurate sodium salt, N-methylacylalanine sodium salt, sodium α-olefinsulfonate, and sodium dioctyl sulfosuccinate; N-alkylaminoglycerols, such as N-lauryldiaminoethylglycerol; These include cerol and N-myristyldiaminoethyl glycerol, N-alkyl-N-carboxymethylammonium betaine and 2-alkyl-1-hydroxyethyl imidazoline betaine sodium; polyoxyethylene alkyl ether, polyoxyethylene alkylaryl ether, polyoxyethylene lanolin alcohol, polyoxyethylene glyceryl mono-fatty acid ester, polyoxyethylene sorbitol fatty acid ester, polyoxyethylene fatty acid ester, higher fatty acid glycerin ester, sorbitan fatty acid ester, Pluronic surfactant, and polyoxyethylene sorbitan fatty acid ester such as polyoxyethylene sorbitan monooleate and polyoxyethylene sorbitan monolaurate.The emulsifier surfactant known to those skilled in the art should be used in the composition of the present invention.

[0273] Another aspect of the present invention is a dermatologically acceptable carrier. Such a suitable carrier is suitable for topical use. It is not only compatible with the active ingredients described herein, but also does not cause any toxicity or safety issues. An effective and safe carrier may range from about 50% to about 99% by weight of the composition of the present invention, more preferably from about 75% to about 99% by weight of the composition, and most preferably from about 85% to about 95% by weight of the composition.

[0274] The choice of pharmaceutical excipient, biological agent, or cosmetic agent to use is often controlled or influenced by the type of inflammatory disease or related condition being treated. For example, when the composition of the present invention is used to treat dermatitis associated with athlete's foot, jock itch, or diaper rash, talc is the preferred pharmaceutical excipient, and antifungal agents are the preferred biological agents. When the composition of the present invention is used to treat scalp eczema, emulsifiers and oils are the preferred pharmaceutical excipients.

[0275] kit The present invention provides a kit that can simplify the administration of the agents described herein. An exemplary kit of the present invention includes a composition described herein in unit dosage form. In one embodiment, the unit dosage form is a container, such as a pre-filled syringe, which may be sterile and contains an agent described herein and a pharmaceutically acceptable carrier, diluent, excipient, or vehicle. The kit may further include a label or printed instructions for use of the agent described herein. The kit may also include a lid speculum, a local anesthetic, and an administration site cleanser. The kit may also further include one or more additional agents described herein. In one embodiment, the kit includes a container containing an effective amount of the composition of the present invention and an effective amount of another composition, such as one described herein.

[0276] The present invention further includes kits comprising one or more of the following: (i) a peptide described herein; (ii) an antigen-binding site of the present invention; (iii) an anti-inflammatory agent as described herein; (iii) A pharmaceutical composition of the present invention.

[0277] In an embodiment, the kit may further comprise a detection means, eg, linked to a peptide of the invention.

[0278] In the case of a kit for therapeutic / prophylactic use, the kit may further comprise a pharmaceutically acceptable carrier.

[0279] Kits of the invention may be packaged with instructions for use in the methods described herein, according to the examples.

[0280] It will be understood that the invention disclosed and defined herein extends to all alternative combinations of two or more of the individual features mentioned or apparent from the text or drawings, all of which different combinations constitute various alternative aspects of the invention. [Example]

[0281] Example 1: Generation of peptides suitable for use in the present invention Synthesis of RP23 (SEQ ID NO: 12). H-KKKK-CH2CH2OCH2CH2OCH2CO-MTPGTQSPFFLLLLLTVLTVV-OH RP23 (peptide 12) was synthesized on a 12.5 μmol scale by automated SPPS on a Chemmatrix® CTC resin, with coupling reactions carried out at 50°C. The pseudoproline dipeptide Fmoc-L-Leu-L-Thr[ψMe,MePro]-OH was incorporated at both Leu-Thr bonds. The peptide was cleaved from the resin with TFA:iPrSiH:thioanisole:HO (90:2.5:2.5:5 v / v / v / v). After preparative HPLC purification and lyophilization, RP23 (peptide 12) was obtained as a white solid (9.6 mg, 26% yield). LRMS calculated: [M+2H] 2+ = 1475.38, [M+3H] 3+ = 983.92, [M+4H] 4+ = 738.19; Measurements (ESI+) 1145.90, 764.10. C 140 H 239 N 31 O 35HRMS calculated for S: [M+H]+ = 2946.760; Found (MALDI-TOF): 2946.758.

[0282] Example 2: Effect of peptide administration on local suppression of the immune system Contact hypersensitivity model - methods The abdomens of 8-week-old female C57BL / 6 mice (Australian BioResources) were roughly shaved. At least 24 hours later, RP23 (SEQ ID NO: 12) was delivered by subcutaneous injection in the abdomen in 50 μl of phosphate-buffered saline. Seven days later, mice were topically sensitized to 1-fluoro-2,4-dinitrobenzene (DNFB, 0.5 vol%) in olive oil / acetone (1:4, v / v, 30 μl) via application to the shaved abdomen. Five days after sensitization, mice were challenged in the ears with topical DNFB (0.25 vol%) in olive oil / acetone (1:4, v / v, 15 μl per ear). Ear thickness was measured daily using manual calipers from immediately before challenge until 96 hours after challenge. Positive controls for ear swelling consisted of mice injected with PBS instead of RP23. Negative or irritation controls consisted of mice that received DNFB challenge rather than sensitization. To account for nonspecific irritation due to the application of DNFB, olive oil and acetone, the mean value of the ear thickness change from this control group was subtracted from the ear thickness change of the other samples.

[0283] Imiquimod-induced psoriasis model - Methods The backs of 8-week-old female C57BL / 6 mice (Australian BioResources) were roughly shaved. At least 24 hours later, 10 nmol of RP23 was delivered by subcutaneous injection in the lumbar region in 200 μl of phosphate-buffered saline (PBS). A group of mice serving as a positive control for disease development received PBS injections alone. One or seven days later, 5% imiquimod cream (Aldara, Inova Pharmaceuticals) was administered topically to the shaved back (57.5 mg) and one ear (5 mg, divided into dorsal and ventral regions) to induce psoriasis-like inflammation. The contralateral ear served as a negative control for imiquimod-induced inflammation. Imiquimod application was repeated daily for four consecutive days. C57BL / 6 mice experience rapid weight loss (up to 20%) following application of imiquimod cream. Supportive care was provided in the form of nutrient supplementation initiated 2 days before imiquimod application and intraperitoneal injections of 300 μl of sterile PBS on days 2 and 3 of imiquimod application. Disease progression was monitored for 5 days by recording weight loss and grading disease on the back skin using the modified Psoriasis Area and Severity Index (PASI) clinical scoring system, which assesses disease severity only. In addition, erythemameter readings (DSM II ColorMeter, Cortex Technology, Denmark) were obtained in four quadrants of the back skin, and skin backfold thickness measurements were recorded using manual calipers. Disease progression on the ear skin was determined by recording ear thickness measured by manual calipers. As a positive control for immunosuppressive therapy, groups of mice received daily topical application of betamethasone dipropionate 0.05% cream (MSD) (30 mg on the back and 10 mg on the ears). Mice receiving steroid therapy experienced a greater degree of weight loss and a significant reduction in spleen weight, indicating systemic immunosuppression.

[0284] We first tested RP23 (SEQ ID NO: 12) as a vaccine antigen in a skin tumor mouse model. Surprisingly, we did not observe a reduction in skin tumor growth in mice treated with RP23; rather, we observed a slight increase in tumor growth (Figure 1), suggesting an immunosuppressive effect. Next, we evaluated whether RP23 could reduce inflammation in a mouse model of contact dermatitis using a contact hypersensitivity assay (Figure 2a). A single nanomolar injection of RP23 before sensitization significantly suppressed the induction of inflammation, as measured by a decrease in ear thickness (Figure 2b) and skin infiltration of inflammatory myeloid cells (Figure 2c). This immunosuppression is specific to the RP23 sequence, as a peptide with the same amino acids in a scrambled order did not significantly reduce ear swelling (Figure 3a). RP23 significantly increased synthetic yield without significantly altering immunosuppression compared to the effect of SEQ ID NO: 3 (MUC1SP) (Figure 3b). Using RP23, we performed a dose-response assay to titrate immunosuppression. Using three doses in the low nanomolar range (2, 5, or 10 nmol, approximately 0.3, 0.8, and 1.6 mg / kg), we observed a dose-dependent reduction in ear swelling up to 100% compared to controls (Figure 3c). Including GM-CSF in the assay abolished much of the suppression provided by RP23 (Figure 3d). Furthermore, in an imiquimod-induced psoriasis mouse model (Figure 4a), a single injection of RP23 reduced local erythema, skin thickness, and scaling (Figures 4b and 4c). This immunomodulatory effect was limited to the area where RP23 was delivered, unlike topical application of glucocorticoids, which caused undesirable systemic immunosuppression (Figure 4d). No side effects of any kind were observed in RP23-injected mice, even when a dose of 10 nmol was injected subcutaneously daily for 6 days (data not shown), supporting the nontoxic and nonirritant nature of this peptide.

[0285] We used an in vitro culture system to investigate whether RP23 can suppress the activity of specific proinflammatory immune cells. In primary mouse splenic dendritic cells, RP23 reduced the secretion of inflammatory chemokines (data not shown) and the expression of surface activation markers (Figure 5a). Culture of mouse bone marrow-derived macrophages with RP23 resulted in reduced release of IL-6, MCP-1, and TNF after LPS stimulation (Figure 5b). Similarly, primary human blood-derived macrophages reduced the release of IL-6 and IL-12 / 23 after stimulation with either LPS or imiquimod (Figure 5c). RP23 was able to induce these changes in macrophages without causing cytostatic or cytotoxic effects on the cells (Figure 6), demonstrating that the peptide is nontoxic. Intradermal injection of RP23 into healthy human skin explants reduced spontaneous activation of dermal dendritic cells (Figure 7).

[0286] In conclusion, the peptides described herein offer potential as novel, locally acting peptide-based therapies for patients seeking improved management of inflammatory skin diseases.

[0287] Furthermore, the modified immune pathways, particularly the reduction of IL-12 / 23, IL-6, and TNF (Figure 5), as well as the reduced activation state of proinflammatory immune cells such as macrophages and DCs (Figures 5, 7, and 15), suggest the broad applicability of RP23 beyond inflammatory skin conditions. The RP23-induced reduction of TCRαβ+ T cells in the skin of psoriatic mice (Figure 11A) and the increase in FoxP3+ CD4+ T cells in skin-draining lymph nodes (Figure 11B) also suggest its suitability for use in T cell-mediated inflammatory conditions. This may include inflammatory conditions further characterized by T helper 1 or 17 inflammation, such as RA and IBD, which may be initiated by or further exacerbated through proinflammatory innate immune cells.

[0288] Example 3: Effect of mutants of RP23 (SEQ ID NO: 12) on inflammatory responses Next, we conducted further in vivo studies using a contact hypersensitivity model to determine the moiety responsible for the immunomodulatory activity of RP23. Seven days before skin sensitization with a contact irritant (DNFB), mice were subcutaneously injected with the peptide (10 nmol) once in the abdomen. Five days later, mice were challenged with DNFB at the distal ear site. Ear thickness was measured over 72 hours. The thickness is proportional to the degree of inflammatory response.

[0289] Compared to mice injected with saline (PBS), SEQ ID NO:12 (RP23), a fluorescently labeled version of SEQ ID NO:12 (SEQ ID NO:21 = AF594-RP23), a recombinantly producible version of SEQ ID NO:12 (SEQ ID NO:22 = K4G3MUC1SP), and a C-terminal fragment of the peptide (SEQ ID NO:2 = MUC1SP C-fragment) all resulted in a reduced inflammatory response (Figure 8).

[0290] [Table 6]

[0291] Example 4: Effects of additional peptide variants on inflammatory responses The efficacy of RP23 (SEQ ID NO: 12) was confirmed in a mouse model of contact dermatitis using a number of additional peptide variants, labeled herein as SEQ ID NO: 32 and SEQ ID NO: 35. The efficacy of these peptides was compared to a scrambled version of SEQ ID NO: 35.

[0292] Mice received a single subcutaneous injection in the abdomen with PBS, RP23 (10 nmol), or one of the peptide variants shown in the table below (equimolar amount of RP23).

[0293] [Table 7]

[0294] Where indicated, photoleucine replaced leucine, and these molecules were used in photocrosslinking studies. Seven days later, the abdominal skin was sensitized with a contact irritant (DNFB), and 5 days later, mice were challenged with DNFB at a site distal to the ear.

[0295] As shown in Figure 9A, mice administered the RP23 mutant, which retained the peptide sequence, showed a reduced specific increase in ear thickness. In contrast, mice administered the control PBS injection or the scrambled mutant of SEQ ID NO: 35 did not show a reduction in ear inflammation. This reiterates the importance of the SEQ ID NO: 30 amino acid sequence for immunosuppressive effects and indicates that specific chemical modifications do not affect the potency of RP23. Therefore, modified RP23 SEQ ID NOs: 32 and 35 can be used instead of RP23 to evaluate RP23-induced immunomodulation.

[0296] Example 5: Effect of peptides described herein in combination with corticosteroid administration on inflammatory responses Psoriasis was induced in C57BL / 6 female mice by daily application (days 0–4) of Aldara cream (5% imiquimod, 57.5 mg) to the backs. Clinical scores (PASI; modified psoriasis area and severity index) were determined for mice receiving prophylactic PBS or RP23 (day -1) with or without topical application of glucocorticoid (0.05% betamethasone dipropionate, 30 mg) to the back on day 0 (Figure 10A). At early time points, i.e., days 1 and 2, a slight decrease in clinical score was observed in mice receiving both RP23 and BD compared with RP23 alone or BD alone. Notably, as the disease progressed, the protective effect of BD alone was lost, whereas protection was maintained when both RP23 and BD were used. However, at later time points, there was no benefit from the combination compared with RP23 alone.

[0297] RP23 and BD therapy were also tested when administered synergistically at the time of treatment (day 1). Psoriasis was induced as described above, but clinical scores were measured daily for 10 days after imiquimod application was stopped on day 5 to record the rate and extent of psoriatic disease resolution (Figure 10B). Again, RP23 + BD provided an early benefit in disease reduction (day 2) and resulted in improvements compared to BD-only treatment in terms of the rate and extent of disease resolution. This suggests that RP23 can be used to enhance the efficacy of therapeutic BD, including when only a single dose of BD or RP23 is used.

[0298] Example 6: RP23 treatment suppresses TCRαβ expression in affected skin + Decreased T cells and regulatory CD4 in skin-draining lymph nodes + Increases T cells C57BL / 6 female mice received PBS or RP23 injections on day -1, then Aldara cream (5% imiquimod, 57.5 mg) was applied daily (days 0–4) to the backs of the mice to induce psoriasis. On day 15, skin and inguinal lymph nodes were analyzed by flow cytometry to quantify T cell subsets as a percentage of total leukocytes (CD45+) in the psoriatic skin of the back (Figure 11A). RP23 treatment reduced the percentage of total CD4+ and CD8+ T cells in the back skin, suggesting a decrease in the proliferation or recruitment of proinflammatory T cells to the skin in the setting of psoriasis.

[0299] CD3+CD4+ TCRαβ T cells in skin-draining lymph nodes (inguinal) were phenotyped by transcription factor staining and flow cytometry to identify cells with a regulatory phenotype (i.e., FoxP3+). In RP23-treated mice, the number of FoxP3-expressing cells was significantly increased, regardless of the co-expression of T-bet or Rorγt, transcription factors that define Th1 or Th17, respectively (Figure 11B). This suggests that RP23 treatment promotes responses that regulate T cell responses, including Th1 or Th17-type inflammation, supporting the utility of RP23 in inflammatory indications where this type of inflammation is pathogenic, including psoriasis, contact dermatitis, IBD (e.g., ulcerative colitis or Crohn's disease), and rheumatoid arthritis.

[0300] Example 7: Peptide formulations for topical application RP23 AF594 was formulated for topical application in a compatible stock emulsion consisting of dimethicone, liquid paraffin, 4-chloro-3-methylphenol, cetrimide, and octadecane-1-ol. It was applied topically to full-thickness human skin explants (7 mg / cm). 2 , 10 nmol RP23) for 24 hours. Cryosections were prepared and analyzed under a fluorescence microscope. AF594 was detectable in the epidermis (E) and beneath the basement membrane (line) in the dermis (D), indicating that RP23, when topically applied to human skin, is able to penetrate the stratum corneum and basement membrane and is found in close association with or captured by cells in both the epidermal and dermal layers of the skin (Figure 12).

[0301] Example 8: Effect of peptide administration on local suppression of the immune system: RP23 treatment reduces inflammation in a mouse model of atopic dermatitis The ears and shaved backs of C57BL / 6 female mice were sensitized (day 0) and challenged (days 7, 10, 14, 17, 19, 21, 23, 25, and 27) with topical application of the irritant oxazolone or vehicle alone. Mice were injected with RP23 on day -1 or both days -7 and -1, compared with injections of PBS alone (days -7 and -1) or topical application of a steroid (0.01% betamethasone dipropionate, 30 mg) applied to the back after each oxazolone application. Ear thickness was measured with manual calipers. RP23-treated mice had reduced ear inflammation compared with PBS-injected control mice, suggesting that RP23 reduced the priming of the inflammatory response to the local irritant.

[0302] Example 9: RP23 interacts with innate immune cells in the skin and lung AF594-RP23+ cells were identified by flow cytometry 24 hours after administration to C57BL / 6 mice. When RP23 was injected subcutaneously in the lumbar region, the peptide was captured in the skin primarily by neutrophils (CD11b+ Ly6G+) or CD11b+ myeloid cell subsets, including macrophages and DCs (Figure 14A). Similarly, when RP23 was delivered to the lung mucosa by intranasal instillation, the peptide was captured primarily by neutrophils, alveolar macrophages, and DCs (Figure 14B).

[0303] Mucosal delivery of RP23 also reduced the expression of activation markers on antigen-presenting cells in the lung. Naive C57BL / 6 mice (n = 5) were administered PBS or RP23 by intranasal instillation, and then 24 h later, cells were collected from bronchoalveolar lavage and perfused lungs. The median fluorescence intensity of MHCII on CD11c+ dendritic cells was determined by flow cytometry. MHCII expression was substantially reduced on DCs in lung tissue (Figure 15A). To determine whether this effect was maintained once robust lung inflammation was induced, C57BL / 6 mice (n = 4) were administered PBS, scrambled RP23 peptide, or RP23 by intranasal instillation. 24 h later, Pam2Cys-SK4-PEG(OH) was administered intranasally to induce an inflammatory state. RP23-treated mice again showed reduced MHCII expression on lung DCs and on DCs in the bronchoalveolar space (Figure 15B). This suggests that RP23 is internalized by APCs and then regulates their function to be less pro-inflammatory.

[0304] The present application also includes the following aspects. [Aspect 1] the amino acid sequence TVLTVV (SEQ ID NO: 1) or the amino acid sequence LLLLLTVLTVV (SEQ ID NO: 2) or a functional variant or fragment thereof 1. An anti-inflammatory peptide comprising: [Aspect 2] - the amino acid sequence TVLTVV (SEQ ID NO: 1) or the amino acid sequence LLLLLTVLTVV (SEQ ID NO: 2) or a functional variant or fragment thereof; - one or more further amino acids, with the proviso that said one or more further amino acids do not define the sequence MTPGTQSPFF in the N-terminal position relative to the sequence of SEQ ID NO: 1 or 2; or the sequence TGSGHASSTP in the C-terminal position relative to the sequence of SEQ ID NO: 1 or 2. and an anti-inflammatory peptide comprising at least 15 amino acid residues, preferably 15 to 1000 residues. [Aspect 3] - the amino acid sequence TVLTVV (SEQ ID NO: 1) or the amino acid sequence LLLLLTVLTVV (SEQ ID NO: 2) or a functional variant or fragment thereof; - one or more further amino acids, - optionally one or more modifications to facilitate solubilization of the peptide in biological fluids, tissues or preparations, to facilitate entry of the peptide into cells, or to enable detection of the peptide; and an anti-inflammatory peptide comprising at least 15 amino acid residues, preferably 15 to 1000 residues. [Aspect 4] 4. The anti-inflammatory peptide according to aspect 3, wherein the peptide comprises or consists of the amino acid sequence of any one of SEQ ID NOs: 1, 2, 3, 30, or 50 to 63, or a functional variant or fragment thereof. [Aspect 5] 5. The anti-inflammatory peptide according to aspect 4, wherein the peptide consists of the sequence of SEQ ID NO: 1, 2, 3, 30, or 50 to 63. [Aspect 6] 5. The anti-inflammatory peptide of embodiment 4, wherein the peptide consists of the sequence of SEQ ID NO:3. [Aspect 7] 5. The anti-inflammatory peptide of embodiment 4, wherein the peptide consists of the sequence of SEQ ID NO: 30. [Aspect 8] The peptide further comprises: - modifications or moieties to enhance the solubilization of the peptide in biological fluids, tissues or formulations; - a moiety for facilitating entry of the peptide into a cell, and - modifications or moieties to enable detection of said peptides 8. The anti-inflammatory peptide according to any one of aspects 1 to 7, comprising one or more of: [Aspect 9] The anti-inflammatory peptide of embodiment 8, wherein the peptide comprises a modification or moiety to enhance solubilization of the peptide in a biological fluid, tissue, or formulation. [Aspect 10] 10. The anti-inflammatory peptide of embodiment 9, wherein the modification or moiety is selected from the group consisting of one or more charged amino acid residues, a polymer or polymer fragment to increase the solubility of the peptide, and combinations thereof. [Aspect 11] 11. The anti-inflammatory peptide of embodiment 10, wherein said modification or moiety comprises one or more charged amino acid residues, preferably said amino acid residues are cationic amino acid residues. [Aspect 12] 12. The anti-inflammatory peptide of embodiment 10 or 11, wherein the moiety comprises polyethylene glycol (PEG). [Aspect 13] 13. The anti-inflammatory peptide of any one of aspects 8 to 12, wherein the modification or moiety that enhances solubilization of the peptide is at the N-terminus of the peptide. [Aspect 14] 14. The anti-inflammatory peptide according to any one of aspects 8 to 13, wherein the peptide comprises a moiety for promoting entry of the peptide into cells. [Aspect 15] 15. The anti-inflammatory peptide of embodiment 14, wherein the portion is a cell-penetrating peptide (CPP) sequence. [Aspect 16] 16. The anti-inflammatory peptide according to any one of aspects 8 to 15, wherein the peptide comprises a moiety that allows for detection of the peptide. [Aspect 17] 17. The anti-inflammatory peptide of embodiment 16, wherein the moiety that allows detection of the peptide is a fluorescent label, a synthetic amino acid, biotin, or a metal conjugate. [Aspect 18] 18. The anti-inflammatory peptide according to any one of aspects 1 to 17, further comprising a linker. [Aspect 19] 19. The anti-inflammatory peptide of any one of aspects 1 to 18, comprising a further anti-inflammatory peptide conjugated to said peptide. [Aspect 20] 20. The peptide of any one of aspects 1 to 19, wherein the functional variant or fragment thereof has at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% amino acid identity with the peptide and retains the function of preventing or treating inflammation. [Aspect 21] 21. The peptide of embodiment 20, wherein the functional variant comprises one or more synthetic isomers of an amino acid derivative. [Aspect 22] 9. The peptide of embodiment 8, wherein the peptide is as defined in any one of SEQ ID NOs: 1 to 66. [Aspect 23] A pharmaceutical composition comprising the anti-inflammatory peptide according to any one of aspects 1 to 22 and a pharmaceutically acceptable carrier, diluent or excipient, or a pharmaceutically acceptable salt thereof. [Aspect 24] 24. The pharmaceutical composition according to embodiment 23, wherein said peptide comprises or consists of the sequence of SEQ ID NO: 3 or 30, or is as defined in SEQ ID NO: 12, 32 or 35. [Aspect 25] 25. The pharmaceutical composition of embodiment 23 or 24, wherein the composition does not comprise granulocyte-macrophage colony-stimulating factor (GM-CSF). [Aspect 26] 26. The pharmaceutical composition according to any one of aspects 23 to 25, further comprising an anti-inflammatory agent. [Aspect 27] 27. The pharmaceutical composition of embodiment 26, wherein the additional anti-inflammatory agent is selected from the group consisting of corticosteroids, antigen binding proteins for inhibiting pro-inflammatory cytokines, DMARDs, or other agents for inhibiting inflammatory processes. [Aspect 28] 28. The pharmaceutical composition of embodiment 27, wherein said additional agent is an antigen-binding protein, preferably a monoclonal antibody, capable of inhibiting the activity of a pro-inflammatory molecule selected from the group consisting of interleukin 6 (IL-6), interleukin 17 (IL-17), interleukin 12 (IL-12), interleukin 23 (IL-23), interleukin 36 (IL-36), and tumor necrosis factor (TNF). [Aspect 29] 29. The pharmaceutical composition of any one of aspects 23 to 28, wherein the composition is suitable for oral, intravenous, topical, intradermal, subcutaneous, intraarticular, intranasal, or inhalation administration. [Aspect 30] A nucleic acid encoding the peptide according to any one of aspects 1 to 14, or a fragment thereof. [Aspect 31] A vector or plasmid comprising the nucleic acid or fragment thereof according to embodiment 30. [Aspect 32] A method for preventing or treating inflammation in an individual, the method comprising administering to the individual a peptide according to any one of aspects 1 to 22, or a pharmaceutical composition according to any one of aspects 23 to 29, thereby treating or preventing inflammation in the individual. [Aspect 33] 33. The method of embodiment 32, further comprising administering an additional anti-inflammatory therapy or agent selected from the group consisting of corticosteroids, antigen binding proteins, preferably monoclonal antibodies, for inhibiting the activity of inflammatory molecules, DMARDs, phototherapy, or other agents for inhibiting inflammation in the individual. [Aspect 34] A method according to embodiment 32 or 33, wherein a further anti-inflammatory agent or therapy is administered simultaneously with the peptide according to any one of embodiments 1 to 22 or the pharmaceutical composition according to any one of embodiments 23 to 29. [Aspect 35] A method according to embodiment 32 or 33, wherein the further anti-inflammatory agent or therapy is administered sequentially with the peptide according to any one of embodiments 1 to 22 or the pharmaceutical composition according to any one of embodiments 23 to 29. [Aspect 36] 23. Use of a peptide according to any one of aspects 1 to 22 in the manufacture of a medicament for the prevention or treatment of inflammation in an individual. [Aspect 37] 37. The use according to embodiment 36, wherein the medicament comprises a further anti-inflammatory agent for the treatment of inflammation. [Aspect 38] 38. The use of embodiment 37, wherein the additional anti-inflammatory agent is selected from the group consisting of a corticosteroid, an antigen binding protein, preferably a monoclonal antibody, for inhibiting the activity of an inflammatory molecule, a DMARD, or other agent for inhibiting inflammation in the individual. [Aspect 39] A method or use according to any one of aspects 32 to 38, wherein the inflammation is associated with an immune response. [Aspect 40] 40. A method or use according to any one of aspects 32 to 39, wherein the inflammation is associated with TNF, IL-6 or IL-12 / 23 production. [Aspect 41] 41. The method or use according to any one of aspects 32 to 40, wherein the inflammation is present in or on a tissue selected from mucosal or skin tissue, preferably dermal tissue, musculoskeletal tissue, lung tissue or intestinal tissue. [Aspect 42] 42. The method or use of any one of aspects 32 to 41, wherein the inflammation is a symptom of a disease or condition selected from the group consisting of an inflammatory skin disease or disorder, a musculoskeletal disease or disorder, a pulmonary disease or disorder, or an intestinal disease or disorder. [Aspect 43] Aspect 43. A method or use according to any one of aspects 32 to 42, wherein the inflammatory disease or disorder is rheumatoid arthritis, contact dermatitis, atopic dermatitis, allergic dermatitis, or psoriasis. [Aspect 44] Aspect 44. A method or use according to any one of aspects 32 to 43, wherein the inflammatory disease is an inflammatory skin disease or disorder, and the method or medicament minimizes one or more symptoms selected from the group consisting of erythema, skin thickness, scaling, pruritus, and inflammatory edema. [Aspect 45] 30. A method for modulating an immune response in an individual, the method comprising administering to an individual in need thereof a peptide according to any one of aspects 1 to 21, or a pharmaceutical composition according to any one of aspects 23 to 29, thereby modulating the immune response in the individual. [Aspect 46] 22. Use of a peptide according to any one of aspects 1 to 21 in the manufacture of a medicament for modulating an immune response in an individual. [Aspect 47] 47. A method or use according to any one of aspects 32 to 46, wherein the method or medicament minimizes the release of pro-inflammatory cytokines, preferably TNF, IL-6 and IL-12 / 23 and IL-17. [Aspect 48] 40. The method or use of embodiment 39, wherein the immune response is primarily an adaptive immune response. [Aspect 49] 40. The method or use of embodiment 39, wherein the immune response is predominantly an innate immune response. [Aspect 50] 50. The method or use according to embodiment 49, wherein the innate immune response is associated with the activity of antigen-presenting cells, preferably macrophages or dendritic cells, at the site of inflammation. [Aspect 51] 51. The method or use according to embodiment 50, wherein administration of the peptide or composition minimizes the activity of antigen-presenting cells, preferably macrophages or dendritic cells, at the site of inflammation. [Aspect 52] 30. The peptide according to any one of aspects 1 to 21 or the pharmaceutical composition according to any one of aspects 23 to 29 for use in the prevention or treatment of inflammation, preferably inflammation associated with abnormal immune activation, more preferably rheumatoid arthritis, contact dermatitis, atopic dermatitis, allergic dermatitis or psoriasis. [Aspect 53] A kit comprising the composition according to any one of aspects 23 to 29. [Aspect 54] 54. The kit of embodiment 53, further comprising an anti-inflammatory agent. [Aspect 55] 54. The kit of embodiment 53, wherein the additional anti-inflammatory agent is selected from the group consisting of a corticosteroid, an antigen-binding protein, and preferably a monoclonal antibody, wherein the monoclonal antibody is capable of inhibiting the activity of a pro-inflammatory molecule selected from the group consisting of interleukin 6 (IL-6), interleukin 17 (IL-17), interleukin 12 (IL-12), interleukin 23 (IL-23), interleukin 36 (IL-36), and tumor necrosis factor (TNF).

Claims

1. - an amino acid sequence comprising TVLTVV (SEQ ID NO: 1), LLLLLTVLTVV (SEQ ID NO: 2), or a functional variant or fragment thereof, wherein the functional variant or fragment has at least 80% amino acid identity to SEQ ID NO: 1 or SEQ ID NO: 2 and retains the function of preventing or treating inflammation, and - one or more modifications or moieties to enhance solubilization of the peptide in biological fluids, tissues or formulations, wherein the one or more modifications or moieties are in the form of one or more positively charged amino acids. An anti-inflammatory peptide comprising: the peptide is 7 to 50 amino acids in length; The anti-inflammatory peptide.

2. 2. The anti-inflammatory peptide of claim 1, wherein the one or more positively charged amino acids comprise arginine, lysine, histidine, or a combination thereof.

3. 3. The anti-inflammatory peptide of claim 1 or claim 2, wherein the one or more positively charged amino acids are polylysine, and the polylysine comprises at least four lysine amino acids.

4. The anti-inflammatory peptide of any one of claims 1 to 3, wherein the amino acid sequence comprises or consists of the amino acid sequence of any one of SEQ ID NOs: 1, 2, 3, 30, 36-38, and 50-63, or a functional variant or fragment thereof.

5. the amino acid sequence consists of the sequence of SEQ ID NO: 3, or The amino acid sequence consists of the sequence of SEQ ID NO:

30. The anti-inflammatory peptide of claim 4.

6. The peptide further comprises: - further modifications or moieties to enhance solubilization of the peptide in biological fluids, tissues or formulations; - a moiety for facilitating the entry of the peptide into cells, and - modifications or moieties to enable detection of said peptides The anti-inflammatory peptide according to any one of claims 1 to 5, comprising one or more of:

7. Further modifications or moieties to enhance the solubilization include: is a polymer or polymer fragment; containing polyethylene glycol (PEG); and / or Located at the N-terminus of the amino acid sequence The anti-inflammatory peptide of claim 6.

8. The anti-inflammatory peptide of claim 6, wherein the portion that promotes cell entry of the peptide is a cell penetrating peptide (CPP) sequence.

9. Contains D- and L-amino acids; further comprising a linker; and / or and a further anti-inflammatory peptide conjugated to said peptide. The anti-inflammatory peptide according to any one of claims 1 to 8.

10. the functional variant or fragment has at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% amino acid identity and retains the function of preventing or treating inflammation; and / or the functional variant comprises one or more synthetic isomers of an amino acid derivative; The anti-inflammatory peptide according to any one of claims 1 to 9.

11. 11. The anti-inflammatory peptide of any one of claims 1 to 10, wherein the peptide is as defined in any one of SEQ ID NOs: 4-6, 10-12, 19-29, 31-43, and 65-66.

12. A pharmaceutical composition comprising the anti-inflammatory peptide or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 11, and a pharmaceutically acceptable carrier, diluent or excipient.

13. the composition comprises an additional anti-inflammatory agent, wherein the additional anti-inflammatory agent is selected from the group consisting of corticosteroids, antigen binding proteins for inhibiting the activity of pro-inflammatory cytokines, e.g., monoclonal antibodies, DMARDs, and other agents for inhibiting inflammation; and / or The composition is suitable for oral, intravenous, topical, intradermal, subcutaneous, intraarticular, intranasal, or inhalation administration. The pharmaceutical composition of claim 12.

14. A nucleic acid encoding the anti-inflammatory peptide according to any one of claims 1 to 11, or a fragment thereof, or a vector or plasmid comprising a nucleic acid encoding the anti-inflammatory peptide according to any one of claims 1 to 11, or a fragment thereof.

15. An anti-inflammatory peptide according to any one of claims 1 to 11 or a pharmaceutical composition according to claim 12 or claim 13 for preventing or treating inflammation in an individual.

16. 12. Use of an anti-inflammatory peptide according to any one of claims 1 to 11 in the manufacture of a medicament for the prevention or treatment of inflammation in an individual.

17. 16. The anti-inflammatory peptide or pharmaceutical composition of claim 15, wherein the anti-inflammatory peptide or pharmaceutical composition is administered together with an additional anti-inflammatory agent selected from the group consisting of corticosteroids, antigen binding proteins for inhibiting the activity of pro-inflammatory molecules, e.g., monoclonal antibodies, DMARDs, and other agents for inhibiting inflammation in the individual; or 17. The use of claim 16, wherein the medicament comprises or is administered in combination with an additional anti-inflammatory agent for the treatment of inflammation, wherein the additional anti-inflammatory agent is selected from the group consisting of corticosteroids, antigen binding proteins for inhibiting the activity of pro-inflammatory molecules, e.g., monoclonal antibodies, DMARDs, and other agents for inhibiting inflammation in the individual.

18. The inflammation is associated with an inflammatory disorder or disease; and / or associated with aberrant immune activation; and / or associated with TNF, IL-6 or IL-12 / 23 production; and / or Located in or on a tissue selected from mucosal tissue and skin tissue An anti-inflammatory peptide or pharmaceutical composition according to claim 15, or a use according to claim 16, or an anti-inflammatory peptide, pharmaceutical composition or use according to claim 17.

19. The anti-inflammatory peptide or pharmaceutical composition of claim 15, or the use of claim 16, or the anti-inflammatory peptide, pharmaceutical composition or use of claim 17, or the anti-inflammatory peptide, pharmaceutical composition or use of claim 18, wherein the inflammation is associated with rheumatoid arthritis, contact dermatitis, atopic dermatitis, alopecia, allergic dermatitis, or psoriasis.

20. A kit comprising the anti-inflammatory peptide of any one of claims 1 to 11, or the pharmaceutical composition of claim 12 or 13.

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