Modulators of protease-activated receptor 2

Abstract

To provide peptides comprising a mutated fragment of a wild type protease-activated receptor 2 (PAR2).SOLUTION: Disclosed herein is a peptide comprising a hydrophobic moiety which can penetrate the cell membrane and act as an antagonist of PAR2. Also provided herein are a composition and a cell comprising the peptide and methods for using the peptide.SELECTED DRAWING: None

Description

[Technical field]

[0001] Related Applications This application claims priority under 35 USC § 119(e) to U.S. provisional application USSN 62 / 255,334, filed November 13, 2015, which is incorporated herein by reference.

[0002] Government support This invention was made with Government support under Grant No. R42DK101240 awarded by the National Institutes of Health-National Institute of Diabetes and Digestive and Kidney Diseases. The Government has certain rights in this invention. [Background technology]

[0003] background A variety of hormones, neurotransmitters and biologically active substances control, regulate or modulate the functions of living organisms through specific receptors located in the cell membrane. In eukaryotes, including yeast and mammals, many of these receptors mediate the transmission of intracellular signals by activating guanine nucleotide-binding proteins (G proteins) to which the receptors are coupled. Such receptors are generally referred to as G protein-coupled receptors (GPCRs), also known as G protein-linked receptors (GPLRs) or seven transmembrane domain receptors (GPCRs). Binding of specific signaling molecules, i.e. ligands, to GPCRs can cause conformational changes in the receptor resulting in a form that can bind and activate G proteins, thereby triggering a cascade of intracellular events that ultimately result in a biological response. Typically, GPCRs interact with G proteins to regulate the synthesis of intracellular second messengers such as cyclic AMP, inositol phosphates, diacylglycerol and calcium ions.

[0004] Known but uncharacterized GPCRs have become prime targets for drug action and development due to their association with many diseases (Jacoby et al., Chem. Med. Chem. 2006, 1:760-782). GPCRs typically share a common structural motif of seven transmembrane helical domains (TM1-TM7) connected by three intracellular (IL-1 / i1-IL-3 / i3) and three extracellular (EL-1 / e1-EL-3 / e3) loops. The seven transmembrane helices form a barrel-like cavity in the cell membrane, and it is the conformational change in this structure triggered by extracellular interaction with ligands that further activates the domain for G protein coupling inside the cell. GPCRs play essential roles in signaling processes that control cell metabolism, fibrosis, tissue remodeling, cell proliferation and motility, adhesion, inflammation, neuronal signaling, and blood coagulation.

[0005] GPCRs, together with G proteins and effectors (intracellular enzymes and proteins regulated by G proteins, as well as channels), are components of modular signaling systems that connect the state of intracellular second messengers to extracellular inputs (Pierce et al., Nature Rev Mole Cell Bio 2002, 3, 639-650). The superfamily of GPCRs is large, with sequencing of the human genome revealing 850 genes that encode them (Hopkins and Groom Nature Reviews Drug Discovery 2002, 1, 727-730). GPCRs can be divided into six classes based on sequence homology and functional similarity (Foord et al., Pharmacol Rev 2005, 57(2): 279-88); class A (or 1) (rhodopsin-like), class B (or 2) (secretin receptor family), class C (or 3) (metabotropic glutamate / pheromone), class D (or 4) (fungal mating pheromone receptors), class E (or 5) (cyclic AMP receptors), and class F (or 6) (Frizzled / Smoothened).

[0006] Among the rhodopsin-like GPCRs (class A or 1), there are the protease-activated receptors (PARs), a subfamily of seven-transmembrane GPCRs that are activated through the cleavage of a portion of their extracellular domain and act as sensors of extracellular protease gradients, allowing cells to respond to proteolytic microenvironments during tissue remodeling in fibrosis, cancer, blood coagulation, and countless other processes, such as those involved in acute and chronic inflammation. Members of the PAR family act as sensors of extracellular protease gradients, allowing cells to respond to proteolytic microenvironments during a wide range of physiological activities, such as tissue remodeling. To date, four different PARs have been identified: PAR1, PAR2, PAR3, and PAR4. They are expressed throughout the human body. Proteases such as trypsin, thrombin, Xa, VIIa, matriptase, hepsin, tryptase and MMP-1 cleave the N-terminal extracellular domain of individual PAR members, thereby unmasking the tethered ligand, which binds to the outer surface of the receptor and activates transmembrane signaling to intracellular G proteins. PAR1 was originally discovered in platelets and serves as the prototype for this specialized class of GPCRs. PAR1 is activated when it is cleaved by thrombin between residues R41-S42 located in the N-terminal extracellular domain of the receptor. PAR3 and PAR4 are also activated by thrombin, while PAR2 is best known as the trypsin / tryptase / Xa / VIIa receptor. Proteolytic cleavage exposes a new N-terminus, which binds to the body of the receptor in an unusual intracellular mode. PARs (e.g., SFLLRN PAR1 (SEQ ID NO:80), TFLLRN PAR1 (SEQ ID NO:71), PRSFLLRN PAR1 (SEQ ID NO:72), SLIGRL PAR2 (SEQ ID NO:73), AYPGKF PAR4A synthetic peptide corresponding to the first new amino acid at the newly truncated N-terminus of (SEQ ID NO:74) can also function as a selective, soluble, intermolecular agonist for PAR.

[0007] PAR1, the major thrombin receptor, is known to affect a wide range of physiological and pathological processes in the cardiovascular system, including endothelial barrier function, vasoreactivity, intimal hyperplasia, inflammation, and homeostasis (Ossovskaya et al., Physiol Rev 2004, 84:579-621). PAR1 is a mediator of endothelial cell proliferation and migration in vitro and is essential for angiogenesis in developing mice. PAR1-deficient mice suffer from half-embryo death at mid-gestation (E9.5) due to defective angiogenesis. Surprisingly, PAR1-deficient mice do not have a phenotypic change in platelet function, which led to the discovery of PAR4. Unlike in humans, PAR4 is the major thrombin receptor in mouse platelets, and PAR4-deficient mice do not signal to thrombin. PAR2, a cell surface receptor for trypsin-like proteases, is widely expressed in inflammatory cells, mesenchymal cells (e.g., fibroblasts, myofibroblasts, smooth muscle cells), stromal cells, endothelium, hepatocytes, astrocytes, keratinocytes, pancreatic cells, neural cells, cardiac cells, and epithelia, including lung, intestine, and hepatobiliary tract. PAR2 plays an important role in many acute and chronic inflammatory diseases of the skin, joints, lung, brain, gastrointestinal tract and liver, and vasculature, and has been associated with the progression of liver, lung, kidney and other fibrotic diseases, atopic dermatitis, chronic and acute pain conditions, pruritus, and pulmonary arterial hypertension. The functional role of PAR3 is unclear, and the synthetic PAR3-tethered ligand TFRGAP (SEQ ID NO: 75) does not stimulate detectable downstream signaling. PARs have also been shown to form functional homodimers / oligomers, and heterodimers / oligomers. PAR1 and PAR3 can serve as cofactors for PAR4, and PAR1 can transactivate PAR2 (Kaneider et al., Nat. Immunol. 2007, 8:1303-12).

[0008] Each PAR couples to a distinct subset of G proteins. For example, PAR1 couples to the Gα-subunit G q , G i and G 12 / 13 These are coupled to the G and H subunits, which are activated separately by different proteases. Thrombin can activate all three heterotrimeric subunits simultaneously, whereas MMP-1 more selectively activates G 12 / 13 Activates signal transduction. PAR1-G q Ca 2+ It stimulates the production of phospholipase C-β by lnsP3, which mobilizes phospholipase C-β, and diacylglycerol (DAG), which activates protein kinase C-α (PKCα), which in turn activates phospholipase A2 and phospholipase D. 12 / 13 PAR1 plays a key role in cell shape change, migration, and rho-dependent oncogenesis. PAR2 can stimulate Gq, Gi, and beta-arrestin signaling. Previously, we reported that PAR1-PAR2 heterodimers mediate the Gq-mediated signaling pathway. 12 / 13 From G i It has been shown that the switch in G protein signaling to G is involved in maintaining endothelial barrier function. i is involved in the activation of rac, PI3K, and the inhibition of adenylate cyclase and cAMP. How PAR1 and PAR2 regulate members of the MAP kinase cascade, such as ERK1 / 2, is still poorly understood. Summary of the Invention

[0009] Cell-penetrating peptides, called PEPDUCINS™, have been devised by attaching membrane-penetrating hydrophobic moieties to peptides derived from wild-type GPCRs, thereby generating artificial agonists and / or antagonists for specific receptor-G protein signaling pathways (Covic L. et al. 2002, PNAS 99: 643-48; U.S. Patent Nos. 6,864,229; 7,696,168; and 8,389,480; each of which is incorporated herein by reference). These lipidated peptides or polypeptides ("lipopeptides") have the ability to rapidly flip or cross membranes and disrupt receptor-G protein signaling in a highly specific manner, i.e., with high selectivity for their cognate receptors by an allosteric mechanism. Lipopeptides for PARs, such as PAR1, PAR2 and PAR4, cholecystokinin A and B (CCKA, CCKB), somatostatin-2 (SSTR2), melanocortin-4 (MC4R), glucagon-like peptide-1 receptor (GLP-1R), and P2Y 12 ADP receptors are made and act as agonists and / or antagonists of the receptors they are derived from.These compositions are useful for activating or inhibiting the activity of a wide range of GPCRs, including the protein family PAR.Human PARs include PAR1 (Genbank Accession No. AF019616), PAR2 (Genbank Accession No. XM003671), PAR3 (Genbank Accession No. NM004101), and PAR4 (Genbank Accession No. NM003950.1), which are incorporated herein by reference.

[0010] While PEPDUCINs™ have been used as effective antagonists without significant and substantial agonist effects on members of the PAR family, there is still a need for more effective PAR2 antagonists to further study the mechanism of receptor-G protein coupling and its involvement in selective contact between receptors and G proteins, and to create therapeutic agents for the treatment and / or prevention of various diseases and conditions in which GPCRs are associated. Candidate pepducins are constructed by attaching a hydrophobic second domain to a first domain that includes a GPCR segment that is more or less likely to be responsible for any of the interface contacts, such as the i3 loop, and its adjacent regions including TM6 and TM5, and / or potentially replacing or inserting into the analogous loop / TM segments in the receptor, thereby modifying receptor-G protein signaling.

[0011] The present disclosure provides new peptides based on the modification of full-length PAR2 or its fragments.In some embodiments, the peptides are chimeric polypeptides.The new peptides are useful for targeting the signal transduction events regulated by PAR2, and for treating and / or preventing diseases and conditions associated with PAR2.For example, the peptides and compositions herein are used to treat diseases or conditions associated with increased or abnormal PAR2 activity or signal transduction, or associated with increased or abnormal PAR2 protease activity.The peptides and compositions herein can also be used to treat constitutive PAR2 activity.

[0012] In one aspect, provided are peptides and compositions thereof that have a substantial antagonistic effect and no substantial agonistic effect on PAR2. In certain embodiments, the peptide comprises one or more point mutations in a region of the peptide corresponding to amino acid positions 270-290 of the human PAR2 sequence. The point mutations may be either substitutions, additions, or deletions, where the additions or deletions include additions or deletions of up to 8 consecutive residues in the wild-type sequence at the point of the mutation. The wild-type human PAR2 sequence is provided below as SEQ ID NO:69. As used herein, the phrase "wild-type human PAR2" refers to the human PAR2 sequence of SEQ ID NO:69.

number

[0013] In another aspect, the peptide is a lipopeptide that includes a hydrophobic moiety (eg, a lipid moiety, an acyl moiety, a steroid moiety, or an amino acid moiety) that enables passage of the peptide across a cell membrane.

[0014] In one embodiment, the peptide has the sequence: X4X5X6X7X8SEX 11 X 12 X 13 X 14 X 15 X 16 X 17 KX 19 (SEQ ID NO: 43), wherein X4 to X 19 The variables are as defined herein and correspond to amino acid residues 273-288 of the wild-type human PAR2 sequence. Table 1 provided herein lists exemplary peptide sequences. In some embodiments, the peptide comprises an amino acid sequence selected from SEQ ID NOs: 1-41. In some embodiments, the peptide comprises a sequence that is about 50% to about 99% homologous to the amino acid sequence of SEQ ID NOs: 1-41. In some embodiments, the peptide comprises a sequence that is about 50% to about 99% identical to the amino acid sequence of SEQ ID NOs: 1-41.

[0015] In some embodiments, the peptide comprises a sequence that is about 50% to about 99% homologous to the amino acid sequence of SEQ ID NO: 43. In some embodiments, the peptide comprises a sequence that is about 50% to about 99% identical to the amino acid sequence of SEQ ID NO: 43.

[0016] In one embodiment, the peptide comprises the sequence of SEQ ID NO:41, wherein the amino acid sequence is at positions X5 and X 15 and wherein the peptide is at least 15 amino acids in length.

[0017] In one embodiment, the peptide comprises the sequence of SEQ ID NO:41, wherein the amino acid sequence is at positions X5 and X 15 Mutations in, and X 19 and X 20 wherein X 20 is a hydrophobic amino acid or a D-amino acid.

[0018] In one embodiment, the peptide comprises the sequence of SEQ ID NO:41, wherein the amino acid sequence is at positions X5 and X 15 Mutations in, and X 19 and X 20 wherein X 14 and X 20 is a D-amino acid.

[0019] In another aspect, the peptide comprises a mutated fragment of wild-type PAR2, wherein the peptide shares at least two sections of at least two consecutive amino acid residues in sequence with the wild-type PAR2 sequence. In one embodiment, the at least two consecutive amino acid residues are found at positions in the peptide corresponding to amino acid positions 270-290 of the human PAR2 sequence, wherein at least one mutation in the mutated fragment of PAR2 is at an amino acid position corresponding to positions 272, 273, 274, 275, 276, 277, 280, 282, 283, 284, 285, 286, 288, and / or 289 of the human PAR2 sequence. Additional sections of at least two consecutive amino acids are also contemplated. For example, the peptide may have three sections of at least 2 consecutive amino acids; one section of at least 2 and at least 3 consecutive amino acids; two sections of at least 3 consecutive amino acids; three sections of at least 3 consecutive amino acids; two sections of at least 3 consecutive amino acids and one section of at least 2 consecutive amino acids; one section of at least 3 consecutive amino acids and one section of at least 4 consecutive amino acids; one section of at least 3 consecutive amino acids, one section of at least 4 consecutive amino acids, and one section of at least 2 consecutive amino acids; two sections of at least 4 consecutive amino acids; one section of at least 4 consecutive amino acids, one section of at least 6 consecutive amino acids. The sections of consecutive amino acids are separated by at least 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acid residues.

[0020] In one embodiment, the peptide comprises a mutated fragment of wild-type protease-activated receptor 2 (PAR2), wherein the peptide has at least three consecutive amino acid residues in its sequence in common with amino acid positions 270-290 of the human PAR2 sequence, and wherein at least one mutation in said mutated fragment of PAR2 is at an amino acid position corresponding to positions 272, 273, 274, 275, 276, 277, 280, 282, 283, 284, 285, 286, 288, and / or 289 of the human PAR2 sequence.

[0021] In another aspect, a pharmaceutical composition is provided comprising the peptide described herein and a pharma- ceutical acceptable excipient. In some embodiments, the pharmaceutical composition is suitable for oral, aerosol, nasal, topical, rectal, vaginal or parenteral administration, or for intravenous, subcutaneous, intradermal or intramuscular injection.

[0022] In yet another aspect, methods are provided for treating various conditions or disorders using the peptides and compositions described herein.In some embodiments, a therapeutically effective amount of peptide or its composition is administered to a subject in need thereof to treat and / or prevent disorders or conditions as described herein.For example, the peptides and compositions herein are used to treat diseases or conditions that are associated with increased or abnormal PAR2 activity or signal transduction, or associated with increased or abnormal PAR2 protease activity.The peptides and compositions herein can also be used to treat constitutive PAR2 activity. Exemplary disorders or conditions include non-alcoholic steatohepatitis (NASH), idiopathic pulmonary fibrosis (IPF), atopic dermatitis (AD, eczema), renal fibrosis, alcoholic steatohepatitis, organ fibrosis, renal fibrosis, myelofibrosis, pulmonary arterial hypertension (PAH), pulmonary fibrosis, pruritis (itch), pancreatitis, chronic kidney disease, nephritis, multiple sclerosis, cancer, leukemia, melanoma, inflammatory disorders and conditions, sepsis, inflammation-associated CNS disorders, bronchitis, asthma, diabetes, complications of diabetes and NASH, obesity, metabolic syndrome, fibrotic diseases, cardiac fibrosis, pulmonary fibrosis, inflammatory bowel disease, Crohn's disease, irritable bowel syndrome, cirrhosis, arthritis, joint fibrosis, keloids, myelofibrosis, systemic fibrosis, scleroderma, psoriasis, urticaria, impetigo, rash, and rosacea.

[0023] The peptides can be combined with other pharmaceutical agents (eg, peptides, small molecules) for use in the treatment of a variety of conditions or disorders.

[0024] In some embodiments, the peptides described herein may be combined with glucagon-like peptide (GLP)-1 receptor agonists.Such combinations are useful for treating, for example, NASH, diabetes, complications of diabetes and NASH, and other metabolic disorders, or as anti-inflammatory agents to suppress the side effects of pancreatitis or inflammation.Non-limiting examples of GLP-1 receptor agonists include liraglutide (VICTOZA®), lixisenatide, and exenatide.In some embodiments, the peptides described herein may be combined with liraglutide, lixisenatide, or exenatide.

[0025] In some embodiments, the peptides described herein may be used in combination with pirfenidone or nintedanib. Such combinations are useful, for example, for treating idiopathic pulmonary fibrosis (IPF) or other fibrotic disorders.

[0026] In a further aspect, provided herein is a kit comprising a peptide or composition as described herein.

[0027] The details of one or more aspects of the present invention are described in the accompanying description below. Other features, objects and advantages of the present invention will become clear from the description. In the specification and the appended claims, the singular form also includes the plural, unless the context clearly dictates otherwise. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the present invention belongs. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods and examples are merely illustrative and are not intended to be limiting. All patents and publications cited herein are incorporated by reference herein to the extent permitted by applicable law. [Brief description of the drawings]

[0028] [Figure 1] Figures 1A-1B. Male obese-diabetic (db / db) mice, 6-8 weeks of age, were purchased from Charles River Labs. db / db mice are leptin-deficient animals that serve as a model for diabetes, obesity, and dyslipidemia. Animals were randomized by blood glucose and body weight on day 2 and fed regular chow ad libitum. Treatment began on day 0, with animals receiving subcutaneous (SC) injections (5 mL / kg) daily from 7-10 AM. HbA1c was monitored on days 0, 7, 14, and 21. Mice (n=10) were treated daily with either 10 mg / kg N-palmitoylated SEQ70 (PZ-235) or vehicle ("Veh"). Food intake (8±2 g / day / mouse) and weight gain (40±1 g on day 0, increasing to 47±1 g on day 21) did not differ significantly between treatment groups over the 3-week period. Quite unexpectedly, diabetic mice treated with PZ-235 had a highly significant (P<0.001) reduction in mean glycated hemoglobin (HbA1c) levels of 0.8% compared to the vehicle group at the 3-week endpoint. Animals in the vehicle group had a mean increase in HbA1c from 4.7% to 7.4% over the 3-week period in vehicle-treated mice, whereas HbA1c levels in PZ-235-treated animals only increased from 4.8% to 6.6%. HbA1c levels reflect long-term blood glucose levels. This unexpected result with PZ-235 would indicate that inhibition of PAR2 by the i3-loop derived pepducin has a profound effect on lowering average glucose levels in the setting of severe diabetes over a relatively short period of time (e.g., 3 weeks of treatment).

[0029] [Diagram 2]Figure 2. Morning blood glucose in diabetic mice from the experiments in Figures 1A and 1B was measured three times per week. As shown in Figure 2, mean baseline (morning) glucose levels gradually increased in the vehicle control group (19 mmol / L increasing to 26 mmol / L). In the PZ-235-treated cohort, morning blood glucose levels were relatively reduced, which is consistent with the beneficial effect on HbA1c observed in Figures 1A and 1B.

[0030] [Diagram 3] Figure 3. Blood glucose profiles of diabetic mice were monitored on days 1, 8, and 22. As shown in Figure 3, mean blood glucose was consistently lowered by 2 mmol / L during the first 6 hours in PZ-235 treated animals compared to controls.

[0031] [Figure 4] Figures 4A-4B. Insulin resistance is a key pathogenic abnormality that defines the metabolic syndrome with respect to type 2 diabetes mellitus (T2DM). Obesity-induced insulin resistance is the dominant factor underlying both metabolic syndrome and T2DM. Obese-diabetic db / db mice rapidly become severely insulin resistant, reflected by a large increase in plasma insulin and a concomitant increase in glucose. At the end of the 3-week experiment, plasma from whole blood was collected from diabetic mice. As shown in Figures 4A and 4B, there was a significant 45% decrease in mean plasma insulin (Figure 4A) and a 22% increase in glucagon levels (Figure 4B) in PZ-235-treated animals compared to the control group. This unexpected result is the first to show that PAR2 inhibitors, such as those exemplified by PZ-235, improve insulin levels in diabetic animals and that similar PAR2 pepducins may have beneficial effects in improving insulin resistance in diabetic humans.

[0032] [Diagram 5]Figures 5A-5B. Plasma triglyceride levels in diabetic mice were measured at baseline and after 3 weeks of treatment with PZ-235 (vs. vehicle). As shown in Figures 5A and 5B, there was a highly significant (P<0.01) reduction in mean plasma triglycerides (TG) of 0.7 mmol / L in PZ-235-treated animals compared to control animals.

[0033] [Figure 6] Figure 6. Liver triglyceride levels in diabetic mice were measured after 3 weeks of treatment with PZ-235 (vs. vehicle). At the end of the 3-week experiment, livers were isolated from diabetic mice, flash frozen, and liver triglycerides were measured. As shown in Figure 6, after 3 weeks of treatment of diabetic mice with PZ-235, there was a significant (P<0.05) 15% reduction in liver triglycerides compared to control animals. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0034] definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. The following definitions are provided to aid in the interpretation of the disclosure and claims of this application. If the definitions in this section do not match those in other places, the definitions set forth in this section shall control.

[0035] As used herein, the terms "about" or "approximately" when used in conjunction with a number refers to any number within 1, 3, 5, 10 or 15% of the referenced number.

[0036] As used herein, "juxtamembrane" means close to the membrane.

[0037] As used herein, the term "peptide" or "polypeptide" refers to a molecule consisting of monomers (amino acids) linked in a linear chain by amide bonds (also known as peptide bonds). The term "polypeptide" refers to any chain of two or more amino acids and does not refer to a specific length of the product. A "peptide" or "polypeptide" as used herein may be derived from a natural biological source or produced by recombinant technology, but is not necessarily translated from a specified nucleic acid sequence. It may be produced in any manner, including by chemical synthesis. One or more of the amino acids in a polypeptide of the invention may be modified, for example, by the addition of a chemical entity such as a carbohydrate group, a phosphate group, a farnesyl group, an isofarnesyl group, a fatty acid group, an acyl group (e.g., an acetyl group), a linker for conjugation, functionalization, or other known protecting or blocking groups. In some embodiments, modification of the peptide results in a more stable peptide (e.g., a longer half-life in vivo).

[0038] The term "amino acid" refers to a molecule that contains both an amino group and a carboxyl group. In some embodiments, the amino acid is an alpha-amino acid. In some embodiments, the amino acid is a natural amino acid. In some embodiments, the amino acid is an unnatural amino acid. There are many known unnatural amino acids, any of which can be included in the peptides of the invention. See, for example, S. Hunt, The Non-Protein Amino Acids, in Chemistry and Biochemistry of the Amino Acids (ed. GC Barrett, Chapman and Hall, 1985).

[0039] Exemplary amino acids include, without limitation, alpha-amino acids, such as D- and L-isomers of the 20 commonly naturally occurring alpha amino acids found in peptides, natural amino acids that are not the 20 commonly naturally occurring amino acids, and unnatural alpha-amino acids. The amino acids used in the construction of the peptides of the invention may be prepared by organic synthesis or obtained by other routes, such as degradation of natural sources or isolation from natural sources. The amino acids may be commercially available or synthesized. Amino acids having hydrophobic side chains include Gly, Pro, Ala, Ile, Leu, Val, Phe, Met, Trp, and Tyr. In some embodiments, amino acids having hydrophobic side chains include Gly, Pro, Ala, Ile, Leu, Val, and Phe. In some embodiments, amino acids having hydrophobic side chains include Ala, Ile, Leu, and Val. Amino acids having polar side chains include Gln, Asn, His, Ser, Thr, Tyr, Cys, Met, Trp. In some embodiments, amino acids having polar side chains include Asn, Cys, Gln, Met, Ser, and Thr. Amino acids having aromatic side chains include Phe, Trp, Tyr, and His. Amino acids having hydrophobic aromatic side chains include Phe, Typ, and Tyr. Amino acids having charged side chains include Asp, Glu, Arg, His, and Lys. Negatively charged side chains include Asp and Glu. Positively charged side chains include Arg, His, and Lys. Neutral amino acids are selected from the group consisting of Ala, Ser, Val, Leu, Ile, Pro, Phe, Trp, Met, Gly, Thr, Cys, Tyr, Asn, and Gln.

[0040] As used herein, the terms "administration", "administering" and the like, when used in reference to providing a pharmaceutical composition to a subject, generally refer to providing one or more pharmaceutical compositions comprising an agent, such as an agonist or antagonist of the PAR2 signaling pathway, in combination with a suitable delivery vehicle, to a subject by any means such that the administered compound achieves one or more of the intended biological effects for which the compound is administered.As non-limiting examples, the composition can be administered via parenteral, subcutaneous, intravenous, intracoronary, rectal, intramuscular, intraperitoneal, transdermal, or buccal delivery routes.

[0041] In one embodiment, "administration" of an agent, such as an agonist or antagonist of the PAR2 signaling pathway, to a patient requires controlled release, i.e., release of the active ingredient from the formulation in a sustained and regulated manner over an extended period of time than an immediate release formulation containing the same amount of active ingredient would release during the same period of time. The dosage administered will depend on the recipient's age, health, weight, and / or thrombotic disease status, and / or other relevant risk factors, type of concurrent treatment, if any, frequency of treatment, and / or the nature of the desired effect.

[0042] As used herein, "agonist" refers to any natural or synthetic peptide, molecule, or combination thereof, which when used in standard bioassays or in vivo or at a therapeutically effective dose, increases biological activity by at least about 1.5-fold, about 1.8-fold, about 2-fold, about 3-fold, about 4-fold, about 5-fold, about 7-fold, about 10-fold, about 20-fold, about 50-fold, or about 100-fold or more over baseline in a control sample (e.g., a buffer or sample without the peptide agonist).

[0043] A "partial agonist" refers to any natural or synthetic peptide, molecule, or combination thereof that increases biological activity by at least about 1.2-fold, about 1.3-fold, about 1.4-fold, or about 1.5-fold or more above baseline in a control sample when used in a standard bioassay or in vivo or at a therapeutically effective dose.

[0044] "Antagonist" or "inhibitor" may be used interchangeably herein and refer to any natural or synthetic peptide, molecule, or combination thereof that interferes with the biological activity of a target by at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 85%, about 90%, about 95%, about 98%, or about 100% when used in standard bioassays or in vivo or at a therapeutically effective dose.

[0045] As used herein, "modulate" means to act as an antagonist, i.e., to inhibit, reduce, alleviate, block or prevent, partially or completely; or to enhance or stimulate, i.e., to act as an agonist, partial agonist or inverse agonist. Modulation may be direct or indirect or allosteric.

[0046] Human wild-type PAR2 has Genbank Accession No. XM-003671, which is hereby incorporated by reference. The sequence of human PAR2 is provided as SEQ ID NO:33.

[0047] In this disclosure, reference to PAR family members in general, or to any individual member of the PAR family members, such as PAR2, is understood to refer to all splice variants, mutants (including but not limited to deletions, insertions, or polymorphisms, or amino acid substitutions), isoforms, and homologs thereof.

[0048] The term "patient" or "subject" as used herein refers to any individual organism. For example, the organism may be a mammal, such as a primate (i.e., for example, a human). Furthermore, the organism may be a domestic animal (i.e., for example, a cat, a dog, etc.), a livestock animal (i.e., for example, a cow, a horse, a pig, a sheep, a goat, etc.), or a research animal (i.e., for example, a mouse, a rabbit, a rat, a guinea pig, etc.).

[0049] The phrase "pharmacologically acceptable" is employed herein to refer to compounds, materials, compositions and / or formulations that are suitable for use in contact with the tissues of human beings and animals, within the scope of sound medical judgment, without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0050] As used herein, "PEPDUCIN™" and "lipopeptide" are used interchangeably and are cell-penetrating peptides that act as intracellular inhibitors of signal transfer from receptors to G proteins. Lipopeptides utilize lipidated fragments of intracellular G protein-coupled receptor loops to modulate the action of GPCRs in targeted cell-signaling pathways. Lipopeptides contain short peptides derived from GPCR intracellular loops tethered to hydrophobic moieties. This structure allows lipopeptides to be tethered in the cell membrane lipid bilayer and target the GPCR / G protein interface via a unique intracellular allosteric mechanism. Examples of PEPDUCIN™ lipopeptides are described in PCT Patent Application No. WO2012 / 139137, and in U.S. Patent Nos. 6,864,229; 8,324,172; 8,354,378; 8,389,480; 8,440,627; and 8,563,519, each of which is incorporated by reference herein.

[0051] The term "therapeutically effective amount," as used herein, means an amount of an active peptide or composition thereof that elicits the biological or medical response desired by a researcher, veterinarian, physician, or other clinician in a tissue, system, animal, or human.

[0052] As used herein, "treating" or "treatment" covers the treatment of thrombotic disease conditions in mammals, particularly humans, including, but not limited to: (a) preventing the disease condition from occurring in a mammal (particularly where such mammal is susceptible to, but has not yet been diagnosed as suffering from, the disease condition); (b) inhibiting the disease condition, i.e., halting its development; and / or (c) alleviating the disease condition, i.e., causing regression of the disease condition.

[0053] The term "homologous" as used herein is an art-recognized term that refers to highly related nucleic acids or proteins at the level of nucleotide or amino acid sequence. Nucleic acids or proteins that are homologous to each other are called homologs. Homologous may refer to the degree of sequence similarity between two sequences (i.e., nucleotide sequence or amino acid). The percentage homology numbers referred to herein reflect the maximum possible homology between two sequences, i.e., the percent homology when the two sequences are aligned to have the maximum number of matching (homologous) positions. Homology can be readily calculated by known methods, such as those described in: Computational Molecular Biology, edited by Lesk, AM, Oxford University Press, New York, 1988; Biocomputing: Informatics and Genome Projects, edited by Smith, DW, Academic Press, New York, 1993; Sequence Analysis in Molecular Biology, von Heinje, G., Academic Press, 1987; Computer Analysis of Sequence Data, Part I, edited by Griffin, AM and Griffin, HG, Humana Press, New Jersey, 1994; and Sequence Analysis Primer, edited by Gribskov, M. and Devereux, J., M Stockton Press, New York, 1991; each of which is incorporated by reference herein. Commonly employed methods for determining homology between sequences include, but are not limited to, those disclosed in Carillo, H., and Lipman, D., SIAM J Applied Math., 48:1073 (1988), which is incorporated herein by reference. Techniques for determining homology are codified in publicly available programs.Exemplary computer software for determining the homology between two sequences include, but are not limited to, the GCG program package, Devereux, J., et al., Nucleic Acids Research, 12(1), 387 (1984)), BLASTP, BLASTN and PASTA Atschul, SF et al., J Molec. Biol., 215, 403 (1990)).

[0054] The term "homologous" refers to a comparison between two sequences. Two nucleotide sequences are considered to be homologous if the polypeptides they encode are at least about 50-60% identical, preferably about 70% identical, for at least one stretch of at least 20 amino acids. Preferably, homologous nucleotide sequences are also characterized by their ability to encode a stretch of at least 4-5 uniquely specified amino acids. Both the identity and the approximate spacing of these amino acids relative to one another must be considered in order for nucleotide sequences to be considered homologous. For nucleotide sequences less than 60 nucleotides in length, homology is determined by the ability to encode a stretch of at least 4-5 uniquely specified amino acids.

[0055] As used herein, the term "identity" refers to the overall relatedness between polymer molecules, for example, between nucleic acid molecules (e.g., DNA and / or RNA molecules), and / or between polypeptide molecules. The calculation of the percent identity of two amino acid sequences can be performed, for example, by aligning the two sequences for optimal comparison (e.g., gaps may be introduced for optimal alignment in one or both of the first and second amino acid sequences, and non-identical sequences may be ignored for comparison). In some embodiments, the length of the sequence aligned for comparison purposes is at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or 100% of the length of the reference sequence. The amino acids at corresponding positions are then compared. If a position in the first sequence is occupied by the same amino acid as the corresponding position in the second sequence, the molecules are identical at that position. The percent identity between two sequences is a function of the number of identical positions shared by the sequences, which takes into account the number of gaps that need to be introduced for optimal alignment of two sequences, and the length of each gap.Comparing sequences and determining the percent identity between two sequences can be achieved using mathematical algorithms.For example, percent identity can be calculated by optimal alignment of sequences using a similarity scoring matrix, such as the Blosum62 matrix described in Henikoff S. and Henikoff JG, PNAS USA 1992, 89: 10915-10919.The percent identity and optimal alignment of two sequences using the Blosum62 similarity matrix and the Needleman and Wunsch algorithm (J. Mol. Biol. 1970, 48: 443-453) can be calculated using the GAP program of Genetics Computer Group (GCG, Madison, Wl, USA), using the default parameters of the program.Specific parameters for calculating percent identity for protein and nucleic acid sequences related to the present invention are set forth below.

[0056] "Rigidifier" or "helix breaker" moieties disrupt the conformational regularity of the alpha-helical backbone. Natural and unnatural amino acids can be rigidifiers / helix breakers. Non-limiting examples of rigidifiers / helix breakers can be amino acids such as Pro, Gly, Trp and Asn; proline analogues with 4-, 5-, 6- or 7-membered rings replacing the proline side chain, such as cyclobutane, cyclopentane, cyclohexane, cycloheptane; amino acids with methyl-amino groups in peptide bonds; 1-aminocyclopropanecarboxylic acid (ACC); para-aminobenzoic acid (Paba); alpha-substituted tyrosine analogues.

[0057] Detailed Description PEPDUCIN™ lipopeptides are cell-penetrating peptides or polypeptides that have been developed to inhibit or activate GPCRs (see, for example, U.S. Patent Nos. 6,864,229 and 7,696,168). In some embodiments, lipopeptides inhibit GPCRs at the inner surface of the lipid bilayer. Provided herein are new peptides and lipopeptides that target the protease-activated receptor 2 (PAR2) transmembrane receptor. Provided herein are new peptides and lipopeptides that contain new mutations that have not previously been taught in other PEPDUCIN™ documents. Studies have implicated PAR2 as playing a role in a wide range of diseases, including asthma (Schmidlin et al., J Immunol 2002, 169: 5315-5321), arthritis (Ferrell et al., 2010), hyperalgesia (Vergnolle et al., 2001), neuropathic and cancer pain (Lam et al., 2010), cancer invasion (Shi et al., Mol Cancer Res 2004, 2: 395-402), nonalcoholic steatohepatitis (NASH), pulmonary arterial hypertension (PAH), atopic dermatitis (AD), pancreatitis, and IBD. A wide range of diseases involve PAR2 signaling, including many that involve inflammatory, fibrotic, and metabolic responses.

[0058] In some embodiments, provided herein is a chimeric polypeptide comprising: (a) a first domain comprising a mutated full-length or fragment of human protease-activated receptor 2 (PAR2); and (b) a second domain attached to said first domain, wherein said second domain comprises a hydrophobic moiety; wherein said chimeric polypeptide is an effective PAR2 antagonist. In some embodiments, the hydrophobic moiety is naturally occurring or non-naturally occurring.

[0059] In some embodiments, the peptide described herein comprises hydrophobic moiety.As used herein, the peptide of the present invention that comprises at least one hydrophobic moiety is referred to as lipopeptide.For example, the hydrophobic moiety attached to peptide herein can be lipid moiety, acyl moiety, steroid moiety, or amino acid moiety, which are further described herein.The peptide and lipopeptide described herein typically target the intracellular surface of PAR2, resulting in the regulation of signal transduction.

[0060] In one embodiment, the peptide has the sequence: X4X5X6X7X8X9X 10 X 11 X 12 X 13 X 14 X 15 X 16 X 17 X 18 X 19 X 20 (SEQ ID NO:42) Contains, where: X4 is absent, A, G, P, or an N-terminal linker; X5 is M, G, P, I, L, V, norleucine (J), methionine sulfoxide (M(SO)), or methionine sulfone (M(SO2)), or absent when X4 is absent; X6 is absent if D, E, H, or X4-X5 are absent; X7 is D, E, H, or absent if X4-X6 are absent; X8 is N, D, or E; X9 is any amino acid; X 10 is any amino acid; X 11 is any amino acid or its D-amino acid, 2-aminoisobutyric acid (B), hydroxyproline (Hyp), P, a proline homologue, G, or a stiffener / helix breaker moiety; X 12 is K, R, P, or absent; X 13 is any amino acid or citrulline (Cit); X 14 is K or X 13 and X 14 any amino acid that renders the peptide bond between uncleavable or reduces the positive charge; X 15 is any amino acid or beta-A; X 16 is A, S, T, G, Q, beta-A, 2-aminoisobutyric acid (B), or absent; X 17 is I, A, L or V; X 18 is K, I, or F; and X 19 is a hydrophobic amino acid, its D-amino acid, or absent.

[0061] In one embodiment, the peptide has the sequence: X4X5X6X7X8SEX 11 X 12 X 13 X 14 X 15 X 16 X 17 KX 19 (SEQ ID NO:43) Contains, where: X4 is absent, A, G, P, or an N-terminal linker; X5 is M, G, P, I, L, V, norleucine (J), methionine sulfoxide (M(SO)), or methionine sulfone (M(SO2)), or absent when X4 is absent; X6 is absent if D, E, H, or X4-X5 are absent; X7 is D, E, H, or absent if X4-X6 are absent; X8 is N, D, or E; X 11is any amino acid or its D-amino acid, 2-aminoisobutyric acid (B), hydroxyproline (Hyp), P, a proline homologue, G, or a stiffener / helix breaker moiety; X 12 is K, R, P, or absent; X 13 is any amino acid or citrulline (Cit); X 14 is K or X 13 and X 14 any amino acid that renders the peptide bond between uncleavable or reduces the positive charge; X 15 is any amino acid or beta-A; X 16 is A, S, T, G, Q, beta-A, 2-aminoisobutyric acid (B), or absent; X 17 is I, A, L or V; and X 19 is a hydrophobic amino acid, its D-amino acid, or absent.

[0062] In one embodiment, the peptide comprises the sequence of SEQ ID NO:43, wherein: X4 is absent, A, or an N-terminal linker; X5 is M, G, I, L, norleucine (J), M(SO), M(SO2), or absent when X4 is absent; X6 is absent if D, E, H, or X4-X5 are absent; X7 is D, E, H, or absent if X4-X6 are absent; X8 is N, D, or E; X 11 is K, P, dP, 2-aminoisobutyric acid (B), hydroxyproline (Hyp), a proline homologue, or a stiffener / helix breaker moiety; X 12 is K or absent; X 13 is R, F, W, Y, citrulline (Cit), or another amino acid; X 14 is K, dK, or another amino acid; X 15 is Q, S, or beta-A; X 16 is A, S, T, G, beta-A, 2-aminoisobutyric acid (B), or absent; X 17 is I or A; X 19 is a hydrophobic amino acid, its D-amino acid; and X 20 is a hydrophobic amino acid, its D-amino acid.

[0063] In one embodiment, the peptide has the sequence: AIX6X7X8SEX 11 KX 13 X 14 X 15 X 16 X 17 X 18 X 19 (SEQ ID NO:44) where X (variable) is defined herein.

[0064] In one embodiment, the peptide has the sequence: AMX6X7X8SEX 11 KX 13 X 14 X 15 X 16 X 17 X 18 X 19 (SEQ ID NO:45) where X (variable) is defined herein.

[0065] In one embodiment, the peptide has the sequence: GLX6X7X8SEX 11 KX 13 X 14 X 15 X 16 X 17 X 18 X 19 (SEQ ID NO:46) where X (variable) is defined herein.

[0066] In one embodiment, the peptide has the sequence: GDENSEX 11 KX 13 X 14 X 15 X 16 X 17 X 18 X 19 (SEQ ID NO:47) where X (variable) is defined herein.

[0067] In one embodiment, the peptide has the sequence: GDENX9EX 11 KX 13 X 14 X 15 X 16 X 17 X 18 X 19 (SEQ ID NO:48) where X (variable) is defined herein.

[0068] In one embodiment, the peptide has the sequence: GLHHDX9EX 11 KX 13 X 14 X 15 X 16 X 17 X 18 X 19 (SEQ ID NO:49) where X (variable) is defined herein.

[0069] In one embodiment, the peptide has the sequence: GLDENX9EX 11 KX 13 X 14 X 15 X 16 X 17 X 18 X 19 (SEQ ID NO:50) where X (variable) is defined herein.

[0070] In one embodiment, the peptide has the sequence: GLX6X7X8X9EX 11 KX 13 X 14 X 15 AIKX 19 (SEQ ID NO:51) where X (variable) is defined herein.

[0071] In one embodiment, the peptide has the sequence: GLHHDX9EX 11 KX 13 X 14 X 15 AIKX 19 (SEQ ID NO:52) where X (variable) is defined herein.

[0072] In one embodiment, the peptide has the sequence: GLDENX9EX 11 KX 13 X 14 X 15 AIKX 19 (SEQ ID NO:53) where X (variable) is defined herein.

[0073] In one embodiment, the peptide has the sequence: GLHHDSEX 11 KX 13 X 14 X 15 X 16 X 17 X 18 X 19 (SEQ ID NO:54) where X (variable) is defined herein.

[0074] In one embodiment, the peptide has the sequence: GLDENSEX 11 KX 13 X 14 X 15 X 16 X 17 X18 X 19 (SEQ ID NO:55) where X (variable) is defined herein.

[0075] In one embodiment, the peptide has the sequence: GLX6X7X8SEX 11 KX 13 X 14 X 15 AIKX 19 (SEQ ID NO:56) where X (variable) is defined herein.

[0076] In one embodiment, the peptide has the sequence: GLHHDSEX 11 KX 13 X 14 X 15 AIKX 19 (SEQ ID NO:57) where X (variable) is defined herein.

[0077] In one embodiment, the peptide has the sequence: GLDENSEX 11 KX 13 X 14 X 15 AIKX 19 (SEQ ID NO:58) where X (variable) is defined herein.

[0078] In one embodiment, the peptide has the sequence: X4X5HHDX9X 10 X 11 X 12 X 13 X 14 X 15 AIKX 19 (SEQ ID NO:59) where X (variable) is defined herein.

[0079] In one embodiment, the peptide has the sequence: X4X5HHDSEX11 KX 13 X 14 X 15 X 16 X 17 X 18 X 19 (SEQ ID NO:60) where X (variable) is defined herein.

[0080] In one embodiment, the peptide has the sequence: X4X5DENSEX 11 X 12 X 13 X 14 X 15 X 16 X 17 X 18 X 19 (SEQ ID NO:61) where X (variable) is defined herein.

[0081] In one embodiment, the peptide has the sequence: X4X5DENSEKX 12 X 13 X 14 X 15 X 16 X 17 X 18 X 19 (SEQ ID NO:62) where X (variable) is defined herein.

[0082] In one embodiment, the peptide has the sequence: X4X5X6X7X8X9X 10 X 11 X 12 X 13 X 14 X 15 AIKX 19 (SEQ ID NO:63) where X (variable) is defined herein.

[0083] In one embodiment, the peptide has the sequence: X4X5X6X7X8SEX 11 X 12 X13 X 14 X 15 AIKX 19 (SEQ ID NO:64) where X (variable) is defined herein.

[0084] In one embodiment, the peptide has the sequence: X4X5X6X7X8SEX 11 X 12 X 13 X 14 X 15 X 16 X 17 X 18 X 19 (SEQ ID NO:65) where X (variable) is defined herein.

[0085] In one embodiment, the peptide has the sequence: X4X5X6X7X8X9X 10 KKRKX 15 X 16 X 17 X 18 X 19 (SEQ ID NO:66) where X (variable) is defined herein.

[0086] In one embodiment, the peptide has the sequence: X4X5DX7X8X9X 10 X 11 KX 13 X 14 X 15 AIKX 19 (SEQ ID NO:67) where X (variable) is defined herein.

[0087] In one embodiment, the peptide has the sequence: X4LX6X7X8SEX 11 KX 13 X 14 X 15 X 16 IKX 19 (SEQ ID NO:68) where X (variable) is defined herein.

[0088] In one embodiment, the peptide comprising SEQ ID NOs: 42 to 68 is 19 and / or X 20 In one embodiment, the peptide comprising SEQ ID NO: 70 further comprises an additional amino acid residue located at X 19 and / or X 20 In some embodiments, the peptide further comprises an additional amino acid residue located at X. In some embodiments, the peptide further comprises X3, which is located N-terminal to X. In some embodiments, the peptide further comprises X2X3. In some embodiments, the peptide further comprises X1X2X3. X1, X2 and X3 are as defined herein. In some embodiments, the peptide further comprises X. 20 If absent, X 19 Does not include X 19 X 18 In one embodiment, the peptide is located C-terminal to X 19 X 20 X 19 and X 20 is as defined herein.

[0089] In some embodiments, the peptide comprises at least one insertion between any of the amino acid positions. In some embodiments, the peptide comprises at least two insertions between any of the amino acid positions. In some embodiments, the insertion is P. In some embodiments, the peptide comprises at least two insertions of P between any of the amino acid positions. In some embodiments, the peptide comprises at least two insertions of P between any of the amino acid positions. In some embodiments, the peptide comprises at least two insertions of P between any of the amino acid positions. In some embodiments, the peptide comprises at least two insertions of P between any of the amino acid positions. 11 and X 12 ; or X 12 and X 13 In some embodiments, the insertion is P. In some embodiments, the peptide comprises amino acids X5 and X6; 11 and X 12 ; or X 12 and X 13 Including the insertion of P between

[0090] As generally defined herein, X1 is located N-terminal to X2 and may be R or K. In some embodiments, X1 is R.

[0091] As generally defined herein, X2 is located N-terminal to X3 and may be S or T. In some embodiments, X2 is S.

[0092] As generally defined herein, X3 is located N-terminal to X4 and may be S, G, P, an N-terminal linker, or a helix breaker. In some embodiments, X3 is S, G, P. In some embodiments, X3 is an N-terminal linker selected from the group consisting of eK and aminohexanoic acid (Ahx). In some embodiments, X3 is Ahx. In some embodiments, X3 is eK. In some embodiments, X3 is a helix breaker as defined herein. In some embodiments, X3 is P. In some embodiments, X3 is G.

[0093] As generally defined herein, X4 is an N-terminal linker, A, G, P, or absent. In some embodiments, X4 is eK or Ahx. In some embodiments, X4 is A, G, or P. In some embodiments, X4 is G. In some embodiments, X4 is A. In some embodiments, X4 is P. In some embodiments, X4 is absent.

[0094] As generally defined herein, X5 is M, G, P, I, L, V, norleucine (J), methionine sulfoxide (M(SO)), or methionine sulfone (M(SO2)), or absent. In some embodiments, X5 is M, G, P, I, L, or V. In some embodiments, X4 is absent and X5 is M, G, P, I, L, V, norleucine (J), methionine sulfoxide (M(SO)), or methionine sulfone (M(SO2)). In some embodiments, X5 is L or P (i.e., an insertion between X5 and X6). In some embodiments, X5 is L.

[0095] In some embodiments, X4 and X5 are absent. In some embodiments, X4 and X5 are both present. In some embodiments, the peptide comprises the following amino acids: X4 is P and X5 is L. In some embodiments, the peptide comprises the following amino acids: X4 is eK, Ahx, G, or P; and X5 is L. In some embodiments, the peptide comprises the following amino acids: X4 is G and X5 is L or P. In some embodiments, the peptide comprises the following amino acids: X4 is G and X5 is L. In some embodiments, the peptide comprises the following amino acids: X4 is A and X5 is I. In some embodiments, the peptide comprises the following amino acids: X4 is A and X5 is M. In some embodiments, the peptide does not comprise X4 and X5 is M, G, P, I, L, or V.

[0096] As generally defined herein, X6 is H, D, E, or absent when X4-X5 are absent. In some embodiments, X6 is H. In some embodiments, X6 is D. In some embodiments, X6 is E.

[0097] As generally defined herein, X7 is H, D, E, or absent when X4-X6 are absent. In some embodiments, X7 is H. In some embodiments, X7 is D. In some embodiments, X7 is E.

[0098] In some embodiments, the peptide does not include X1-X7.

[0099] As generally defined herein, X8 is D, E, or N. In some embodiments, X8 is N. In some embodiments, X8 is D. In some embodiments, X8 is E.

[0100] In some embodiments, the peptide comprises the following amino acids: X6 is H, D, or E, X7 is H or E, and X8 is D or E. In some embodiments, X6 is D, X7 is H, and X8 is N. In some embodiments, the peptide comprises the following amino acids: X6 is H, X7 is H, and X8 is D. In some embodiments, X6 is H, X7 is H, and X8 is E. In some embodiments, X6 is H, X7 is H, and X8 is N. In some embodiments, X6 is D, X7 is E, and X8 is N. In some embodiments, X6 is H, X7 is E, and X8 is N.

[0101] As generally defined herein, X9 is any amino acid. In some embodiments, X9 is S, T, H, R, and K. In some embodiments, X9 is S or H. In some embodiments, X9 is S or T. In some embodiments, X9 is R or L.

[0102] As generally defined herein, X 10 is any amino acid. 10 is E or D. In some embodiments, X 10 is E. In some embodiments, X 10 is D.

[0103] As generally defined herein, X 11may be any amino acid or its D-amino acid, 2-aminoisobutyric acid (B), hydroxyproline (Hyp), P, a proline homologue, G, or a stiffening / helix breaker moiety. 11 is K. In some embodiments, X 11 is 2-aminoisobutyric acid (B). In some embodiments, X 11 is a stiffening / helix breaker moiety. In some embodiments, X 11 is P or a proline homologue. 11 is P. In some embodiments, X 11 is G. In some embodiments, X 11 is dP or Hyp.

[0104] As generally defined herein, X 12 may be K, R, P, or absent. 12 is K. In some embodiments, X 12 is R. In some embodiments, X 12 is P. In some embodiments, X 12 is absent (i.e., deleted).

[0105] In some embodiments, the peptide comprises the following amino acid: 11 is K and X 12 is K. In some embodiments, the peptide comprises the following amino acid: X 11 is B;X 12 is K. In some embodiments, the peptide comprises the following amino acid: X 11 is P, dP, or Hyp; X 12 is K. In some embodiments, the peptide comprises the following amino acid: X 11 is K, P, dP, or Hyp; X 12 is absent.

[0106] As generally defined herein, X 13may be any amino acid or citrulline (Cit). 13 is citrulline. 13 is any neutral aromatic amino acid. 13 is R. In some embodiments, X 13 is F, W, or Y. In some embodiments, X 13 is Y. In some embodiments, X 13 is F. In some embodiments, X 13 is W.

[0107] As generally defined herein, X 14 K, or X by proteases 13 and X 14 and X are any amino acids that render it impossible to cleave the peptide bond between them, or any amino acid that reduces the positive charge (i.e., neutral or negatively charged amino acids). 14 is K. Amino acids that render the peptide bond incapable of cleavage by a protease include D-amino acids or amino acids that have an N-methyl in the peptide bond. In some embodiments, X 14 is any D-amino acid. 14 is an amino acid having an N-methyl at the peptide bond. 14 is any amino acid that reduces the positive charge (i.e., a neutral or negatively charged amino acid). 14 In one embodiment, X 14 is L, I, or V. In some embodiments, X 14 In some embodiments, X is dL, dI, or dV. 14 is any amino acid that reduces the positive charge (i.e., a neutral or negatively charged amino acid). 14 is a neutral amino acid. 14 is a negatively charged amino acid. 14is a negatively charged side chain, such as D and E. In some embodiments, X 14 is a neutral charged side chain selected from A, S, V, L, I, P, F, W, M, G, T, C, Y, N, and Q.

[0108] As generally defined herein, X 15 is any amino acid, or beta-A. 15 is a polar amino acid, such as Q, N, H, S, T, Y, C, M, or W. In some embodiments, X 15 is a polar amino acid, such as Q or S. In some embodiments, X 15 is W, Y, or F. In some embodiments, X 15 is Q, S, or W. In some embodiments, X 15 is Beta-A.

[0109] As generally defined herein, X 16 is A, S, T, G, Q, N, beta-A, 2-aminoisobutyric acid (B), or absent (i.e., deleted). 16 In some embodiments, X is absent. 16 is A, B or Q. In some embodiments, X 16 is A. In some embodiments, X 16 is B. In some embodiments, X 16 is Q.

[0110] As generally defined herein, X 17 is I, A, L or V. In some embodiments, X 17 is I or A. In some embodiments, X 17 is A. In some embodiments, X 17 is I.

[0111] As generally defined herein, X 18 is K, I, or F. In some embodiments, X 18 is K or I. In some embodiments, X18 is I. In some embodiments, X 18 is K. In some embodiments, X 18 is F. In some embodiments, X 18 is not F.

[0112] As generally defined herein, X 19 is a hydrophobic amino acid, its D-amino acid, or X by proteases. 18 and X 19 or the absence of any amino acid that renders it impossible to cleave the peptide bond between X 19 is a protease-induced X 18 and X 19 In one embodiment, X is any amino acid that makes it impossible to cleave the peptide bond between 19 is G, P, A, I, L, V, F, or a D-amino acid thereof. 19 is L, I, V, dL, dI, or dV.

[0113] As generally defined herein, X 20 X 19 It is located on the C-terminal side of the hydrophobic amino acid, its D-amino acid, and X by proteases. 19 and X 20 or the absence of any amino acid that renders it impossible to cleave the peptide bond between X 20 is a protease-induced X 19 and X 20 In one embodiment, X is any amino acid that makes it impossible to cleave the peptide bond between 20 is G, P, A, I, L, V, F, or a D-amino acid thereof. 20 is L, I, V, dL, dI, or dV.

[0114] Position X1~X 20Any of the foregoing and subsequent embodiments and claimed embodiments recited for amino acids in are applicable to the amino acids corresponding to positions 280-289 of the human PAR2 sequence, and vice versa.

[0115] The wild-type PAR2 referred to herein can be from any source. In some embodiments, the wild-type protease-activated receptor 2 (PAR2) is from primates or rodents. In some embodiments, the wild-type PAR2 is from monkeys. In some embodiments, the wild-type PAR2 is from mice or rats. In some embodiments, the wild-type PAR2 is from humans.

[0116] In certain embodiments, the peptide comprises amino acids selected from SEQ ID NOs: 1-68 and 70. Table 1 lists exemplary peptide sequences. Positions X1-X 20 corresponds to positions 270-289 of the human PAR2 sequence. The amino acids found at positions 274-287 of wild-type human PAR2 correspond to positions X5-X6 of SEQ ID NO:41 as provided in Table 1. 18 As used herein, eK is epsilon lysine, Ahx is aminohexanoic acid, B is 2-aminoisobutyric acid, Hyp is hydroxyproline, Cit is citrulline, βA is beta-alanine, and J is norleucine. D-amino acids are indicated with a lower case d before the one-letter abbreviation (e.g., dK, dI, dP). A dash (-) indicates a deletion. [Table 1-1] [Table 1-2]

[0117] In certain embodiments, the peptide comprises a sequence that is at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% homologous to any of the amino acid sequences of SEQ ID NOs: 1 to 68 and 70. In certain embodiments, the peptide comprises a sequence that is about 50% to about 99%, about 60% to about 99%, about 70% to about 99%, about 75% to about 99%, about 80% to about 99%, about 85% to about 99%, about 90% to about 99%, or about 95% to about 99% homologous to any of the amino acid sequences of SEQ ID NOs: 1 to 68 and 70.

[0118] In some embodiments, the peptide is at least 13 amino acids long. In some embodiments, the peptide is at least 15 amino acids long. In some embodiments, the peptide is at least 20 amino acids long. In some embodiments, the peptide is at least 25 amino acids long. In some embodiments, the peptide is 13-25 amino acids long. In some embodiments, the peptide is 13-20 amino acids long. In some embodiments, the peptide is 13-18 amino acids long. In some embodiments, the peptide is 15-25 amino acids long. In some embodiments, the peptide is 15-20 amino acids long. In some embodiments, the peptide is 15-18 amino acids long. In some embodiments, the peptide is 16-18 amino acids long. In some embodiments, the peptide is 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 amino acids long.

[0119] In some embodiments, the peptide comprises at least one unnatural amino acid. In some embodiments, the peptide comprises one or two unnatural amino acids. In some embodiments, the peptide comprises at least one D-amino acid. In some embodiments, the peptide comprises one or two D-amino acids. In some embodiments, the peptide comprises one to five D-amino acids. In some embodiments, the peptide comprises one to ten D-amino acids. In some embodiments, the peptide comprises all D-amino acids. In some embodiments, the peptide has a molecular weight of at least 2000 Da.

[0120] In certain embodiments, the peptide comprises a sequence that is at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% identical to any of the amino acid sequences of SEQ ID NOs: 1 to 68 and 70. In certain embodiments, the peptide comprises a sequence that is about 50% to about 99%, about 60% to about 99%, about 70% to about 99%, about 75% to about 99%, about 80% to about 99%, about 85% to about 99%, about 90% to about 99%, or about 95% to about 99% identical to any of the amino acid sequences of SEQ ID NOs: 1 to 68 and 70.

[0121] In some embodiments, the peptide comprises the amino acid sequence of SEQ ID NOs: 1-68 and 70 with 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acid changes (e.g., amino acid substitutions, deletions and / or additions). In some embodiments, the amino acid changes are amino acid substitutions in which 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids are mutated to another amino acid. In some embodiments, the amino acid changes are additions or deletions, including additions or deletions of up to 1, 2, 3, 4, 5, 6, 7 or 8 residues at the point of the mutation in the wild type sequence. The added or deleted residues may be consecutive or non-consecutive residues.

[0122] In another aspect, the peptide comprises a mutated fragment of wild-type PAR2, wherein the peptide shares at least two sections of at least two consecutive amino acid residues in sequence with the wild-type PAR2 sequence. In certain embodiments, the at least two consecutive amino acid residues are found at amino acid positions in wild-type PAR2 corresponding to amino acid positions 270-290 of the human PAR2 sequence, wherein at least one mutation in said mutated fragment of PAR2 is at an amino acid position corresponding to positions 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, and / or 289 of the human PAR2 sequence. Additional sections of at least two consecutive amino acids are also contemplated. In some embodiments, the peptide comprises three sections of at least two consecutive amino acids; one section of at least two and at least three consecutive amino acids; two sections of at least three consecutive amino acids; three sections of at least three consecutive amino acids; two sections of at least three consecutive amino acids and one section of at least two consecutive amino acids; one section of at least three consecutive amino acids and one section of at least four consecutive amino acids; one section of at least three consecutive amino acids, one section of at least four consecutive amino acids, and one section of at least two consecutive amino acids; two sections of at least four consecutive amino acids; or one section of at least four consecutive amino acids and one section of at least six consecutive amino acids, together with the wild-type PAR2 sequence. The consecutive amino acid sections are separated by at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 amino acid residues. In some embodiments, the amino acid residues separating the contiguous sections of amino acids are the same residues as those found at the corresponding positions in the sequence of the wild-type PAR2 sequence. In some embodiments, the amino acid residues separating the contiguous sections of amino acids are different from the residues found at the corresponding positions in the sequence of the wild-type PAR2 sequence.

[0123] In one embodiment, the peptide comprises a mutated fragment of wild-type protease-activated receptor 2 (PAR2), wherein the peptide has at least three consecutive amino acid residues in common with wild-type PAR2 amino acids at positions corresponding to positions 270-290 of the human PAR2 sequence, and wherein at least one mutation in the mutated fragment of PAR2 is at an amino acid position corresponding to position 282 of the human PAR2 sequence.

[0124] In one embodiment, the peptide comprises a mutated fragment of wild-type protease-activated receptor 2 (PAR2), wherein the peptide shares at least three consecutive amino acid residues in a sequence corresponding to positions 270-290 of the human PAR2 sequence with wild-type PAR2, and wherein at least one mutation in the mutated fragment of PAR2 is at an amino acid position corresponding to position 280 of the human PAR2 sequence.

[0125] In some embodiments, the at least one mutation at an amino acid position is at an amino acid position corresponding to position 280 or 282 of the human PAR2 sequence, but not at both positions 280 or 282. In some embodiments, the at least one mutation at an amino acid position is at an amino acid position corresponding to both positions 280 and 282 of the human PAR2 sequence.

[0126] In one embodiment, at least one mutation in said mutated fragment of PAR2 is at an amino acid position corresponding to position 273 of the human PAR2 sequence.

[0127] In certain embodiments, at least one mutation in said mutated fragment of PAR2 is at an amino acid position corresponding to positions 275, 276, and / or 277 of the human PAR2 sequence.

[0128] In certain embodiments, at least one mutation in said mutated fragment of PAR2 is at an amino acid position corresponding to positions 273, 274, 282, and / or 284 of the human PAR2 sequence.

[0129] In certain embodiments, at least one mutation in said mutated fragment of PAR2 is at an amino acid position corresponding to position 274, 275, 276, 277, and / or 284 of the human PAR2 sequence. In certain embodiments, the peptide further comprises a mutation at an amino acid position corresponding to position 280 of the human PAR2 sequence.

[0130] In certain embodiments, the peptides described herein further comprise a mutation at an amino acid position corresponding to position 289 of the human PAR2 sequence.

[0131] In certain embodiments, the peptides described herein further comprise a mutation at an amino acid position corresponding to position 288 of the human PAR2 sequence.

[0132] In certain embodiments, the peptides described herein further comprise a mutation at an amino acid position corresponding to position 283 of the human PAR2 sequence.

[0133] In certain embodiments, the peptides described herein further comprise a mutation at an amino acid position corresponding to position 285 of the human PAR2 sequence.

[0134] In certain embodiments, the peptides described herein further comprise a mutation at an amino acid position corresponding to position 282 of the human PAR2 sequence.

[0135] In some embodiments, a mutated fragment of PAR2 refers to a PAR2 sequence having amino acids corresponding to positions 270-290 of the human PAR2 sequence. In some embodiments, a peptide described herein comprises at least one mutation in said mutated fragment of PAR2 at a position corresponding to positions 273, 274, 275, 276 and / or 277 of the human PAR2 sequence. In some embodiments, a peptide described herein comprises at least one mutation in said mutated fragment of PAR2 at a position corresponding to positions 274, 284 or 287 of the human PAR2 sequence. In some embodiments, a peptide comprises the sequence AIHHD (SEQ ID NO: 76) at a position corresponding to positions 273-277 of the human PAR2 sequence. In some embodiments, a peptide comprises the sequence AIDEN (SEQ ID NO: 77) at a position corresponding to positions 273-277 of the human PAR2 sequence. In some embodiments, a peptide comprises the sequence GLHHD (SEQ ID NO: 78) at a position corresponding to positions 273-277 of the human PAR2 sequence. In some embodiments, the peptide comprises the sequence GLDEN (SEQ ID NO: 79) at a position corresponding to positions 273-277 of the human PAR2 sequence. In some embodiments, the peptide described herein comprises at least one mutation in said mutated fragment of PAR2 at a position corresponding to positions 273, 274, 282, and / or 284 of the human PAR2 sequence. In some embodiments, the peptide described herein comprises at least one mutation in said mutated fragment of PAR2 at a position corresponding to positions 274, 275, 276, 277, and / or 284 of the human PAR2 sequence. In some embodiments, the peptide described herein comprises at least one mutation in said mutated fragment of PAR2 at a position corresponding to positions 275, 276, and / or 277 of the human PAR2 sequence. In some embodiments, the peptide described herein comprises at least one mutation in said mutated fragment of PAR2 at a position corresponding to position 287 of the human PAR2 sequence.In some embodiments, the peptides described herein comprise, in said mutated fragment of PAR2, a mutation at a position corresponding to position 287 of the human PAR2 sequence, and a mutation at a position corresponding to position 274 or position 284. In some embodiments, the peptides described herein further comprise a mutation at a position corresponding to position 289 of the human PAR2 sequence. In some embodiments, the peptides described herein further comprise a mutation at a position corresponding to position 280 of the human PAR2 sequence.

[0136] In some embodiments, the section of consecutive amino acid residues of the described peptides is found in the third intracellular (i3) loop, the fifth transmembrane helix (TM5), and / or the sixth transmembrane helix (TM6) of PAR2. In some embodiments, the section of consecutive amino acid residues of the described peptides is located at a position in wild-type PAR2 that corresponds to an amino acid residue in positions 270-290 of the human PAR2 sequence. In some embodiments, the peptide has three or more consecutive amino acid residues in common with the i3 loop of wild-type human PAR2. In some embodiments, the peptide has four or more consecutive amino acid residues in common with the i3 loop of wild-type human PAR2, five or more consecutive amino acid residues in common with the i3 loop of wild-type human PAR2, or six or more consecutive amino acid residues in common with the TM6 of wild-type human PAR2. In some embodiments, the peptide shares 4 or more consecutive amino acid residues, 5 or more consecutive amino acid residues, or 6 or more consecutive amino acid residues in sequence with TM6 of wild-type human PAR2. In some embodiments, the peptide shares 2 or more consecutive amino acid residues in sequence with TM5 of wild-type human PAR2. In some embodiments, the peptide shares 3 or more consecutive amino acid residues, 4 or more consecutive amino acid residues, 5 or more consecutive amino acid residues, or 6 or more consecutive amino acid residues in sequence with TM5 of wild-type human PAR2.

[0137] In certain embodiments, the peptide shares at least 4, 5, 6, 7, 8, 9, or 10 consecutive amino acid residues in sequence with the human PAR2 sequence.

[0138] In some embodiments, the peptide comprises 2-15 mutations compared to the wild-type PAR2 fragment from which the peptide is derived. In some embodiments, the peptide has 2-10 mutations compared to the wild-type PAR2 fragment. In some embodiments, the peptide has 5-10 mutations compared to the wild-type PAR2 fragment. In some embodiments, the peptide has 5 mutations compared to the wild-type PAR2 fragment. In some embodiments, the peptide has 6 mutations compared to the wild-type PAR2 fragment. In some embodiments, the peptide has 7 mutations compared to the wild-type PAR2 fragment. In some embodiments, the peptide has 8 mutations compared to the wild-type PAR2 fragment. In some embodiments, the peptide has 9 mutations compared to the wild-type PAR2 fragment. In some embodiments, the peptide has 10 mutations compared to the wild-type PAR2 fragment. In some embodiments, the peptide has 11 mutations compared to the wild-type PAR2 fragment. In some embodiments, the peptide has 12 mutations compared to the wild-type PAR2 fragment. In some embodiments, the peptide has 6-12 mutations compared to the wild-type PAR2 fragment.

[0139] In some embodiments, the peptide comprises an S and an E at positions in the peptide corresponding to positions 278 and 279, respectively, of the human PAR2 sequence. In some embodiments, the peptide comprises an H and an E at positions in the peptide corresponding to positions 278 and 279, respectively, of the human PAR2 sequence. In some embodiments, the peptide comprises a K at a position in the peptide corresponding to position 287 of the human PAR2 sequence. In some embodiments, the peptide comprises an S and an E at positions in the peptide corresponding to positions 278 and 279, respectively, of the human PAR2 sequence, and a K at a position in the peptide corresponding to position 287 of the human PAR2 sequence.

[0140] In some embodiments, at least one mutation at the amino acid position corresponding to position 273 of the human PAR2 sequence is A, G, P, or an N-terminal linker. As used herein, N-terminal linkers include, but are not limited to, eK, aminohexanoic acid (Ahx), proline, and glycine. In some embodiments, the peptide does not include an amino acid corresponding to position 273 when the starting residue at the N-terminus of the peptide is the amino acid corresponding to position 274 of the human PAR2 sequence.

[0141] In certain embodiments, at least one mutation at an amino acid position corresponding to position 274 of the human PAR2 sequence is M, G, P, I, L, norleucine (J), methionine sulfoxide (M(SO)), or methionine sulfone (M(SO2)). In certain embodiments, the peptide does not include amino acids corresponding to positions 273 and 274 when the starting residue at the N-terminus of the peptide is the amino acid corresponding to position 275 of the human PAR2 sequence.

[0142] In some embodiments, at least one mutation at an amino acid position corresponding to position 275 of the human PAR2 sequence is D, E, H. In some embodiments, when the starting residue at the N-terminus of the peptide is the amino acid corresponding to position 276 of the human PAR2 sequence, the peptide does not include amino acids corresponding to positions 273, 274, and 275.

[0143] In some embodiments, at least one mutation at the amino acid position corresponding to position 276 of the human PAR2 sequence is D, E, H. In some embodiments, the N-terminus of the peptide is the amino acid corresponding to position 277. In some embodiments, when the starting residue at the N-terminus of the peptide is the amino acid corresponding to position 277 of the human PAR2 sequence, the peptide does not include amino acids corresponding to positions 273, 274, 275, and 276.

[0144] In certain embodiments, at least one mutation at an amino acid position corresponding to position 277 of the human PAR2 sequence is N, D, or E.

[0145] In some embodiments, the at least one mutation at the amino acid position corresponding to position 278 of the human PAR2 sequence is any amino acid. In some embodiments, the at least one mutation at the amino acid position corresponding to position 278 of the human PAR2 sequence is S, T, H, R, and K.

[0146] In some embodiments, at least one mutation at the amino acid position corresponding to position 279 of the human PAR2 sequence is any amino acid. In some embodiments, at least one mutation at the amino acid position corresponding to position 279 of the human PAR2 sequence is N, D, or E.

[0147] In some embodiments, the at least one mutation at an amino acid position corresponding to position 280 of the human PAR2 sequence is any amino acid or a D-amino acid thereof. In some embodiments, the at least one mutation at an amino acid position corresponding to position 280 of the human PAR2 sequence is K, P, dP, 2-aminoisobutyric acid (B), hydroxyproline (Hyp), a proline homologue, G, or a stiffening / helix breaker moiety.

[0148] In some embodiments, at least one mutation at an amino acid position corresponding to position 282 of the human PAR2 sequence is any amino acid or citrulline (Cit). In some embodiments, at least one mutation at an amino acid position corresponding to position 282 of the human PAR2 sequence is R, F, W, Y, or citrulline (Cit).

[0149] In some embodiments, at least one mutation at an amino acid position corresponding to position 284 of the human PAR2 sequence is any amino acid or beta-alanine (beta-A; β-A). In some embodiments, at least one mutation at an amino acid position corresponding to position 284 of the human PAR2 sequence is Q, S, or beta-alanine (beta-A; β-A).

[0150] In certain embodiments, at least one mutation at an amino acid position corresponding to position 289 of the human PAR2 sequence is I, V, L, A, or a D-amino acid thereof.

[0151] In certain embodiments, the peptides described herein comprise at least one mutation in said mutated fragment of PAR2 at positions corresponding to positions 275, 276, and / or 277 of the human PAR2 sequence. In certain embodiments, the peptide comprises the sequence HHD, which corresponds to positions 275-277 of the human PAR2 sequence. In certain embodiments, the peptide comprises H at positions corresponding to positions 275 and 276 of the human PAR2 sequence, and a negatively charged amino acid (e.g., N, D, E) at position corresponding to position 277 of the human PAR2 sequence. In certain embodiments, the peptide comprises the sequence DEN, which corresponds to positions 275-277 of the human PAR2 sequence. In certain embodiments, the peptide comprises EN at positions corresponding to positions 276 and 277, and D or H at position corresponding to position 275 of the human PAR2 sequence. In certain embodiments of any of the embodiments, the peptide further comprises amino acids 278 (i.e., X9 of SEQ ID NO: 42) and 279 (i.e., X of SEQ ID NO: 42) of the human PAR2 sequence. 10 ) contains an SE at a position corresponding to

[0152] In one embodiment, the peptide is located at position 287 of wild-type human PAR2 (i.e., X of SEQ ID NO:42). 18In some embodiments, the peptide does not contain a K to F mutation at a position corresponding to position 287 of wild type human PAR2. In some embodiments, the peptide does not contain a K to A mutation at a position corresponding to position 274 of wild type human PAR2 (i.e., X5 of SEQ ID NO:42). In some embodiments, the peptide does not contain a M to A mutation at a position corresponding to position 274 of wild type human PAR2 (i.e., X5 of SEQ ID NO:42). In some embodiments, the peptide does not contain a M to G mutation at a position corresponding to position 284 of wild type human PAR2 (i.e., X 15 ) in the position corresponding to R to S. In some embodiments, the peptide does not contain an R to Q mutation in the position corresponding to wild-type human PAR2 position 284.

[0153] As described herein, the peptide comprises an amino acid addition, deletion, or substitution compared to wild-type PAR2. In one embodiment, the peptide comprises an amino acid addition, deletion, or substitution at position 281 of the human PAR2 sequence (i.e., X of SEQ ID NO: 42). 12 In one embodiment, the peptide comprises a deletion at a position corresponding to position 285 of the human PAR2 sequence (i.e., X 16 In some embodiments, the peptide comprises a deletion or substitution of a methionine (M) with another residue at a position corresponding to position 274 of the human PAR2 sequence. In some embodiments, the peptide comprises a deletion or substitution of an arginine (R) with another residue at a position corresponding to position 284 of the human PAR2 sequence. In some embodiments, the peptide comprises a substitution of an arginine (R) with another residue having a shorter side chain at a position corresponding to position 284 of the human PAR2 sequence. In some embodiments, the peptide comprises a substitution or deletion of a lysine (K) at a position corresponding to position 287 of the human PAR2 sequence.

[0154] In one embodiment, the peptide comprises the sequence of SEQ ID NO:41, wherein the amino acid sequence is at positions X5 and X 15wherein X5 is M, G, P, I, L, norleucine (J), M(SO), M(SO2), and wherein the peptide is at least 15 amino acids in length. In one embodiment, the peptide comprises the sequence of SEQ ID NO: 41, wherein the amino acid sequence is at positions X5 and X 15 wherein X5 is M, G, P, I, L, norleucine (J), M(SO), M(SO2), and wherein X 15 is S or Q, and wherein the peptide is at least 15 amino acids long. In certain embodiments, the peptide is at least 20 amino acids long. In certain embodiments, the peptide is at least 25 amino acids long. In certain embodiments, the peptide is 15-25 amino acids long. In certain embodiments, the peptide is 15-20 amino acids long. In certain embodiments, the peptide is 15-18 amino acids long. In certain embodiments, the peptide is 16-18 amino acids long. In certain embodiments, the peptide is 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 amino acids long. In certain embodiments, the peptide comprises at least one unnatural amino acid. In certain embodiments, the peptide comprises one or two unnatural amino acids. In certain embodiments, the peptide comprises at least one D-amino acid. In certain embodiments, the peptide comprises one or two D-amino acids. In certain embodiments, the peptide comprises one to five D-amino acids. In certain embodiments, the peptide comprises one to ten D-amino acids. In some embodiments, the peptide comprises all D-amino acids. In some embodiments, the peptide has a molecular weight of at least 2000 Da. In some embodiments, the peptide exhibits 70% or at least 80% inhibition of PAR2 as assessed by calcium flux using 10 μM peptide and 8 μM SLIGRL (SEQ ID NO: 73) agonist in cells. In some embodiments, the peptide exhibits at least 40% or at least 50% inhibition of PAR2 as assessed by calcium flux using 3 μM peptide and 8 μM SLIGRL (SEQ ID NO: 73) agonist in cells.

[0155] In one embodiment, the peptide comprises the sequence of SEQ ID NO:41, wherein the amino acid sequence is at positions X5 and X 15 and containing a mutation at positions X4 and X 19 In one embodiment, the peptide comprises the sequence of SEQ ID NO: 41, wherein the amino acid sequence is at positions X5 and X 15 containing mutations at positions X4, X 19 and X 20 In some embodiments, the additional amino acid at position X4 is A. In some embodiments, the additional amino acid at position X4 is an N-terminal linker selected from the group consisting of eK, aminohexanoic acid (Ahx), proline, or glycine, in some embodiments, the additional amino acid at position X4 is eK. In some embodiments, the additional amino acid at position X4 is Ahx. In some embodiments, the additional amino acid at position X4 is proline. In some embodiments, the additional amino acid at position X4 is glycine. ... 19 The additional amino acid at position X is a hydrophobic amino acid, a D-amino acid thereof, or is absent. 19 In one embodiment, the additional amino acid at position X is a hydrophobic amino acid. 19 The additional amino acid at position X is A, I, L, F, V, P or G. 19 In one embodiment, the additional amino acid at position X is L. 19 In one embodiment, the additional amino acid at position X is a D-amino acid that is a hydrophobic amino acid. 19 In one embodiment, the additional amino acid at position X is a D-amino acid. 19 The additional amino acid in is a D-amino acid of V. 19 The aspect of X 20 In one embodiment, the peptide comprises the sequence of SEQ ID NO: 41, wherein the amino acid sequence is at positions X5 and X 15 and containing a mutation at positions X4 and X 19wherein X is G and X further comprises an additional amino acid. 19 is a hydrophobic amino acid or a D-amino acid thereof. In some embodiments, the peptide is at least 20 amino acids long. In some embodiments, the peptide is at least 25 amino acids long. In some embodiments, the peptide is at least 15-25 amino acids long. In some embodiments, the peptide is at least 15-20 amino acids long. In some embodiments, the peptide is at least 15-18 amino acids long. In some embodiments, the peptide is at least 16-18 amino acids long. In some embodiments, the peptide is at least 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 amino acids long. In some embodiments, the peptide comprises at least one unnatural amino acid. In some embodiments, the peptide comprises one or two unnatural amino acids. In some embodiments, the peptide comprises at least one D-amino acid. In some embodiments, the peptide comprises one or two D-amino acids. In some embodiments, the peptide comprises one to five D-amino acids. In some embodiments, the peptide comprises one to ten D-amino acids. In some embodiments, the peptide comprises all D-amino acids. In some embodiments, the peptide has a molecular weight of at least 2000 Da. In some embodiments, the peptide exhibits 70% or at least 80% inhibition of PAR2 as assessed by calcium flux using 10 μM peptide.

[0156] In one embodiment, the peptide comprises the sequence of SEQ ID NO:41, wherein the amino acid sequence is at positions X5 and X 15 and at least one mutation selected from E or H at X6; D or H at X7; and D or E at X8. In one embodiment, the peptide comprises the sequence of SEQ ID NO: 41, wherein the amino acid sequence is at positions X5 and X6. 15and at least one mutation selected from H at X6; H at X7; and D at X8. In an embodiment, the aforementioned peptide comprises H at X6; H at X7; and D at X8. In an embodiment, the peptide is at least 20 amino acids long. In an embodiment, the peptide is at least 25 amino acids long. In an embodiment, the peptide is 15-25 amino acids long. In an embodiment, the peptide is 15-20 amino acids long. In an embodiment, the peptide is 15-18 amino acids long. In an embodiment, the peptide is 16-18 amino acids long. In an embodiment, the peptide is 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 amino acids long. In an embodiment, the peptide comprises at least one unnatural amino acid. In an embodiment, the peptide comprises one or two unnatural amino acids. In an embodiment, the peptide comprises at least one D-amino acid. In an embodiment, the peptide comprises one or two D-amino acids. In some embodiments, the peptide comprises 1-5 D-amino acids. In some embodiments, the peptide comprises 1-10 D-amino acids. In some embodiments, the peptide comprises all D-amino acids. In some embodiments, the peptide has a molecular weight of at least 2000 Da. In some embodiments, the peptide exhibits 70% or at least 80% inhibition of PAR2 as assessed by calcium flux using 10 μM peptide.

[0157] In one embodiment, the peptide comprises the sequence of SEQ ID NO:41, wherein the amino acid sequence is at positions X5 and X 15 A mutation in X 14 In one embodiment, the peptide comprises a sequence of SEQ ID NO: 41, wherein the amino acid sequence is at positions X5 and X 15 A mutation in X 19 wherein X 19 is a D-amino acid. In one embodiment, the peptide comprises the sequence of SEQ ID NO: 41, wherein the amino acid sequence at positions X5 and X 15A mutation in X 19 and X 20 wherein X 20 is a hydrophobic amino acid or a D-amino acid. In one embodiment, the peptide comprises the sequence of SEQ ID NO: 41, wherein the amino acid sequence at positions X5 and X 15 A mutation in X 19 wherein X 14 and X 19 is a D-amino acid. In one embodiment, the peptide comprises the sequence of SEQ ID NO: 41, wherein the amino acid sequence at positions X5 and X 15 A mutation in X 19 and X 20 wherein X 14 and X 20 is a D-amino acid.

[0158] In one embodiment, the peptide comprises the sequence of SEQ ID NO:41, wherein the amino acid sequence is at positions X5 and X 15 and wherein the peptide is at least 15 amino acids in length. In some embodiments, the peptide is up to 20 amino acids in length. In some embodiments, the peptide is at least 2000 Da in length. In some embodiments, the peptide exhibits 70% or at least 80% inhibition of PAR2 as assessed by calcium flux using 10 μM peptide and 8 μM SLIGRL (SEQ ID NO: 73) agonist in cells. In some embodiments, the peptide exhibits at least 40% or at least 50% inhibition of PAR2 as assessed by calcium flux using 3 μM peptide and 8 μM SLIGRL (SEQ ID NO: 73) agonist in cells.

[0159] In some embodiments, the peptide comprises at least one D-amino acid. In some embodiments, the aforementioned peptide comprises one or two D-amino acids. In some embodiments, the peptide comprises at position X 13 It also contains a mutation in X 13The embodiments for are described herein. In an embodiment, the peptide further comprises a mutation at position X11. The embodiments for X11 are described herein. In an embodiment, the aforementioned peptide comprises a mutation at either X11 or X13, but not at both positions.

[0160] In some embodiments, the peptide comprises a hydrophobic moiety. In some embodiments, the peptide comprises at least two hydrophobic moieties. In some embodiments, the peptide comprises at least three hydrophobic moieties. The hydrophobic moiety may be attached to the N-terminus, the C-terminus, and / or to an amino acid residue between the N-terminus and the C-terminus. The hydrophobic moiety allows the peptide to cross a cell membrane. In some embodiments, the hydrophobic moiety is naturally occurring. In some embodiments, the hydrophobic moiety is not naturally occurring. In some embodiments, the hydrophobic moiety comprises a lipid moiety, an acyl moiety, a steroid moiety, or an amino acid moiety. In some embodiments, the hydrophobic moiety comprises a phospholipid, cholesterol, steroid, sphingosine, ceramide, octylglycine, 2-cyclohexylalanine, benzoylphenylalanine, or a C1 or C2 acyl group. In some embodiments, the hydrophobic moiety comprises a steroid moiety. In some embodiments, the steroid moiety is deoxycholic acid, lithocholic acid, or a salt thereof. The steroid moiety may be coupled to a free amino group on the peptide, such as on the N-terminus or on an amino acid side chain.

[0161] The lipid moiety may be a straight chain fatty acid. In some embodiments, the lipid moiety is selected from the group consisting of capryloyl (C8); nonanoyl (C9); capryl (C 10 );Undecanoyl(C 11 );Lauroyl(C 12 ); Tridecanoyl (C 13 );Myristoyl(C 14 ); Pentadecanoyl (C 15 ); Palmitoyl (C 16 );Phytanoyl (methyl substituted C 16 );Heptadecanoyl(C 17);Stearoyl(C 18 ); Nonadecanoyl (C 19 ); Arachidoyl (C 20 ); Heneicosanoyl (C 21 );Behenoyl(C 22 );trucisanoyl(C 23 ); and lignoceroyl (C 24 In some embodiments, the lipid moiety is myristoyl (C 14 ), pentadecanoyl (C 15 ), or palmitoyl (C 16 In some embodiments, the hydrophobic moiety is palmitoyl.

[0162] The hydrophobic moiety may be attached to the peptide through an amide bond, an ester bond, an ether bond, a carbon-carbon bond, a carbon-nitrogen bond, a carbon-oxygen bond, or a sulfur-sulfur bond. The hydrophobic moiety may be attached to the peptide using groups on the peptide, including, but not limited to, sulfhydryl, amine, alcohol, and phenolic groups. Other peptide groups and types of bonds useful for attaching hydrophobic moieties are known in the art. Other cell-penetrating and / or membrane-tethering hydrophobic moieties include cholesterol, phospholipids, steroids, sphingosine, ceramide, octyl-glycine, 2-cyclohexylalanine, benzoylphenylalanine, C1 or C2 acyl groups, or C3-C8 fatty acids.

[0163] In certain embodiments, the hydrophobic moiety is attached to the N-terminus, the C-terminus, both the N-terminus and the C-terminus of the peptide, or to an internal residue of the peptide (i.e., an amino acid between the C-terminal amino acid and the N-terminal amino acid).

[0164] In some embodiments, the hydrophobic moiety is attached to the N-terminus of the peptide. In some embodiments, the hydrophobic moiety is attached to the C-terminus of the peptide. In some embodiments, the hydrophobic moiety is attached to an internal residue of the peptide that is not located at the N-terminus or C-terminus. In some embodiments, the hydrophobic moiety is attached to a residue that is within 3 residues of the N-terminus. In some embodiments, the hydrophobic moiety is attached to a residue that is within 5 residues of the N-terminus. In some embodiments, the hydrophobic moiety is attached to a residue that is within 8 residues of the N-terminus. In some embodiments, the hydrophobic moiety is attached to a residue that is within 3 residues of the C-terminus. In some embodiments, the hydrophobic moiety is attached to a residue that is within 5 residues of the C-terminus. In some embodiments, the hydrophobic moiety is attached to a residue that is within 8 residues of the C-terminus. The positions of the hydrophobic moieties of the peptides described above are applicable to any of the peptides and various embodiments described herein.

[0165] In some embodiments, the peptide is about 10-30 amino acids long. In some embodiments, the peptide is about 10-20 amino acids long. In some embodiments, the peptide is about 10-15 amino acids long. In some embodiments, the peptide is about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 amino acids long. In some embodiments, the peptide is at least 13 amino acids long. In some embodiments, the peptide is at least 15 amino acids long. In some embodiments, the peptide is at least 20 amino acids long. In some embodiments, the peptide is at least 25 amino acids long. In some embodiments, the peptide is 13-25 amino acids long. In some embodiments, the peptide is 13-20 amino acids long. In some embodiments, the peptide is 13-18 amino acids long. In some embodiments, the peptide is 15-25 amino acids long. In some embodiments, the peptide is 15-20 amino acids in length. In some embodiments, the peptide is 15-18 amino acids in length. In some embodiments, the peptide is 16-18 amino acids in length. In some embodiments, the peptide is 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 amino acids in length. The aforementioned peptide lengths are applicable to any of the peptides and various embodiments described herein.

[0166] In some embodiments, the peptide comprises a hydrophobic portion having a molecular weight range of about 1500 Da to about 2500 Da. In some embodiments, the peptide comprises a hydrophobic portion having a molecular weight range of about 1700 Da to about 2300 Da. In some embodiments, the peptide comprises a hydrophobic portion having a molecular weight range of about 2000 Da to about 2300 Da. The aforementioned molecular weight ranges are applicable to any of the peptides and various embodiments described herein.

[0167] In certain embodiments, the peptide has a solubility in aqueous solution of up to about 30 mg / mL, about 40 mg / mL, about 50 mg / mL, about 60 mg / mL, about 100 mg / mL, or about 120 mg / mL.

[0168] The peptides described herein may comprise L-amino acids, D-amino acids, or a combination thereof. In some embodiments, all residues in a peptide are L-amino acids. In some embodiments, all residues in a peptide are D-amino acids. In some embodiments, the residues in a peptide are a combination of L-amino acids and D-amino acids. In some embodiments, a peptide comprises 1-5 residues that are D-amino acids. In some embodiments, at least 5% of the peptide sequences comprise D-amino acids. In some embodiments, at least 10% of the peptide sequences comprise D-amino acids. In some embodiments, at least 20% of the peptide sequences comprise D-amino acids. In some embodiments, up to 15% of the peptide sequences comprise D-amino acids. In some embodiments, up to 20% of the peptide sequences comprise D-amino acids. In some embodiments, up to 50% of the peptide sequences comprise D-amino acids. In some embodiments, up to 60% of the peptide sequences comprise D-amino acids. In some embodiments, up to 80% of the peptide sequences comprise D-amino acids. In some embodiments, up to 90% of the peptide sequences comprise D-amino acids. In some embodiments, about 5-15% of the peptide sequences include D-amino acids. In some embodiments, about 5-20% of the peptide sequences include D-amino acids. In some embodiments, about 5-50% of the peptide sequences include D-amino acids.

[0169] In some embodiments, the peptide is a PAR2 antagonist. In some embodiments, the peptide shows at least 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90% or 95% inhibition of PAR2. In some embodiments, the peptide shows at least 70% inhibition of PAR2. In some embodiments, the peptide shows at least 80% inhibition of PAR2. Various methods are known for measuring antagonist activity. For example, antagonist activity can be measured by calcium flux experiments using, for example, 3 μM or 10 μM peptide. A decrease in the maximum calcium signal or slope indicates antagonist activity.

[0170] In some embodiments, the peptides exhibit a substantial antagonist effect and no substantial agonist effect, hi some embodiments, the peptides exhibit less than about 30%, 25%, 20%, 15%, 10%, or 5% of the agonist effect.

[0171] Methods of Use and Treatment PAR2, a cell surface receptor for trypsin-like proteases, is widely expressed in inflammatory cells, mesenchymal cells (e.g. fibroblasts, myofibroblasts, smooth muscle cells), interstitial cells, endothelium, hepatocytes, astrocytes, keratinocytes, pancreatic cells, neural cells, cardiac cells, and epithelia, including lung, intestine, and hepatobiliary tract.PAR2 plays a role in many acute and chronic inflammatory diseases of skin, joints, lung, brain, gastrointestinal tract and liver, and vasculature, and has been implicated in the progression of liver, lung, kidney, and other fibrotic diseases, atopic dermatitis, chronic and acute pain conditions, pruritus, and pulmonary arterial hypertension.

[0172] In addition to its well-recognized role in vascular biology, PAR has also been proposed to be involved in regulating survival, apoptosis, and tumor growth (e.g., Yang et al., Cancer Res 2009, 69:6223-31).PAR2 is important in tumor cell biology in melanoma (Tellez C, et al., Oncogene 2003, 22:3130-37), in hepatocellular carcinoma, and in the processes of invasion and metastasis in breast, ovarian, colon, and pancreatic cancer.

[0173] PAR2 mediates many (patho)physiological pathways involved in acute and chronic inflammation, arthritis, allergic responses, sepsis, inflammatory pain, and cancer cell invasion and metastasis. The pleiotropic downstream pathways activated by PAR2 include calcium mobilization, phospholipase C-β-dependent generation of inositol phosphates and diacylglycerol, activation of Rho and Rac, MAPK and beta-arrestin signaling, and gene transcription (Ossovskaya et al., 2004, supra).

[0174] As a cell surface sensor of proteases, PAR2 confers the ability to respond or over-respond to the rapidly changing proteolytic microenvironment that occurs during inflammation.PAR2-deficient mice show reduced granulocyte infiltration and tissue damage, as well as suppressed inflammatory cytokines in models of intestinal inflammation, autoimmunity, and encephalomyelitis (Noorbakhsh, et al., J Exp Med 2006, 203:425-35; Cenac et al., Am J Pathol 2002, 161:1903-15).In PAR2-deficient mice, reduced cardiac ischemia / reperfusion injury was also observed, which correlated with reduced inflammatory mediators (Antoniak et al., Arterioscler Thromb Vasc Biol. 2010, 30: 2136-42). Conversely, overstimulation of PAR2 can result in severe edema, granulocyte infiltration, increased tissue permeability, tissue damage and hypotension (Vergnolle et al., Br J Pharmacol 1999, 127: 1083-90; Cenac et al., 2002, supra). Agonists of PAR2, including trypsin and synthetic SLIGRL (SEQ ID NO: 73) peptide, also cause the release of calcitonin and substance P from sensory neurons, which causes neutrophil infiltration, edema, hyperalgesia and cancer pain (Vergnolle et al., Nat Med 2001, 7: 821-26; Lam et al., Pain 2010, 149: 263-72). PAR2 has been linked to arthritis, as evidenced by a significant reduction in joint inflammation in PAR2-deficient mice (Ferrell et al. J Clin Invest 2003, 111: 35-41) and up-regulation of receptor expression in synovial tissue in osteoarthritis and rheumatoid arthritis (Ferrell et al., Ann Rheum Dis. 2010, 69: 2051-2054).Sievert and colleagues (Knight V, Tchongue J, Lourensz D, Tipping P, Sievert W. Protease-activated receptor 2 promotes experimental liver fibrosis in mice and activates human hepatic stellate cells. Hepatology 2012;55:879-87) showed that PAR2-deficient mice provided significant protection against liver fibrosis in a mouse model. A recent report by Ruf and Samad (Badeanlou L, Furlan-Freguia C, Yang G, Ruf W, Samad F. Tissue factor-protease-activated receptor 2 signaling promotes diet-induced obesity and adipose inflammation. Nat Med 2011;17:1490-7) provides evidence for a role for macrophage-derived tissue factor (TF)-mediated PAR-2 ​​signaling in diet-induced obesity and adipose tissue inflammation.

[0175] Atopic dermatitis (AD) or severe eczema is the most common chronic inflammatory skin disease, affecting approximately 18 million people in the United States. Typical clinical findings include multiple inflammatory lesions, erosions with lichenification, fibrous papules, and severe dry skin with increased susceptibility to infection. The main uncontrolled symptoms are intense itching and excessive scratching, which can cause further peeling, erosion, and infection. Current prescription-based drug treatments consist of topical or systemic corticosteroids, or calcineurin inhibitors for the most severely affected patients, which can exhibit severe side effects and are generally not suitable for long-term treatment. PAR2 has been identified as a key mediator in the pathogenesis of AD (Steinhoff, M., CU Corvera, MS Thoma, W. Kong, BE McAlpine, GH Caughey, JC Ansel, and NW Bunnett. Proteinase-activated receptor-2 in human skin: tissue distribution and activation of keratinocytes by mast cell tryptase. (1999) Exp Dermatol 8: 282-94;Lee, SE, SK Jeong, and SH Lee. Protease and protease-activated receptor-2 signaling in the pathogenesis of atopic dermatitis. (2010) Yonsei Med J 51: 808-22). PAR2 is upregulated in multiple skin cell types, including keratinocytes, inflammatory cells and sensory nerve endings, during the progression from acute to chronic dermatitis (Frateschi, S., E. Camerer, G. Crisante, S. Rieser, M. Membrez, RP Charles, F. Beermann, JC Stehle, B. Breiden, K. Sandhoff, S. Rotman, M. Haftek, A. Wilson, S. Ryser, M. Steinhoff, S.R. Coughlin, and E. Hummler. PAR2 absence completely rescues inflammation and ichthyosis caused by altered CAP1 / Prss8 expression in mouse skin. (2011) Nat Commun 2: 161;Seeliger, S., C.K. Derian, N. Vergnolle, N.W. Bunnett, R. Nawroth, M. Schmelz, P.Y. Von Der Weid, J. Buddenkotte, C. Sunderkotter, D. Metze, P. Andrade-Gordon, E. Harms, D. Vestweber, T.A. Luger, and M. Steinhoff. Proinflammatory role of proteinase-activated receptor-2 in humans and mice during cutaneous inflammation in vivo. (2003) FASEB J 17: 1871-85;Rattenholl, A. and M. Steinhoff. Proteinase-activated receptor-2 in the skin: receptor expression, activation and function during health and disease. (2008) Drug News Perspect 21: 369-81;Buddenkotte, J., C. Stroh, I.H. Engels, C. Moormann, V.M. Shpacovitch, S. Seeliger, N. Vergnolle, D. Vestweber, T.A. Luger, K. Schulze-Osthoff, and M. Steinhoff.Agonists of proteinase-activated receptor-2 stimulate upregulation of intercellular cell adhesion molecule-1 in primary human keratinocytes via activation of NF-kappa B. (2005) J Invest Dermatol 124: 38-45). Increased protease activity in the skin from environmental sources (e.g., DerP / F from house dust mites) and local inflammatory proteases such as mast cell tryptase (Kawakami, T., T. Ando, ​​M. Kimura, BS Wilson, and Y. Kawakami. Mast cells in atopic dermatitis. (2009) Curr Opin Immunol 21: 666-78), kallikrein-5, and cathepsin S (Viode, C., O. Lejeune, V. Turlier, A. Rouquier, C. Casas, V. Mengeaud, D. Redoules, and AM Schmitt. Cathepsin S, a new pruritus biomarker in clinical dandruff / seborrhoeic dermatitis evaluation. (2014) Exp Dermatol 23: 274-5) contribute to aberrant PAR2 signaling and activation of inflammatory responses and pruritus (Briot, A., C. Deraison, M. Lacroix, C. Bonnart, A. Robin, C. Besson, P. Dubus, and A. Hovnanian. Kallikrein 5 induces atopic dermatitis-like lesions through PAR2-mediated thymic stromal lymphopoietin expression in Netherton syndrome. (2009) J Exp Med 206: 1135-47; de Veer, SJ, L. Furio, JMHarris, and A. Hovnanian. Proteases: common culprits in human skin disorders. (2014) Trends Mol Med 20: 166-178). Ablation of PAR2 stimulates overexpression of thymic stromal lymphopoietin (TSLP) (Duchatelet, S. and A. Hovnanian. Genetics of Atopic Dermatitis: Beyond Filaggrin-the Role of Thymic Stromal Lymphopoietin in Disease Persistence. (2014) JAMA Dermatol 150: 248-50), which mediates AD pathogenesis and itch through a subset of C-fibers (Wilson, SR, L. The, LM Batia, K. Beattie, GE Katibah, SP McClain, M. Pellegrino, DM Estandian, and DM Bautista. The epithelial cell-derived atopic dermatitis cytokine TSLP activates neurons to induce itch. (2013) Cell 155: 285-95).

[0176] Idiopathic pulmonary fibrosis (IPF), the most common of the interstitial lung diseases (ILDs), occurs in approximately 128,000 people in the United States, with 48,000 new cases diagnosed each year. The typical clinical course is a progressive fibrotic disease characterized by scarring and "honeycombing" of the lungs, which causes irreversible loss of the tissue's ability to transport oxygen. Co-morbid pulmonary hypertension is commonly observed in patients with IPF and is responsible for a worse clinical prognosis. IPF eventually robs patients of the ability to breathe, resulting in a mortality rate of 66% at 5 years after diagnosis. This corresponds to an unacknowledged high number of deaths per year (n=40,000), which is roughly the same annual rate as the number of deaths attributable to breast cancer. Current treatments have focused primarily on blocking the proliferation of lung fibroblasts. Nintedanib, a pan-tyrosine kinase inhibitor, appears to have some benefit in patient-important outcomes (slower disease progression), but no significant effect on mortality was detected in three clinical trials. Pirfenidone, an IPF drug with unknown mechanism of action, has shown a small reduction in mortality and a slower rate of forced vital capacity (FVC) decline. Despite the advent of these two newly approved drugs, there is still no truly effective treatment for IPF (especially for subjects with more advanced disease), and both drugs cause significant GI and liver toxicity, and pirfenidone induces rash / photosensitivity.PAR2 has recently been identified as a key mediator in the pathogenesis of IPF (Wygrecka, M., G. Kwapiszewska, E. Jablonska, S. von Gerlach, I. Henneke, D. Zakrzewicz, A. Guenther, KT Preissner, and P. Markart. Role of protease-activated receptor-2 in idiopathic plumonary fibrosis. (2011) Am J Respir Crit Care Med 183: 1703-14;Wygrecka, M., BK Dahal, D. Kosanovic, F. Petersen, B. Taborski, S. von Gerlach, M. Didiasova, D. Zakrzewicz, KT Preissner, RT Schermuly, and P. Markart. Mast cells and fibroblasts work in concert to aggravate plumonary fibrosis: role of transmembrane SCF and the PAR-2 / PKC-alpha / Raf-1 / p44 / 42 signaling pathway. (2013) Am J Pathol 182: 2094-108;Park, YS, CM Park, HJ Lee, JM Goo, DH Chung, SM Lee, JJ Yim, YW Kim, SK Han, and CG Yoo. Clinical implication of protease-activated receptor-2 in idiopathic plumonary fibrosis. (2012) Respir Med 107: 256-62).PAR2 is upregulated in lung epithelium, fibroblasts, and inflammatory cells during the progression of IPF, and IPF patients with high PAR2 expression in the lung have worse overall survival and pulmonary honeycombing.Increased procoagulant protease (factor VIIa / Xa / TF) activity and local inflammatory proteases such as mast cell tryptase in the lung induce aberrant PAR2 signaling and activation of the fibrotic response. The number of mast cells in the lungs of patients with fibrotic lung disease is also increased and correlates with the severity of fibrosis. Proteolytic cleavage of PAR2 stimulates overexpression of TGF-β, which induces fibroblast lesion formation and αSMA production through the ERK1 / 2 pathway. Thus, effective PAR2 blockade prevents the chronic tryptase-driven positive feedback mechanism and the procoagulant protease activity of PAR2 in lung epithelium, fibroblasts and inflammatory cells, suppressing the fibrotic response in IPF patients acting as a unique disease-modifying agent.

[0177] Increased expression of PAR2 has also been demonstrated in the pancreas and in experimental rat liver fibrosis and has been shown to correlate with the degree of interstitial fibrosis in IgA nephropathy (Michael, ES, A. Kuliopulos, L. Covic, ML Steer, and G. Perides. Pharmacological inhibition of PAR2 with the pepducin P2pal-18S protects mice against acute experimental biliary pancreatitis. (2013) Am J Physiol Gastrointest Liver Physiol 304: G516-26;Grandaliano, G., P. Pontrelli, G. Cerullo, R. Monno, E. Ranieri, M. Ursi, A. Loverre, L. Gesualdo, and FP Schena. Protease-activated receptor-2 expression in IgA nephropathy: a potential role in the pathogenesis of interstitial fibrosis. (2003) J Am Soc Nephrol 14: 2072-83;Ikeda, O., H. Egami, T. Ishiko, S. Ishikawa, H. Kamohara, H. Hidaka, S. Mita, and M. Ogawa. Expression of proteinase-activated receptor-2 in human pancreatic cancer: a possible relation to cancer invasion and induction of fibrosis. (2003) Int J Oncol 22: 295-300).In lung diseases, high expression of PAR2 has been reported in PAH (Kwapiszewska, G., P. Markart, BK Dahal, B. Kojonazarov, LM Marsh, RT Schermuly, C. Taube, A. Meinhardt, HA Ghofrani, M. Steinhoff, W. Seeger, KT Preissner, A. Olschewski, N. Weissmann, and M. Wygrecka. PAR-2 ​​inhibition reverses experimental pulmonary hypertension. (2012) Circ Res 110: 1179-91), bronchopulmonary dysplasia and childhood respiratory distress syndrome (Cederqvist, K., C. Haglund, P. Heikkila, MD Hollenberg, R. Karikoski, and S. Andersson. High expression of pulmonary proteinase-activated receptor 2 in acute and chronic lung injury in preterm infants. (2005) Pediatr Res 57: 831-6) have been observed. PAR2 was localized to hyperplastic ATII cells and fibroblasts / myofibroblasts in fibrotic lungs. In addition, fibroblasts isolated from IPF lungs showed significantly higher PAR2 levels than fibroblasts extracted from donor lungs. TGF-β, a cytokine known to be critically involved in the pathogenesis of IPF, potently induced PAR2 synthesis in donor lung fibroblasts. Quiescent tissue fibroblasts constitutively express lower levels of PAR2, but conditions that promote fibroblast activation markedly increase PAR2 expression.Thus, transformation of low-PAR2 to high-PAR2 positive fibroblasts occurs in wound models and in normal and hypertrophic human scars (Materazzi, S., S. Pellerito, C. Di Serio, M. Paglierani, A. Naldini, C. Ardinghi, F. Carraro, P. Geppetti, G. Cirino, M. Santucci, F. Tarantini, and D. Massi. Analysis of protease-activated receptor-1 and -2 in human scar formation. (2007) J Pathol 212: 440-9). Collectively, these data support a mechanism in which tissue injury / injury triggers induction and activation of PAR2 by extracellular proteases, which drives physiological tissue repair versus pathological tissue responses, resulting in fibrosis in patients.

[0178] Tryptase, a major proinflammatory serine protease, can also cleave and activate PAR2. Local or systemic release of high levels of mast cell-derived tryptase can have life-threatening consequences, including acute asthma, systemic mastocytosis, and anaphylaxis (Caughey, Immunol Rev 2007, 217: 141-54), and can cause idiopathic pulmonary fibrosis. A specific and effective pharmacological inhibitor of PAR2 therefore has the potential to provide beneficial anti-inflammatory effects and reduce the harmful activity of mast cells, neutrophils, monocytes and / or macrophages, T cells, and other lymphocytes that overexpress PAR2 and cause tissue damage.

[0179] The new peptides, including versions of lipopeptides, are useful for targeting signal transduction events regulated by PAR2, as well as their upstream or downstream effects. For example, the peptides, lipopeptides and compositions herein are used to treat diseases or conditions associated with increased or abnormal PAR2 activity or signal transduction, or associated with increased or abnormal PAR2 protease activity. The peptides and compositions herein can also be used to treat constitutive PAR2 activity. Provided herein is a method for treating a disease or condition associated with PAR2 in a subject in need thereof, comprising administering to the subject an effective amount of a peptide or lipopeptide as described herein. Provided herein is a method for treating a disease or condition associated with PAR2 in a subject in need thereof, comprising instructing the subject to take an effective amount of a peptide as described herein for the subject. Also provided herein is a peptide for use in treating a disease or condition associated with PAR2 in a subject in need thereof.

[0180] In certain embodiments, the peptides and lipopeptides herein are useful in methods of treatment for a variety of PAR2 disorders, including, but not limited to, non-alcoholic steatohepatitis (NASH), idiopathic pulmonary fibrosis (IPF), atopic dermatitis (AD, eczema), renal fibrosis, alcoholic steatohepatitis, organ fibrosis, renal fibrosis, myelofibrosis, pulmonary arterial hypertension (PAH), pulmonary fibrosis, pruritus, pancreatitis, chronic kidney disease, nephritis, multiple sclerosis, cancer, leukemia, melanoma, inflammatory disorders and conditions, sepsis, inflammation-associated CNS disorders, bronchitis, asthma, diabetes, complications of diabetes and NASH, obesity, metabolic syndrome, fibrotic diseases, cardiac fibrosis, pulmonary fibrosis, inflammatory bowel disease, Crohn's disease, irritable bowel syndrome, cirrhosis, arthritis, joint fibrosis, keloids, myelofibrosis, systemic fibrosis, scleroderma, psoriasis, urticaria, impetigo, rash, and rosacea. In some embodiments, the disorder is alcoholic steatohepatitis (NASH), idiopathic pulmonary fibrosis (IPF), atopic dermatitis (AD, eczema), renal fibrosis, alcoholic steatohepatitis, organ fibrosis, renal fibrosis, bone marrow fibrosis, pulmonary arterial hypertension (PAH), pulmonary fibrosis, pruritus (itch), pancreatitis, chronic kidney disease, nephritis, multiple sclerosis, cancer, leukemia, melanoma, inflammatory disorders and conditions, sepsis, inflammation-related CNS disorders, bronchitis, asthma, diabetes, complications of diabetes and NASH, obesity, metabolic syndrome, fibrotic disease, cardiac fibrosis, pulmonary fibrosis, inflammatory bowel disease, Crohn's disease, irritable bowel syndrome, cirrhosis, arthritis, joint fibrosis, keloid, bone marrow fibrosis, systemic fibrosis, scleroderma, psoriasis, urticaria, impetigo, rash, or rosacea.In some embodiments, the PAR2 disorder is NASH. In some embodiments, the disorder is NASH.In some embodiments, the disorder is diabetes.In some embodiments, the PAR2 disorder is a cancer selected from the group consisting of colon, skin, melanocyte, breast, prostate, central nervous system, brain, immune system, pancreas, head and neck, esophagus, kidney, reproductive system, ovary, endometrium, and cervix cancer.In some embodiments, the disorder is a cancer selected from the group consisting of colon, skin, melanocyte, breast, prostate, central nervous system, brain, immune system, pancreas, head and neck, esophagus, kidney, reproductive system, ovary, endometrium, and cervix cancer.

[0181] In some embodiments, the peptides and lipopeptides herein are useful in methods of treatment for conditions involving inflammation. In some embodiments, the peptides and lipopeptides herein are useful in methods of treatment for pancreatitis, asthma, rheumatoid arthritis, osteoarthritis, cancer, chronic pain, visceral pain, cancer pain, multiple sclerosis, inflammatory bowel disease, irritable bowel syndrome, mast cell disease, mastocytosis, gout, sepsis, arterial restenosis, atherosclerosis, inflammatory diseases of the airways and gastrointestinal tract, pruritus, ichthyoses, pruritus, inflammatory skin diseases, psoriasis, and Alzheimer's disease.

[0182] In some embodiments, the peptides and lipopeptides herein are useful in methods of treatment to reduce glycated hemoglobin (HbA1c) levels by about 0.5% to about 1.0% in subjects treated with the peptides and lipopeptides, as compared to a vehicle control group not treated with the peptides and lipopeptides. In some embodiments, the peptides and lipopeptides herein are useful to reduce HbA1c levels by about 0.4% to about 1.0%, about 0.5% to about 1.0%, about 0.6% to about 1.0%, about 0.8% to about 1.0%, or about 0.9% to about 1.0%. In some embodiments, the peptides and lipopeptides herein are useful to reduce HbA1c levels by about 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, or 0.9%. In some embodiments, the peptides and lipopeptides herein are useful in methods of treatment to reduce insulin levels by about 40% to about 60% in subjects treated with the peptides and lipopeptides compared to a vehicle control group not treated with the peptides and lipopeptides. In some embodiments, the peptides and lipopeptides herein are useful to reduce insulin levels by about 40% to about 60%, about 45% to about 60%, about 45% to about 50%, about 50% to about 60%, about 50% to about 55%, or about 55% to about 60%. In some embodiments, the peptides and lipopeptides herein are useful to reduce insulin levels by about 40%, about 45%, about 50%, about 55%, or about 60%. In some embodiments, the subject is a mammal. In some embodiments, the subject is a human. In some embodiments, the subject is a non-human animal.

[0183] Pharmaceutical Compositions Provided herein is a pharmaceutical composition comprising a peptide as described herein and a pharmaceutical acceptable excipient. Pharmaceutical compositions include compositions for therapeutic use. Such compositions may comprise one or more additional therapeutic active agents. Antagonist peptides can be administered to mammals in need of treatment, including humans, alone or in combination with pharmaceutical acceptable carriers, excipients or diluents in pharmaceutical compositions according to standard pharmaceutical practice. Compounds can be administered orally or parenterally, including intravenous, intramuscular, intraperitoneal, subcutaneous, rectal and topical routes of administration. The phrase "active ingredient" or "agent" generally refers to peptide as described herein.

[0184] Although the description of pharmaceutical compositions provided herein is directed primarily to pharmaceutical compositions for administration to humans, those skilled in the art will appreciate that such compositions are generally suitable for administration to animals of all kinds. Modifications of pharmaceutical compositions for administration to a variety of animals are well understood, and a veterinary pharmacologist of ordinary skill can design and / or implement such modifications, if necessary, with no more than routine experimentation.

[0185] The pharmaceutical compositions described herein can be prepared by any method known or hereafter developed in the field of pharmacology.In general, such preparation methods include the step of bringing the active ingredient into association with an excipient and / or one or more other accessory ingredients, and then, if necessary and / or desirable, shaping and / or packaging the product into a desired single-dose or multiple-dose unit.

[0186] The pharmaceutical composition of the present invention can be prepared, packaged, and / or sold in bulk, as a single unit dose, or as a plurality of single unit doses.As used herein, a "unit dose" is a discrete amount of pharmaceutical composition that contains a predetermined amount of active ingredient.The amount of active ingredient is generally equal to the dosage of active ingredient administered to a subject, and / or a convenient fraction of such a dosage, such as half or a third of such a dosage.

[0187] The relative amounts of active ingredient, pharma- ceutically acceptable excipient, and / or any additional ingredients in a pharmaceutical composition of the invention will vary depending on the identity, size, and / or disorder of the subject being treated, and also depending on the route by which the composition is to be administered. By way of example, the composition may contain from 0.1% to 100% (w / w) active ingredient.

[0188] As used herein, pharma- ceutically acceptable excipients include any and all solvents, suspending media, diluents or other liquid vehicles, dispersing or suspending aids, surfactants, isotonicity agents, thickening or emulsifying agents, preservatives, solid binders, lubricants, etc., that are suitable for the particular administration form desired. Remington's The Science and Practice of Pharmacy, 21st Edition, AR Gennaro, (Lippincott, Williams & Wilkins, Baltimore, MD, 2006) discloses a variety of excipients used in formulating pharmaceutical compositions, and known techniques for their preparation. Use of any conventional carrier medium is contemplated within the scope of the present invention, except insofar as it is incompatible with the substance or its derivatives, such as producing any undesirable biological effects or otherwise interacting in a deleterious manner with any other component of the pharmaceutical composition.

[0189] In some embodiments, the pharma- ceutically acceptable excipient is at least 95%, 96%, 97%, 98%, 99%, or 100% pure. In some embodiments, the excipient is approved for human use and for veterinary use. In some embodiments, the excipient is approved by the U.S. Food and Drug Administration. In some embodiments, the excipient is pharmaceutical grade. In some embodiments, the excipient meets the standards of the United States Pharmacopoeia (USP), the European Pharmacopoeia (EP), the British Pharmacopoeia, and / or the International Pharmacopoeia.

[0190] Pharmaceutically acceptable excipients used in the manufacture of pharmaceutical compositions include, but are not limited to, inert diluents, dispersing and / or granulating agents, surfactants and / or emulsifying agents, disintegrating agents, binding agents, preservatives, buffering agents, lubricants and / or oils. Such excipients may be optionally included in the formulations of the present invention. Excipients such as cocoa butter and suppository waxes, coloring agents, coating agents, sweetening agents, flavoring agents and perfuming agents may be present in the composition according to the judgment of the formulator.

[0191] The pharmaceutical composition containing the active ingredient may be in the form suitable for oral use, for example, as a tablet, troche, lozenge, aqueous or oily suspension, dispersible powder or granule, emulsion, hard or soft capsule, or syrup or elixir. The composition intended for oral use can be prepared according to any method known in the art for the manufacture of pharmaceutical compositions, and such compositions may contain one or more agents selected from the group consisting of sweeteners, flavoring agents, coloring agents, and preservatives, in order to provide a medicament elegant and palatable preparation. The tablet contains the active ingredient mixed with non-toxic pharmaceutically acceptable excipients suitable for the manufacture of tablets. These excipients may be, for example, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents such as microcrystalline cellulose, croscarmellose sodium, corn starch or alginic acid; binders such as starch, gelatin, polyvinylpyrrolidone or gum arabic, and lubricants such as magnesium stearate, stearic acid or talc. Tablets may be uncoated or coated by known techniques to mask unpleasant tastes of the drug or to delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained action over a longer period of time. For example, water-soluble taste masking agents such as hydroxypropylmethylcellulose or hydroxypropylcellulose, or time-delay materials such as ethylcellulose, cellulose acetate butyrate may be employed.

[0192] Formulations for oral use may be presented as hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent, such as calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules in which the active ingredient is mixed with a water-soluble carrier, such as polyethylene glycol, or an oil medium, such as peanut oil, liquid paraffin or olive oil.

[0193] Aqueous suspensions contain the active material mixed with excipients suitable for the manufacture of aqueous suspensions.Such excipients can be suspending agents, such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum arabic;dispersing or wetting agents can be naturally occurring phosphatides, such as lecithin, or condensation products of alkylene oxides with fatty acids, such as polyoxyethylene stearate, or condensation products of ethylene oxide with long-chain aliphatic alcohols, such as heptadecaethylene-oxetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitols, such as polyoxyethylene sorbitol monolaurate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, such as polyethylene sorbitan monolaurate. The aqueous suspensions may also contain one or more preservatives, for example ethyl, or n-propyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents, and one or more sweetening agents, such as sucrose, saccharin, or aspartame.

[0194] Oily suspensions can be formulated by suspending the active ingredient in vegetable oil, such as peanut oil, olive oil, sesame oil or coconut oil, or in mineral oil, such as liquid paraffin.Oily suspensions can contain thickening agents, such as beeswax, hard paraffin or cetyl alcohol.In order to provide a palatable oral preparation, sweeteners, such as those mentioned above, and flavoring agents can be added.These compositions can be preserved by adding antioxidants, such as butylated hydroxyanisole or alpha-tocopherol.

[0195] Dispersible powders and granules suitable for preparing aqueous suspension by adding water provide active ingredient mixed with dispersing or wetting agent, suspending "agent" and one or more preservatives.Suitable dispersing or wetting agent and suspending agent are exemplified by those already mentioned above.Additional excipients, such as sweeteners, flavorings and colorings, may also be present.These compositions can be preserved by adding antioxidants such as ascorbic acid.

[0196] The pharmaceutical composition of the present invention may also be in the form of an oil-in-water emulsion.The oil phase may be vegetable oil, such as olive oil or peanut oil, or mineral oil, such as liquid paraffin, or a mixture thereof.Suitable emulsifiers may be naturally occurring phosphatides, such as soybean lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides, such as sorbitan monolaurate, and the condensation products of said partial esters with ethylene oxide, such as polyoxyethylene sorbitan monolaurate.Emulsions may also contain sweeteners, flavorings, preservatives and antioxidants.

[0197] Syrups and elixirs can be formulated with sweetening agents, for example glycerol, propylene glycol, sorbitol or sucrose, and such formulations may also contain a demulcent, a preservative, flavouring and colouring agents and antioxidant.

[0198] The pharmaceutical compositions may be in the form of a sterile injectable aqueous solution.Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution.

[0199] The sterile injectable preparation may also be a sterile injectable oil-in-water microemulsion in which the active ingredient is dissolved in the oily phase. For example, the active ingredient may be first dissolved in a mixture of soybean oil and lecithin. The oily solution may then be introduced into a mixture of water and glycerol and processed to form a microemulsion.

[0200] The injectable solution or microemulsion may be introduced into the patient's bloodstream by local bolus injection. Alternatively, it may be advantageous to administer the solution or microemulsion in such a manner as to maintain a constant circulating concentration of the compound. To maintain such a constant concentration, a continuous intravenous delivery device may be utilized. One example of such a device is the Deltec CADD-PLUS™ model 5400 intravenous pump.

[0201] The pharmaceutical compositions may be in the form of a sterile injectable aqueous or oleagenous suspension for intramuscular and subcutaneous administration. The suspensions may be formulated according to known techniques using those suitable dispersing or wetting agents and suspending agents mentioned above. The sterile injectable preparations may also be sterile injectable solutions or suspensions in non-toxic parenterally acceptable diluents or solvents, for example as solutions in 1,3-butanediol. In addition, sterile, fixed oils are conventionally employed as solvents or suspending media. For this purpose, any bland, fixed oil may be employed, including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables.

[0202] The compounds for the present invention can be administered in intranasal form, through topical use of suitable intranasal vehicles and delivery devices, or via transdermal routes, using the form of transdermal patches well known to those skilled in the art. To be administered in the form of a transdermal delivery system, the administration of the dosage is of course continuous rather than intermittent throughout the entire administration regimen. The compounds of the present invention can also be delivered as suppositories employing bases such as cocoa butter, glycerinated gelatin, hydrogenated vegetable oils, mixtures of polyethylene glycols of various molecular weights, and fatty acid esters of polyethylene glycol. The compounds of the present invention can also be administered in the form of liposomal delivery systems, such as small unilamellar vesicles, large unilamellar vesicles, and multilamellar vesicles. Liposomes can be formed from various phospholipids, such as cholesterol, stearylamine, or phosphatidylcholine. The compounds of the present invention can also be delivered by using monoclonal antibodies as individual carriers to which the compound molecules are coupled. The compounds of the present invention can also be coupled with soluble polymers as targetable drug carriers. Such polymers may include polyvinylpyrrolidone, pyran copolymers, polyhydroxypropylmethacrylamide-phenol, polyhydroxy-ethylaspartamide-phenol, or polyethyleneoxide-polylysine substituted with palmitoyl residues. Additionally, the compounds of the present invention may be coupled to classes of biodegradable polymers useful in achieving controlled release of drugs, such as polylactic acid, polyglycolic acid, copolymers of polylactic acid and polyglycolic acid, polyepsiloncaprolactone, polyhydroxybutyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates, and block copolymers of crosslinked or amphiphilic hydrogels. When the compositions according to the invention are administered to human subjects, the prescribing physician will determine the daily dosage, with the dosage usually varying according to the age, weight and response of the individual patient, as well as the severity of the patient's condition. In one embodiment, a suitable amount of the "agent" is administered to the mammal undergoing treatment for thrombosis. Administration is in an amount of about 0.1 mg to about 60 mg of the "agent" per kg of body weight per day, or 0.5 mg to about 40 mg of the "agent" per kg of body weight per day. Another therapeutic dosage comprising the composition of the invention includes about 0.01 mg to about 1000 mg of the agent. In another embodiment, the dosage includes about 1 mg to about 5000 mg of the agent.

[0203] kit In another aspect, the kit includes one or more of the compositions (e.g., those including or for producing the peptides provided). A "kit" as used herein typically defines a package or collection that includes one or more of the compositions of the invention and / or other compositions related to the invention, such as those described herein. The kit may include isolated or purified peptides, lipopeptides, polynucleotides, vectors encoding the peptides defined, and / or cells expressing or capable of expressing the peptides provided, and combinations thereof. Each of the compositions of the kit may be provided in liquid form (e.g., in solution), in solid form (e.g., dry powder), or in suspension, such as a frozen suspension of cells. In some cases, some of the compositions may be constitutable or otherwise processable (e.g., into an active form), for example, by the addition of suitable solvents or other species (which may or may not be provided with the kit). Examples of other compositions or components relevant to the present invention include, but are not limited to, solvents, surfactants, diluents, salts, buffering agents, chelating agents, fillers, antioxidants, binders, bulking agents, preservatives, desiccants, antimicrobial agents, needles, syringes, packaging materials, test tubes, bottles, flasks, beakers, dishes, frits, filters, rings, clamps, wraps, patches, containers, etc., for using, administering, modifying, assembling, storing, packaging, preparing, mixing, diluting, and / or storing the composition components for a particular application, e.g., to a sample and / or subject. The kit may, in some cases, include instructions in any form provided in association with the composition of the present invention, in such a manner that a person skilled in the art would understand that the instructions should be associated with the composition of the present invention.For example, the instructions may include instructions for using, modifying, activating, mixing, diluting, preserving, administering, assembling, storing, packaging and / or preserving the composition and / or other compositions associated with the kit.In some cases, the instructions may also include instructions for delivery and / or administration of the composition, for example for a specific application, for example to materials and / or subjects.

[0204] example In order that the invention described herein may be more fully understood, the following examples are set forth. It should be understood that these examples are for illustrative purposes only and are not to be construed as limiting the invention in any manner.

[0205] The peptides found in Table 1 were prepared with acyl chain fatty acids. In particular, N-palmitoylated peptides were synthesized by standard fmoc solid phase synthesis methods using C-terminal amides as previously described (see Covic, et al. (2002) Pepducin-based intervention of thrombin-receptor signaling and systemic platelet activation. Nat. Med. 8:1161-1165).

[0206] Palmitic acid was dissolved in 50% N-methylpyrrolidone / 50% methylene chloride and coupled to the deprotected N-terminal amide of the peptide overnight. After cleavage from the resin, the palmitoylated peptide was purified by C18, C5 or C4 reverse phase chromatography to a purity of >95%.

[0207] Table 2 provides antagonist activity for the resulting palmitoylated peptides of Table 1 (SEQ ID NO: 1-11, 13-41 and 70). Table 2 also provides antagonist activity for the resulting myristoylated peptide of Table 1 (SEQ ID NO: 12). Antagonist activity of peptides was measured using calcium flux assay with SW620 human colon adenocarcinoma cells that endogenously express PAR2. Calcium signal of 100 μM SLIGRL (SEQ ID NO: 73) after 1 minute pretreatment with 3 μM or 10 μM palmitoylated peptide. The final concentration of DMSO vehicle was 0.2%. Antagonist activity of 3 μM and 10 μM peptide against 8 μM SLIGRL (SEQ ID NO: 73; a known PAR2 agonist) was measured by maximum calcium flux. Experiments were repeated at least 2-3 times each with similar results. [Table 2]

[0208] Equivalents and Scope In the claims, articles such as "a," "an," and "the" can mean one or more than one, unless indicated to the contrary or otherwise clear from the context. A claim or description containing "or" between one or more members of a group is considered satisfied if one, more than one, or all of the group members are present in, employed in, or otherwise relevant to a given product or process, unless indicated to the contrary or otherwise clear from the context. The invention includes embodiments in which exactly one member of a group is present in, employed in, or otherwise relevant to a product or process. The invention includes embodiments in which more than one, or all of the members of a group are present in, employed in, or otherwise relevant to a product or process.

[0209] Moreover, the present invention encompasses all variations, combinations, and permutations in which one or more limitations, elements, clauses, and descriptive terms from one or more of the enumerated claims are introduced into another claim. For example, any claim that is dependent on another claim can be modified to include one or more limitations found in any other claim that is dependent on the same base claim. When elements are presented as a list, for example in Markush group format, each subgroup of elements is also disclosed, and any element can be removed from the group. In general, when the invention, or aspects of the invention, are referred to as comprising certain elements and / or features, it should be understood that an embodiment or aspect of the invention consists of or consists essentially of such elements and / or features. For the sake of brevity, these embodiments have not been specifically recited in words herein. It is also noted that the terms "comprising" and "containing" are intended to be open and allow for the inclusion of additional elements or steps. When ranges are presented, the endpoints are included. Additionally, unless otherwise indicated or otherwise apparent from the context and the understanding of one of ordinary skill in the art, values ​​expressed as ranges can assume any specific value or subrange within the ranges described in different embodiments of the invention, down to the tenth of the unit of the lower limit of that range, unless the context clearly dictates otherwise.

[0210] This application refers to various issued patents, published patent applications, journal articles, and other publications, all of which are incorporated herein by reference. In the event of a conflict between any of the incorporated references and this specification, this specification shall control. In addition, any aspect of the present invention that falls within the scope of the prior art may be expressly excluded from one or more of the claims. Such aspects may be excluded even if the exclusion is not expressly set forth herein, since they are considered to be known to those skilled in the art. Any aspect of the present invention may be excluded from any claim for any reason, whether or not related to the existence of prior art.

[0211] Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments described herein. The scope of the embodiments described herein is not intended to be limited to the above description, but rather is set forth in the appended claims. Those skilled in the art will recognize that various changes and modifications to this description may be made without departing from the spirit or scope of the invention as defined in the following claims.

[0212] Any patents, patent applications, publications or other disclosure materials identified in the specification are incorporated herein by reference in their entirety. Any material said to be incorporated by reference herein, or portions thereof, that conflicts with existing definitions, statements or other disclosure materials set forth herein, is incorporated only to the extent that no conflict arises between the incorporated material and the material of the present disclosure.

Claims

1. The following sequence: X ４ X ５ X ６ X ７ X ８ X ９ X １０ X １１ X １２ X １３ X １４ X １５ X １６ X １７ X １８ X １９ X ２０ (Array number 42), Where: X ４ is absent, A, G, P, or an N-terminal linker; X ５ is M, G, P, I, L, V, norleucine (J), methionine sulfoxide (M(SO)), or methionine sulfone (M(SO ２ )) or X ４ is absent if absent; X ６ is D, E, H, or X ４ ~X ５ is absent if absent; X ７ is D, E, H, or X ４ ~X ６ is absent if absent; X ８ is N, D, or E; X ９ is any amino acid; X １０ is any amino acid; X １１ is any amino acid or a D-amino acid thereof, 2-aminoisobutyric acid (B), hydroxyproline (Hyp), P, a proline homologue, G, or a stiffener / helix breaker moiety; X １２ is K, R, P, or absent; X １３ is any amino acid or citrulline (Cit); X １４ is K or X １３ and X １４ any amino acid that renders the peptide bond between X １５ is any amino acid or beta-A; X １６ is A, S, T, G, Q, beta-A, 2-aminoisobutyric acid (B), or absent; X １７ is I, A, L or V; X １８ is K, I, or F; and X １９ is a hydrophobic amino acid, a D-amino acid thereof, or is absent; A peptide comprising:

2. X ９ are S, T, H, R, and K; X １０ is E or D; and X １８ The peptide of claim 1, wherein is K, I, or F.

3. The following sequence: X ４ X ５ X ６ X ７ X ８ SEX １１ X １２ X １３ X １４ X １５ X １６ X １７ K.X. １９ (SEQ ID NO:43), Where: X ４ is absent, A, G, P, or an N-terminal linker; X ５ is M, G, P, I, L, V, norleucine (J), methionine sulfoxide (M(SO)), or methionine sulfone (M(SO ２ )) or X ４ is absent if absent; X ６ is D, E, H, or X ４ ~X ５ is absent if absent; X ７ is D, E, H, or X ４ ~X ６ is absent if absent; X ８ is N, D, or E; X １１ is any amino acid or a D-amino acid thereof, 2-aminoisobutyric acid (B), hydroxyproline (Hyp), P, a proline homologue, G, or a stiffener / helix breaker moiety; X １２ is K, R, P, or absent; X １３ is any amino acid or citrulline (Cit); X １４ is K or X １３ and X １４ any amino acid that renders the peptide bond between X １５ is any amino acid or beta-A; X １６ is A, S, T, G, Q, beta-A, 2-aminoisobutyric acid (B), or absent; X １７ is I, A, L or V; and X １９ is a hydrophobic amino acid, a D-amino acid thereof, or is absent; The peptide of claim 2 or 3, comprising:

4. X １３ The peptide according to any one of claims 1 to 3, wherein is R, F, W, Y, or citrulline (Cit).

5. X １３ The peptide according to any one of claims 1 to 4, wherein is R.

6. X １３ The peptide according to any one of claims 1 to 5, wherein is F.

7. X １１ The peptide according to any one of claims 1 to 6, wherein is hydroxyproline (Hyp).

8. X １１ The peptide according to any one of claims 1 to 6, wherein is K.

9. X １１ The peptide according to any one of claims 1 to 6, wherein

10. X ４ 10. The peptide of claim 1, wherein is an N-terminal linker selected from the group consisting of eK, aminohexanoic acid (Ahx), proline, or glycine.

11. X ４ The peptide according to any one of claims 1 to 9, wherein is G.

12. X ４ The peptide according to any one of claims 1 to 9, wherein is A.

13. X ４ The peptide according to any one of claims 1 to 9, wherein:

14. X at the N-terminus ３ wherein X ３ The peptide of any one of claims 1 to 12, wherein is S, G, P, an N-terminal linker, or a helix breaker.

15. X ３ 15. The peptide of claim 14, wherein is an N-terminal linker selected from the group consisting of eK, aminohexanoic acid (Ahx), proline, or glycine.

16. X ３ The peptide of claim 15, wherein is eK.

17. X at the N-terminus ２ wherein X ２ The peptide according to any one of claims 14 to 16, wherein is S.

18. X at the N-terminus １ wherein X １ is R.

19. At the N-terminus of the peptide, X ３ ;X ２ X ３ or X １ X ２ X ３ 19. The peptide according to any one of claims 1 to 18, comprising: X ３ is S, G, P, an N-terminal linker, or a helix breaker; X ２ is S or T; and X １ is R or K.

20. 20. The peptide of claim 19, wherein the N-terminal linker is eK or aminohexanoic acid (Ahx).

21. X ６ The peptide according to any one of claims 1 to 20, wherein is D.

22. X ６ The peptide according to any one of claims 1 to 20, wherein is H.

23. X ６ The peptide according to any one of claims 1 to 20, wherein is E.

24. X ７ The peptide according to any one of claims 1 to 23, wherein is E.

25. X ７ The peptide according to any one of claims 1 to 23, wherein is H.

26. X ８ The peptide according to any one of claims 1 to 25, wherein is N.

27. X ８ The peptide according to any one of claims 1 to 25, wherein is D.

28. X ６ X ７ X ８ The peptide according to any one of claims 1 to 20, wherein said peptide is HHD.

29. X ６ X ７ X ８ The peptide according to any one of claims 1 to 20, wherein is DEN.

30. X １５ The peptide according to any one of claims 1 to 29, wherein is Q.

31. X １５ The peptide according to any one of claims 1 to 29, wherein is S.

32. X １５ The peptide according to any one of claims 1 to 29, wherein is beta-A.

33. X １６ The peptide according to any one of claims 1 to 32, wherein is A.

34. X １６ The peptide according to any one of claims 1 to 32, wherein is B.

35. X １５ If is beta-A, then X １６ The peptide of any one of claims 1 to 32, wherein:

36. X １７ The peptide according to any one of claims 1 to 35, wherein is I.

37. position ４ X ５ X ６ X ７ The peptide of any one of claims 1 to 9 or 26 to 36, wherein the amino acid residue in is absent.

38. X １９ A peptide according to any one of claims 1 to 37, wherein is I, V, L, A or a D-amino acid thereof.

39. X １９ is a D-amino acid of I, V, L or A.

40. X １９ is dL or dV.

41. X １９ is L.

42. X １９ The peptide of any one of claims 1 to 37, wherein: is absent.

43. X ２０ 42. The peptide according to any one of claims 1 to 41, comprising at the C-terminus: ２０ is a hydrophobic residue or a D-amino acid thereof.

44. X ２０ is I, V, L, A or a D-amino acid thereof.

45. X ２０ is a D-amino acid of I, V, L or A.

46. X ２０ The peptide of claim 45, wherein is dI or dV.

47. X １４ 47. The peptide of any one of claims 1 to 46, wherein is a neutral or negatively charged amino acid.

48. X １４ The peptide according to any one of claims 1 to 46, wherein is K or dK.

49. X ４ is an N-terminal linker; X ５ is G, I or L; X １５ is S; and X ２０ is I, V, L, A or a D-amino acid thereof.

50. 50. The peptide of claim 49, wherein the N-terminal linker is eK, aminohexanoic acid (Ahx), proline, or glycine.

51. X ４ is G; X ５ is L; X １５ is S; ２０ is I, V, L, A or a D-amino acid thereof.

52. X ５ and X ６ ;X １１ and X １２ or X １２ and X １３ 52. The peptide of any one of claims 1 to 51, further comprising an amino acid insertion between:

53. 53. The peptide of any one of claims 1 to 52, wherein the amino acid insertion is P.

54. A peptide comprising an amino acid sequence selected from SEQ ID NOs: 1-68 and 70.

55. A peptide comprising a sequence that is about 50% to about 99% homologous to the amino acid sequence of SEQ ID NOs: 1-68 and 70.

56. A peptide comprising a sequence that is about 50% to about 99% identical to the amino acid sequence of SEQ ID NOs: 1-68 and 70.

57. 57. The peptide of any one of claims 54 to 56, comprising an amino acid sequence comprising SEQ ID NOs: 1 to 68 and 70 having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acid changes, wherein the changes are amino acid substitutions, deletions and / or additions.

58. 54. The peptide of any one of claims 1 to 53, comprising a sequence that is about 50% to about 99% homologous to the amino acid sequence of SEQ ID NO:

43.

59. 54. The peptide of any one of claims 1 to 53, comprising a sequence that is about 75% to about 99% identical to the amino acid sequence of SEQ ID NO:

43.

60. A peptide comprising the sequence of SEQ ID NO: 41, the amino acid sequence being at position X ５ and X １５ and wherein the peptide is at least 15 amino acids in length.

61. A peptide comprising the sequence of SEQ ID NO: 41, the amino acid sequence being at position X ５ and X １５ A mutation in X １９ and X ２０ and further comprising an additional amino acid in the formula: １９ and X ２０ are independently a hydrophobic amino acid or a D-amino acid.

62. X １４ and X ２０ 62. The peptide of claim 61, wherein at least one of is a D-amino acid.

63. position １３ 63. The peptide of any one of claims 60 to 62, further comprising a mutation in

64. position １１ 64. The peptide of any one of claims 60 to 63, further comprising a mutation in

65. position ４ and X １９ 65. The peptide of any one of claims 60 to 64, further comprising additional amino acids at

66. X ４ 66. The peptide of any one of claims 60 to 65, wherein is an N-terminal linker selected from the group consisting of A, or eK, aminohexanoic acid (Ahx), proline, or glycine.

67. X １９ is a hydrophobic amino acid, a D-amino acid thereof.

68. position ２０ 68. The peptide of any one of claims 60 to 67, further comprising additional amino acids at

69. X ２０ is a D-amino acid.

70. X ５ is M, G, P, I, L, V, norleucine (J), M(SO), or M(SO ２ 70. The peptide according to any one of claims 60 to 69,

71. X １５ is S or Q.

72. X ６ D, E, or H in ７ D, E, or H in ８ The peptide of any one of claims 60 to 71, further comprising at least one mutation selected from D, E, or N in the formula:

73. X １４ 73. The peptide of any one of claims 60 to 72, further comprising a D-amino acid in the formula:

74. A peptide comprising a mutated fragment of wild-type protease-activated receptor 2 (PAR2), wherein at least three consecutive amino acid residues in the sequence corresponding to positions 270-290 of the human PAR2 sequence are shared with wild-type PAR2, and wherein at least one mutation in the mutated fragment of PAR2 is at an amino acid position corresponding to position 282 of the human PAR2 sequence.

75. 75. The peptide of claim 74, wherein the amino acid position corresponding to position 280 of the human PAR2 sequence is not mutated.

76. A peptide comprising a mutated fragment of wild-type protease-activated receptor 2 (PAR2), wherein at least three consecutive amino acid residues in a sequence corresponding to amino acid positions 270-290 of the human PAR2 sequence are shared with wild-type PAR2, and wherein at least one mutation in said mutated fragment of PAR2 is at an amino acid position corresponding to position 280 of the human PAR2 sequence.

77. 77. The peptide of claim 76, wherein the amino acid position corresponding to position 282 of the human PAR2 sequence is not mutated.

78. A peptide comprising a mutated fragment of wild-type protease-activated receptor 2 (PAR2), wherein the sequence shares with wild-type PAR2 at least three consecutive amino acid residues in amino acid positions corresponding to positions 270-290 of the human PAR2 sequence, and wherein at least one mutation in the mutated fragment of PAR2 is at the amino acid position corresponding to position 273 of the human PAR2 sequence.

79. A peptide comprising a mutated fragment of wild-type protease-activated receptor 2 (PAR2), wherein at least three consecutive amino acid residues in a sequence corresponding to amino acid positions 270-290 of the human PAR2 sequence are shared with wild-type PAR2, and wherein at least one mutation in said mutated fragment of PAR2 is at an amino acid position corresponding to position 275, 276, or 277 of the human PAR2 sequence.

80. A peptide comprising a mutated fragment of wild-type protease-activated receptor 2 (PAR2), wherein at least three consecutive amino acid residues in a sequence corresponding to amino acid positions 270-290 of the human PAR2 sequence are shared with wild-type PAR2, and wherein at least one mutation in said mutated fragment of PAR2 is at an amino acid position corresponding to positions 273, 274, or 284 of the human PAR2 sequence.

81. 81. The peptide of claim 80, wherein the amino acid at position 273 is an N-terminal linker selected from A, eK, aminohexanoic acid (Ahx), proline and glycine.

82. 1. A peptide comprising a mutated fragment of wild-type protease-activated receptor 2 (PAR2), wherein the sequence shares with wild-type PAR2 at least three consecutive amino acid residues in amino acid positions corresponding to positions 270-290 of the human PAR2 sequence, and wherein at least one mutation in the mutated fragment of PAR2 is at an amino acid position corresponding to positions 274, 275, 276, 277, or 284 of the human PAR2 sequence.

83. The amino acid at position 274 is M, G, P, I, L, V, norleucine (J), M(SO), or M(SO ２ 83. The peptide according to any one of claims 80 to 82,

84. 84. The peptide of any one of claims 79 to 83, wherein the amino acid at position 275 is D, E, or H.

85. 85. The peptide of any one of claims 79 to 84, wherein the amino acid at position 276 is D, E, or H.

86. 86. The peptide of any one of claims 79 to 85, wherein the amino acid at position 277 is D, E, or N.

87. 87. The peptide of any one of claims 80 to 86, wherein the amino acid at position 284 is any amino acid or beta-A.

88. 88. The peptide of any one of claims 80 to 87, further comprising a mutation at an amino acid position corresponding to position 280 of the human PAR2 sequence.

89. 90. The peptide of claim 88, wherein the amino acid at position 280 is any amino acid, 2-aminoisobutyric acid (B), hydroxyproline (Hyp), a proline homolog, or a stiffening / helix breaker moiety.

90. 90. The peptide of any one of claims 80 to 89, further comprising a mutation at an amino acid position corresponding to position 282 of the human PAR2 sequence.

91. 91. The peptide of claim 90, wherein the amino acid at position 282 is any amino acid or citrulline (Cit).

92. 92. The peptide of claim 91, wherein the amino acid at position 282 is R, F, W, Y, or citrulline (Cit).

93. 93. The peptide of any one of claims 74 to 92, further comprising a mutation at an amino acid position corresponding to position 289 of the human PAR2 sequence.

94. 94. The peptide of claim 93, wherein the amino acid at the position corresponding to position 289 is a hydrophobic amino acid or a D-amino acid thereof.

95. 94. The peptide of claim 93, wherein the amino acid at the position corresponding to position 289 is I, V, L, A or a D-amino acid thereof.

96. 96. The peptide of any one of claims 74 to 95, further comprising a mutation at an amino acid position corresponding to position 288 of the human PAR2 sequence.

97. 97. The peptide of claim 96, wherein the amino acid at the position corresponding to position 288 is a hydrophobic amino acid or a D-amino acid thereof.

98. The amino acid at the position corresponding to position 288 is X １９ is I, V, L, A or a D-amino acid thereof.

99. 100. The peptide of any one of claims 1 to 99, which does not contain a mutation at a position corresponding to both positions 288 and 289 of the human PAR2 sequence.

100. 100. The peptide of any one of claims 74 to 99, further comprising a mutation at an amino acid position corresponding to position 283 of the human PAR2 sequence.

101. 101. The peptide of claim 100, wherein the amino acid at position 283 is any amino acid that renders the peptide bond between amino acid positions 282 and 283 uncleavable or any amino acid that reduces the positive charge.

102. A peptide according to any one of claims 74 to 101, further comprising a mutation at an amino acid position corresponding to position 285 of the human PAR2 sequence.

103. 103. The peptide of claim 102, wherein the amino acid at the position corresponding to position 285 is A, S, T, G, Q, N, beta-A, 2-aminoisobutyric acid (B), or absent.

104. A peptide according to any one of claims 74 to 103, further comprising a mutation at an amino acid position corresponding to position 272 of the human PAR2 sequence.

105. 105. The peptide of claim 104, wherein the amino acid at position 282 is an N-terminal linker selected from S, eK, aminohexanoic acid (Ahx), proline or glycine.

106. A peptide comprising a mutant fragment of wild-type PAR2, wherein at least one section of at least two consecutive amino acid residues and at least one section of at least three consecutive amino acid residues in the sequence are common to the wild-type PAR2 sequence.

107. A peptide comprising a mutated fragment of wild-type PAR2, wherein the peptide shares at least two sections of at least two consecutive amino acid residues in sequence with the wild-type PAR2 sequence.

108. A peptide comprising a mutated fragment of wild-type PAR2, wherein the peptide shares at least two sections of at least three consecutive amino acid residues in sequence with the wild-type PAR2 sequence.

109. 109. The peptide of any one of claims 106 to 108, wherein at least one further section of at least two consecutive amino acid residues in the sequence is shared with the wild-type PAR2 sequence.

110. A peptide comprising a mutated fragment of wild-type PAR2, wherein the peptide shares at least three sections of at least three consecutive amino acid residues in sequence with the wild-type PAR2 sequence.

111. A peptide comprising a mutant fragment of wild-type PAR2, wherein at least one section of at least three amino acids and at least one section of at least four consecutive amino acid residues in the sequence are common to the wild-type PAR2 sequence.

112. 112. The peptide of claim 111, wherein at least one more section of at least two consecutive amino acid residues in the sequence is shared with the wild-type PAR2 sequence.

113. A peptide comprising a mutated fragment of wild-type PAR2, wherein the peptide shares at least two sections of at least four consecutive amino acid residues in sequence with wild-type PAR2.

114. A peptide comprising a mutant fragment of wild-type PAR2, wherein the peptide has one section of at least four consecutive amino acid residues and one section of at least six consecutive amino acid residues in common with wild-type PAR2.

115. A peptide according to any one of claims 106 to 114, comprising at least one mutation at an amino acid position corresponding to position 273, 274, 275, 276 or 277 of the human PAR2 sequence.

116. 116. The peptide of claim 115, comprising: The amino acid position corresponding to position 273 is A, or an N-terminal linker selected from eK, aminohexanoic acid (Ahx), proline, and glycine; The amino acid position corresponding to position 274 is M, G, P, I, L, V, norleucine (J), M(SO), or M(SO ２ ) and The amino acid positions corresponding to position 275 are D, E, H; the amino acid position corresponding to position 276 is D, E, or H; and The amino acid position corresponding to position 277 is D, E, or N; The peptide.

117. A peptide according to any one of claims 106 to 114, comprising at least one mutation at an amino acid position corresponding to position 273, 274, 282 or 284 of the human PAR2 sequence.

118. 116. The peptide of claim 115, comprising: The amino acid position corresponding to position 273 is A, or an N-terminal linker selected from eK, aminohexanoic acid (Ahx), proline, and glycine; The amino acid position corresponding to position 274 is M, G, P, I, L, V, norleucine (J), M(SO), or M(SO ２ ) and The amino acid position corresponding to position 282 is R, F, W, Y, or citrulline (Cit); and The amino acid position corresponding to position 284 is Q, S, or beta-A; The peptide.

119. 118. The peptide of any one of claims 106 to 117, comprising at least one mutation at an amino acid position corresponding to position 274, 275, 276, 277 or 284 of the human PAR2 sequence.

120. 120. The peptide of any one of claims 106 to 119, further comprising a mutation at an amino acid position corresponding to position 289 of the human PAR2 sequence.

121. 121. The peptide of claim 120, wherein the amino acid position corresponding to position 289 is a hydrophobic amino acid or a D-amino acid thereof.

122. 122. The peptide of claim 121, wherein the amino acid position corresponding to position 289 is I, V, L, A or a D-amino acid thereof.

123. 13. The peptide of any one of claims 106 to 122, further comprising a mutation at an amino acid position corresponding to position 280 of the human PAR2 sequence.

124. 124. The peptide of claim 123, wherein the amino acid position corresponding to position 280 is any amino acid, 2-aminoisobutyric acid (B), hydroxyproline (Hyp), a proline homolog, or a stiffening / helix breaker moiety.

125. A peptide according to any one of claims 106 to 124, wherein the section of consecutive amino acid residues corresponds to amino acid residues within positions 270 to 290 of the human PAR2 sequence.

126. 126. The peptide of any one of claims 74 to 125, wherein the sequence shares at least 4, 5, 6, 7, 8, 9 or 10 consecutive amino acid residues with the wild-type PAR2 sequence.

127. 127. The peptide of any one of claims 1 to 126, comprising 2 to 15 mutations compared to the wild-type PAR2 fragment.

128. 128. A peptide according to any one of claims 1 to 127, comprising the consensus sequence HHD at positions corresponding to positions 275 to 277 of the human PAR2 sequence.

129. A peptide according to any one of claims 1 to 127, comprising the consensus sequence DEN at positions corresponding to positions 275 to 277 of the human PAR2 sequence.

130. 128. A peptide according to any one of claims 1 to 127, comprising the consensus sequence GLHHD (SEQ ID NO: 78) at positions corresponding to positions 273 to 277 of the human PAR2 sequence.

131. A peptide according to any one of claims 1 to 127, comprising the consensus sequence GLDEN (SEQ ID NO: 79) at positions corresponding to positions 273 to 277 of the human PAR2 sequence.

132. The peptide according to any one of claims 74 to 131, wherein the wild-type PAR2 is human PAR2.

133. 133. A peptide according to any one of claims 74 to 132, wherein the sections of consecutive amino acids are separated by at least one amino acid residue.

134. 134. A peptide according to any one of claims 1 to 133, comprising a hydrophobic portion.

135. 135. A peptide according to any one of claims 1 to 134, comprising at least two hydrophobic moieties.

136. 136. The peptide of claim 134 or 135, wherein the hydrophobic moiety comprises a lipid moiety, an acyl moiety, a steroid moiety, or an amino acid moiety.

137. The lipid portion is: capryloyl (C ８ ); nonanoyl (C ９ ); Capryl (C １０ ); undecanoyl (C １１ ); Lauroyl (C １２ ); tridecanoyl (C １３ ); Myristoyl (C １４ ); pentadecanoyl (C １５ Palmitoyl (C １６ Phytanoyl (methyl substituted C １６ ); heptadecanoyl (C １７ ); stearoyl (C １８ ); nonadecanoyl (C １９ ); Arachidoyl (C ２０ Henicosanoyl (C ２１ ); behenoyl (C ２２ ); tolucisanoyl (C ２３ ); and lignoceroyl (C ２４ 137. The peptide of claim 136, selected from the group consisting of:

138. The lipid portion is myristoyl (C １４ ), pentadecanoyl (C １５ ), or palmitoyl (C １６ ). The peptide of claim 137,

139. The lipid portion is myristoyl (C １４ ). The peptide of claim 138,

140. The lipid portion is palmitoyl (C １６ ). The peptide of claim 138,

141. 138. The peptide of claim 137, wherein the steroid moiety is deoxycholic acid or lithocholic acid.

142. 142. The peptide of any one of claims 134 to 141, wherein the hydrophobic moiety is attached to the N-terminus, the C-terminus, or to an internal residue of the peptide.

143. 143. The peptide of claim 142, wherein the hydrophobic moiety is attached to the N-terminus of the peptide.

144. 144. A peptide according to any one of claims 1 to 143, which is 10 to 30 amino acids in length.

145. 145. The peptide of claim 144, which is 10 to 20 amino acids in length.

146. 145. The peptide of claim 144, which is 10 to 15 amino acids in length.

147. 145. The peptide of claim 144, which is 15 to 20 amino acids in length.

148. 145. The peptide of claim 144, which is 13, 14, 15, 16, 17, 18, 19 or 20 amino acids in length.

149. 149. A peptide according to any one of claims 134 to 148, comprising a hydrophobic portion having a molecular weight range of about 1500 Da to about 2500 Da.

150. 149. A peptide according to any one of claims 134 to 148, comprising a hydrophobic portion having a molecular weight range of about 1700 Da to about 2300 Da.

151. 149. A peptide according to any one of claims 134 to 148, comprising a hydrophobic portion having a molecular weight range of about 2000 Da to about 2300 Da.

152. 149. A peptide according to any one of claims 134 to 148, comprising a hydrophobic portion having a molecular weight range of at least about 2000 Da.

153. 153. The peptide of any one of claims 1 to 152, having a solubility in aqueous solution of up to about 30 mg / mL, about 40 mg / mL, about 50 mg / mL, about 60 mg / mL, about 100 mg / mL, or about 120 mg / mL.

154. 154. The peptide of any one of claims 1 to 153, comprising D-amino acids.

155. 155. A peptide according to any one of claims 1 to 154, comprising 1, 2, 3, 4 or 5 D-amino acids.

156. 156. A peptide according to any one of claims 55 to 155, which does not contain a K to F or K to A mutation at a position corresponding to position 287 of wild-type human PAR2.

157. A peptide according to any one of claims 55 to 156, which does not contain an M to A or M to G mutation at a position corresponding to position 274 of wild-type human PAR2.

158. 158. A peptide according to any one of claims 55 to 157, which does not contain an R to S or R to Q mutation at a position corresponding to position 284 of wild-type human PAR2.

159. 159. The peptide of any one of claims 55 to 158, which comprises an amino acid addition, deletion, or substitution compared to wild-type PAR2.

160. 160. A peptide according to any one of claims 1 to 159 which is a PAR2 antagonist.

161. 161. The peptide of any one of claims 1 to 160, which exhibits at least 40% inhibition of PAR2 using a calcium flux assay with 3 μM peptide and 8 μM SLIGRL (SEQ ID NO: 73) agonist in cells.

162. A pharmaceutical composition comprising a peptide according to any one of claims 1 to 161.

163. 163. The pharmaceutical composition of claim 162, comprising a pharma- ceutically acceptable excipient, carrier or diluent.

164. 164. A pharmaceutical composition according to claim 162 or 163 suitable for oral, nasal, topical, rectal, vaginal or parenteral administration, or for intravenous, subcutaneous or intramuscular injection.

165. A method for treating a disorder, comprising administering to a subject in need thereof a peptide according to any one of claims 1 to 161.

166. 166. The method of claim 165, wherein the disorder is non-alcoholic steatohepatitis (NASH), idiopathic pulmonary fibrosis (IPF), atopic dermatitis (AD, eczema), renal fibrosis, alcoholic steatohepatitis, organ fibrosis, renal fibrosis, myelofibrosis, pulmonary arterial hypertension (PAH), pulmonary fibrosis, pruritus (itch), pancreatitis, chronic kidney disease, nephritis, multiple sclerosis, cancer, leukemia, melanoma, inflammatory disorders and conditions, sepsis, inflammation-related CNS disorders, bronchitis, asthma, diabetes, complications of diabetes and NASH, obesity, metabolic syndrome, fibrotic diseases, cardiac fibrosis, pulmonary fibrosis, inflammatory bowel disease, Crohn's disease, irritable bowel syndrome, cirrhosis, arthritis, joint fibrosis, keloids, myelofibrosis, systemic fibrosis, scleroderma, psoriasis, urticaria, impetigo, rash, or rosacea.

167. 166. The method of claim 165, wherein the disorder is a cancer selected from the group consisting of colon, skin, melanocyte, breast, prostate, central nervous system, brain, immune system, pancreatic, head and neck, esophageal, kidney, reproductive system, ovarian, endometrial, and cervical cancer.

168. 166. The method of claim 165, wherein the disorder is diabetes.

169. 162. A method for reducing glycated hemoglobin (HbA1c) levels by about 0.5% to about 1.0% in a subject in need thereof, comprising administering to a subject in need thereof a peptide described in any one of claims 1 to 161.

170. 170. The method of claim 169, wherein glycated hemoglobin (HbA1c) levels are reduced by about 0.8%.

171. 162. A method for reducing insulin levels by about 40% to about 60%, comprising administering to a subject in need thereof a peptide according to any one of claims 1 to 161.

172. 172. The method of claim 171, wherein insulin levels are reduced by about 45%.

173. The method of any one of claims 169 to 172, wherein the subject is a human.

174. A peptide according to any one of claims 1 to 161 for use in treating a disorder.

175. The peptide of claim 174, wherein the disorder is non-alcoholic steatohepatitis (NASH), idiopathic pulmonary fibrosis (IPF), atopic dermatitis (AD, eczema), renal fibrosis, alcoholic steatohepatitis, organ fibrosis, renal fibrosis, myelofibrosis, pulmonary arterial hypertension (PAH), pulmonary fibrosis, pruritus (itch), pancreatitis, chronic kidney disease, nephritis, multiple sclerosis, cancer, leukemia, melanoma, inflammatory disorders and conditions, sepsis, inflammation-related CNS disorders, bronchitis, asthma, diabetes, complications of diabetes and NASH, obesity, metabolic syndrome, fibrotic diseases, cardiac fibrosis, pulmonary fibrosis, inflammatory bowel disease, Crohn's disease, irritable bowel syndrome, cirrhosis, arthritis, joint fibrosis, keloids, myelofibrosis, systemic fibrosis, scleroderma, psoriasis, urticaria, impetigo, rash, and rosacea.

176. The peptide of claim 174, wherein the disorder is a cancer selected from the group consisting of colon, skin, melanocyte, breast, prostate, central nervous system, brain, immune system, pancreas, head and neck, esophagus, kidney, reproductive system, ovarian, endometrial, and cervical cancer.

177. 175. The peptide of claim 174, wherein the disorder is diabetes.

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