KR20120105429A - Methods of treating inflammation - Google Patents

Abstract

In some embodiments, disclosed herein is a method of treating a MIF mediated disease. In some embodiments, the method comprises (i) binding MIF to CXCR2 and CXCR4, and / or (ii) MIF activation of CXCR2 and CXCR4; (iii) the ability of MIF to form homomers; Or administering an agent that inhibits a combination thereof.

Description

Inflammation treatment method {METHODS OF TREATING INFLAMMATION}

Cross-reference

This application discloses US Provisional Application No. 61 / 245,214, filed Sep. 23, 2009; And US Provisional Application No. 61 / 319,039, filed March 30, 2010, both of which are incorporated herein by reference.

Inflammatory diseases, diseases, conditions and symptoms are characterized in part by the migration of lymphocytes and monocytes into affected tissue. Migration of lymphocytes and monocytes leads to tissue damage and exacerbates inflammatory diseases, diseases, conditions and symptoms. MIF increases and decreases in leukocytes following affected tissue. In general, MIF interacts with CXCR2 and CXCR4 receptors on white blood cells to induce and maintain white blood cell migration. Many leukocytes and monocytes follow the MIF gradient and eventually lead to affected tissue. In general, MIF induces and maintains leukocyte and monocyte migration by interacting with CXCR2 and CXCR4 receptors on leukocytes and monocytes.

There is a need for new methods of treating inflammatory diseases, diseases, conditions (eg, atherosclerosis) and symptoms that do not interfere with (a) non-inflammatory processes or (b) desired inflammatory processes. The inventors have found that unwanted harmful inflammation can be treated by inhibiting the ability of MIF to bind CXCR2, CXCR4, CD44, and CD74. In addition, we believe that targeting the correct regions of MIF and CXCR2, CXCR4, CD44, and CD74 will not affect other (eg, desired beneficial) interactions of MIF, CXCR2, CXCR4, CD44, and CD74. It has been found that this would inhibit MIF's ability to bind CXCR2, CXCR4, CD44, and CD74, thereby preventing unwanted inflammation.

In certain embodiments, herein are the following domains of MIF: N-terminal / pseudo ELR motif / domain, alpha-helix # 1 motif / domain, MIF N-loop motif / domain, loop-barrel-loop motif / Peptides that competitively bind to a binding partner of one of a domain, a C-terminal motif / domain, or a combination thereof are disclosed. In some embodiments, the peptide has the following domains: N-terminal tail, pseudo ELR-loop, alpha-helix # 1 motif / domain, PPQ-loop, PDQ-loop, IGK-loop, NRS-helix, SPDR-loop, Competitively binds to the binding partner of the C-terminal tail, or a combination thereof. In some embodiments, the peptide binds competitively with a binding partner of the N-loop domain. In some embodiments, the peptide comprises an amino acid that competitively binds to the binding partner of MIF leu47. In some embodiments, the peptide binds competitively with a binding partner of the pseudo ELR domain. In some embodiments, the peptide comprises LMAFGGSSEP (SEQ ID NO: 18); LMAFGGSS (SEQ ID NO: 20); Cyclic CLMAFGGSSEPCALC (SEQ ID NO: 423); VHVVPDQLMA (SEQ ID NO: 465); QLMAFGGSSE (SEQ ID NO: 468); VNTNVPRASVPDG (SEQ ID NO: 437); NVPRASVPDG (SEQ ID NO: 172); Cyclic CNVPRASVPDGC (SEQ ID NO: 440); NVPRASVPD (SEQ ID NO: 82); Cyclic CLMAFGGSSEP [Abu] ALC (SEQ ID NO: 429), wherein Abu is isosteric L-amino acid, alpha-aminobutyric acid; Or cyclic CLMAFGGSSEPSALC (SEQ ID NO: 469).

In certain embodiments, disclosed herein are peptides that competitively bind to the binding partner of one motif / domain of CXCR2. In some embodiments, the peptide binds competitively with a binding partner of one of the following domains: CXCR2 extracellular loop 1, CXCR2 extracellular loop 2, CXCR2 extracellular loop 3, or CXCR2 N-terminal / domain.

In certain embodiments, disclosed herein are peptides that competitively bind to the binding partner of one motif / domain of CXCR4. In some embodiments, the peptide is SEADDRYICDRFYPNDLWVVV; Or competitively with a binding partner of DDRYICDRFYPNDLW.

In certain embodiments, disclosed herein are peptides that competitively bind to the binding partner of one motif / domain of CD44.

In certain embodiments, disclosed herein are peptides that competitively bind to a binding partner of one motif / domain of CD74.

In certain embodiments, the application herein includes (a) a first peptide that competitively binds to a binding partner of the N-loop motif of MIF; And (b) a second peptide that competitively binds to the binding partner of the pseudo ELR motif of MIF, wherein the first peptide and the second peptide maintain their activity in the fusion peptide. . In some embodiments, the fusion peptide comprises (a) a first peptide that competitively binds to the binding partner of the N-loop motif of MIF; (b) a second peptide that competitively binds to the binding partner of the pseudo ELR motif of MIF; And (c) a third peptide that competitively binds to the binding partner of the pseudo ELR motif of MIF, wherein the first peptide and the second peptide maintain their activity in the fusion peptide. In some embodiments, the fusion peptide is LMAFGGSSEP (SEQ ID NO: 18); LMAFGGSS (SEQ ID NO: 20); Cyclic CLMAFGGSSEPCALC (SEQ ID NO: 423); VHVVPDQLMA (SEQ ID NO: 465); QLMAFGGSSE (SEQ ID NO: 468); VNTNVPRASVPDG (SEQ ID NO: 437); NVPRASVPDG (SEQ ID NO: 172); Cyclic CNVPRASVPDGC (SEQ ID NO: 440); NVPRASVPD (SEQ ID NO: 82); Cyclic CLMAFGGSSEP [Abu] ALC (SEQ ID NO: 429), wherein Abu is isosteric L-amino acid, alpha-aminobutyric acid; Or a peptide selected from cyclic CLMAFGGSSEPSALC (SEQ ID NO: 469). In some embodiments, the fusion peptide is represented by formula (IV):

In some embodiments, the fusion peptide is represented by formula (V):

In some embodiments, the linker includes alkyl, heteroalkyl, alkylene, alkenylene, alkynylene, heteroalkylene, carbocycle, heterocycle, aromatic ring, nonaromatic ring, substituted ring, monocyclic ring, polish Click rings, or combinations thereof.

In certain embodiments, the disclosure includes (a) an antibody, (b) a peptide described herein, and (c) a linker that binds the peptide to the Fab region of the antibody, wherein the peptide and antibody are present in a peptibody Peptibodies are disclosed that maintain their activity. In some embodiments, the linker binds said peptide to an antigen binding site. In some embodiments, the linker includes alkyl, heteroalkyl, alkylene, alkenylene, alkynylene, heteroalkylene, carbocycle, heterocycle, aromatic ring, nonaromatic ring, substituted ring, monocyclic ring, polish Click rings, or combinations thereof. In some embodiments, the antibody is IgA, IgD, IgE, IgG, or IgM.

In certain embodiments, disclosed herein is the use of a substance composition described herein to treat an inflammatory disease, disease or condition. In some embodiments, the inflammatory disease, disease or condition is atherosclerosis; Abdominal aortic aneurysm; Acute disseminated encephalomyelitis; Moyamoya disease; Takayasu's disease; Acute coronary syndrome; Cardiac allograft angiopathy; Lung inflammation; Acute respiratory distress syndrome; Pulmonary fibrosis; Acute disseminated encephalomyelitis; Addison's disease; Ankylosing spondylitis; Antiphospholipid antibody syndrome; Autoimmune hemolytic anemia; Autoimmune hepatitis; Autoimmune inner ear disease; Bullous whey; Shaga's disease; Chronic obstructive pulmonary disease; Celiac disease; Dermatitis; Type 1 diabetes; Type 2 diabetes; Endometriosis; Goodpasture syndrome; Graves' disease; Guillain-Barré syndrome; Hashimoto's disease; Idiopathic thrombocytopenic purpura; Interstitial cystitis; Systemic lupus erythematosus (SLE); Metabolic syndrome; Multiple sclerosis; Myasthenia gravis; myocarditis; Narcolepsy; obesity; Vulgaris vulgaris; Pernicious anemia; Polymyositis; Primary biliary cirrhosis; Rheumatoid arthritis; Schizophrenia; Scleroderma; Sjogren's syndrome; Vasculitis; Vitiligo; Wegener's granulomatosis; Allergic rhinitis; Prostate cancer; Non-small cell lung carcinoma; Ovarian cancer; Breast cancer; Melanoma; Stomach cancer; Colon cancer; Brain cancer; Metastatic bone disease; Pancreatic cancer; Bladder cancer; Hepatocellular carcinoma; Liver cancer; Lung adenocarcinoma; Esophageal squamous cell carcinoma; CNS tumors; Blood tumors; Lymphoma; Nasal polyps; Gastrointestinal cancer; Ulcerative colitis; Crohn's disease; Collagen colitis; Lymphocytic colitis; Ischemic colitis; Converting colitis; Behcet's syndrome; Infectious colitis; Indeterminate colitis; Inflammatory liver disease; Endotoxic shock; Septic shock; Rheumatoid spondylitis; Ankylosing spondylitis; Gout arthritis; Rheumatic polymyalgia; Alzheimer's disease; Parkinson's disease; epilepsy; AIDS dementia; asthma; Adult respiratory distress syndrome; bronchitis; Cystic fibrosis; Acute leukocyte-mediated lung injury; Distal proctitis; Wegener's granulomatosis; Fibromyalgia; bronchitis; Uveitis; conjunctivitis; psoriasis; eczema; dermatitis; Smooth muscle hyperplasia disease; meningitis; Shingles; encephalitis; Nephritis; Tuberculosis; Retinitis; Atopic dermatitis; Pancreatitis; Periodontal gingivitis; Coagulation necrosis; Liquefied necrosis; Fibrotic necrosis; Neointimal hyperplasia; Myocardial infarction; stroke; Organ transplant rejection; Influenza, or combinations thereof. In some embodiments, the inflammatory disease, disease or condition is prostate cancer; Non-small cell lung carcinoma; Ovarian cancer; Breast cancer; Melanoma; Stomach cancer; Colon cancer; Brain cancer; Metastatic bone disease; Pancreatic cancer; Bladder cancer; Hepatocellular carcinoma; Liver cancer; Lung adenocarcinoma; Esophageal squamous cell carcinoma; CNS tumors; Blood tumors; Lymphoma; Or a combination thereof. In some embodiments, the inflammatory disease, disease or condition is rheumatoid arthritis. In some embodiments, the inflammatory disease, disease or condition is acute respiratory distress syndrome. In some embodiments, the inflammatory disease, disease or condition is glomerulonephritis. In some embodiments, the inflammatory disease, disease or condition is an inflammatory bowel disease. In some embodiments, the inflammatory disease, disease or condition is an abdominal aortic aneurysm disease. In some embodiments, the inflammatory disease, disease or condition is chronic obstructive pulmonary disease. In some embodiments, the inflammatory disease, disease or condition is asthma. In some embodiments, the inflammatory disease, disease or condition is lupus. In some embodiments, the inflammatory disease, disease or condition is sepsis.

In certain embodiments, disclosed herein is the use of a substance composition described herein for treating, preventing and reducing angiogenesis.

In certain embodiments, disclosed herein is a pharmaceutical composition for treating an inflammatory disease, disease, condition or condition in an individual in need thereof for treating an inflammatory disease, disease, condition or condition, comprising a substance composition described herein.

The novel features of the invention are set forth in detail in the appended claims. The features and advantages of the present invention will be further understood by reference to the following detailed description describing exemplary embodiments utilizing the principles of the present invention, and to the accompanying drawings:
1 shows the crystal structure of MIF trimer. The pseudo ELR motif / domain forms a ring in the trimer, while the N-loop motif / domain extends outward from the pseudo ELR ring.
2 shows the nucleotide sequences of annotated MIF showing sequences corresponding to N-loop motifs / domains and pseudo ELR motifs / domains.
3 shows the nucleic acid sequences of human MIF and corresponding MIF motifs / domains.
4 shows that the peptide of SEQ ID NO: 18 blocks chemotaxis in human peripheral blood mononuclear cells (PBMC).
5 shows that the peptide of SEQ ID NO: 423 significantly antagonizes MIF inducible chemotaxis in PMBC.
FIG. 6 shows that the peptide of SEQ ID NO: 423 significantly antagonizes MIF inducible chemotaxis in PMBC in a dose dependent manner.
FIG. 7 shows that the peptides disclosed herein block (under flow) MIF mediated monocyte arrest on human aortic endothelial cells. 4 nM MIF was used. 5 uM peptide was used. * p <0.05; ** p <0.01; *** = p <0.005.
FIG. 8 shows that the peptide of SEQ ID NO: 18 blocks MIF mediated monocyte stop HuAoEC in a dose dependent manner. 4 nM MIF was used. * p <0.05; ** p <0.01; *** = p <0.005.
9 shows the results of probing the MIF interface using peptide SPOT arrays. 1 uM of MIF was incubated overnight. Streptavidin-POD (1: 10,000) was incubated for 2 hours at room temperature.
10 shows the results of probing the MIF CXCR2 interface using a peptide SPOT array. 1 uM of MIF was incubated overnight. Streptavidin-POD (1: 10,000) was incubated for 2 hours at room temperature.
Figure 11 shows the use of peptide SPOT arrays using wild-type MIF (1 μM Biotin-MonoQ-MIF: streptavidin-POD) and mutant MIF (1 μM Biotin-R11A-D44A-MIF streptavidin-POD). The results of probing the MIF CXCR2 interface are presented. Streptavidin-POD (1: 10,000) was incubated for 2 hours at room temperature. Wild-type MIF and mutant MIF were incubated overnight at room temperature.
FIG. 12 shows the results of probing the MIF CXCR4 interface using a peptide SPOT array using wild type MIF (1 μM Biotin-MIF: Streptavidin-POD). Streptavidin-POD (1: 10,000) was incubated for 2 hours at room temperature. Wild type MIF was incubated overnight at 4 ° C.
FIG. 13 shows that SEQ ID NO: 18 inhibits the attachment of leukocytes to the carotid artery of Apoe-deficient mice. Deficiency controls of the same age who received Western diet for 8 wk. 5 of each legion; Quantification from 10 10 high power fields (HPF) throughout the carotid artery 1 × injection of SEQ ID NO. 18 or scrambled SEQ ID NO. 18 for 3 days prior to adhesion experiment & IVM.
FIG. 14 shows that SEQ ID NO 18 decreases TNFα and MCP-1 in a mouse peritonitis model. Thioglycolate (TG): (3% IP). Vehicle, SEQ ID NO: 18, SEQ ID NO: 85 and Dex: 30 min before and 30 min after TG challenge. 2 hr after TG challenge, peritoneal lavage for cell count and chemokine.
FIG. 15 shows that SEQ ID NO: 18 reduces MCP-1 and monocyte levels in a mouse peritonitis model. Thioglycolate (TG): (3% IP). Vehicle, SEQ ID NO: 422, SEQ ID NO: 421, SEQ ID NO: 451 and Dex: 30 min before and 30 min after TG challenge. 2 hr after TG challenge, peritoneal lavage for cell count and chemokine.
16 shows the proposed MIF signaling adjustment mechanism.
FIG. 17 shows a schematic of the structure and surface exposure of MIF N-loops and a two site binding model for MIF / CXCR2. A, 3D architectural homology between CXCL8 and MIF focused on receptor interaction motifs. Binding of the regular ligand CXCL8 to CXCR2 includes N-loops and ELR motifs. MIF includes N-like loops (SEQ ID NO: 47-56) and pseudo (E) LR motifs (constituting amino acids R12 and D45, 3D-ELR motifs). For reasons of clarity only the monomer structures of CXCL8 and MIF are shown. B, diagram showing the structure of MIF. C, two-site binding model application for the binding of chemokine / chemokine receptors to MIF. Interaction interface between proposed MIF and CXCR2 (site 1: interaction between N-like loop of MIF and receptor N-terminus; site 2: between pseudo (E) LR-motif of MIF and receptor exoloops EL2 and 3 Interaction). D, trimer structure of MIF shown in surface mode.
18 is a peptide spot array assay identifying the site of interaction between MIF and the extracellular domain of CXCR2 by peptide spot array assay. Short 15-mer peptides representing full-length human MIF (A) and CXCR2 extracellular domains (B and C) were synthesized directly on amino cellulose membranes. CXCR2 peptides correspond to N-terminal and extracellular loops (EL) 1-3. Peptide strips were incubated with 1 μM Biotin-MIF (A and B) or Biotin-R12A / D45A-MIF (C) and detected using streptavidin-POD. A, MIF strip developed using biotin-MIF. B, CXCR2-strip developed using biotin-MIF. C, CXCR2-strip developed using Biotin-R12A / D45A-MIF.
FIG. 19 shows that MIF N-loop peptides inhibit the interaction between MIF and CXCR2. The effect of the peptide was examined by a competitive receptor binding assay, which measures the reversal of the inhibitory effect by N-loop peptide on MIF on tracer binding. HEK293 cells stably expressing CXCR2 were incubated with 1 μM human MIF and 100 μM, the specified N-like loop peptide of MIF as a competitor, with the radioiodinated I125CXCL8 tracer. Plot shows specific% I125-CXCL8 binding. Tracer binding in the absence of MIF and peptide (in buffer) was set to 100% and the competitive effect of MIF in the absence of peptide was set to 0%. The data is presented as mean ± SEM obtained from three independent experiments, each of which was performed in duplicate measurements (* = p <0.01).
20 is a model depicting the interaction at the MIF / CXCR2 interface following a general two site binding mechanism.
21 is a model depicting a peptibody.

In some embodiments, disclosed herein are peptides, small molecules, antibodies, and peptibodies (collectively, "composition of matter") for treating inflammatory diseases, diseases, conditions, and symptoms. Further disclosed herein are pharmaceutical compositions and methods for treating inflammatory diseases, diseases, conditions and symptoms. In some embodiments, the inflammatory disease, disease, condition or symptom is characterized by unwanted MIF signaling. In some embodiments, the inflammatory disease, disease, condition or symptom is characterized by MIF mediated leukocyte recruitment.

In certain embodiments, herein are the following domains of MIF: N-terminal / pseudo ELR motif / domain, alpha-helix # 1 motif / domain, MIF N-loop motif / domain, loop-barrel-loop motif / domain, C -Peptides that competitively bind with a binding partner of one of the terminal motifs / domains, or combinations thereof. In some embodiments, the peptide has the following domains: N-terminal tail, pseudo ELR-loop, alpha-helix # 1 motif / domain, PPQ-loop, PDQ-loop, IGK-loop, NRS-helix, SPDR-loop, Competitively binds to the binding partner of the C-terminal tail, or a combination thereof. In some embodiments, the peptide binds competitively with a binding partner of the N-loop domain. In some embodiments, the peptide comprises an amino acid that competitively binds to the binding partner of MIF leu47. In some embodiments, the peptide binds competitively with a binding partner of the pseudo ELR domain. In some embodiments, the peptide comprises LMAFGGSSEP (SEQ ID NO: 18); LMAFGGSS (SEQ ID NO: 20); Cyclic CLMAFGGSSEPCALC (SEQ ID NO: 423); VHVVPDQLMA (SEQ ID NO: 465); QLMAFGGSSE (SEQ ID NO: 468); VNTNVPRASVPDG (SEQ ID NO: 437); NVPRASVPDG (SEQ ID NO: 172); Cyclic CNVPRASVPDGC (SEQ ID NO: 440); NVPRASVPD (SEQ ID NO: 82); Cyclic CLMAFGGSSEP [Abu] ALC (SEQ ID NO: 429), wherein Abu is isosteric L-amino acid, alpha-aminobutyric acid; Or cyclic CLMAFGGSSEPSALC (SEQ ID NO: 469).

In certain embodiments, disclosed herein are peptides that competitively bind to the binding partner of one motif / domain of CXCR2. In some embodiments, the peptide binds competitively with a binding partner of one of the following domains: CXCR2 extracellular loop 1, CXCR2 extracellular loop 2, CXCR2 extracellular loop 3, or CXCR2 N-terminal / domain.

In certain embodiments, disclosed herein are peptides that competitively bind to the binding partner of one motif / domain of CXCR4. In some embodiments, the peptide is SEADDRYICDRFYPNDLWVVV; Or competitively with a binding partner of DDRYICDRFYPNDLW.

In certain embodiments, disclosed herein are peptides that competitively bind to the binding partner of one motif / domain of CD44.

In certain embodiments, disclosed herein are peptides that competitively bind to a binding partner of one motif / domain of CD74.

In certain embodiments, the application herein includes (a) a first peptide that competitively binds to a binding partner of the N-loop motif of MIF; And (b) a second peptide that competitively binds to the binding partner of the pseudo ELR motif of MIF, wherein the first peptide and the second peptide maintain their activity in the fusion peptide. . In some embodiments, the fusion peptide comprises (a) a first peptide that competitively binds to the binding partner of the N-loop motif of MIF; (b) a second peptide that competitively binds to the binding partner of the pseudo ELR motif of MIF; And (c) a third peptide that competitively binds to the binding partner of the pseudo ELR motif of MIF, wherein the first peptide and the second peptide maintain their activity in the fusion peptide. In some embodiments, the fusion peptide is LMAFGGSSEP (SEQ ID NO: 18); LMAFGGSS (SEQ ID NO: 20); Cyclic CLMAFGGSSEPCALC (SEQ ID NO: 423); VHVVPDQLMA (SEQ ID NO: 465); QLMAFGGSSE (SEQ ID NO: 468); VNTNVPRASVPDG (SEQ ID NO: 437); NVPRASVPDG (SEQ ID NO: 172); Cyclic CNVPRASVPDGC (SEQ ID NO: 440); NVPRASVPD (SEQ ID NO: 82); Cyclic CLMAFGGSSEP [Abu] ALC (SEQ ID NO: 429), wherein Abu is isosteric L-amino acid, alpha-aminobutyric acid; Or a peptide selected from cyclic CLMAFGGSSEPSALC (SEQ ID NO: 469). In some embodiments, the fusion peptide is represented by formula (IV):

In some embodiments, the fusion peptide is represented by formula (V):

In some embodiments, the linker includes alkyl, heteroalkyl, alkylene, alkenylene, alkynylene, heteroalkylene, carbocycle, heterocycle, aromatic ring, nonaromatic ring, substituted ring, monocyclic ring, polish Click rings, or combinations thereof.

In certain embodiments, the disclosure includes (a) an antibody, (b) a peptide described herein, and (c) a linker that binds the peptide to the Fab region of the antibody, wherein the peptide and antibody are present in a peptibody Peptibodies are disclosed that maintain their activity. In some embodiments, the linker binds said peptide to an antigen binding site. In some embodiments, the linker includes alkyl, heteroalkyl, alkylene, alkenylene, alkynylene, heteroalkylene, carbocycle, heterocycle, aromatic ring, nonaromatic ring, substituted ring, monocyclic ring, polish Click rings, or combinations thereof. In some embodiments, the antibody is IgA, IgD, IgE, IgG, or IgM.

In certain embodiments, disclosed herein is the use of a substance composition described herein to treat an inflammatory disease, disease or condition. In some embodiments, the inflammatory disease, disease or condition is cancer. In some embodiments, the inflammatory disease, disease or condition is atherosclerosis; Abdominal aortic aneurysm; Acute disseminated encephalomyelitis; Moyamoya disease; Takayasu's disease; Acute coronary syndrome; Cardiac allograft angiopathy; Lung inflammation; Acute respiratory distress syndrome; Pulmonary fibrosis; Acute disseminated encephalomyelitis; Addison's disease; Ankylosing spondylitis; Antiphospholipid antibody syndrome; Autoimmune hemolytic anemia; Autoimmune hepatitis; Autoimmune inner ear disease; Bullous whey; Shaga's disease; Chronic obstructive pulmonary disease; Celiac disease; Dermatitis; Type 1 diabetes; Type 2 diabetes; Endometriosis; Goodpasture syndrome; Graves' disease; Guillain-Barré syndrome; Hashimoto's disease; Idiopathic thrombocytopenic purpura; Interstitial cystitis; Systemic lupus erythematosus (SLE); Metabolic syndrome; Multiple sclerosis; Myasthenia gravis; myocarditis; Narcolepsy; obesity; Vulgaris vulgaris; Pernicious anemia; Polymyositis; Primary biliary cirrhosis; Rheumatoid arthritis; Schizophrenia; Scleroderma; Sjogren's syndrome; Vasculitis; Vitiligo; Wegener's granulomatosis; Allergic rhinitis; Prostate cancer; Non-small cell lung carcinoma; Ovarian cancer; Breast cancer; Melanoma; Stomach cancer; Colon cancer; Brain cancer; Metastatic bone disease; Pancreatic cancer; Bladder cancer; Hepatocellular carcinoma; Liver cancer; Lung adenocarcinoma; Esophageal squamous cell carcinoma; CNS tumors; Blood tumors; Lymphoma; Nasal polyps; Gastrointestinal cancer; Ulcerative colitis; Crohn's disease; Collagen colitis; Lymphocytic colitis; Ischemic colitis; Converting colitis; Behcet's syndrome; Infectious colitis; Indeterminate colitis; Inflammatory liver disease; Endotoxic shock; Septic shock; Rheumatoid spondylitis; Ankylosing spondylitis; Gout arthritis; Rheumatic polymyalgia; Alzheimer's disease; Parkinson's disease; epilepsy; AIDS dementia; asthma; Adult respiratory distress syndrome; bronchitis; Cystic fibrosis; Acute leukocyte-mediated lung injury; Distal proctitis; Wegener's granulomatosis; Fibromyalgia; Bronchitis; uveitis; conjunctivitis; psoriasis; eczema; dermatitis; Smooth muscle hyperplasia disease; meningitis; Shingles; encephalitis; Nephritis; Tuberculosis; Retinitis; Atopic dermatitis; Pancreatitis; Periodontal gingivitis; Coagulation necrosis; Liquefied necrosis; Fibrotic necrosis; Neointimal hyperplasia; Myocardial infarction; stroke; Organ transplant rejection; Influenza, or a combination thereof.

In certain embodiments, disclosed herein is the use of a substance composition described herein to treat, prevent, or reduce angiogenesis.

In certain embodiments, disclosed herein is a pharmaceutical composition for treating an inflammatory disease, disease, condition or condition in an individual in need thereof for treating an inflammatory disease, disease, condition or condition, comprising a substance composition described herein.

Justice

The terms "individual," "subject", or "patient" are used interchangeably. As used herein, these terms mean any mammal (ie, taxonomic animal system: vertebrate: vertebrate: any species of the neck, family, and genus belonging to a mammal). In some embodiments, the mammal is a human. In some embodiments, the mammal is nonhuman. In some embodiments, the mammal is a member of the taxa: a primate (eg, lemur, lorid, galago, tarsier, monkey, ape and human); Rodents (eg, mice, rats, squirrels, squirrels and hind mice); Rabbits (eg hares, rabbits, and hares); Hedgehog (eg, hedgehog and furry hedgehog); Pythons (eg, pythons, moles and solenodons); Bat neck (eg, bat); Whales (eg whales, dolphins, and dolphins); Carnivorous trees (eg, cats, limbs and other cats; dogs, bears, weasels and seals); Bases (eg, horses, zebras, macs and radishes); Cows (eg, pigs, camels, cattle and deer); Equipment (eg elephants); Manatee (eg manatees, dugongs and walruses); Armor cuff (eg, armadillo); Driftwood (eg, anteater and sloth); Opossum (eg, american opossum); Marsupials (eg, shrew opossums); Chinchilla (eg, Monito del Monte); Moles (eg moles); Pocket cats (eg, marsupial predators); Bandikurmok (eg, bandikut and bilbi); Or kangaroos (eg, wombat, koala, foursome, squirrel, kangaroo, wallaroo and wallaby). In some embodiments, the animal is a reptile (ie, species of any neck, family, and genus belonging to the taxonomic family: vertebrate: vertebrate: reptile). In some embodiments, the animal is a bird (ie, animal kingdom: vertebrate: vertebrate: avian). None of these terms require a situation characterized by the management (eg, continuous or intermittent) of a healthcare worker (eg, a physician, registered nurse, skilled nurse, medical assistant, hospital handyman, or hospice worker), or It is not limited to this.

When referring to the interaction between a binding molecule (ie, an agent; eg, a peptide or peptide mimetic) and a protein or polypeptide or epitope, the phrase “specifically binds” is typically of interest. A binding molecule that binds detectably and specifically detects with high affinity to a target. Preferably, under defined or physiological conditions, the specified antibody or binding molecule binds to a particular polypeptide, protein or epitope but does not bind in significant or unwanted amounts to other molecules present in the sample. In other words, the specified antibody or binding molecule does not unwanted cross-react with non-target antigens and / or epitopes. Various immunoassay formats can be used to select antibodies or other binding molecules that are immunoreactive with a particular polypeptide and have the desired specificity. For example, solid phase ELISA immunoassays, BIA cores, flow cytometry and radioimmunoassays can be used to select monoclonal antibodies with the desired immunoreactivity and specificity. For a description of immunoassay formats and conditions used to measure or assess immunoreactivity and specificity, see Harlow, 1988, Antibodies, A LABORATORY MANUAL, Cold Spring Harbor Publications, New York (hereinafter referred to as "Harlow"). See

"Selective binding," "selectivity," and the like refer to the preference of an agent to interact with one molecule compared to another. Preferably, the interaction between the agents and proteins disclosed herein is specific and selective. Note that in some embodiments, the agent is designed to “specifically bind” and “selectively bind” to two different but similar targets without binding to other unwanted targets.

The terms "polypeptide," "peptide" and "protein" are used interchangeably herein and refer to polymers of amino acid residues. The term applies to naturally occurring amino acid polymers as well as to amino acid polymers in which one or more amino acid residues are non-naturally occurring amino acids (eg, amino acid analogs). The term includes amino acid chains (ie antigens) of any length, including full length proteins, wherein the amino acid residues are linked by covalent peptide bonds.

The terms "motif" and "domain" are used interchangeably herein. As used herein, a separate, contiguous or non-adjacent portion that is folded independently of the rest of the polypeptide and possesses the function of that asset.

The term "destruction" means impeding function. For example, destroying a motif / domain means interrupting the function of the motif / domain.

The term "antigen" refers to a substance capable of inducing antibody production. In some embodiments, the antigen is a substance that specifically binds to an antibody variable region.

The terms "antibody" and "antibodies" refer to monoclonal antibodies, polyclonal antibodies, bispecific antibodies, multispecific antibodies, graft antibodies, human antibodies, humanized antibodies, synthetic antibodies, chimeric antibodies, camelized antibodies, Single-chain Fv (scFv), single chain antibody, Fab fragment, F (ab ') fragment, disulfide-linked Fv, intrabody and anti-idiotype (antiId) antibodies and An antigen binding fragment of any of the above. In particular, antibodies include immunoglobulin molecules and immunologically active fragments of immunoglobulin molecules, ie molecules containing antigen binding sites. Depending on the amino acid sequences of the constant motifs / domains of these heavy chains, immunoglobulins can be classified into different classes. The heavy chain constant domains (Fc) corresponding to different classes of immunoglobulins are named α, δ, ε, γ, and μ, respectively. Subunit structures and tertiary structures of different classes of immunoglobulins are well known. Immunoglobulin molecules can be of any type (eg, IgG, IgE, IgM, IgD, IgA, and IgY), class (eg, IgG ₁ , IgG ₂ , IgG ₃ , IgG ₄ , IgA _1, and IgA ₂ ) or It is a subclass. The terms "antibody" and "immunoglobulin" are used interchangeably in a broad sense. In some embodiments, an antibody is part of a macromolecule formed by intrinsic or non-covalent binding of an antibody with one or more other proteins or peptides.

The term "variable motif / domain" in connection with an antibody refers to the variable motif / domain of the antibody used for binding and specificity of each particular antibody to that particular antigen. However, the variability is not evenly distributed throughout the antibody's variable motifs / domains. Rather, it is concentrated in three segments called hypervariable regions (also known as CDRs) in both the light and heavy chain variable motifs / domains. The more highly conserved portions of variable motifs / domains are called "framework regions" or "FRs." The variable motifs / domains of the unmodified heavy and light chains each comprise four FRs (FR1, FR2, FR3 and FR4), which are β sheet structures interspersed with three CDRs forming a generally linked loop, in some cases β Adopt part of the sheet structure. The CDRs of each chain are held very closely together by the FRs and, together with the CDRs from the other chain, contribute to forming the antigen-binding site of the antibody (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed.Public Health Service, National Institutes of Health, Bethesda, Md. (1991), pages 647-669).

As used herein, the terms "hypervariable region" and "CDR" refer to amino acid residues of an antibody that is responsible for antigen-binding. CDRs bind in a complementary manner to the antigen and comprise amino acid residues from three sequence regions known for CDR1, CDR2 and CDR3 for the V _H and V _L chains, respectively. In light chain variable motifs / domains, CDRs are described by Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)] typically correspond to approximately residues 24-34 (CDRL1), 50-56 (CDRL2) and 89-97 (CDRL3), and in heavy chain variable motifs / domains, the CDRs are typically approximately residues 31-35 (CDRH1), 50-65 (CDRH2) and 95-102 (CDRH3). It is understood that amino acid numbering may vary since the CDRs of different antibodies may contain inserts. The Kabat numbering scheme describes these insertions as a numbering strategy that utilizes letters appended to specific residues to reflect any insertion in numbering between different antibodies (eg, 27A, 27B, 27C, 27D of CDRL1 in the light chain). , 27E, and 27F). Alternatively, in light chain variable motifs / domains, CDRs are described by Chothia and Lesk, J. Mol. Biol., 196: 901-917 (1987), typically correspond approximately to residues 26-32 (CDRL1), 50-52 (CDRL2) and 91-96 (CDRL3) and in heavy chain variable motifs / domains, Typically corresponds approximately to residues 26-32 (CDRH1), 53-55 (CDRH2) and 96-101 (CDRH3).

The constant motif / domain (Fc) of the antibody does not directly participate in the binding of the antigen to the antibody but instead instead participates in antibody in antibody dependent cytotoxicity through interaction with various effector functions, eg Fc receptor (FcR). Indicates. Fc motifs / domains may also increase the bioavailability of the antibody in the circulation after administration to the patient.

As used herein, the term "affinity" refers to the equilibrium constant for the reversible binding of two agents and denoted by Kd. The affinity of the binding protein for the ligand, eg, the antibody to the epitope, is for example about 100 nanomolar (nM) to about 0.1 nM, about 100 nM to about 1 picomolar (pM), or about 100 nM to about 1 femtomol (fM). As used herein, the term "binding force" means resistance to dissociation after dilution of a complex consisting of two or more agents.

The term “peptibody” refers to a molecule comprising peptide (s) fused directly or indirectly to an antibody or one or more antibody motifs / domains (eg, the Fc motif / domain of an antibody), wherein the peptide portion Specifically binds to the desired target. Such peptide (s) may be fused to an Fc region or inserted into an Fc loop, a modified Fc molecule. The term “peptibodies” does not include Fc-fusion proteins (eg, full length proteins fused to Fc motifs / domains).

The terms "isolated" and "purified" mean a substance that is substantially or essentially isolated or concentrated from its natural environment. For example, an isolated nucleic acid is one that is separated from at least some or other nucleic acids or components (proteins, lipids, etc.) of nucleic acids normally flanked by it in a sample. In another example, a polypeptide is purified when substantially removed or concentrated from its natural environment. Methods of purifying and isolating nucleic acids and proteins have been reported in detail by methodologies. Embodiments of “substantially” include at least 20%, at least 40%, at least 50%, at least 75%, at least 85%, at least 90%, at least 95%, or at least 99%.

As used herein, the terms “treat,” “treating,” or “treatment” and other grammatical synonyms refer to alleviation, inhibition or reduction of symptoms, reduction or inhibition of the severity of a disease or condition symptom. Reducing the incidence, treating prophylactically, reducing or suppressing recurrence, preventing, delaying, delaying recurrence, weakening or improving recurrence, improving the underlying metabolic cause of symptoms Suppressing the disease or condition, for example, stopping the development of the disease or condition, alleviating the disease or condition, causing regression of the disease or condition, alleviating the symptoms caused by the disease or condition Or stopping the symptoms of the disease or condition. The term also includes achieving a therapeutic benefit. By therapeutic benefit is meant that an improvement is observed in the subject through the removal or amelioration of the underlying disease to be treated, and / or the removal or amelioration of one or more of the physiological symptoms associated with the underlying disease.

As used herein, the terms "prevent," "preventing" or "prevention" and other grammatical synonyms refer to preventing additional symptoms, preventing the underlying metabolic cause of symptoms, or inhibiting a disease or condition. And, for example, to stop the development of a disease or condition, and to include a prophylactic. The term further includes achieving a prophylactic benefit. For prophylactic benefit, the composition is optionally administered to an individual at risk of developing a particular disease, to an individual for whom one or more of the physiological symptoms of the disease have been reported, or to an individual at risk of developing the disease.

As used herein, the term "effective amount" or "therapeutically effective amount" refers to a sufficient amount of one or more agents administered to achieve the desired result, eg, to relieve to some extent one or more symptoms of the disease or condition being treated. Means quantity. In certain cases, the result is a reduction and / or alleviation of the signs, symptoms or causes of the disease, or any other desired alteration of the biological system. In certain instances, the result is a reduction in the growth, death, and induction of apoptosis in at least one abnormally proliferating cell, eg, cancer stem cells. In certain cases, an “effective amount” for therapeutic use is an amount of a composition comprising an agent described herein that is needed to significantly reduce the disease clinically. In any individual case, the appropriate “effective amount” is determined using any suitable technique, eg, dose escalation studies.

As used herein, the terms “administer,” “administering,” “administration,” and the like refer to methods used to deliver an agent or composition to a desired site of biological action. Such methods include, but are not limited to, oral route, intraduodenal route, parenteral injection (including intravenous, subcutaneous, intraperitoneal, intramuscular, endovascular or infusion), topical and rectal administration. Dosing techniques optionally used in conjunction with the agents and methods described herein are described, for example, in Goodman and Gilman, The Pharmacological Basis of Therapeutics, current ed .; Pergamon; and Remington's, Pharmaceutical Sciences (current edition), Mack Publishing Co., Easton, Pa. In some embodiments, the formulations and compositions described herein are administered orally.

As used herein, the term "pharmaceutically acceptable" does not destroy the biological activity or properties of the formulations described herein, but rather is relatively nontoxic (ie, the toxicity of the material is significantly greater than the benefits of the material). Means. In some cases, pharmaceutically acceptable substances are administered to a subject without significant or undesirably significant biological effects or with no significant interaction in any harmful manner with any component of the composition comprising them.

I. Migration Inhibitory Factor (MIF)

In some embodiments, disclosed herein are peptides, small molecules, antibodies, and peptibodies (collectively, "material compositions") for treating inflammatory diseases, diseases, conditions, and symptoms. Further disclosed herein are pharmaceutical compositions and methods for treating inflammatory diseases, diseases, conditions and symptoms.

MIF is an anti-inflammatory cytokine. In certain cases, it is secreted by activated immune cells (eg, lymphocytes (T-cells)) in response to infection, inflammation, or tissue damage. In certain cases, MIF is a ligand for receptors CXCR2, CXCR4, CD44, and CD74. In some embodiments, the material compositions, methods, and / or pharmaceutical compositions disclosed herein (partially or completely) inhibit the activity of CXCR2 CXCR4, CD44, and / or CD74.

In certain cases, MIF induces chemotaxis in peripheral white blood cells (eg, lymphocytes, granulocytes, monocytes / macrophages, and TH-17 cells) according to the MIF gradient. In some embodiments, the material compositions, methods, and / or pharmaceutical compositions disclosed herein interfere with chemotaxis according to the MIF gradient, or reduce chemotaxis according to the MIF gradient. In certain cases, MIF induces chemotaxis of white blood cells (eg, lymphocytes, granulocytes, monocytes / macrophages, and TH-17 cells) for sites of infection, inflammation, or tissue damage. In some embodiments, the material compositions, methods, and / or pharmaceutical compositions disclosed herein interfere with or reduce the chemotaxis of white blood cells to sites of infection, inflammation, or tissue damage. In certain cases, chemotaxis of white blood cells (eg, lymphocytes, granulocytes, monocytes / macrophages, and TH-17 cells) according to a MIF gradient causes inflammation, inflammation or tissue damage. In some embodiments, the material compositions, methods, and / or pharmaceutical compositions disclosed herein treat inflammation at the site of infection, inflammation, or tissue damage. In certain cases, chemotaxis of monocytes according to the RANTES gradient causes monocyte arrest (ie monocyte deposition on epithelium) at the site of injury or inflammation. In some embodiments, the material compositions, methods, and / or pharmaceutical compositions disclosed herein interfere with or reduce monocyte arrest at the site of injury or inflammation. In some embodiments, the substance compositions, methods, and / or pharmaceutical compositions disclosed herein inhibit and treat lymphocyte mediated diseases. In some embodiments, the material compositions, methods, and / or pharmaceutical compositions disclosed herein treat granulocyte mediated diseases. In some embodiments, the material compositions, methods, and / or pharmaceutical compositions disclosed herein treat macrophage mediated diseases. In some embodiments, the substance compositions, methods, and / or pharmaceutical compositions disclosed herein treat a Th-17 mediated disease. In some embodiments, the methods and / or compositions disclosed herein treat pancreatic beta cell mediated diseases.

In certain cases, MIF is inducible by glucocorticoids, which are inducible by mechanisms involved in the acceleration of atherosclerosis associated with many diseases requiring glucocorticoid therapy. Thus, in some embodiments, the compositions and methods described herein inhibit the induction of MIF expression by glucocorticoids.

Human MIF peptides are encoded by nucleotide sequences located on chromosome 22 in cell formation band 22q11.23.

In certain cases, the mature MIF protein is a homotrimer comprising three polypeptides of about 114 amino acids: Methionine 1 was removed from each MIF peptide monomer during translation.

In certain cases, human MIF peptides comprise nucleic acid sequence SEQ ID NO: 422:

Is coded by

In certain cases, human MIF peptides include SEQ ID NO 1:

Is coded by

In certain cases, the porcine MIF peptide is SEQ ID NO: 2.

Is coded by

In certain cases, the bovine MIF peptide is SEQ ID NO: 3:

Is coded by

In certain cases, the murine MIF peptide is SEQ ID NO: 4:

Is coded by

In certain cases, the rat MIF peptide is SEQ ID NO 5:

Is coded by

In some embodiments, a peptide disclosed herein comprises a sequence that competitively binds to a binding partner of a MIF pseudo ELR motif / domain. Pseudo ELR motifs / domains are not contiguous, but include properly spaced residues (see Arg12 and Asp45 & FIG. 11). Pseudo ELR motifs / domains comprise the amino acid sequence of amino acids 12 to 45 (the numbering comprises a methionine residue number one). This is equivalent to the pseudo ELR motif / domain of amino acids 11-44, wherein the methionine residue at No. 1 was not counted (in this case, the pseudo ELR motif / domain includes Arg 11 and Asp 44). In some embodiments, the material compositions, methods, and / or pharmaceutical compositions disclosed herein inhibit the binding of pseudo ELR motifs / domains to CXCR2 and / or CXCR4. In some embodiments, the material compositions, methods, and / or pharmaceutical compositions disclosed herein treat inflammatory diseases, diseases, conditions and symptoms by inhibiting binding of the pseudo ELR motif / domain to CXCR2 and / or CXCR4.

MIF peptides include N-terminal loop motifs / domains (N-loops). In certain cases, MIF N-loops mediate binding to CXCR2 and / or CXCR4 receptors. In certain cases, the N-loop motif / domain of MIF comprises consecutive residues 44-57 of the MIF (ie, P45 D45 Q46 L47 M48 A49 F50 G51 G52 S53 S54 E55 P56 C57; see FIG. 11), wherein 1 Burn methionine was not included. This is equivalent to amino acids 43-56 and wherein methionine residue no. 1 was not counted. In certain cases, the N-loop motif / domain of MIF comprises amino acids 45-60, wherein methionine 1 was not included. In certain cases, the N-loop motif / domain of MIF comprises amino acids 44-61, wherein methionine 1 was included. In certain cases, the N-loop motif / domain of MIF comprises amino acids 43-62. In certain cases, the N-loop motif / domain of MIF comprises amino acids 42-63, wherein methionine 1 was included. In certain cases, the N-loop motif / domain of MIF comprises amino acids 41-64, wherein methionine 1 was included. In certain cases, the N-loop motif / domain of MIF comprises amino acids 40-65, wherein methionine 1 was included. In certain cases, the N-loop motif / domain of MIF comprises amino acids 46-59, wherein methionine 1 was included. In certain cases, the N-loop motif / domain of MIF comprises amino acids 47-59, wherein methionine 1 was included. In certain cases, the N-loop motif / domain of MIF comprises amino acids 48-59, wherein methionine 1 was included. In certain cases, the N-loop motif / domain of MIF comprises amino acids 50-59, wherein methionine 1 was included. In certain cases, the N-loop motif / domain of MIF comprises amino acids 47-58, wherein methionine 1 was included. In certain cases, the N-loop motif / domain of MIF comprises amino acids 47-57, wherein methionine 1 was included. In certain cases, the N-loop motif / domain of MIF comprises amino acids 47-56, wherein methionine 1 was included. In certain cases, the N-loop motif / domain of MIF comprises amino acids 48-58, wherein methionine 1 was included. In some embodiments, the N-loop motif / domain comprises amino acids 48-57, wherein methionine 1 is included. In some embodiments, the material compositions, methods, and / or pharmaceutical compositions disclosed herein inhibit the binding of N-loop motifs / domains to CXCR2 and / or CXCR4. In some embodiments, the material compositions, methods, and / or pharmaceutical compositions disclosed herein treat inflammatory diseases, diseases, conditions and symptoms by inhibiting the binding of N-loop motifs / domains to CXCR2 and / or CXCR4.

MIF polypeptides include the following motifs / domains: N-terminal / pseudo ELR motifs / domains (MIF _1-17 ), alpha-helix # 1 motifs / domains (ie, MIF _{18 -31} ), MIF N-loop motifs / domains (ie , MIF _32-60 ), loop-barrel-loop motifs / domains (ie, MIF _{64 -93} ), and C-terminal motifs / domains (ie, MIF _{90 -114} ). Alternatively, MIF polypeptide to motif / domain: N- terminal tail (i.e., MIF _{1 -7),} Pseudomonas ELR- loop (i.e., MIF _{7 -17),} an alpha-helix # 1 motif / domain (i.e., MIF _{18 -31} ), PPQ-loop (ie, MIF _{32 -38} ), PDQ-loop (ie, MIF _{43 -56} ), IGK-loop (ie, MIF _{64 -71} ), NRS-helix (ie, MIF _{72 -89} ), SPDR-loop (ie, MIF _{90 -94} ), and C-terminal tail (ie, MIF _101-114 ). In some embodiments, the peptides disclosed herein to a domain of: N- terminus / pseudo ELR motif / domain (MIF1 _-17), an alpha-helix # 1 motif / domain (i.e., MIF _{18 -31),} N- MIF loop Motif / domain (ie MIF _{32 -60} ), loop-barrel-loop motif / domain (ie MIF _{64 -93} ), C-terminal motif / domain (ie MIF _{90 -114} ), or as mentioned above Competitively binds to a binding partner of any combination of domains. In some embodiments, the domain for the peptide disclosed herein: N- terminal tail (i.e., MIF _{1 -7),} Pseudomonas ELR- loop (i.e., MIF _{7 -17),} an alpha-helix # 1 motif / domain (i. E. , MIF _{18 -31} ), PPQ-loop (ie MIF _{32 -38} ), PDQ-loop (ie MIF _{43 -56} ), IGK-loop (ie MIF _{64 -71} ), NRS-helix (ie, MIF _{72 -89} ), SPDR-loop (ie, MIF _{90 -94} ), C-terminal tail (ie, MIF _{101 -114} ), or a binding partner of a combination of any of the aforementioned domains. In some embodiments, the peptides disclosed herein bind competitively with a binding partner of MIF ₄₇ (Leucine).

In some embodiments, the material compositions, methods, and / or pharmaceutical compositions disclosed herein comprise (1) binding of N-loop motifs / domains to CXCR2 and / or CXCR4; And (2) inhibits binding of the pseudo ELR motif / domain to CXCR2 and / or CXCR4. In some embodiments, the material compositions, methods, and / or pharmaceutical compositions disclosed herein comprise (1) binding of N-loop motifs / domains to CXCR2 and / or CXCR4; And (2) inhibiting the binding of the pseudo ELR motif / domain to CXCR2 and / or CXCR4 to treat inflammatory diseases, diseases, conditions and symptoms.

II. Active agent

In some embodiments, disclosed herein are peptides, small molecules, antibodies, and peptibodies (collectively, "material compositions") for treating inflammatory diseases, diseases, conditions, and symptoms. Further disclosed herein are pharmaceutical compositions and methods for treating inflammatory diseases, diseases, conditions and symptoms.

In some embodiments, further disclosed herein are material compositions, methods, and pharmaceutical compositions that inhibit the ability of MIF to bind CXCR2, CXCR4, CD44, CD74, or a combination thereof. Further disclosed herein are substance compositions, methods, and pharmaceutical compositions for treating inflammatory diseases, diseases, conditions and symptoms by inhibiting the ability of MIF to bind CXCR2, CXCR4, CD44, CD74, or a combination thereof. In certain cases, occupying, masking, or otherwise disrupting motifs / domains on MIF is mediated by other agonists / ligands, such as CXCR2, CXCR4, CD44 and / or CD74 signaling (eg, IL-8). / CXCL8, GROβ / CXCL2, GROα, GROγ, ENA78, CXCR2 interaction with NAP2; and Stromal Cell-Derived Factor-1a (SDF-1a) / CXCL12, and CXCR4 interaction with GP120 ) Does not affect.

Any amino acid of any of the peptides disclosed herein corresponds to an effective substituent for the original amino acid and acts as such but does not substantially reduce binding or bioactivity compared to the parent peptide sequence. It may be substituted by. The phrase “does not substantially reduce binding or bioactivity as compared to the parent peptide sequence” means that the modified peptide sequence is about 60%, about 65%, about 70%, about 75%, about 75% of the same activity of the parent peptide sequence. 80%, about 85%, about 90%, or about 95%. Non-natural amino acids include D amino acids such as D-phenylalanine (D-F) and D-cyclohexyl alanine (D-CA); Norleucine (NLe), L-cyclohexyl alanine (L-CA), and L-amino acid, alpha-aminobutyric acid (Abu). In some embodiments, amino acids of the peptides disclosed herein are substituted with non-natural amino acids. In some embodiments, amino acids of peptides disclosed herein comprise N and / or C-terminal chemical modifications that improve ADME-PK.

MIF  Destruction of Motif / Domain

In some embodiments, the substance composition inhibits the ability of MIF to interact with CXCR2, CXCR4, CD74, CD44, or a combination thereof. In some embodiments, inflammatory diseases, diseases, conditions and symptoms are treated, diagnosed, or monitored by destroying the ability of MIF to interact with CXCR2, CXCR4, CD74, CD44 or a combination thereof. In some embodiments, the ability of MIF to interact with CXCR2, CXCR4, CD74, CD44, or a combination thereof, occupies, masks, or masks motifs / domains on MIF to which CXCR2, CXCR4, CD74, and / or CD44 bind, Or otherwise destroyed. In some embodiments, the ability of MIF to interact with CXCR2, CXCR4, CD74, CD44, or a combination thereof may include N-terminal / pseudo ELR motifs / domains (MIF _{1 -17} ), alpha-helix # 1 motifs / domains ( Ie MIF _{18 -31} ), MIF N-loop motif / domain (ie MIF _{32 -60} ), loop-barrel-loop motif / domain (ie MIF _{64 -93} ), and C-terminal motif / domain (ie , MIF _{90 -114} ), which is inhibited by occupying, masking, or otherwise destroying a motif / domain monomer or part thereof. In some embodiments, the ability of MIF is CXCR2, CXCR4, CD74, CD44 or combinations thereof can serve as the cross-N- terminal tail (i.e., MIF _{1 -7),} Pseudomonas ELR- loop (i.e., MIF _{7 -17)} , Alpha-helix # 1 motif / domain (ie MIF _{18 -31} ), PPQ-loop (ie MIF _{32 -38} ), PDQ-loop (ie MIF _{43 -56} ), IGK-loop (ie MIF _{64) -71} ) all or a motif / domain selected from _NRS- helix (ie, MIF _{72 -89} ), SPDR-loop (ie, MIF _{90 -94} ), and C-terminal tail (ie, MIF _101-114 ) It is inhibited by occupying, shielding, or otherwise destroying some. In some embodiments, the ability of MIF to interact with CXCR2, CXCR4, CD74, CD44, or a combination thereof is inhibited by occupying, masking, or otherwise destroying MIF ₄₇ (leucine).

In some embodiments, the ability of MIF to interact with CXCR2, CXCR4, CD74, CD44 or a combination thereof may occupy or mask one or more motifs / domains on the MIF to which CXCR2, CXCR4, CD74 and / or CD44 bind Or is otherwise inhibited by small molecules, peptides, antibodies, and / or peptibodies that disrupt. In certain instances, occupying, masking, or otherwise disrupting one or more motifs / domains on MIF is derived from other agonists / ligands (eg, IL-8 / CXCL8, GRObeta / CXCL2, and / or stromal cells). It does not affect CXCR2 and CXCR4 signaling mediated by factor 1a (SDF-1a) / CXCL12).

In certain cases, the pseudo ELR motif / domain of MIF mediates binding of ligands (eg, CD44, CD74, CXCR2, CXCR4) to MIF. In some embodiments, the binding of the small molecule, peptide, antibody, and / or peptibody to all or a portion of the pseudo ELR motif / domain of the MIF will bind the MIF to CXCR2, CXCR4, CD74, CD44, or a combination thereof. Curb your ability to do so. In some embodiments, the small molecule, peptide, antibody, and / or a peptibody, N- terminal tail (i.e., MIF _{1 -7)} a combination of the entire or a portion thereof and / or pseudo ELR- loop (i.e., MIF _{7 -17} ) binding to all or a portion thereof inhibits the ability of MIF to bind CXCR2, CXCR4, CD74, CD44 or a combination thereof.

In certain cases, N-loop motifs / domains of MIF mediate the binding of ligands (eg, CD44, CD74, CXCR2, CXCR4) to MIF. In some embodiments, binding of the small molecule, peptide, antibody, and / or peptibody to the N-loop motif / domain, or a portion thereof, of the MIF binds the MIF to CXCR2, CXCR4, CD74, CD44, or a combination thereof. Curb your ability to do so. In some embodiments, binding of small molecules, peptides, antibodies, and / or peptibodies to all or a portion of a PPQ-loop (ie, MIF _{32 -38} ), and / or a PDQ-loop (ie, MIF _{43- 56} ) Binding to all or a portion thereof inhibits the ability of MIF to bind CXCR2, CXCR4, CD74, CD44 or a combination thereof. In some embodiments, conformational changes in MIF where the binding of small molecules, peptides, antibodies, and / or peptibodies to all or a portion of the N-loop motif / domain of MIF interferes with receptor or substrate interaction Will result. In some embodiments, binding of small molecules, peptides, antibodies, and / or peptibodies to all or a portion of a PPQ-loop (ie, MIF _{32 -38} ), and / or a PDQ-loop (ie, MIF _{43 −) 56} ) Binding to all or a portion thereof results in conformational changes in MIF that interfere with receptor or substrate interaction.

In certain cases, amino acids 65-94 of MIF (eg, IGKIGGAQNRSYSKLLCGLLAERLRISPDR (SEQ ID NO: 8); numbering comprises methionine 1) mediate binding of CXCR2 to MIF. In some embodiments, binding of the small molecule, peptide, antibody, and / or peptibody to all or part of amino acids 65-94 of MIF inhibits the ability of MIF to bind CXCR2. In some embodiments, binding of the peptide to all or part of amino acids 65-94 of MIF inhibits the ability of MIF to bind CXCR2. In some embodiments, binding of the antibody to all or part of amino acids 65-94 of MIF inhibits the ability of MIF to bind CXCR2. In some embodiments, the binding of peptibodies to all or a portion of amino acids 65-94 of MIF inhibits the ability of MIF to bind CXCR2. In some embodiments, the binding of the small molecule to all or part of amino acids 65-94 of the MIF inhibits the ability of the MIF to bind CXCR2.

In certain instances, amino acids 80-95 of MIF (eg, LCGLLAERLRISPDRV (SEQ ID NO: 9); the numbering comprises methionine 1) mediate binding of the ligand to MIF. In some embodiments, binding of the small molecule, peptide, antibody, and / or peptibody to all or part of amino acids 80-95 of MIF inhibits the ability of MIF to bind to the ligand. In some embodiments, binding of the peptide to all or a portion of amino acids 80-95 of the MIF inhibits the ability of the MIF to bind a ligand. In some embodiments, binding of the antibody to all or part of amino acids 80-95 of MIF inhibits the ability of MIF to bind a ligand. In some embodiments, the binding of peptibodies to all or a portion of amino acids 80-95 of MIF inhibits the ability of MIF to bind a ligand. In some embodiments, binding of the MIF to all or a portion of amino acids 80-95 of the MIF inhibits the ability of the MIF to bind a ligand.

In some embodiments, an inflammatory disease, disease, condition, and condition comprises a peptide that occupies, masks, or otherwise destroys all or part of a motif / domain on the MIF to which CXCR2, CXCR4, CD74 and / or CD44 binds. By administration, treatment, diagnosis, or monitoring. In some embodiments, the peptide specifically binds to all or a portion of the pseudo ELR motif / domain of MIF. In some embodiments, the peptide specifically binds to all or a portion of the N-loop motif / domain of MIF. In some embodiments, the peptide specifically binds to all or part of both pseudo ELR and N-loop motifs.

In some embodiments, the formulation comprises a peptide that specifically binds to all or a portion of a peptide sequence as follows: VNTNVPPRASVPDGFLSELTQQLAQATGKPPQYIAVHVVPDQL (SEQ ID NO: 10) and the corresponding feature / domain of at least one of a MIF monomer or MIF trimer; A peptide that specifically binds to all or a portion of a peptide sequence as follows: PDQLMAFGGSSEPCALCSL (SEQ ID NO: 11) and the corresponding feature / domain of at least one of a MIF monomer or MIF trimer; A peptide that specifically binds to all or a portion of a peptide sequence as follows: VNTNVPPRASVPDGFLSELTQQLAQATGKPPQYIAVHVVPDQLMAFGGSSEPCALCSL (SEQ ID NO: 12) and the corresponding feature / domain of at least one of a MIF monomer or MIF trimer; A peptide that specifically binds to all or a portion of a peptide sequence as follows: PDQLMAFGGSSEPCALCSLHSI (SEQ ID NO: 13) and the corresponding feature / domain of at least one of a MIF monomer or MIF trimer.

In some embodiments, an inflammatory disease, disease, condition, and condition comprises an antibody that occupies, masks, or otherwise destroys all or a portion of the motif / domain on the MIF to which CXCR2, CXCR4, CD74 and / or CD44 binds. By administration, treatment, diagnosis, or monitoring. In some embodiments, the antibody specifically binds to all or a portion of the pseudo ELR motif / domain of MIF. In some embodiments, the antibody specifically binds to all or a portion of the N-loop motif / domain of MIF. In some embodiments, the antibody specifically binds to all or part of both pseudo ELR and N-loop motifs.

In some embodiments, the inflammatory disease, disease, condition, and symptom is a peptibody that occupies, masks, or otherwise destroys all or part of the motif / domain on the MIF to which CXCR2, CXCR4, CD74 and / or CD44 binds. It is treated, diagnosed, or monitored by administering. In some embodiments, the peptibody specifically binds to all or a portion of the pseudo ELR motif / domain of MIF. In some embodiments, the peptibody specifically binds to all or a portion of the N-loop motif / domain of MIF. In some embodiments, the peptibody specifically binds to all or part of both pseudo ELR and N-loop motifs.

In some embodiments, the inflammatory disease, disease, condition and symptom is a small molecule that occupies, masks, or otherwise destroys all or part of the motif / domain on the MIF to which CXCR2, CXCR4, CD74 and / or CD44 binds. It is treated, diagnosed, or monitored by administering. In some embodiments, the small molecule specifically binds to all or a portion of the pseudo ELR motif / domain of MIF. In some embodiments, the small molecule specifically binds to all or a portion of the N-loop motif / domain of MIF. In some embodiments, the small molecule specifically binds to all or part of both pseudo ELR and N-loop motifs.

CXCR2  And CXCR4  Destruction of Motif / Domain

In some embodiments, the material composition destroys all or a portion of the motifs / domains on CXCR2 to which CXCR4, MIF, CD44 and / or CD74 bind. In some embodiments, an inflammatory disease, disease, condition, and condition comprises an agent that occupies, masks, or otherwise destroys all or part of the motif / domain on CXCR2 to which CXCR4, MIF, CD44, and / or CD74 binds. By administration, treatment, diagnosis, or monitoring.

In some embodiments, the substance composition destroys all or a portion of the motifs / domains on CXCR4 to which CXCR2, MIF, CD44 and / or CD74 bind. In some embodiments, an inflammatory disease, disease, condition, and condition comprises an agent that occupies, masks, or otherwise destroys all or part of a motif / domain on CXCR4 to which CXCR2, MIF, CD44, and / or CD74 binds. By administration, treatment, diagnosis, or monitoring.

In some embodiments, the agent that inhibits binding of CXCR4, MIF, CD74, and / or CD44 to CXCR2 is a peptide. In some embodiments, the agent that inhibits binding of CXCR2, MIF, CD74, and / or CD44 to CXCR4 is a peptide.

In some embodiments, the agent that inhibits binding of CXCR4, MIF, CD74 and / or CD44 to CXCR2 is an antibody. In some embodiments, the agent that inhibits binding of CXCR2, MIF, CD74, and / or CD44 to CXCR4 is an antibody.

In some embodiments, the agent that inhibits binding of CXCR4, MIF, CD74, and / or CD44 to CXCR2 is a peptibody. In some embodiments, the agent that inhibits binding of CXCR2, MIF, CD74, and / or CD44 to CXCR4 is a peptibody.

In some embodiments, the agent that inhibits binding of MIF, CD74 and / or CD44 to CXCR2 and / or CXCR4 is a derivative of hydroxycinnamate, a Schiff base tryptophan analog, or an iminoquinone metabolite of acetaminophen. .

In some embodiments, an agent that inhibits the binding of MIF, CD74 and / or CD44 to CXCR2 and / or CXCR4 may comprise glyburide, provenidide, DIDS (4,4-diisothiocyanatostilben-2,2 -Disulfonic acid), bumetanide, furosemide, sulfobromophthalein, diphenylamine2-carboxylic acid, flufenamic acid, or a combination thereof.

In some embodiments, an agent that inhibits binding of MIF, CD74, and / or CD44 to CXCR2 may comprise CXCL8 ( _3-74 ) K11R / G31P; IL-8 ( _4-72 ); IL-8 ( _6-72 ); Recombinant IL-8 (rIL-8); Recombinant IL-8, NMeLeu (rhIL-8 with N-methylated leucine at position 25); (AAR) IL-8 (IL-8 with N-terminal Ala4-Ala5 instead of Glu4-Leu5); GRO alpha ( _1-73 ) (also known as CXCL1); GRO alpha ( _4-73 ); GRO alpha ( _5-73 ); GRO alpha ( _6-73 ); Recombinant GRO (rGRO); (ELR) PF4 (PF4 with an ELR sequence at the N-terminus); Recombinant PF4 (rPF4); Antirukinate; Sch527123 (-hydroxy-N, N-dimethyl-3- {2-[[(R) -1- (5-methyl-furan-2-yl) -propyl] amino] -3,4-dioxo-cyclo But-1-enylamino} -benzamide); N- (3- (aminosulfonyl) -4-chloro-2-hydroxyphenyl) -N '-(2,3-dichlorophenyl) urea; SB-517785-M (GSK); SB 265610 (N- (2-bromophenyl) -N '-(7-cyano-1H-benzotriazol-4-yl) urea); SB225002 (N- (2-bromophenyl) -N '-(2-hydroxy-4-nitrophenyl) urea); SB455821 (GSK), SB272844 (GSK); DF2162 (4-[(1R) -2-amino-1-methyl-2-oxoethyl] phenyl trifluoromethanesulfonate); Lepariccin; Or a combination thereof.

In some embodiments, an agent that inhibits the binding of MIF, CD74 and / or CD44 to CXCR4 may be selected from ALX40-4C (N-alpha acetyl-nona-D-arginine amide acetate); AMD-070 (AMD 11070, AnorMED); Flicsapor (AMD3100); AMD3465 (AnorMED); AMD8664 (1-pyridin-2-yl-N- [4-(, 4,7-triazacyclotetracanan-4-ylmethyl) benzyl] methanamine); KRH-1636 from Kureha Chemical Industry Co. Limited; KRH-2731 from Kureha Chemical Industry Co. Limited; KRH-3955 from Kureha Chemical Industry Co. Limited; KRH-3140 from Kureha Chemical Industry Co. Limited; T134 (L-Citrulline 16 TW70, wherein the C-terminal amide is substituted with carboxylic acid); ^{T22 ([Tyr 5, 12,} Lys 7] - poly silent page II); TW70 (Des- [Cys8,13, Tyr9,12]-[D-Lys10, Pro11] -T22); T140 (H-Arg-Arg-Nal-Cys-Tyr-Arg-Lys-DLys-Pro-Tyr-Arg-Cit-Cys-Arg-OH); TC14012 (RR-Nal-CY- (L) Cit-K- (D) Cit-PYR- (L) citrulline-CR-NH2, where Nal = L-3- (2-naphthylalanine), Cit = citrulline Arg-Nal (2) -Cys ( - POL3026 (Arg (*); ( the vMIP-II _(11-71 added with D amino acids in the N- terminal)) RCP168; and peptides to cyclize) with a cysteine; TN14003 1x) -Tyr-Gln-Lys- (d-Pro) -Pro-Tyr-Arg-Cit-Cys (1x) -Arg-Gly- (d-Pro) (*)); POL2438; Compound 3 (N- ( 1-methyl-1-phenylethyl) -N-[((3S) -1- {2- [5- (4H-1,2,4-triazol-4-yl) -1H-indol-3-yl ] Ethyl} pyrrolidin-3-yl) methyl] amine); isothiourea 1a-1u (for information about isothiourea 1a-1u, see Gebhard Thoma, et al., Orally Bioavailable Isothioureas Block Function of the Chemokine Receptor CXCR4 In Vitro and In Vivo, J. Med. Chem., Article ASAP (2008), which is incorporated herein by reference in its entirety); or combinations thereof.

In some embodiments, agents that inhibit binding of MIF, CD74 and / or CD44 to CXCR2 and / or CXCR4 include COR100140 from Genzyme Corp / Cortical Pty Ltd .; Iso-1 ((S, R) -3- (4-hydroxyphenyl) -4,5-dihydro-5-isoxazole acetic acid, methyl ester); 4-IPP (4-iodo-6-phenylpyrimidine); Or a combination thereof.

CD74  Destruction of Motif / Domain

In some embodiments, the material composition destroys all or part of the motif / domain on CD74 to which MIF, CD44, CXCR2, and / or CXCR4 bind. In some embodiments, an inflammatory disease, disease, condition, and condition is an agent that occupies, masks, or otherwise destroys all or part of the motif / domain on CD74 to which MIF, CD44, CXCR2, and / or CXCR4 binds. It is treated, diagnosed, or monitored by administering.

In some embodiments, the agent that inhibits binding of MIF, CD44, CXCR2, CXCR4, or a combination thereof to CD74 is a peptide.

In some embodiments, an agent that inhibits binding of MIF, CD44, CXCR2, CXCR4, or a combination thereof to CD74 is an antibody. Agents that inhibit binding of MIF, CD44, CXCR2, CXCR4, or a combination thereof to CD74 are M-B741, 555538 (BD Pharmingen).

In some embodiments, the agent that inhibits binding of MIF, CD44, CXCR2, CXCR4, or a combination thereof to CD74 is a peptibody.

In some embodiments, an agent that inhibits binding of MIF, CD44, CXCR2, CXCR4, or a combination thereof to CD74 is a small molecule.

In certain instances, occupying, masking, or otherwise destroying all or part of the motifs / domains on the MIF may result in other agonists / ligands (eg, IL-8 / CXCL8, GRObeta / CXCL2 and / or substrates). It does not affect CD74 signaling mediated by cell derived factor 1a (SDF-1a) / CXCL12).

CD44  Destruction of Motif / Domain

In some embodiments, the material composition destroys all or part of the motif / domain on CD44 to which MIF, CD74, CXCR2, and / or CXCR4 bind. In some embodiments, an inflammatory disease, disease, condition, and condition is an agent that occupies, masks, or otherwise destroys all or part of the motif / domain on CD44 to which MIF, CD74, CXCR2, and / or CXCR4 binds. It is treated, diagnosed, or monitored by administering.

In some embodiments, the agent that inhibits binding of MIF, CD74, CXCR2, CXCR4, or a combination thereof to CD44 is a peptide.

In some embodiments, an agent that inhibits binding of MIF, CD74, CXCR2, CXCR4, or a combination thereof to CD44 is an antibody.

In some embodiments, the agent that inhibits binding of MIF, CD74, CXCR2, CXCR4, or a combination thereof to CD44 is a peptibody.

In some embodiments, an agent that inhibits binding of MIF, CD74, CXCR2, CXCR4, or a combination thereof to CD44 is a small molecule.

MIF Mimetics

In some embodiments, the substance composition destroys the ability of MIF to bind CXCR2, CXCR4, CD74, CD44, or a combination thereof. In some embodiments, the substance composition is a peptide that competitively binds to a binding partner of a MIF motif / domain (eg, pseudo ELR, or N-loop motif / domain). In some embodiments, inflammatory diseases, diseases, conditions and symptoms are treated, diagnosed, or monitored by destroying the ability of MIF to bind CXCR2, CXCR4, CD74, CD44 or a combination thereof. In some embodiments, an inflammatory disease, disorder, condition, and condition is a MIF motif / domain (eg, pseudo ELR, or N) in an individual in need of treatment, diagnosis, or monitoring for an inflammatory disease, disorder, condition, and condition. Treatment, diagnosis, or monitoring by administering a peptide that competitively binds to the binding partner of the loop motif / domain). In some embodiments, the peptide binds to CXCR2, CXCR4, CD74, CD44 or a combination thereof, thereby preventing CXCR2, CXCR4, CD44 or CD74 from binding to MIF.

In some embodiments, the peptide adopts structural or functional features similar to the N-loop motif / domain of MIF. In some embodiments, the peptide comprises the sequence of formula (I):

Where

X ¹ is selected from the group consisting of threonine, glycine, proline and alanine;

X ² is selected from the group consisting of glycine, asparagine, aspartic acid, and serine;

X ³ is selected from the group consisting of methionine, isoleucine, leucine, alanine, proline, lysine, glutamine, arginine and lysine;

X ⁴ is selected from the group consisting of methionine, isoleucine and leucine;

X ⁵ is selected from the group consisting of alanine, threonine, methionine, serine and valine;

X ⁶ is selected from the group consisting of phenylalanine, histidine, arginine and lysine;

X ⁷ is selected from the group consisting of aspartic acid, glutamic acid, threonine, glycine and alanine;

X ⁸ is selected from the group consisting of serine, threonine, lysine and arginine;

X ⁹ is selected from the group consisting of serine, asparagine, glycine, threonine, aspartic acid, glutamic acid, glutamine and histidine;

X ¹⁰ is selected from the group consisting of aspartic acid, glutamic acid, alanine and asparagine;

X ¹¹ is selected from the group consisting of cysteine, alanine, serine, threonine and valine.

In some embodiments, X ¹ is proline. In some embodiments, X ² is aspartic acid. In some embodiments, X ³ is leucine. In some embodiments, X ⁴ is methionine. In some embodiments, X ⁵ is alanine. In some embodiments, X ⁶ is phenylalanine. In some embodiments, X ⁷ is glycine. In some embodiments, X ⁸ is serine. In some embodiments, X ⁹ is serine. In some embodiments, X ¹⁰ is glutamic acid. In some embodiments, X ¹¹ is serine cysteine.

In some embodiments, the peptide comprises a contiguous amino acids of at least three (and the numbering including 1 methionine) of human MIF _{44 -57.} In some embodiments, the peptide comprises three or more consecutive amino acids of murine MIF _{44 -57} . In some embodiments, the peptide comprises three or more consecutive amino acids of porcine MIF _{44 -57} . In some embodiments, the peptide comprises three or more consecutive amino acids of bovine MIF _{44 -57} . In some embodiments, the peptide comprises three or more consecutive amino acids of rat MIF _{44 -57} .

In some embodiments, the peptide is selected from Table 1 below. Any amino acid of any of the peptides disclosed herein corresponds to an effective substituent for the original amino acid and acts as such but does not substantially reduce binding or bioactivity compared to the parent peptide sequence. It may be substituted by. Non-natural amino acids include D amino acids such as D-phenylalanine (D-F) and D-cyclohexyl alanine (D-CA); Norleucine (NLe), L-cyclohexyl alanine (L-CA), and L-amino acid, alpha-aminobutyric acid (Abu). In some embodiments, amino acids of the peptides disclosed herein are substituted with non-natural amino acids. In some embodiments, amino acids of peptides disclosed herein comprise N and / or C-terminal chemical modifications that improve ADME-PK.

[Table 1]

In some embodiments, the peptide comprises cyclic: CLMAFGGS SEPC (SEQ ID NO: 422); CLMAFGGSSEPCALC (SEQ ID NO: 423); CGLMAFGGSSEPGC (SEQ ID NO: 424); CGGLMAFGGS SEPGGC (SEQ ID NO: 425); CGGSLMAFGGSSEPSGGC (SEQ ID NO: 426); CGGSGLMAFGGSSEPGSGGC (SEQ ID NO: 427); CGGSGGLMAFGGSSEPGGSGGC (SEQ ID NO: 428); CLMAFGGSSEP [Abu] ALC (SEQ ID NO: 429), wherein Abu is isosteric L-amino acid, alpha-aminobutyric acid; CFGGSSEPCALC (SEQ ID NO: 441); CSSEPCALC (SEQ ID NO: 443); CFGGSSEPCC (SEQ ID NO: 444); CFGGSSEPC (SEQ ID NO: 445); CGSSEPCALCC (SEQ ID NO: 446); CAFGGSSEPCAC (SEQ ID NO: 449); CAFGGSSC (SEQ ID NO: 450); CLMAFGGSSEC (SEQ ID NO: 463); Or cyclic CLMAFGGSSEPSALC (SEQ ID NO: 469).

In some embodiments, the peptide is linear: CLMAFGGSSEPCALC (SEQ ID NO: 442); Linear CAFGGSSC (SEQ ID NO: 447); CAFGGSSEPCAC (SEQ ID NO: 448); CLMAFGGSSEC (SEQ ID NO: 464).

In some embodiments, the peptide comprises LMA [NLe] AFGGSSEPC [NLe] (SEQ ID NO: 430), where NLe is norleucine; LMA [ _L -CA] AFGGSSEPC [ _L -CA] (SEQ ID NO: 431), where _L -CA is _L -cyclohexylalanine; LMA [ _D -CA] AFGGSSEPC [ _D -CA] (SEQ ID NO: 432), wherein _D -CA is _D -cyclohexylalanine; LMA [ _D- F] AFGGSSEPC [ _D- F] (SEQ ID NO: 433), wherein _D- F is _D -phenylalanine; ( _D ) -MAFGGSSEPC (SEQ ID NO: 434); ( _D ) -CPESSGGFAML (SEQ ID NO: 435); ( _L ) -CPESSGGFAML (SEQ ID NO: 436); CLMAFGGSSEPCACG (SEQ ID NO: 452); CLMAFGGSSEPCCGG (SEQ ID NO: 453); CLMAFGGSSEPCGGG (SEQ ID NO: 454); CLMAFGGS SECGGGG (SEQ ID NO: 455); CLMAFGGSSCGGGGG (SEQ ID NO: 456); CLMAFGGSCGGGGG (SEQ ID NO: 457); CLMAFGGCGGGGGGG (SEQ ID NO: 458); CLMAFGCGGGGGGGG (SEQ ID NO: 459); Or CLMAFGGSSEPCALG (SEQ ID NO: 460).

In some embodiments, the peptides disclosed herein bind competitively with a binding partner of MIF _{40 -49} (ie, the peptide has the sequence VHVVPDQLMA (SEQ ID NO: 465)). In some embodiments, the peptides disclosed herein bind competitively with a binding partner of MIF _{42 -51} (ie, the peptide has the sequence VVPDQLMAFG (SEQ ID NO: 466)). In some embodiments, the peptides disclosed herein bind competitively with all or a portion of MIF _{45 -57} (ie, the peptide has the sequence DQLMAFGGSSEPC (SEQ ID NO: 467)). In some embodiments, the peptides disclosed herein are combined with competitive binding of MIF _{46 -55} partner (i.e., the peptide has a sequence QLMAFGGSSE (SEQ ID NO: 468)). In some embodiments, the peptide has the sequence: VHVVPDQLMA (SEQ ID NO: 421), VVPDQLMAFG (SEQ ID NO: 461), DQLMAFGGSSEPC (SEQ ID NO: 462), or QLMAFGGSSE (SEQ ID NO: 69).

In some embodiments, the peptide comprises the sequence of formula (II):

Where

X ¹ is selected from the group consisting of valine, isoleucine, threonine, phenylalanine and leucine;

X ² is selected from the group consisting of asparagine, arginine, aspartic acid, glutamic acid, serine and alanine;

X ³ is selected from the group consisting of valine, isoleucine, arginine, lysine and leucine;

X ⁴ is selected from the group consisting of proline, alanine, cysteine and leucine;

X ⁵ is selected from the group consisting of arginine, lysine, glutamine, serine, alanine, aspartic acid, glutamic acid and asparagine;

X ⁶ is selected from the group consisting of alanine, aspartic acid, glutamic acid, asparagine, serine and glutamine;

X ⁷ is selected from the group consisting of serine, glutamic acid, aspartic acid, asparagine, arginine, glycine, lysine and arginine;

X ⁸ is selected from the group consisting of valine, isoleucine and phenylalanine;

X ⁹ is selected from the group consisting of aspartic acid, glutamic acid, valine, serine and threonine;

X ¹⁰ is selected from the group consisting of glycine, alanine, threonine, aspartic acid and glutamic acid.

In some embodiments, X ¹ is valine. In some embodiments, X ² is asparagine. In some embodiments, X ³ is valine. In some embodiments, X ⁴ is proline. In some embodiments, X ⁵ is arginine. In some embodiments, X ⁶ is alanine. In some embodiments, X ⁷ is serine. In some embodiments, X ⁸ is valine. In some embodiments, X ⁹ is aspartic acid. In some embodiments, X ¹⁰ is glycine.

In some embodiments, the peptide comprises a contiguous amino acids of at least three (and the numbering including 1 methionine) of human MIF _{1 -45.} In some embodiments, the peptide comprises three or more consecutive amino acids of murine MIF _{1 -45} . In some embodiments, the peptide comprises three or more consecutive amino acids of porcine MIF _{1 -45} . In some embodiments, the peptide comprises three or more consecutive amino acids of bovine MIF _{1 -45} . In some embodiments, the peptide comprises three or more consecutive amino acids of rat MIF _{1 -45} .

In some embodiments, the peptide is selected from Table 2 below. Any amino acid of any of the peptides disclosed herein corresponds to an effective substituent for the original amino acid and acts as such but does not substantially reduce binding or bioactivity compared to the parent peptide sequence. It may be substituted by. Non-natural amino acids include D amino acids such as D-phenylalanine (D-F) and D-cyclohexyl alanine (D-CA); Norleucine (NLe), L-cyclohexyl alanine (L-CA), and L-amino acid, alpha-aminobutyric acid (Abu). In some embodiments, amino acids of the peptides disclosed herein are substituted with non-natural amino acids. In some embodiments, amino acids of peptides disclosed herein comprise N and / or C-terminal chemical modifications that improve ADME-PK.

[Table 2]

In some embodiments, the peptide comprises cyclic: CPRASVPDGC (SEQ ID NO: 438), CGGSGGPRASVPDGGGSGGC (SEQ ID NO: 439); Or CNVPRASVPDGC (SEQ ID NO: 440).

In some embodiments, the peptide adopts a structural or functional feature similar to amino acid residues 65-94 (the numbering comprises methionine number 1). In some embodiments, the peptide comprises a sequence of formula (III):

Where

X ¹ is selected from the group consisting of lysine, arginine, cysteine, serine and alanine;

X ² is selected from the group consisting of isoleucine, valine and phenylalanine;

X ³ is selected from the group consisting of glycine, asparagine and serine;

X ⁴ is selected from the group consisting of glycine, proline, alanine, aspartic acid and glutamic acid;

X ⁵ is selected from the group consisting of alanine, proline, lysine, arginine, asparagine, aspartic acid and glutamic acid;

X ⁶ is selected from the group consisting of glutamine, valine, lysine, arginine, leucine, aspartic acid and glutamic acid;

X ⁷ is selected from the group consisting of lysine, arginine, asparagine, isoleucine and valine;

X ⁸ is selected from the group consisting of serine, asparagine, glutamine, aspartic acid, glutamic acid, lysine and arginine;

X ⁹ is selected from the group consisting of tyrosine, histidine and asparagine;

X ¹⁰ is selected from the group consisting of serine, threonine and alanine;

X ¹¹ is selected from the group consisting of lysine, aspartic acid, glutamic acid, alanine, serine and glycine;

X ¹² is selected from the group consisting of leucine, glutamine, lysine, arginine, leucine, serine and alanine;

X ¹³ is selected from the group consisting of cysteine, tyrosine, phenylalanine, serine, alanine and threonine;

X ¹⁴ is selected from the group consisting of glycine, aspartic acid, glutamic acid, lysine and arginine;

X ¹⁵ is selected from the group consisting of leucine, glutamine, isoleucine, histidine and phenylalanine;

X ¹⁶ is selected from the group consisting of leucine, methionine, isoleucine and cysteine;

X ¹⁷ is selected from the group consisting of alanine, threonine, serine, arginine, lysine, alanine, glutamine and glycine;

X ¹⁸ is selected from the group consisting of glutamic acid, aspartic acid, lysine and arginine;

X ¹⁹ is selected from the group consisting of arginine, histidine, glutamine, aspartic acid, glutamic acid, glycine, threonine and lysine;

X ²⁰ is selected from the group consisting of arginine, histidine, glycine, asparagine, lysine, arginine, aspartic acid and glutamic acid;

X ²¹ is selected from the group consisting of serine, aspartic acid, glutamic acid, lysine, arginine and proline;

X ²² is selected from the group consisting of proline, alanine, lysine, arginine and glycine;

X ²³ is selected from the group consisting of aspartic acid, glutamic acid, asparagine and alanine;

X ²⁴ is selected from the group consisting of histidine, tyrosine, lysine and arginine.

In some embodiments, X ¹ is lysine. In some embodiments, X ² is isoleucine. In some embodiments, X ³ is glycine. In some embodiments, X ⁴ is glycine. In some embodiments, X ⁵ is alanine. In some embodiments, X ⁶ is glutamine. In some embodiments, X ⁷ is arginine. In some embodiments, X ⁸ is serine. In some embodiments, X ⁹ is tyrosine. In some embodiments, X ¹⁰ is serine. In some embodiments, X ¹¹ is lysine. In some embodiments, X12 is leucine. In some embodiments, X ¹³ is cysteine. In some embodiments, X ¹⁴ is glycine. In some embodiments, X ¹⁵ is leucine. In some embodiments, X ¹⁶ is leucine. In some embodiments, X ¹⁷ is alanine. In some embodiments, X ¹⁸ is glutamic acid. In some embodiments, X ¹⁹ is arginine. In some embodiments, X ²⁰ is arginine. In some embodiments, X ²¹ is serine. In some embodiments, X ²² is proline. In some embodiments, X ²³ is aspartic acid. In some embodiments, X ²⁴ is arginine.

In some embodiments, the peptide comprises three or more consecutive amino acids of human MIF _{65 -94} . In some embodiments, the peptide comprises three or more consecutive amino acids of murine MIF _{65 -94} . In some embodiments, the peptide comprises three or more consecutive amino acids of porcine MIF _{65 -94} . In some embodiments, the peptide comprises three or more consecutive amino acids of bovine MIF _{65 -94} . In some embodiments, the peptide comprises three or more consecutive amino acids of rat MIF _{65 -94} .

In some embodiments, the peptide is selected from Table 3 below. Any amino acid of any of the peptides disclosed herein corresponds to an effective substituent for the original amino acid and acts as such but does not substantially reduce binding or bioactivity compared to the parent peptide sequence. It may be substituted by. Non-natural amino acids include D amino acids such as D-phenylalanine (D-F) and D-cyclohexyl alanine (D-CA); Norleucine (NLe), L-cyclohexyl alanine (L-CA), and L-amino acid, alpha-aminobutyric acid (Abu). In some embodiments, amino acids of the peptides disclosed herein are substituted with non-natural amino acids. In some embodiments, amino acids of peptides disclosed herein comprise N and / or C-terminal chemical modifications that improve ADME-PK.

[Table 3]

In some embodiments, the peptide is: CVHVVPDQLMAC (SEQ ID NO: 451).

CD74 Mimetics

CD74 is a transmembrane protein that binds to MIF. In some embodiments, CD74 is a receptor for MIF. In some embodiments, the material compositions, methods, and / or pharmaceutical compositions disclosed herein inhibit the binding of CD74 to CXCR2, CXCR4, MIF, CD44, or a combination thereof. In some embodiments, the material compositions, methods, and / or pharmaceutical compositions disclosed herein treat inflammatory diseases, diseases, conditions and symptoms by inhibiting binding of CD74 to CXCR2, CXCR4, MIF, CD44, or a combination thereof.

In some embodiments, the inflammatory disease, disease, condition and condition is a CD74 motif / domain (eg, C-terminal / cell) to an individual in need of treatment, diagnosis, or monitoring for an inflammatory disease, condition, condition and condition. Treatment, diagnosis, or monitoring is by administering a peptide that competitively binds to a binding partner of an exogenous (lumen) motif / domain). In some embodiments, the peptide binds competitively with MIF, CD44, CXCR2, and / or CXCR4, thereby preventing CD74 from binding to MIF, CD44, CXCR2, and / or CXCR4.

In some embodiments, the peptide adopts a structural or functional feature similar to CD74.

In some embodiments, the peptide comprises three or more consecutive amino acids of human CD74. In some embodiments, the peptide comprises three or more consecutive amino acids of bovine CD74. In some embodiments, the peptide comprises three or more consecutive amino acids of porcine CD74. In some embodiments, the peptide comprises three or more consecutive amino acids of murine CD74. In some embodiments, the peptide comprises three or more consecutive amino acids of rat CD74.

In some embodiments, the peptides are selected from Table 4 below. Any amino acid of any of the peptides disclosed herein corresponds to an effective substituent for the original amino acid and acts as such but does not substantially reduce binding or bioactivity compared to the parent peptide sequence. It may be substituted by. Non-natural amino acids include D amino acids such as D-phenylalanine (D-F) and D-cyclohexyl alanine (D-CA); Norleucine (NLe), L-cyclohexyl alanine (L-CA), and L-amino acid, alpha-aminobutyric acid (Abu). In some embodiments, amino acids of the peptides disclosed herein are substituted with non-natural amino acids. In some embodiments, amino acids of peptides disclosed herein comprise N and / or C-terminal chemical modifications that improve ADME-PK.

[Table 4]

CXCR2 Of CXCR4 Mimetics

In some embodiments, the material compositions, methods, and / or pharmaceutical compositions disclosed herein inhibit the binding of CXCR2 to CXCR4, MIF, CD44, CD74, or a combination thereof. In some embodiments, the material compositions, methods, and / or pharmaceutical compositions disclosed herein treat inflammatory diseases, diseases, conditions, and symptoms by inhibiting binding of CXCR2 to CXCR4, MIF, CD44, CD74, or a combination thereof. In some embodiments, the material compositions, methods, and / or pharmaceutical compositions disclosed herein inhibit the binding of CXCR4 to CXCR2, MIF, CD44, CD74, or a combination thereof. In some embodiments, the material compositions, methods, and / or pharmaceutical compositions disclosed herein treat inflammatory diseases, diseases, conditions and symptoms by inhibiting the binding of CXCR4 to CXCR2, MIF, CD44, CD74, or a combination thereof.

In some embodiments, the peptides disclosed herein bind competitively with a binding partner of the CXCR2 domain / motif. In some embodiments, an inflammatory disease, condition, condition, and condition comprises a peptide that competitively binds a binding partner of the CXCR2 motif / domain to an individual in need of treatment, diagnosis, or monitoring for an inflammatory disease, condition, condition, and condition. By administration, treatment, diagnosis, or monitoring. In some embodiments, the peptide binds to MIF, CD74 and / or CD44, thereby preventing CXCR2 from binding to MIF, CD74 and / or CD44.

In some embodiments, the peptides disclosed herein are CXCR2 extracellular loop 1 (i.e., CXCR2 _108-120), the extracellular loop 2 (i.e., CXCR2 _{184 -212),} and / or extracellular loop 3 (i.e., CXCR2 _{286 -} Competitive coupling with a coupling partner of ₃₀₀ ). In some embodiments, the peptides disclosed herein competitively bind with binding partners of extracellular loop 2 (ie, CXCR2 _184-212 ), and / or extracellular loop 3 (ie, CXCR2 _286-300 ). In some embodiments, the inflammatory disease, disease, condition and condition is determined by the subject in need of treatment, diagnosis, or monitoring for inflammatory disease, disease, condition and condition, CXCR2 extracellular loop 1 (ie, CXCR2 _108-120 ), extracellular loop 2 (i.e., CXCR2 _184-212), and / or extracellular loop 3 is competitively treatment, diagnosis, or monitoring the administration of the peptides that bind with a binding partner (i.e., CXCR2 _{286 -300).} In some embodiments, the inflammatory disease, disorder, neuropathy and symptoms 2 to a subject in need of treatment, diagnosis, or monitoring of inflammatory diseases, disorders, complications and conditions extracellular loop (i.e., CXCR2 _{184 -212),} and / or extracellular loop 3 is competitively treatment, diagnosis, or monitoring the administration of the peptides that bind with a binding partner (i.e., CXCR2 _{286 -300).}

In some embodiments, the peptides disclosed herein bind competitively with a binding partner of CXCR2 N-terminal / domain (ie, CXCR2 _1-39 ). In some embodiments, the inflammatory disease, disorder, neuropathy and symptoms of inflammatory diseases, disorders, CXCR2 to a subject in need of treatment, diagnosis, or monitoring of the symptoms, complications, and N- terminus / domain (i.e., CXCR2 _{1 -39)} Treatment, diagnosis, or monitoring is by administering a peptide that competitively binds to its binding partner.

In some embodiments, an inflammatory disease, condition, condition, and condition comprises a peptide that competitively binds a binding partner of the CXCR4 motif / domain to an individual in need of treatment, diagnosis, or monitoring for an inflammatory disease, condition, condition, and condition. By administration, treatment, diagnosis, or monitoring. In some embodiments, the inflammatory disease, disease, condition, and symptom is a combination of CXCR4 extracellular loop 1 and / or extracellular loop 2 to an individual in need of treatment, diagnosis, or monitoring for the inflammatory disease, condition, condition and condition. Treatment, diagnosis, or monitoring is by administering a peptide that competitively binds with the partner. In some embodiments, the inflammatory disease, disease, condition and condition is competitive with a binding partner of CXCR4 amino acids 182-202 (SEADDRYICDRFYPNDLWVVV) to an individual in need of treatment, diagnosis, or monitoring for an inflammatory disease, condition, condition and condition. It is treated, diagnosed, or monitored by administering a binding peptide. In some embodiments, the inflammatory disease, disease, condition and condition is competitive with a binding partner of CXCR4 amino acids 185-199 (DDRYICDRFYPNDLW) to an individual in need of treatment, diagnosis, or monitoring for an inflammatory disease, condition, condition and condition. It is treated, diagnosed, or monitored by administering a binding peptide. In some embodiments, the peptide binds to MIF, CD74 and / or CD44, thereby preventing CXCR4 from binding to MIF, CD74 and / or CD44.

In some embodiments, the peptide comprises three or more consecutive amino acids of human CXCR2. In some embodiments, the peptide comprises three or more consecutive amino acids of bovine CXCR2. In some embodiments, the peptide comprises three or more consecutive amino acids of porcine CXCR2. In some embodiments, the peptide comprises three or more consecutive amino acids of murine CXCR2. The peptide comprises three or more consecutive amino acids of rat CXCR2.

In some embodiments, the peptide comprises three or more consecutive amino acids of human CXCR4. In some embodiments, the peptide comprises three or more consecutive amino acids of bovine CXCR4. In some embodiments, the peptide comprises three or more consecutive amino acids of porcine CXCR4. In some embodiments, the peptide comprises three or more consecutive amino acids of murine CXCR4. The peptide comprises three or more consecutive amino acids of rat CXCR4.

In some embodiments, the peptide is selected from Table 5 below. Any amino acid of any of the peptides disclosed herein corresponds to an effective substituent for the original amino acid and acts as such but does not substantially reduce binding or bioactivity compared to the parent peptide sequence. It may be substituted by. Non-natural amino acids include D amino acids such as D-phenylalanine (D-F) and D-cyclohexyl alanine (D-CA); Norleucine (NLe), L-cyclohexyl alanine (L-CA), and L-amino acid, alpha-aminobutyric acid (Abu). In some embodiments, amino acids of the peptides disclosed herein are substituted with non-natural amino acids. In some embodiments, amino acids of peptides disclosed herein comprise N and / or C-terminal chemical modifications that improve ADME-PK.

[Table 5]

CD44 Mimetics

CD44 is a cell surface glycoprotein involved in cell cellular interactions, cell adhesion and migration. In certain instances, human CD44 has the following sequence:

을 갖는다.

Has

In certain cases, murine CD44 has the following sequence:

을 갖는다.

Has

In certain cases, CD44 forms a complex with CD74. In some embodiments, inhibiting (partially or completely) binding of CD44 to CD74 reduces or inhibits inflammation. In some embodiments, inhibiting binding of CD44 and MIF (partially or completely) reduces or inhibits inflammation.

In some embodiments, the material compositions, methods, and / or pharmaceutical compositions disclosed herein inhibit the binding of CD44 to CXCR2, CXCR4, MIF, CD74, or a combination thereof. In some embodiments, the material compositions, methods, and / or pharmaceutical compositions disclosed herein treat inflammatory diseases, diseases, conditions and symptoms by inhibiting binding of CD44 to CXCR2, CXCR4, MIF, CD74, or a combination thereof.

In some embodiments, an inflammatory disease, condition, condition, and condition comprises a peptide that competitively binds a binding partner of the CD44 motif / domain to an individual in need of treatment, diagnosis, or monitoring for an inflammatory disease, condition, condition, and condition. By administration, treatment, diagnosis, or monitoring. In some embodiments, the peptide binds to MIF, CXCR2, CXCR4, CD74, or a combination thereof.

In some embodiments, the peptide comprises three or more consecutive amino acids of human CD44. In some embodiments, the peptide comprises three or more consecutive amino acids of bovine CD44. In some embodiments, the peptide comprises three or more consecutive amino acids of porcine CD44. In some embodiments, the peptide comprises three or more consecutive amino acids of murine CD44. The peptide comprises three or more consecutive amino acids of rat CD44.

In some embodiments, the peptides are selected from Table 6 below. Any amino acid of any of the peptides disclosed herein corresponds to an effective substituent for the original amino acid and acts as such but does not substantially reduce binding or bioactivity compared to the parent peptide sequence. It may be substituted by. Non-natural amino acids include D amino acids such as D-phenylalanine (D-F) and D-cyclohexyl alanine (D-CA); Norleucine (NLe), L-cyclohexyl alanine (L-CA), and L-amino acid, alpha-aminobutyric acid (Abu). In some embodiments, amino acids of the peptides disclosed herein are substituted with non-natural amino acids. In some embodiments, amino acids of peptides disclosed herein comprise N and / or C-terminal chemical modifications that improve ADME-PK.

TABLE 6

F. Fusion Peptides

In some embodiments, the substance composition destroys the ability of MIF to bind CXCR2, CXCR4, CD74, or a combination thereof. In some embodiments, the material composition is a fusion peptide that binds to both N-loop motifs / domains of MIF and pseudo ELR motifs / domains of MIF. In some embodiments, the inflammatory disease, disease, condition, or symptom is treated by destroying the ability of MIF to bind CXCR2, CXCR4, CD74, or a combination thereof. In some embodiments, the inflammatory disease, disease, condition, or condition is an N-loop motif / domain of MIF and pseudo ELR of MIF in an individual in need of treatment, diagnosis, or monitoring for an inflammatory disease, disease, condition, and condition. Treatment, diagnosis, or monitoring is by administering a fusion peptide that binds to the motif / domain.

In some embodiments, the peptide comprising the fusion peptide is derived from human MIF, bovine MIF, porcine MIF, murine MIF, rat MIF, or a combination thereof. In some embodiments, the peptide comprising the fusion peptide is artificially constructed.

In some embodiments, the fusion peptide comprises at least one peptide that competitively binds to the binding partner of the N-loop motif / domain of MIF, and at least one peptide that competitively binds to the binding partner of the pseudo ELR motif / domain of MIF. do. In some embodiments, the fusion peptide comprises (a) a first peptide that competitively binds to the binding partner of the N-loop motif / domain of MIF; And (b) a second peptide that competitively binds to the binding partner of the pseudo ELR motif / domain of MIF. In some embodiments, the fusion peptide comprises (a) a first peptide that competitively binds to the binding partner of the N-loop motif / domain of MIF; (b) a second peptide that competitively binds to the binding partner of the pseudo ELR motif / domain of MIF; And (c) a third peptide that competitively binds to the binding partner of the pseudo ELR motif / domain of MIF.

In some embodiments, the fusion peptide comprises (a) a first peptide that competitively binds to the binding partner of the N-loop motif / domain of MIF; And (b) a second peptide that competitively binds to the binding partner of the pseudo ELR motif / domain of MIF, wherein the first peptide and the second peptide are chemically linked. In some embodiments, the fusion peptide comprises (a) a first peptide that competitively binds to the binding partner of the N-loop motif / domain of MIF; (b) a second peptide that competitively binds to the binding partner of the pseudo ELR motif / domain of MIF; And (c) a third peptide that competitively binds to the binding partner of the pseudo ELR motif / domain of MIF, wherein the first peptide and the second peptide are chemically linked.

In some embodiments, the fusion peptide comprises (a) a first peptide having the sequence MAFGGSSEPC; And (b) a second peptide having the sequence NVPRA. In some embodiments, the fusion peptide comprises (a) a first peptide having the sequence MAFGGSSEPC; (b) a second peptide having the sequence NVPRA; And (c) a third peptide having the sequence SVPDG.

In some embodiments, the methods and compositions disclosed herein comprise (a) a first peptide having the sequence LQDP; And (b) a second peptide having the sequence NVPRA.

In some embodiments, the first peptide and the second peptide are directly bonded to each other (eg, via covalent or ionic bonds).

Linker

In some embodiments, the one or more peptides that competitively bind with the binding partner of the N-loop motif / domain of MIF and the one or more peptides that competitively bind with the binding partner of the pseudo ELR motif / domain of MIF For example, indirectly through a linker). In some embodiments, at least one peptide that competitively binds to the binding partner of the N-loop motif / domain of MIF and at least one peptide that competitively binds to the binding partner of the pseudo ELR motif / domain of MIF is bound by a linker It is.

In some embodiments, the linker comprises (a) a first peptide that competitively binds to the binding partner of the N-loop motif / domain of MIF; And (b) a second peptide that competitively binds to the binding partner of the pseudo ELR motif / domain of MIF. In some embodiments, the fusion peptide is a peptide of Formula (IV):

Wherein Peptide 1 and Peptide 2 are selected from any of the peptides disclosed herein.

In some embodiments, the linker comprises (a) a first peptide that competitively binds to the binding partner of the N-loop motif / domain of MIF; (b) a second peptide that adopts structural or functional characteristics similar to the first portion of the pseudo ELR motif / domain of MIF; And (c) a third peptide that adopts structural or functional characteristics similar to the second portion of the pseudo ELR motif / domain of MIF. In some embodiments, the fusion peptide is a peptide of Formula (V):

Wherein, peptide 1, peptide 2, and peptide 3 are selected from any of the peptides disclosed herein.

As used herein, a "linker" is any molecule capable of binding (eg, covalently) to multiple peptides. In some embodiments, the linker binds to the peptide by covalent bonds. In some embodiments, covalent bonds include ether bonds, thioether bonds, amine bonds, amide bonds, carbon-carbon bonds, carbon-nitrogen bonds, carbon-oxygen bonds, or carbon-sulfur bonds.

In some embodiments, the linker is flexible. In some embodiments, the linker is rigid. In some embodiments, the linker is long enough to allow the fusion peptide to bind to both the pseudo ELR and the N-loop motif / domain of MIF.

In some embodiments, the linker binds two peptides. In some embodiments, the linker binds three peptides.

In some embodiments, the linker described herein binds to the C-terminus of one or more peptides that form a fusion peptide. In some embodiments, the linker binds to the N-terminus of one or more peptides that form a fusion peptide. In some embodiments, the linker described herein binds the C-terminus of one or more peptides and the N-terminus of the remaining peptides.

In some embodiments, the linker comprises a linear structure. In some embodiments, the linker comprises a nonlinear structure. In some embodiments, the linker comprises a branched structure. In some embodiments, the linker comprises a cyclic structure.

In some embodiments, the linker is alkyl. In some embodiments, the linker is heteroalkyl.

In some embodiments, the linker is alkylene. In some embodiments, the linker is alkenylene. In some embodiments, the linker is alkynylene. In some embodiments, the linker is heteroalkylene.

"Alkyl" group means an aliphatic hydrocarbon group. The alkyl moiety can be saturated alkyl or unsaturated alkyl. Depending on the structure, the alkyl group can be a monovalent radical or a divalent radical (ie an alkylene group).

The "alkyl" moiety may have from 1 to 10 carbon atoms (when alkyl is mentioned herein, for example, the numerical range "1 to 10" means each integer contained within a given range). For example, "1 to 10 carbon atoms" includes 1, including up to 10 carbon atoms, although the definition also includes the term "alkyl" where no numerical range is specified. Means that an alkyl group may consist of 2 carbon atoms, 2 carbon atoms, 3 carbon atoms, etc. The alkyl group may be “lower alkyl” having 1 to 6 carbon atoms. The group may be specified by “C ₁ -C ₄ alkyl” or a similar name, for example only, “C ₁ -C ₄ alkyl” indicates that 1 to 4 carbon atoms are present in the alkyl chain, ie the alkyl chain is Methyl, ethyl, pro , Iso-propyl, n-butyl, iso-butyl, sec-butyl, and t-butyl Typical alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, Pentyl, hexyl, ethenyl, propenyl, butenyl, and the like.

In some embodiments, the linker comprises a ring structure (eg aryl). As used herein, the term "ring" refers to any covalently cyclized structure. Rings include, for example, carbocycles (eg aryl and cycloalkyl), heterocycles (eg heteroaryl and nonaromatic heterocycles), aromatics (eg aryl and heteroaryl), and non Aromatic (eg, cycloalkyl and non-aromatic heterocycles). The ring may be optionally substituted. The ring may be monocyclic or polycyclic.

As used herein, the term "aryl" refers to an aromatic ring where each atom forming the ring is a carbon atom. Aryl rings may be formed by 5, 6, 7, 8, 9 or more carbon atoms. The aryl group may be optionally substituted. Examples of aryl groups include, but are not limited to, phenyl, naphthalenyl, phenanthrenyl, anthracenyl, fluorenyl, and indenyl. Depending on the structure, the aryl group can be a monovalent radical or a divalent radical (ie an arylene group).

The term "cycloalkyl" means a monocyclic or polycyclic non-aromatic radical, wherein each atom forming the ring (ie, the skeletal atom) is a carbon atom. Cycloalkyls can be saturated or partially unsaturated. Cycloalkyl groups include groups having 3 to 10 ring atoms. Cycloalkyls include, but are not limited to, cyclopropyl, cyclobutyryl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.

In some embodiments, the ring is cycloalkane. In some embodiments, the ring is cycloalkene.

In some embodiments, the ring is an aromatic ring. The term "aromatic" means a planar ring with a nonlocal π-electron system containing 4n + 2π electrons, where n is an integer. Aromatic rings can be formed with 5, 6, 7, 8, 9 or more atoms. The aromatic may be optionally substituted. The term "aromatic" includes both carbocyclic aryl (eg, phenyl) and heterocyclic aryl (or "heteroaryl" or "heteroaromatic") groups (eg, pyridine). The term includes monocyclic or fused ring polycyclic (ie, rings which share adjacent carbon atom pairs) groups.

In some embodiments, the ring is a heterocycle. The term "heterocycle" refers to heteroaromatic and heteroalicyclic groups containing 1 to 4 heteroatoms each selected from O, S and N, wherein each heterocyclic group is 4 to 10 in its ring system. Ring, wherein the ring of the group means heteroaromatic and heteroalicyclic groups which do not contain two adjacent 0 or S atoms. Non-aromatic heterocyclic groups include groups having only 3 atoms in their ring system, but aromatic heterocyclic groups must have at least 5 atoms in their ring system. Heterocyclic groups include benzo fused ring systems. An example of a ternary heterocyclic group is aziridinyl. An example of a four-membered heterocyclic group is azetidinyl (derived from azetidine). An example of a 5 membered heterocyclic group is thiazolyl. An example of a 6 membered heterocyclic group is pyridyl and an example of a 10 membered heterocyclic group is quinolinyl. Examples of non-aromatic heterocyclic groups include pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl, piperidino, morpholi Furnace, thiomorpholino, thioxanyl, piperazinyl, azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxepanyl, thiepanyl, oxazinyl, diazepinyl, tizefinyl, 1,2,3,6-tetrahydropyridinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolanyl, Pyrazolinyl, ditianyl, dithiolayl, dihydropyranyl, dihydrothienyl, dihydrofuranyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, 3-azabicyclo [3.1.0] hexa Nil, 3-azabicyclo [4.1.0] heptanyl, 3H-indolyl, and quinolizinyl. Examples of aromatic heterocyclic groups include pyridinyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, Pyrrolyl, quinolinyl, isoquinolinyl, indolyl, benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolinyl, phthalazinyl, pyridazinyl, triazinyl, isoindoleyl, Putridinyl, furinyl, oxdiazolyl, thiadiazolyl, furazanyl, benzo furazanyl, benzothiophenyl, benzothiazolyl, benzoxazolyl, quinazolinyl, quinoxalinyl, naphthyridinyl, and furopyridinyl There is this. Preferably, the group may be C attached or N attached. For example, groups derived from pyrrole are pyrrole-1-yl (N attachment) or pyrrole-3-yl (C attachment). Additionally, groups derived from imidazole may be imidazol-1-yl or imidazol-3-yl (both N attached) or imidazol-2-yl, imidazol-4-yl or imidazol-5-yl ( All C attached). Heterocyclic groups include benzo fused ring systems and ring systems substituted with one or two oxo (═O) moieties such as pyrrolidin-2-ones. Depending on the structure, the heterocycle group may be a monovalent radical or a divalent radical (ie a heterocycline group).

In some embodiments, the ring is fused. The term "fused" means a structure in which two or more rings inherent one or more bonds. In some embodiments, the ring is a dimer. In some embodiments, the ring is a trimer. In some embodiments, the ring is substituted.

The term "carbocyclic" or "carbocycle" means a ring that is each atom carbon atom forming a ring. Carbocycles include aryl and cycloalkyl. Thus, the term identifies carbocycles and heterocycles (“heterocyclics”) that contain one or more atoms (ie, heteroatoms) whose ring backbone differs from carbon. Heterocycles include heteroaryl and heterocycloalkyl. Carbocycles and heterocycles are optionally substituted.

In some embodiments, the linker is substituted. The term "optionally substituted" or "substituted" means that the groups mentioned are C ₁ -C ₆ alkyl, C 3 -Cgcycloalkyl, aryl, heteroaryl, C ₂ -C ₆ heteroalicyclic, hydroxy, C _1- C ₆ alkoxy, aryloxy, C ₁ -C ₆ alkylthio, arylthio, C ₁ -C ₆ alkylsulfoxide, arylsulfoxide, C ₁ -C ₆ alkylsulfone, arylsulfone, cyano, halo, C _2- From C ₈ acyl, C ₂ -C ₈ acyloxy, nitro, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ fluoroalkyl, and amino (including C ₁ -C ₆ alkylamino), and protected derivatives thereof It means that it can be substituted with one or more additional group (s) selected individually and independently. In one example, any substituent is L ^S R ^S wherein each L ^S is a bond, -0-, -C (= 0)-, -S-, -S (= 0)-, -S (= 0 ) _2- , -NH-, -NHC (= 0)-, -C (= 0) NH-, S (= 0) ₂ NH-, -NHS (= 0) _2- , -OC (= 0) NH -, -NHC (= 0) 0-,-(C ₁ -C ₆ alkyl)-, or-(C ₂ -C ₆ alkenyl)-, each R ^S is H, (C ₁ -C ₄ alkyl), (C ₃ -C ₈ cycloalkyl), heteroaryl, aryl, and C ₁ -C ₆ heteroalkyl). Optionally substituted non-aromatic groups may be substituted with one or more oxo (= 0). Protecting groups capable of forming protective derivatives of such substituents are known to those skilled in the art.

In some embodiments, the linker is an amino acid. In some embodiments, the fusion peptide is a peptide of formula (VI):

Wherein Peptide 1 and Peptide 2 are selected from any of the peptides disclosed herein.

In some embodiments, the linker is an artificial amino acid. In some embodiments, the linker is β amino acid. In some embodiments, the linker is a γ amino acid.

In some embodiments, the linker is polyethylene glycol (PEG). In some embodiments, the linker is diamino acid. In some embodiments, the linker is diaminopropionic acid.

In some embodiments, the linker is hydrolyzable.

In one non-limiting example, the fusion peptide is as follows:

E. MIF Trimerization  Moderator

In some embodiments, an inflammatory disease, disease, condition, or symptom is treated by modulating the ability of MIF to form homomultimers. In some embodiments, an inflammatory disease, disease, condition, or condition is treated by destroying the ability of MIF to form trimers. In some embodiments, the inflammatory disease, disease, condition, or condition is treated by promoting MIF trimerization.

In certain cases, the functionally active (or mature) MIF comprises three MIF peptide sequences (ie, trimers). In certain cases, the pseudo ELR motif / domain of each MIF polypeptide forms a ring in the trimer. In certain cases, the N-loop motif / domain of each MIF polypeptide extends outwards from the pseudo ELR ring (see FIG. 1).

In certain cases, residues 38-44 of one subunit interact with residues 48-50 of another subunit. In certain cases, residues 96-102 of one subunit interact with residues 107-109 of another subunit. In certain cases, the motifs / domains on one subunit formed by N73, R74, S77, K78, C81 (where the numbering includes methionine number 1) are N110, S111, T112 of another subunit (the numbering is 1 And methionine).

In some embodiments, the MIF trimerization disruptor is derived from, or introduces, any or all of amino acid residues 38-44 of MIF (eg, human, bovine, swine, murine, or rat). . In some embodiments, the MIF trimerization disrupting agent is a peptide derived from any or all of amino acid residues 48-50 of MIF (eg, human, bovine, pig, murine, or rat). In some embodiments, the MIF trimerization disruptor is a peptide derived from any or all of amino acid residues 57-66 of MIF (eg, human, bovine, swine, murine, or rat), and / or the amino acid Any or all of residues 57-66 are introduced. In some embodiments, the MIF trimerization disruptor is a peptide derived from any or all of amino acid residues 61-70 of MIF (eg, human, bovine, pig, murine, or rat), and / or the amino acid Any or all of residues 61-70 are introduced. In some embodiments, the MIF trimerization disruptor is a peptide derived from any or all of amino acid residues 96-102 of MIF (eg, human, bovine, pig, murine, or rat), and / or the amino acid Any or all of residues 96-102 are introduced. In some embodiments, the MIF trimerization disruptor is a peptide derived from any or all of amino acid residues 107-109 of MIF (eg, human, bovine, swine, murine, or rat), and / or the amino acid Any or all of the residues 107-109 are introduced. In some embodiments, the MIF trimerization disruptor comprises amino acid residues N73, R74, S77, K78, and C81 of MIF (eg, human, bovine, swine, murine, or rat), wherein the numbering comprises methionine 1 And / or any or all of the amino acid residues N73, R74, S77, K78, and C81, wherein the numbering comprises methionine 1). In some embodiments, the MIF trimerization disruptor is any of amino acid residues N110, S111, and T112 of MIF (eg, human, bovine, pig, murine, or rat), wherein the numbering comprises methionine 1). One or all of them and / or introduce any or all of the amino acid residues N110, S111, and T112 (the numbering comprises methionine 1).

In some embodiments, the MIF trimerization disrupting agent is any one or any of amino acid residues 57-66 of the MIF (eg, human, bovine, swine, murine, or rat), wherein the numbering comprises methionine 1) A peptide derived from all and / or introducing any or all of the amino acid residues 57-66 (the numbering comprises methionine number 1). In some embodiments, the MIF trimerization breaker is a peptide of formula (VII):

Where

X ¹ is selected from the group consisting of cysteine, alanine, serine, and threonine;

X ² is selected from the group consisting of alanine, proline, glycine and cysteine;

X ³ is selected from the group consisting of leucine, valine and phenylalanine;

X ⁴ is selected from the group consisting of cysteine, glycine, threonine and isoleucine;

X ⁵ is selected from the group consisting of serine, valine, glutamine and asparagine;

X ⁶ is selected from the group consisting of leucine, valine, isoleucine and methionine;

X ⁷ is selected from the group consisting of histidine, cysteine, lysine, arginine, and leucine.

In some embodiments, the MIF trimerization breaker comprises three or more consecutive amino acids of human MIF _{57 -66} . In some embodiments, the MIF trimerization _disruptor comprises three or more consecutive amino acids of murine MIF _{57 -66} . In some embodiments, the MIF trimerization breaker comprises three or more consecutive amino acids of porcine MIF _{57 -66} . In some embodiments, the MIF trimerization breaker comprises three or more consecutive amino acids of bovine MIF _{57 -66} . In some embodiments, the MIF trimerization breaker comprises three or more consecutive amino acids of rat MIF _{57 -66} .

In some embodiments, the MIF trimerization disruptor is an antibody that binds to any or all of amino acid residues 38-44 of MIF. In some embodiments, the MIF trimerization disruptor is an antibody that binds to any or all of amino acid residues 48-50 of MIF. In some embodiments, the MIF trimerization disruptor is an antibody that binds to any or all of amino acid residues 57-66 of MIF. In some embodiments, the MIF trimerization disruptor is an antibody that binds to any or all of amino acid residues 61-70 of MIF. In some embodiments, the MIF trimerization disruptor is an antibody that binds to any or all of amino acid residues 96-102 of MIF. In some embodiments, the MIF trimerization disruptor is an antibody that binds to any or all of amino acid residues 107-109 of MIF. In some embodiments, the MIF trimerization disruptor is an antibody that binds to any or all of amino acid residues N73, R74, S77, K78, and C81 of MIF. In some embodiments, the MIF trimerization disruptor is an antibody that binds to any or all of amino acid residues N110, S111, and T112 of MIF.

In some embodiments, the MIF trimerization disruptor is a small molecule that binds to any or all of amino acid residues 38-44 of MIF. In some embodiments, the MIF trimerization disruptor is a small molecule that binds to any or all of amino acid residues 48-50 of MIF. In some embodiments, the MIF trimerization disruptor is a small molecule that binds to any or all of amino acid residues 57-66 of MIF. In some embodiments, the MIF trimerization disruptor is a small molecule that binds to any or all of amino acid residues 61-70 of MIF. In some embodiments, the MIF trimerization disruptor is a small molecule that binds to any or all of amino acid residues 96-102 of MIF. In some embodiments, the MIF trimerization disruptor is a small molecule that binds to any or all of amino acid residues 107-109 of MIF. In some embodiments, the MIF trimerization disruptor is a small molecule that binds to any or all of amino acid residues N73, R74, S77, K78, and C81 of MIF. In some embodiments, the MIF trimerization disruptor is a small molecule that binds to any or all of amino acid residues N110, S111, and T112 of MIF.

In some embodiments, the MIF trimerization disruptor is a peptibody that binds to any or all of amino acid residues 38-44 of MIF. In some embodiments, the MIF trimerization disruptor is a peptibody that binds to any or all of amino acid residues 48-50 of MIF. In some embodiments, the MIF trimerization disruptor is a peptibody that binds to any or all of amino acid residues 57-66 of MIF. In some embodiments, the MIF trimerization breaker is a peptibody that binds to any or all of amino acid residues 61-70 of MIF. In some embodiments, the MIF trimerization breaker is a peptibody that binds to any or all of amino acid residues 96-102 of MIF. In some embodiments, the MIF trimerization disruptor is a peptibody that binds to any or all of amino acid residues 107-109 of MIF. In some embodiments, the MIF trimerization breaker is a peptibody that binds to any or all of amino acid residues N73, R74, S77, K78, and C81 of MIF. In some embodiments, the MIF trimerization disruptor is a peptibody that binds to any or all of amino acid residues N110, S111, and T112 of MIF.

F. Peptides Mimetics

In some embodiments, peptide mimetics are used in place of the polypeptides disclosed herein, including those used to treat or prevent inflammatory diseases.

Peptide mimetics (and peptide-based inhibitors) are developed using, for example, computerized molecular modeling. Peptide mimetics can be selected from -CH ₂ NH-, -CH ₂ S-, -CH ₂ -CH _2- , -CH = CH- (cis and trans), -CH-CF- ( Trans), —CoCH ₂ —, —CH (OH) CH ₂ —, and —CH ₂ SO—, and is designed to include structures having one or more peptide bonds optionally substituted by a binding moiety selected from the group consisting of: trans). In some embodiments, such peptide mimetics have better chemical stability, enhanced pharmacological properties (half life, absorption, efficacy, efficacy, etc.), altered specificity (eg, broad biological activity spectrum), reduced antigenicity More economically. In some embodiments, the peptide mimetic is provided with one or more labels, either directly or via spacers (eg, amide groups), at non-interfering position (s) on analogs predicted through quantitative structural activity data and / or molecular modeling. Or covalent attachment with the conjugate. These non-interfering positions are generally positions that do not form direct contact with the receptor (s) to which the peptide mimetic specifically binds to produce a therapeutic effect. In some embodiments, systematic substitution of one or more amino acids of the consensus sequence with D amino acids of the same type (eg, with D-lysine instead of L-lysine) is used to produce more stable peptides with the desired properties. .

Phage display peptide libraries have emerged as a technique for generating peptide mimetics (Scott, JK et al., (1990) Science 249: 386; Devlin, JJ et al., (1990) Science 249: 404). US Pat. No. 5,223,409, US5,733,731; US5,498,530; US5,432,018; US5,338,665; US5,922,545; WO 96/40987 and WO 98/15833, each of which is herein incorporated by reference. Included)). In this library, random peptide sequences are displayed fused with coat proteins of fibrous phage. Typically, the displayed peptides are eluted according to affinity for the extracellular motif / domain to which the antibody is immobilized (in case of PF4 or RANTES). In some embodiments the peptide mimetics are isolated by biopanning (Nowakowski, G.S, et al., (2004) Stem Cells 22: 1030-1038). In some embodiments, cells that specifically bind phage are isolated by screening the library through the use of FAC using whole cells expressing MIF. Retained phage is concentrated through repeated biopanning and regrowth in successive rounds. The highest binding peptides are sequenced to identify key residues in one or more structurally related peptide families. The peptide sequence also suggests which residues to substitute through mutagenesis or at alanine scanning at the DNA level. In some embodiments, mutagenesis libraries are generated and screened to further optimize the sequence of the highest binding factor (Lowman (1997) Ann. Rev. Biophys. Biomol. Struct. 26: 401-24).

In some embodiments, structural analysis of protein-protein interactions is used to present peptides that competitively bind to binding partners of the polypeptides described herein. In some embodiments, the crystal structure obtained from such an analysis shows the identity and relative orientation of important residues of the polypeptide based on peptide design. See, eg, Takasaki, et al., (1997) Nature Biotech, 15: 1266-70.

In some embodiments, the agent is a peptide or polypeptide. In some embodiments, the peptide comprises a peptide that competitively binds to a binding partner of VNTNVPPRASVPDGFLSELTQQLAQATGKPPQYIAVHVVPDQL (SEQ ID NO: 10); Peptides that competitively bind with a binding partner of PDQLMAFGGSSEPCALCSL (SEQ ID NO: 11); Peptides that competitively bind to a binding partner of VNTNVPPRASVPDGFLSELTQQLAQATGKPPQYIAVHVVPDQLMAFGGSSEPCALCSL (SEQ ID NO: 12); Peptides that competitively bind with a binding partner of PDQLMAFGGSSEPCALCSLHSI (SEQ ID NO: 13); Or a combination thereof.

G. Antibodies

In some embodiments, the inflammatory disease, disease, condition, or symptom is treated by destroying the ability of MIF to bind CXCR2, CXCR4, CD74, or a combination thereof. In some embodiments, an inflammatory disease, condition, condition, or condition is administered by administering an antibody that binds MIF, one or more MIF motifs to an individual in need of treatment, diagnosis, or monitoring for an inflammatory disease, condition, condition, and condition. Is treated. In some embodiments, an inflammatory disease, disease, condition, or condition is treated by administering an antibody that binds CD44 to an individual in need of treatment, diagnosis, or monitoring for an inflammatory disease, disease, condition, and condition. In some embodiments, an inflammatory disease, disease, condition, or condition is treated by administering an antibody that binds to CD74 to an individual in need of treatment, diagnosis, or monitoring for an inflammatory disease, condition, condition, and condition. In some embodiments, an inflammatory disease, condition, condition, or condition is treated by administering an antibody that binds CXCR2 to an individual in need of treatment, diagnosis, or monitoring for an inflammatory disease, condition, condition, and condition. In some embodiments, an inflammatory disease, disease, condition, or condition is treated by administering an antibody that binds CXCR4 to an individual in need of treatment, diagnosis, or monitoring for an inflammatory disease, condition, condition, and condition.

In some embodiments, the antibody is a human antibody or humanized antibody. In some embodiments, the antibody is human IgG. In some embodiments, the antibody is or comprises one or more polypeptides derived from human IgG1, IgG4, IgG2, IgD, IgA or IgM. The antibodies disclosed herein are produced by any suitable method.

Antigen-based Antibody Development

In some embodiments, the antibodies disclosed herein are generated by contacting a host (eg, mouse or rabbit) with an antigen. In some embodiments, the antigen is a MIF monomer. In some embodiments, the antigen is a MIF trimer. In some embodiments, the antigen is a fragment of a full length MIF polypeptide. In some embodiments, the antigen is a polypeptide comprising all or a portion of MIF _{50 -65} . In some embodiments, the antigen is a polypeptide comprising the whole or a part of MIF N- terminus / pseudo ELR motif / domain (MIF _{1 -17).} In some embodiments, the antigen is a polypeptide comprising all or part of a MIF alpha-helix # 1 motif / domain (ie, MIF _{18 -31} ). In some embodiments, the antigen is a polypeptide comprising the whole or a part of MIF N- loop motif / domain (i.e., MIF _{32 -60).} In some embodiments, the antigen is a polypeptide comprising all or part of a MIF loop-barrel-loop motif / domain (ie, MIF _{64 -93} ). In some embodiments, the antigen is a polypeptide comprising all or part of a MIF C-terminal motif / domain (ie, MIF _{90 -114} ). In some embodiments, the antigen is a polypeptide comprising the whole or a part of MIF N- terminal tail (i.e., MIF _{1 -7).} In some embodiments, the antigen is a polypeptide comprising the whole or a part of MIF pseudo ELR- loop (i.e., MIF _{7 -17).} In some embodiments, the antigen is a polypeptide comprising all or part of a MIF PPQ-loop (ie, MIF _32-38 ). In some embodiments, the antigen is a polypeptide comprising all or part of a MIF PDQ-loop (ie, MIF _{43 -56} ). In some embodiments, the antigen is a polypeptide comprising all or part of a MIF IGK-loop (ie, MIF _{64 -71} ). In some embodiments, the antigen is a polypeptide comprising all or part of a MIF NRS-helix (ie, MIF _{72 -89} ). In some embodiments, the antigen is a polypeptide comprising all or part of a MIF SPDR-loop (ie, MIF _{90 -94} ). In some embodiments, the antigen is a polypeptide comprising all or part of a MIF C-terminal tail (ie, MIF _{101 -114} ).

In some embodiments, an antibody disclosed herein is produced by contacting a host (eg, mouse or rabbit) with two or more antigens. In some embodiments, the antigen is a polypeptide comprising all or a portion of a MIF N-terminal / pseudo ELR motif; Polypeptides comprising all or a portion of a MIF N-loop motif; Polypeptides comprising all or part of a MIF loop-barrel-loop motif; Polypeptides comprising all or part of a MIF C-terminal motif; A polypeptide comprising all or a portion of the MIF alpha-helix # 1 motif; Polypeptides comprising all or part of a MIF N-terminal tail; Polypeptides comprising all or a portion of a MIF pseudo ELR motif / domain; Polypeptides comprising all or part of a MIF PPQ-loop; Polypeptides comprising all or part of a MIF PDQ-loop; Polypeptides comprising all or part of a MIF IGK-loop; Polypeptides comprising all or part of a MIF NRS-helix; Polypeptides comprising all or part of a MIF SPDR-loop; Polypeptides comprising all or part of a C-terminal tail; MIF and _{50 -65} is selected from a polypeptide comprising the whole or a portion thereof.

In some embodiments, an antibody disclosed herein is generated by contacting a host (eg, mouse or rabbit) with three or more antigens. In some embodiments, the antigen is a polypeptide comprising all or a portion of a MIF N-terminal / pseudo ELR motif; Polypeptides comprising all or a portion of a MIF N-loop motif; Polypeptides comprising all or part of a MIF loop-barrel-loop motif; Polypeptides comprising all or part of a MIF C-terminal motif; A polypeptide comprising all or a portion of the MIF alpha-helix # 1 motif; Polypeptides comprising all or part of a MIF N-terminal tail; MIF pseudo ELR motif / domain; Polypeptides including all or part thereof; Polypeptides comprising all or part of a MIF PPQ-loop; Polypeptides comprising all or part of a MIF PDQ-loop; Polypeptides comprising all or part of a MIF IGK-loop; Polypeptides comprising all or part of a MIF NRS-helix; Polypeptides comprising all or part of a MIF SPDR-loop; Polypeptides comprising all or part of a C-terminal tail; MIF and _{50 -65} is selected from a polypeptide comprising the whole or a portion thereof.

DNA  Based antibody development

In some embodiments, an antibody disclosed herein is generated by contacting a host with a nucleic acid sequence encoding (or alternatively, a “MIF nucleic acid sequence”) that encodes part or all of a MIF polypeptide.

In some embodiments, the MIF nucleic acid sequence has been cloned into an expression vector (eg, plasmid).

In some embodiments, the host is a mammal. In some embodiments, the host is a mouse, rabbit, or rat. In some embodiments, the host is a mammalian cell. In some embodiments, the host is a bacterial cell.

In some embodiments, the MIF nucleic acid sequence is injected into the host intramuscularly or intradermally to contact the MIF nucleic acid sequence with the host. In some embodiments, the contact further comprises applying a current to the injection site (ie, electroporation). In some embodiments, a gene gun is used to contact the MIF nucleic acid sequence with a host.

In some embodiments, the nucleic acid sequences encoding some or all of the MIF polypeptides are expressed by one host cell (or a plurality of host cells) to produce the expressed MIF polypeptide. In some embodiments, the expressed MIF polypeptide is cysteinylated. In some embodiments, the expressed MIF polypeptide is phosphorylated. In some embodiments, the expressed MIF polypeptide is glycosylated.

In some embodiments, a method of producing an antibody disclosed herein further comprises contacting the host with an adjuvant. In some embodiments, the adjuvant is administered as a nucleic acid sequence. In some embodiments, the adjuvant is administered as a polypeptide or polysaccharide. In some embodiments, the adjuvant is cytokine, lymphokine, or a combination thereof. In some embodiments, the adjuvant is interleukin, tumor necrosis factor, GM-CSF, or a combination thereof. In some embodiments, the adjuvant is B7-1, B7-2, CD40L, or a combination thereof. In some embodiments, an expression vector comprising a MIF nucleic acid sequence further comprises a nucleic acid sequence encoding an adjuvant. In some embodiments, the host is contacted with a second expression vector encoding the adjuvant.

In some embodiments, the nucleic acid sequence encodes a MIF N-terminal tail / pseudo ELR motif. In some embodiments, the nucleic acid sequence encodes MIF _{50 -65} . In some embodiments, the nucleic acid sequence encodes a MIF N-loop motif. In some embodiments, the nucleic acid sequence encodes a MIF loop-barrel-loop motif. In some embodiments, the nucleic acid sequence encodes a MIF C-terminal motif. In some embodiments, the nucleic acid sequence encodes a MIF alpha-helix # 1 motif / domain (ie, TTCCTGAGCGAGCTGACACAGCAGCTGGCCCAGGCCACCGGC). In some embodiments, the nucleic acid sequence encodes a MIF N-terminal tail (ie, CCCATGTTCATCGTGAACACC). In some embodiments, the nucleic acid sequence encodes a MIF pseudo ELR motif / domain (ie, AACGTGCCCAGAGCCAGCGTGCCCGACGGC). In some embodiments, the nucleic acid sequence encodes a MIF PPQ-loop (ie, AAGCCCCCTCAGTAT ATCGCC). In some embodiments, the nucleic acid sequence encodes a MIF PDQ-loop (ie, CCCGACCAGCTGATGGCCTTCGGCGGCAGCAGCGAGCCTTGC). In some embodiments, the nucleic acid sequence encodes a MIF IGK-loop (ie, ATCGGCAAGATCGGCGGAGCCC AG). In some embodiments, the nucleic acid sequence encodes a MIF NRS-helix (ie, AACAGAAGCTACAGCAAGCTGCTGTGCGGCCTGCTGGCCGAGAGACTGAGAATC). In some embodiments, the nucleic acid sequence encodes an SPDR-loop (ie, AGCCCCGACAGAGTGTACATCAACTACTACGAC). In some embodiments, the nucleic acid sequence encodes a C-terminal tail (ie, ATGAACGCCGCCAACGTGGGCTGGAACAACAGCACCTTCGCC).

In some embodiments, an antibody disclosed herein comprises a host comprising a sequence encoding a MIF N-terminal tail / pseudo ELR motif, a sequence encoding a MIF N-loop motif, a sequence encoding a MIF loop-barrel-loop motif, a MIF A sequence encoding a C-terminal motif, a sequence encoding a MIF alpha-helix # 1 motif, a sequence encoding a MIF N-terminal tail, a sequence encoding a MIF pseudo ELR motif / domain, a sequence encoding a MIF PPQ-loop , A sequence encoding a MIF PDQ-loop, a sequence encoding a MIF IGK-loop, a sequence encoding a MIF NRS-helix, a sequence encoding a MIF SPDR-loop, a sequence encoding a MIF C-terminal tail, and a MIF _{50 It is} produced by contacting two or more nucleic acid sequences selected from sequences encoding _-65 . In some embodiments, an antibody disclosed herein comprises a host comprising a sequence encoding a MIF N-terminal tail / pseudo ELR motif, a sequence encoding a MIF N-loop motif, a sequence encoding a MIF loop-barrel-loop motif, a MIF A sequence encoding a C-terminal motif, a sequence encoding a MIF alpha-helix # 1 motif, a sequence encoding a MIF N-terminal tail, a sequence encoding a MIF pseudo ELR motif / domain, a sequence encoding a MIF PPQ-loop , A sequence encoding a MIF PDQ-loop, a sequence encoding a MIF IGK-loop, a sequence encoding a MIF NRS-helix, a sequence encoding a MIF SPDR-loop, a sequence encoding a MIF C-terminal tail, and a MIF _{50 It is} produced by contacting a nucleic acid sequence encoding two or more MIF polypeptide motifs selected from sequences encoding _-65 .

In some embodiments, an antibody disclosed herein comprises a host comprising a sequence encoding a MIF N-terminal tail / pseudo ELR motif, a sequence encoding a MIF N-loop motif, a sequence encoding a MIF loop-barrel-loop motif, a MIF A sequence encoding a C-terminal motif, a sequence encoding a MIF alpha-helix # 1 motif, a sequence encoding a MIF N-terminal tail, a sequence encoding a MIF pseudo ELR motif / domain, a sequence encoding a MIF PPQ-loop , MIF sequence encoding a PDQ- loop, MIF sequence encoding a IGK- loop, MIF NRS- sequence encoding a helix, MIF sequence encoding a SPDR- loop, MIF C- terminal tail, and _{50 -65} MIF coding It is produced by contacting three or more nucleic acid sequences selected from the sequence to be made. In some embodiments, an antibody disclosed herein comprises a host comprising a sequence encoding a MIF N-terminal tail / pseudo ELR motif, a sequence encoding a MIF N-loop motif, a sequence encoding a MIF loop-barrel-loop motif, a MIF A sequence encoding a C-terminal motif, a sequence encoding a MIF alpha-helix # 1 motif, a sequence encoding a MIF N-terminal tail, a sequence encoding a MIF pseudo ELR motif / domain, a sequence encoding a MIF PPQ-loop , A sequence encoding a MIF PDQ-loop, a sequence encoding a MIF IGK-loop, a sequence encoding a MIF NRS-helix, a sequence encoding a MIF SPDR-loop, a sequence encoding a MIF C-terminal tail, and a MIF ₅₀ Generated by contacting a nucleic acid sequence encoding at least three MIF polypeptide motifs selected from the sequences encoding _-65 .

Antibody production

In some embodiments, the antibodies disclosed herein are produced through the use of hybridomas. As used herein, “hybridoma” is an endless proliferating cell that produces antibodies. In some embodiments, the host (eg, mouse or rabbit) is inoculated with the antigen or nucleic acid. In some embodiments, B cells are extracted from the spleen of the host. In some embodiments, hybridomas are produced by fusing (1) the extracted B cells with (2) myeloma cells (ie, hypoxanthine-guanine-phosphoribosyl transferase negative, indefinite myeloma cells). In some embodiments, B cells and myeloma cells are cultured together and exposed to an agent (eg PEG) that makes the cell membrane more permeable.

In some embodiments, the culture comprises a plurality of hybridomas, a plurality of myeloma cells, and a plurality of B cells. In some embodiments, the cells are each individually (eg, incubated with HAT medium) for the culture conditions for selecting hybridomas.

In some embodiments, individual hybridomas (ie clones) are isolated and cultured. In some embodiments, hybridomas are injected into laboratory animals. In some embodiments, the hybridomas are cultured in cell culture.

Humanized antibody

In some embodiments, the methods described herein comprise humanized monoclonal antibodies. In some embodiments, humanized monoclonal antibodies comprise heavy and light chain constant regions derived from human sources and variable regions derived from murine sources.

In some embodiments, humanized immunoglobulins, including humanized antibodies, are constructed by genetic engineering. In some embodiments, a humanized immunoglobulin comprises the same framework as the framework of a particular human immunoglobulin chain (ie, an acceptor or recipient), and three CDRs from a non-human (donor) immunoglobulin chain. In some embodiments, a limited number of amino acids in the framework of the humanized immunoglobulin chain are identified and selected to be identical to the amino acid at that position in the donor instead of the acceptor.

In some embodiments, a framework is used that is derived from certain human immunoglobulins that are homologous to donor immunoglobulins so that they can be humanized. For example, the sequence of the mouse heavy (or light) variable region may be stored in a data bank (e.g., the National Biomedical Research Foundation Protein Identification Resource or the National Biotechnology Information Center (NCBI)). The degree of homology for different human regions can be, for example, from about 40% to about 60%, about 70%, by comparison with the sequences of human heavy (or light chain) variable regions in the Protein Sequence Database of the National Center for Biotechnology Information). It has been shown that more than 80% can vary widely. As an acceptor immunoglobulin, fewer amino acid alterations will appear when transformed from donor immunoglobulin to humanized immunoglobulin by selecting one of the human heavy chain variable regions with the greatest homology with the heavy chain variable region of the donor immunoglobulin. As an acceptor immunoglobulin, fewer amino acid alterations will appear when transformed from donor immunoglobulin to humanized immunoglobulin by selecting one of the light chain variable regions of homology with the donor immunoglobulin having the greatest homology.

In some embodiments, humanized immunoglobulins comprise light and heavy chains derived from the same human antibody as the acceptor sequence. In some embodiments, humanized immunoglobulins comprise light and heavy chains derived from different human antibody germline sequences as acceptor sequences; When such a combination is used, conventional assays (eg ELISA) can be used to readily determine whether VH and VL bind to epitopes of interest. In some embodiments, a human antibody will be selected in which the light and heavy chain variable region sequences together have the largest homology with the donor light and heavy chain variable region sequences overall. In some embodiments, affinity is made higher by selecting a few amino acids in the framework of the humanized immunoglobulin chain that are identical to the amino acids at these positions in the donor instead of the acceptor.

Any suitable method of modifying the framework region is contemplated herein. In some embodiments, the relevant framework amino acid to be altered is selected based on the difference in amino acid framework residues between the donor and acceptor molecules. In some embodiments, the amino acid position to be altered is a residue known to be important for or contribute to the CDR conformation (eg, a regular framework residue is important for CDR conformation and / or structure). In some embodiments, the relevant framework amino acids to be altered are selected based on the frequency of amino acid residues at a particular framework position (eg, by comparing the selected framework with other framework sequences within its subfamily). Or residues present at low frequencies at locations). In some embodiments, related framework amino acids to be altered are selected based on proximity to the CDRs. In some embodiments, the relevant framework amino acids to be altered are selected based on known or predicted proximity at the antigen-CDR interface, or predicted to modulate CDR activity. In some embodiments, the relevant framework amino acid to be altered is a framework residue known to or predicted to form a contact between the heavy (VH) and light (VL) variable region interface. In some embodiments, the relevant framework amino acids to be altered are framework residues that are inaccessible to the solvent.

In some embodiments, amino acid alterations are introduced at some or all of the selected positions into the coding nucleic acid for the acceptor variable region framework and donor CDRs. In some embodiments, altered framework or CDR sequences are prepared individually, tested, or combined and tested sequentially or simultaneously.

In some embodiments, the variability at any or all of the altered positions is from several to a plurality of different amino acid residues, including all 20 naturally occurring amino acids or functional equivalents and analogs thereof. In some embodiments, non-naturally occurring amino acids are also contemplated.

In some embodiments, the humanized antibody sequence is cloned into a vector. In some embodiments, any suitable vector is used. In some embodiments, the vector is a plasmid, virus, eg, appropriate phage, or phagemid. For further details, see, for example, Molecular Cloning: a Laboratory Manual: 2nd edition, Sambrook et al., 1989, Cold Spring Harbor Laboratory Press. Numerous known techniques and protocols for nucleic acid engineering, such as nucleic acid construct preparation, mutagenesis, sequencing, introduction of DNA into cells and gene expression, and protein analysis, are described in Short Protocols in Molecular Biology, Second Edition. , Ausubel et al., Eds., John Wiley & Sons, 1992. The disclosures of Sambrook et al. And Ausubel et al. Are incorporated herein by reference.

In some embodiments, any suitable host cell is transformed with a vector expressing a humanized antibody sequence. In some embodiments, the host cell is a bacterial, mammalian cell, yeast and baculovirus system. Expression of antibodies and antibody fragments in prokaryotic cells such as E. coli is well established in the art. For a review, see, eg, Pluckthun, A. Bio / Technology 9: 545-551 (1991). Expression in eukaryotic cells in culture as an option for the production of antibodies and antigen binding fragments described herein is also available to those skilled in the art and, for recent reviews, see, eg, Raff, ME (1993) Curr. Opinion Biotech. 4: 573-576; (Trill JJ et al., (1995) Curr. Opinion Biotech 6: 553-560)), each of which is incorporated herein by reference. refer to.

In some embodiments a mammalian expression system is used. In some embodiments, the mammalian expression system is dehydrofolate reductase deficient (“dhfr-”) Chinese hamster ovary cells. In some embodiments, dhfr-CHO cells are transfected with an expression vector containing a functional DHFR gene, along with the gene encoding the desired humanized antibody.

In some embodiments, the DNA is transformed by any suitable method. For eukaryotic cells, suitable techniques include, for example, calcium phosphate transfection, DEAE dextran, electroporation, liposome mediated transfection and retroviruses or other viruses such as vaccinia, or baculos, Transduction with rovirus. For bacterial cells, suitable techniques include, for example, calcium chloride transformation, electroporation and transfection with bacteriophages.

In some embodiments, DNA sequences encoding antibodies or antigen binding fragments thereof are prepared synthetically rather than cloning. In some embodiments, the DNA sequence is designed using appropriate codons for an antibody or antigen binding fragment amino acid sequence. In general, when a sequence is used for expression, it will select a preferred codon for the intended host. In some embodiments, a complete sequence is assembled from overlapping oligonucleotides prepared by standard methods and assembled into a complete coding sequence. See, eg, Edge, Nature, 292: 756 (1981); Nambair et al., Science, 223: 1299 (1984); Jay et al., J. Biol. Chem., 259: 6311 (1984), each of which is incorporated herein by reference to the disclosure.

H. Peptibodies

In some embodiments, the substance composition destroys the ability of MIF to bind CXCR2, CXCR4, CD74, or a combination thereof. In some embodiments, the substance composition is a peptibody. In some embodiments, the inflammatory disease, disease, condition, or symptom is treated by destroying the ability of MIF to bind CXCR2, CXCR4, CD74, or a combination thereof. In some embodiments, the inflammatory disease, disease, condition, or condition is treated, diagnosed, or monitored by administering peptibodies to an individual in need thereof for treating, diagnosing, or monitoring the inflammatory disease, condition, condition, and condition.

The term “peptibody” includes peptide (s) directly or indirectly (eg, covalently) bound to an antibody or one or more antibody motifs / domains (eg, the Fc motif / domain of the antibody). Molecule, wherein the peptide moiety specifically binds to a desired target. Such peptide (s) may be fused to an Fc region or inserted into an Fc loop, a modified Fc molecule. The term “peptibodies” does not include Fc-fusion proteins (eg, full length proteins fused to Fc motifs / domains).

In some embodiments, the peptibody comprises (a) an antibody, and (b) a peptide disclosed herein; Here, peptides and antibodies maintain their activity in peptibodies. In some embodiments, the peptide is linked (directly or indirectly) to the antibody. In some embodiments, the peptide is covalently linked (directly or indirectly) to the antibody. In some embodiments, the peptide is bound (directly or indirectly) to the Fab region of an antibody. In some embodiments, the peptide is bound (directly or indirectly) to the antigen binding site of the antibody.

In some embodiments, the peptide is linked to the antibody via a reactive side chain. The reactive side chains may be naturally present or may be placed in the antibody by mutation. The reactive moiety of the antibody combining site may bind to the antibody, for example, when encoded by a nucleic acid present in the identified lymphoid cells that initially produce the antibody. Alternatively, amino acid residues can occur by intentionally mutating the DNA to encode specific residues. The reactive moiety can be a non-natural moiety resulting from biosynthetic introduction using, for example, unique codons, tRNAs, and aminoacyl-tRNAs as discussed herein. In another approach, amino acid residues or reactive functional groups thereof (eg, nucleophilic amino groups or sulfhydryl groups) may be attached to amino acid residues at the antibody combining site.

Catalytic antibodies are one source of antibodies comprising one or more reactive amino acid side chains. Such antibodies include aldolase antibodies, beta lactamase antibodies, esterase antibodies, amidase antibodies and the like.

In some embodiments, the peptide is indirectly bound to the antibody via a linker. In some embodiments, the linker includes alkyl, heteroalkyl, alkylene, alkenylene, alkynylene, heteroalkylene, carbocycle, heterocycle, aromatic ring, nonaromatic ring, substituted ring, monocyclic ring, polish Click rings, or combinations thereof.

In some embodiments, the antibody is IgA, IgD, IgE, IgG, or IgM. In some embodiments, the antibody is a humanized antibody.

In some embodiments, the peptibody is CovX ™ body.

III . MIF  Motif / Domain Destroyer  Analysis to confirm

In some embodiments, agents that bind to the MIF motifs / domains disclosed herein are identified. In some embodiments, agents binding to the MIF motifs / domains disclosed herein do not affect MIF independent signaling events in CXCR2 and CXCR4.

In some embodiments, peptide libraries are generated comprising extracellular N-terminal motifs / domains and / or extracellular loops of CXCR2 and CXCR4. In some embodiments, the peptide ranges in size from about 5 amino acids to about 20 amino acids; About 7 amino acids to about 18 amino acids; From about 10 amino acids to about 15 amino acids. In some embodiments, peptide libraries are screened for inhibition of MIF mediated signaling through CXCR2 and CXCR4 using any suitable method (eg, HTS GPCR screening method). In some embodiments, peptide libraries are further screened for inhibition of IL-8 and / or SDF-1 mediated signaling on CXCR2 and CXCR4. In some embodiments, if the peptide inhibits MIF signaling through CXCR2 and CXCR4 but allows SDF-1 and IL-8 mediated signaling through CXCR2 and CXCR4, the peptide is identified as being a MIF motif / domain disrupting peptide. Can be.

In some embodiments, peptide sequences from extracellular N-terminal motifs / domains and extracellular loops of CXCR2 and CXCR4 are arrayed on the membrane. In some embodiments, peptide sequences from extracellular N-terminal motifs / domains and extracellular loops of CXCR2 and CXCR4 are arrayed on the membrane and probed with full length MIF. In some embodiments, MIF is labeled (eg, isotope labeling, radiolabeling, or fluorophore labeling). In some embodiments, peptide sequences that specifically bind to labeled MIF are analyzed for inhibition of MIF mediated signaling through CXCR2 and CXCR4. In some embodiments, any suitable method (eg, GPCR screening assay) is used to screen peptide sequences that inhibit MIF mediated signaling through CXCR2 and CXCR4.

In some embodiments, any of the peptides and / or polypeptides mentioned above (eg, peptides derived from pseudo ELR motifs / domains of MIF or N-loop motifs / domains of MIF) have a structure-activity relationship (SAR: structure -activity relationship) as a "model" for performing chemistry (as specifically provided herein). In some embodiments, smaller chemistry is obtained through SAR chemistry. In some embodiments, MIF will bind to CXCR2 and / or CXCR4 via a smaller peptide (eg, by determining amino acid residues involved in breaking the ability of MIF to bind CXCR2 and / or CXCR4). Get small molecules that destroy your ability to

IV . MIF Trimerization Destroyer  Analysis to confirm

In some embodiments, MIF trimerization disrupting peptide is identified. In some embodiments, the MIF motif / domain trimerization disrupting peptide does not affect MIF independent signaling events in CXCR2 and CXCR4. In some embodiments, any of the amino acid sequences mentioned above (eg, amino acid residues 38-44 (beta-2 strand) of MIF, amino acid residues 48-50 (beta-3 strand) of MIF, amino acid residue 96 of MIF -102 (beta-5 strands), amino acid residues 107-109 (beta-6 strands) of MIF, amino acid residues N73, R74, S77, K78, and C81 of MIF, and / or amino acid residues N110, S111 of MIF, and Peptides and / or polypeptides derived from T112) are screened for inhibition of MIF mediated signaling via CXCR2 and CXCR4 using any suitable method (eg, GPCR screening technique).

In some embodiments, MIF trimerization disrupting peptide is identified. In some embodiments, the MIF motif / domain trimerization disrupting peptide does not affect MIF independent signaling events in CXCR2 and CXCR4. In some embodiments, any of the amino acid sequences mentioned above (eg, amino acid residues 38-44 (beta-2 strand) of MIF, amino acid residues 48-50 (beta-3 strand) of MIF, amino acid residue 96 of MIF -102 (beta-5 strands), amino acid residues 107-109 (beta-6 strands) of MIF, amino acid residues N73, R74, S77, K78, and C81 of MIF, and / or amino acid residues N110, S111 of MIF, and Peptides and / or polypeptides derived from T112) are used as a "model" for performing structure-activity relationship (SAR) chemistry. In some embodiments, smaller chemistry is obtained through SAR chemistry. In some embodiments, the ability of MIF to form homotrimers through smaller peptides (eg, by identifying amino acid residues involved in destroying the ability of MIF to form homotrimers). Obtain small molecules that destroy

In some embodiments, the MIF small molecule, peptide, and / or antibody antagonist is derived from any or all of amino acid residues 1-45 of SEQ ID NO: 1, and / or any of amino acid residues 1-45 of SEQ ID NO: 1 Or introduce them all. In some embodiments, the MIF small molecule, peptide, and / or antibody antagonist is a peptide derived from any or all of amino acid residues 2-45 of SEQ ID NO: 1, and / or in amino acid residues 2-45 of SEQ ID NO: 1 Introduce one or all of them. In some embodiments, the MIF small molecule, peptide, and / or antibody antagonist is a peptide derived from any or all of amino acid residues 3-45 of SEQ ID NO: 1, and / or in amino acid residues 3-45 of SEQ ID NO: 1 Introduce one or all of them. In some embodiments, the MIF small molecule, peptide, and / or antibody antagonist is a peptide derived from any or all of amino acid residues 4-45 of SEQ ID NO: 1, and / or in amino acid residues 4-45 of SEQ ID NO: 1 Introduce one or all of them. In some embodiments, the MIF small molecule, peptide, and / or antibody antagonist is a peptide derived from any or all of amino acid residues 5-45 of SEQ ID NO: 1, and / or in amino acid residues 5-45 of SEQ ID NO: 1 Introduce one or all of them. In some embodiments, the MIF small molecule, peptide, and / or antibody antagonist is a peptide derived from any or all of amino acid residues 6-45 of SEQ ID NO: 1, and / or in amino acid residues 6-45 of SEQ ID NO: 1 Introduce one or all of them. In some embodiments, the MIF small molecule, peptide, and / or antibody antagonist is a peptide derived from any or all of amino acid residues 7-45 of SEQ ID NO: 1, and / or in amino acid residues 7-45 of SEQ ID NO: 1 Introduce one or all of them. In some embodiments, the MIF small molecule, peptide, and / or antibody antagonist is a peptide derived from any or all of amino acid residues 8-45 of SEQ ID NO: 1, and / or in amino acid residues 8-45 of SEQ ID NO: 1 Introduce one or all of them. In some embodiments, the MIF small molecule, peptide, and / or antibody antagonist is a peptide derived from any or all of amino acid residues 9-45 of SEQ ID NO: 1, and / or in amino acid residues 9-45 of SEQ ID NO: 1 Introduce one or all of them. In some embodiments, the MIF small molecule, peptide, and / or antibody antagonist is a peptide derived from any or all of amino acid residues 10-45 of SEQ ID NO: 1, and / or in amino acid residues 10-45 of SEQ ID NO: 1 Introduce one or all of them.

In some embodiments, any of the amino acid sequences mentioned above (eg, amino acid residues 1-45 of SEQ ID NO: 1; amino acid residues 2-45 of SEQ ID NO: 1; amino acid residues 3-45 of SEQ ID NO: 1; SEQ ID NO: 1) Amino acid residues 4-45 of the amino acid residues 5-45 of SEQ ID NO: 1 amino acid residues 6-45 of SEQ ID NO: 1 amino acid residues 7-45 of SEQ ID NO: 1 amino acid residues 8-45 of SEQ ID NO: 1; Peptides and / or polypeptides derived from amino acid residues 9-45, or amino acid residues 10-45 of SEQ ID NO: 1, are used as "models" to perform structure-activity relationship (SAR) chemistry. In some embodiments, smaller chemistry is obtained through SAR chemistry. In some embodiments, the ability of MIF to form homotrimers through smaller peptides (eg, by determining amino acid residues involved in breaking the ability of MIF to form homotrimers). Obtain small molecules that destroy

In some embodiments, the antagonist of MIF is an antisense molecule and / or siRNA molecule complementary to the MIF gene and / or MIR RNA sequence. In some embodiments, siRNA and / or antisense molecules have a level or half-life of MIF mRNA and / or protein of at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, about 50 At least%, at least about 60%, at least about 80%, at least about 90%, at least about 95%, or substantially 100%.

V. Cell Line

In some embodiments, disclosed herein is a cell line expressing recombinant human CXCR4 + human CD74. In some embodiments, the cell line expressing recombinant human CXCR4 + human CD74 is a human cell line (eg, HEK293). In some embodiments, the cell line expressing recombinant human CXCR4 + human CD74 is a non-human cell line (eg, CHO).

VI. Inflammation

In some embodiments, the methods and compositions described herein treat an inflammatory disease, disease, condition, or symptom (eg, acute or chronic). In some embodiments, the methods and compositions described herein treat an inflammatory disease, disease, condition, or condition resulting from (partially or completely) an infection. In some embodiments, the methods and compositions described herein are (in part or completely) inflammatory diseases, diseases caused by tissue damage (eg, by burns, frostbite, exposure to cytotoxic agents, or trauma). Treat the condition, condition, or symptoms. In some embodiments, the methods and compositions described herein treat (in part or completely) an inflammatory disease, disease, condition, or symptom resulting from an autoimmune disease. In some embodiments, the methods and compositions described herein treat an inflammatory disease, disease, condition, or symptom resulting from the presence of a foreign body (eg, a thorn). In some embodiments, the methods and compositions described herein treat an inflammatory disease, disease, condition, or condition resulting from exposure to toxins and / or chemical stimulants.

As used herein, “acute inflammation” develops over a few minutes to several hours and stops when the irritation is removed (eg, an infectious agent is killed by an immune response, administration of a therapeutic agent, or a foreign body is involved in an immune response or extraction). By inflammation or healing of damaged tissue). The short duration of acute inflammation is due to the short half-life of most inflammatory mediators.

In certain cases, acute inflammation begins with the activation of white blood cells (eg, dendritic cells, endothelial cells, and mast cells). In certain cases, leukocytes release inflammatory mediators (eg, histamine, proteoglycans, serine proteases, icosanoids, and cytokines). In certain cases, inflammatory mediators cause (partially or completely) the symptoms associated with inflammation. For example, in certain cases, inflammatory mediators swell posterior capillary vasculature and increase vascular permeability. In certain cases, subsequent blood flow increase after vasodilation (partially or completely) results in redness and fever. In certain cases, increased vascular permeability leads to exudation of plasma into the tissue resulting in edema. In certain cases, the latter causes leukocytes to migrate to the site of inflammatory stimulation according to the chemotactic gradient. In addition, in certain cases, structural changes occur in blood vessels (eg, capillaries and vasculature). In certain cases, structural changes are induced (partially or completely) by monocytes and / or macrophages. In certain cases, structural changes include, but are not limited to, remodeling of blood vessels, and angiogenesis. In certain cases, angiogenesis causes increased transport of white blood cells, causing chronic inflammation to persist. In addition, in certain cases, histamine and bradykinin stimulate nerve endings resulting in itching and / or pain.

In certain cases, chronic inflammation is the presence of persistent irritants (eg, persistent acute inflammation, bacterial infections (eg, infections caused by mycobacterial tuberculosis)), prolonged exposure to chemical agents (eg, silica or tobacco smoke). From exposure and autoimmune responses (eg, rheumatoid arthritis). In certain cases, persistent stimulation continues to inflame (eg, due to continuous monocyte recruitment, and macrophage proliferation). In certain cases, continuous inflammation further damages the tissue and results in additional recruitment of mononuclear cells to sustain and worsen inflammation. In certain cases, the physiological response to inflammation also includes angiogenesis and fibrosis.

In some embodiments, the methods and compositions described herein treat an inflammatory disease, disease, condition, or symptom. In one non-limiting example, inflammatory diseases, diseases and conditions include atherosclerosis; Abdominal aortic aneurysm; Acute disseminated encephalomyelitis; Moyamoya disease; Takayasu's disease; Acute coronary syndrome; Cardiac allograft angiopathy; Lung inflammation; Acute respiratory distress syndrome; Pulmonary fibrosis; Acute disseminated encephalomyelitis; Addison's disease; Ankylosing spondylitis; Antiphospholipid antibody syndrome; Autoimmune hemolytic anemia; Autoimmune hepatitis; Autoimmune inner ear disease; Bullous whey; Shaga's disease; Chronic obstructive pulmonary disease; Celiac disease; Dermatitis; Type 1 diabetes; Type 2 diabetes; Endometriosis; Goodpasture syndrome; Graves' disease; Guillain-Barré syndrome; Hashimoto's disease; Idiopathic thrombocytopenic purpura; Interstitial cystitis; Systemic lupus erythematosus (SLE); Metabolic syndrome, multiple sclerosis; Myasthenia gravis; Myocarditis, narcolepsy; obesity; Vulgaris vulgaris; Pernicious anemia; Polymyositis; Primary biliary cirrhosis; Rheumatoid arthritis; Schizophrenia; Scleroderma; Sjogren's syndrome; Vasculitis; Vitiligo; Wegener's granulomatosis; Allergic rhinitis; Prostate cancer; Non-small cell lung carcinoma; Ovarian cancer; Breast cancer; Melanoma; Stomach cancer; Colon cancer; Brain cancer; Metastatic bone disease; Pancreatic cancer; Bladder cancer; Hepatocellular carcinoma; Liver cancer; Lung adenocarcinoma; Esophageal squamous cell carcinoma; CNS tumors (eg, glioblastoma and neuroblastoma); Blood tumors; Lymphoma; Nasal polyps; Gastrointestinal cancer; Ulcerative colitis; Crohn's disease; Collagen colitis; Lymphocytic colitis; Ischemic colitis; Converting colitis; Behcet's syndrome; Infectious colitis; Indeterminate colitis; Inflammatory liver disease, endotoxin shock, sepsis shock, rheumatoid spondylitis, ankylosing spondylitis, gouty arthritis, rheumatoid polymyalgia, Alzheimer's disease, Parkinson's disease, epilepsy, AIDS dementia, asthma, adult respiratory distress syndrome, bronchitis, acute leukocyte-mediated lung injury Distal proctitis, Wegener's granulomatosis, fibromyalgia, bronchitis, cystic fibrosis, uveitis, conjunctivitis, psoriasis, eczema, dermatitis, smooth muscle hyperplasia, meningitis, shingles, encephalitis, nephritis, tuberculosis, retinitis, atopic dermatitis, pancreatitis, periodontitis Gingivitis, coagulation necrosis, liquefaction necrosis, fibrotic necrosis, neoendometrial hyperplasia, myocardial infarction; stroke; Organ transplant rejection; Influenza (eg, H1N1 influenza A), or combinations thereof. In some embodiments, the methods and compositions disclosed herein treat, reduce, or prevent angiogenesis.

In some embodiments, the inflammatory disease, disease or condition is cancer. In some embodiments, the inflammatory disease, disease or condition is prostate cancer; Non-small cell lung carcinoma; Ovarian cancer; Breast cancer; Melanoma; Stomach cancer; Colon cancer; Brain cancer; Metastatic bone disease; Pancreatic cancer; Bladder cancer; Hepatocellular carcinoma; Liver cancer; Lung adenocarcinoma; Esophageal squamous cell carcinoma; CNS tumors (eg, glioblastoma and neuroblastoma); Blood tumors; Lymphoma; Or a combination thereof.

In some embodiments, the inflammatory disease, disease or condition is cardiovascular disease. In some embodiments, the inflammatory disease, disease or condition is atherosclerosis, peripheral vascular disease, cerebrovascular disease (ie stroke), hypertension (ie high blood pressure), heart failure, rheumatic heart disease, bacterial endocarditis, cardiomyopathy, pulmonary circulation disease , Venous & lymphatic system diseases, or a combination thereof.

Atherosclerosis

In some embodiments, the methods and compositions described herein treat atherosclerosis. As used herein, “atherosclerosis” means inflammation of the arterial wall and includes all atheromatous stages (eg, lipid deposition, intima-mediated thickening, and intima infiltration of monocytes) and all atherosclerotic lesions. (Type I to VIII lesions). In one embodiment, atherosclerosis results from (partially or completely) accumulation of macrophages. In some embodiments, the methods and compositions described herein prevent the accumulation of macrophages, reduce the number of accumulated macrophages, and / or slow down the rate of macrophage accumulation. In certain cases, atherosclerosis results from (partially or completely) the presence of oxidized LDL. In certain cases, oxidized LDL damages artery walls. In some embodiments, the methods and compositions described herein prevent oxidized LDL induced damage to the arterial wall, reduce a portion of the arterial wall damaged by the oxidized LDL, and / or reduce the severity of damage to the arterial wall / Or slow down the rate at which the artery wall is damaged by oxidized LDL. In certain cases, monocytes respond to damaged arterial walls (ie monocytes follow a chemotactic gradient to the damaged arterial walls). In certain cases, monocytes differentiate macrophages. In certain cases, macrophages endothelially import oxidized-LDL (cells, such as macrophages that endorse LLD, are called "foam cells"). In some embodiments, the methods and compositions described herein prevent the formation of foam cells, reduce the number of foam cells, and / or slow down the rate at which foam cells are formed. In certain cases, the foam cells die and subsequently rupture. In certain cases, rupture of the foam cells deposits oxidized cholesterol on the arterial walls. In some embodiments, the methods and compositions described herein prevent the deposition of oxidized cholesterol deposited on the arterial wall, and / or reduce the amount of oxidized cholesterol deposited on the arterial wall, and / or the oxidized cholesterol Slow down the rate at which it is deposited. In certain cases, arterial walls become inflamed due to damage caused by oxidized LDL. In some embodiments, the methods and compositions described herein prevent arterial wall inflammation, reduce a portion of the arterial wall where inflammation has occurred, and / or reduce the severity of inflammation. In certain cases, inflammation of the arterial wall causes (partially or completely) the expression of matrix metalloproteinase (MMP) -2, CD40 ligand, and tumor necrosis factor (TNF) α. In some embodiments, the methods and compositions described herein prevent the expression of matrix metalloproteinases (MMP) -2), CD40 ligands, and tumor necrosis factor (TNF) α, or expressed matrix metalloproteins. Reduce the amount of Inase (MMP) -2), CD40 ligand, and tumor necrosis factor (TNF) α. In certain cases, the cells form a firm envelope on the site of inflammation. In some embodiments, the methods and compositions described herein prevent firm shell formation, reduce the affected arterial wall portion due to the rigid shell, and / or slow down the rate at which the firm shell is formed. In certain cases, the cell envelope narrows the artery. In some embodiments, the methods and compositions described herein prevent arterial narrowing and / or reduce the portion of the artery being narrowed, reduce the severity of narrowing, and / or slow the rate at which the artery narrows.

In certain cases, atherosclerotic plaques (partially or completely) cause stenosis (ie, vascular narrowing). In certain cases, stenosis causes (partially or completely) blood flow reduction. In some embodiments, the methods and compositions described herein treat stenosis and / or restenosis. In certain cases, physical damage of stenosis atherosclerotic lesions due to percutaneous intervention (eg, balloon angioplasty or stent) leads to the development of endometrial hyperplasia. In certain cases, acute damage of the vascular wall not only leads to acute endothelial deprivation and platelet adhesion, but also to apoptosis of SMC in the mesenteric vascular wall. In certain cases, the accumulation of phenotypically unique SMCs in the lining layer in response to impaired function to preserve the integrity of the arterial vascular wall, however, results in gradual narrowing of the vessels thereafter. In certain cases, monocyte recruitment causes a more persistent and chronic inflammatory response. In some embodiments, the methods and compositions disclosed herein inhibit the accumulation of morphologically inherent SMCs in the intima. In some embodiments, the methods and compositions disclosed herein inhibit the accumulation of morphologically intrinsic SMCs in the endothelial layer in individuals treated with balloon angioplasty or stents.

In certain cases, rupture of the atherosclerotic plaque causes (partially or completely) infarction (eg, myocardial infarction or stroke) in the tissue. In certain cases, myocardial MIF expression is upregulated in the survival of cardiomyocytes and macrophages following acute myocardial ischemic injury. In certain cases, hypoxia and oxidative stress induce MIF secretion from cardiomyocytes through an atypical protein kinase C dependent export mechanism, resulting in extracellular signal-regulated kinase activation. In certain cases, elevated serum levels of MIF are detected in individuals with acute myocardial infarction. In certain cases, MIF contributes to macrophage accumulation in the infarcted area and contributes to the pre-inflammatory role of myocyte-induced damage during infarction. In some embodiments, the methods and compositions described herein treat infarction. In certain cases, reperfusion injury is involved after infarction. In some embodiments, the methods and compositions described herein treat reperfusion injury.

In some embodiments, the antibodies disclosed herein are administered to identify and / or locate atherosclerotic plaques. In some embodiments, the antibody is labeled for imaging. In some embodiments, the antibody is labeled for medical imaging. In some embodiments, the antibody is labeled for radiological imaging, PET imaging, MRI imaging, and fluorescence imaging. In some embodiments, the antibody is localized to a circulatory region with a high concentration of MIF. In some embodiments, the circulatory region with a high concentration of MIF is an atherosclerotic plaque. In some embodiments, the labeled antibody is prepared by any suitable method (eg, gamma camera, MRI, PET scanner, computed tomography (CT), functional magnetic resonance imaging (fMRI)). ) And single photon emission computed tomography (SPECT).

Abdominal aortic aneurysm

In certain cases, atherosclerotic plaques develop (partially or completely) aneurysms. In some embodiments, the methods and compositions described herein are administered for the treatment of an aneurysm. In some embodiments, the methods and compositions described herein are administered to treat an abdominal aortic aneurysm (“AAA”). As used herein, “abdominal aortic aneurysm” is a local hypertrophy of the abdominal aorta characterized by an increase of at least 50% above the normal aortic diameter. In some embodiments, the methods and compositions described herein reduce abdominal aortic hypertrophy.

In certain cases, abdominal aortic aneurysms are caused (partially or completely) due to breakage of structural proteins (eg, elastin and collagen). In some embodiments, the material compositions, methods, and / or pharmaceutical compositions disclosed herein partially or completely inhibit breakage of structural proteins (eg, elastin and collagen). The material compositions, methods and / or pharmaceutical compositions disclosed herein promote the regeneration of structural proteins (eg elastin and collagen). In certain cases, breakage of structural proteins is caused by activated MMPs. In some embodiments, the material compositions, methods, and / or pharmaceutical compositions disclosed herein partially or completely inhibit the activation of MMPs. In some embodiments, the compositions and / or methods disclosed herein inhibit upregulation of MMP-1, MMP-9 or MMP-12. In certain cases, MMPs are activated after abdominal aortic section infiltration by leukocytes (eg, macrophages and neutrophils).

In some embodiments, the methods and compositions described herein reduce infiltration of white blood cells. In certain cases, MIF upregulates early abdominal aortic aneurysm. In certain cases, leukocytes are gradients of the MIF into abdominal aortic sections sensitive to the development of AAA (eg, aortic sections affected by anastomosis, infection, cystic mesenteric necrosis, arteritis, trauma, anastomosis destruction causing pseudoaneurysm) To move accordingly. In some embodiments, the substance compositions, methods, and / or pharmaceutical compositions disclosed herein partially or completely inhibit the activity of MIF. In some embodiments, the material compositions, methods, and / or pharmaceutical compositions disclosed herein partially or completely inhibit the ability of MIF to act as chemokines on macrophages and neutrophils.

In some embodiments, the antibodies disclosed herein are administered to identify and / or locate AAA in an individual in need of identification and / or location of AAA. In some embodiments, an individual in need thereof exhibits one or more risk factors for developing AAA (eg, 60 years of age or older; male; smoking; high blood pressure; high serum cholesterol; diabetes; atherosclerosis). In some embodiments, the antibody is labeled for imaging. In some embodiments, the antibody is labeled for medical imaging. In some embodiments, the antibody is labeled for radiological imaging, PEG imaging, MRI imaging, and fluorescence imaging. In some embodiments, the antibody localizes to a circulatory region with a high concentration of MIF. In some embodiments, the circulatory region with a high concentration of MIF is AAA. In some embodiments, the labeled antibody is prepared by any suitable method (eg, gamma camera, MRI, PET scanner, computed X-ray tomography (CT), functional magnetic resonance imaging (fMRI), and single photon emission computed tomography). (SPECT)).

Other diseases

In some embodiments, the methods and compositions described herein treat T cell mediated autoimmune disease. In certain cases, T cell mediated autoimmune disease is characterized by a T cell mediated immune response against itself (eg, natural cells and tissues). Examples of T cell mediated autoimmune diseases include colitis, multiple sclerosis, arthritis, rheumatoid arthritis, osteoarthritis, juvenile arthritis, psoriatic arthritis, acute pancreatitis, chronic pancreatitis, diabetes, insulin-dependent diabetes mellitus (IDDM) ), Insulin salts, inflammatory bowel disease, Crohn's disease, ulcerative colitis, autoimmune hemolytic syndrome, autoimmune hepatitis, autoimmune neuropathy, autoimmune ovarian insufficiency, autoimmune testicles, autoimmune thrombocytopenia, reactive arthritis, ankylosing spondylitis, Secondary hematological findings of silicone graft-related autoimmune diseases, Sjogren's syndrome, systemic lupus erythematosus (SLE), vasculitis syndrome (eg giant cell arteritis, Behcet's disease & Wegener's granulomatosis), vitiligo, autoimmune disease (eg For example, anemia), drug-induced autoimmunity, Hashimoto's thyroiditis, pituititis, idiopathic thrombocytopenic purpura, metal Conductive autoimmunity, myasthenia gravis, blisters window, autoimmune hearing loss (e. G., Meni Fish Pathology), Good fasteners processing syndrome, Graves' disease, HIV-related autoimmune syndromes and gilraeng - including, Barre disease, and the like.

In some embodiments, the methods and compositions described herein treat pain. Pain includes, but is not limited to, acute pain, acute inflammatory pain, chronic inflammatory pain and neuropathic pain.

In some embodiments, the methods and compositions described herein treat hypersensitivity. As used herein, "hypersensitivity" means an unwanted immune system response. Hypersensitivity is divided into four categories. Type I hypersensitivity includes allergies (eg, atopy, anaphylaxis, or asthma). Type II hypersensitivity is cytotoxic / antibody mediated (e.g., autoimmune hemolytic anemia, thrombocytopenia, fetal fluorosis or goodpasture syndrome). Type III is an immune complex disease (e.g., serum sickness, Artus response or SLE). Type IV is delayed-type hypersensitivity (DTH), a cell-mediated immune memory response and is antibody independent (eg contact dermatitis, tuberculin skin test or chronic transplant rejection).

As used herein, “allergy” is a disease characterized by excessive activation of basophils and mast cells by IgE. In certain cases, excessive activation of mast cells and basophils by IgE causes (in part or completely) an inflammatory response. In certain cases, the inflammatory response is local. In certain cases, the inflammatory response causes narrowing of the airways (ie bronchial contraction). In certain cases, the inflammatory response causes inflammation of the nose (ie rhinitis). In certain cases, the inflammatory response is systemic (ie anaphylaxis).

In some embodiments, the methods and compositions described herein treat angiogenesis. As used herein, “angiogenesis” refers to the formation of new blood vessels. In certain cases, angiogenesis occurs with chronic inflammation. In certain cases, angiogenesis is induced by monocytes and / or macrophages. In some embodiments, the material compositions, methods, and / or pharmaceutical compositions disclosed herein inhibit angiogenesis. In certain cases, MIF is expressed in endothelial progenitor cells. In certain cases, MIF is expressed in tumor associated neovascular structures.

In some embodiments, the invention includes a method of treating neoplasia. In certain cases, neoplastic cells induce an inflammatory response. In certain cases, part of the inflammatory response to neoplastic cells is angiogenesis. In certain cases, angiogenesis promotes the development of neoplasia. In some embodiments, the neoplasia is angiosarcoma, Ewing's sarcoma, osteosarcoma, and other sarcomas, breast carcinoma, caecum carcinoma, colon carcinoma, lung carcinoma, ovarian carcinoma, pharynx carcinoma, rectal S phase colon carcinoma, pancreatic carcinoma, kidney carcinoma Endometrial carcinoma, gastric carcinoma, liver carcinoma, head and neck carcinoma, breast carcinoma and other carcinoma, Hodgkin's lymphoma and other lymphoma, malignant and other melanoma, parotid tumor, chronic lymphocytic leukemia and other leukemia, astrocytoma, glioma, Hemangiomas, retinoblastoma, neuroblastoma, auditory neuroma, neurofibroma, trachoma and purulent granulomas.

In some embodiments, disclosed herein is a method of promoting angiogenesis comprising administering to a subject a MIF or MIF analog.

As used herein, “septicemia” is a disease characterized by systemic inflammation. In certain cases, inhibiting the expression or activity of MIF increases the survival rate of individuals with sepsis. In some embodiments, the methods and compositions described herein treat sepsis. In certain cases, sepsis causes (partially or completely) myocardial dysfunction (eg, myocardial dysfunction). In some embodiments, the methods and compositions described herein treat myocardial dysfunction resulting from sepsis (eg, myocardial dysfunction).

In certain cases, MIF induces kinase activation and phosphorylation in the heart (ie, indicators of cardiac inhibition). In some embodiments, the methods and compositions described herein treat myocardial dysfunction resulting from sepsis (eg, myocardial dysfunction).

In certain cases, LPS induces expression of MIF. In certain cases, MIF is induced by endotoxin during sepsis and acts as an initiation factor in myocardial inflammatory response, cardiomyocyte apoptosis and cardiac dysfunction.

In some embodiments, the methods and compositions described herein inhibit myocardial inflammatory response resulting from endotoxin exposure. In some embodiments, the methods and compositions described herein inhibit cardiomyocyte cell apoptosis resulting from endotoxin exposure. In some embodiments, the methods and compositions described herein inhibit cardiac dysfunction resulting from endotoxin exposure.

In certain cases, inhibition of MIF significantly (partially or completely) increases survival factors (eg, Bcl-2, Bax and phosphorylation-Akt) and improves cardiomyocyte survival and myocardial function. In some embodiments, the methods and compositions described herein increase the expression of Bcl-2, Bax or phosphorylated-Akt.

In certain cases, MIF mediates late and long-term cardiac suppression following burn injury and / or major tissue damage. In some embodiments, the methods and compositions described herein treat long term cardiac inhibition after burn injury. In some embodiments, the methods and compositions described herein treat long term cardiac inhibition after major tissue injury.

In certain cases, MIF is released from the lungs during sepsis.

In certain cases, antibody neutralization of MIF inhibits the development of autoimmune myocarditis and reduces its severity. In some embodiments, the methods and compositions described herein treat autoimmune myocarditis.

VII. Combination

In some embodiments, disclosed herein is (a) (i) binding of MIF to CXCR2 and CXCR4, and / or (ii) MIF activation of CXCR2 and CXCR4; (iii) the ability of MIF to form homomers; Or agents which inhibit combinations thereof; And (b) a synergistic combination of a second agent (“MIF mediated disease agent”) selected from agents that treat inflammatory diseases, diseases, conditions, and symptoms (“MIF mediated disease agents”). .

In some embodiments, disclosed herein is (a) (i) binding of MIF to CXCR2 and CXCR4, and / or (ii) MIF activation of CXCR2 and CXCR4; (iii) the ability of MIF to form homomers; Or agents which inhibit combinations thereof; And (b) a synergistic combination of a second agent selected from agents that treat a disease whose component is inflammation.

In some embodiments, disclosed herein is (a) (i) binding of MIF to CXCR2 and CXCR4, and / or (ii) MIF activation of CXCR2 and CXCR4; (iii) the ability of MIF to form homomers; Or agents which inhibit combinations thereof; And (b) a synergistic combination of a second active agent selected from agents that treat unwanted inflammatory side effects. In certain cases, statins (eg, atorvastatin, lovastatin and simvastatin) induce inflammation. In certain cases, statin administration causes (partially or completely) myositis.

As used herein, the terms “pharmaceutical combination,” “perform additional therapy,” “administering additional therapeutic agent,” and the like refer to pharmaceutical therapies produced through the mixing or combination of more than one active ingredient, and Both fixed and unfixed combinations. The term "fixed combination" means that one or more of the agents described herein, and one or more auxiliary agents, both of which are administered simultaneously to the individual in a single formulation or in the form of a single entity. The term "unfixed combination" means the administration of one or more of the formulations described herein, and one or more adjuvants to the subject as individual entities simultaneously, simultaneously, or continuously with various intervening dates; Wherein, such administration may provide two or more agents at effective levels in the body of the individual. In some cases, the adjuvant is administered once or for a period of time, after which the agent is administered once or for a period of time. In other cases, the adjuvant is administered for a period of time, after which it is administered in a therapy comprising administration of both the adjuvant and both agents. In another embodiment, the formulation is administered once or for a period of time, after which the adjuvant formulation is administered once or for a period of time. This also applies to cocktail therapy, eg the administration of three or more active ingredients.

As used herein, the terms "co-administration," "administered in combination with," and grammatical synonyms, refer to the administration of a selected therapeutic agent to a single individual, and the agent is administered through the same or through different routes of administration or the same. It is intended to include treatment regimens that are administered at different or different times. In some embodiments, the formulations described herein will be co-administered with other formulations. This term includes administering two or more agents to an animal such that both agents and / or metabolites thereof are present in the animal at the same time. This includes simultaneous administration in separate compositions, at different time points in separate compositions, and / or in compositions in which both agents are present. Thus, in some embodiments, the formulations described herein and other formulation (s) are administered in a single composition. In some embodiments, the formulations described herein and other formulation (s) are mixed into the composition.

When considering combination therapy or prophylaxis, the agents described herein are not limited by the specific nature of the combination. For example, the formulations described herein are optionally administered in combination as a simple mixture as well as as a chemical hybrid. An example of the latter is when the agent is covalently bound to the target carrier or active agent. Covalent bonding can be performed in a number of ways, for example, but not by way of limitation, through the use of commercially available crosslinking agents. In addition, the combination treatment is optionally administered separately or incidentally.

In some embodiments, (a) an agent disclosed herein; And (b) co-administration of the second agent (partially or completely), allows the medical professional to increase the prescribed dose of the agent for MIF mediated disease. In certain cases, myositis induced through statins is dose dependent. In some embodiments, the formulation of an agent (partially or completely) allows a medical professional to increase the prescribed dose of statins.

In some embodiments, (a) the formulation; And (b) co-administration of the second agent (partially or completely), the medical professional can prescribe the second agent (ie, co-administration rescues the agent for MIF mediated disease).

In some embodiments, the second agent is an agent that targets HDL levels via indirect means (eg, CETP inhibition). In some embodiments, the non-selective HDL therapy comprises (1) an agent disclosed herein; (2) modulators of interaction between RANTES and platelet factor 4; Or (3) combination with a combination thereof converts the second agent, which targets HDL levels by indirect means, into a more effective therapy.

In some embodiments, the second agent is administered before, after, or concurrent with the administration of the inflammatory modulator.

VIII. Pharmaceutical therapy

In some embodiments, the second agent is niacin, fibrate, statins, Apo-A1 mimetic peptides (eg, DF-4, Novartis), apoA-I transcription upregulators, ACAT inhibitors, CETP modulators, glycoproteins (GP: Glycoprotein) IIb / IIIa receptor antagonists, P2Y12 receptor antagonists, Lp-PLA2-inhibitors, anti-TNF agents, IL-1 receptor antagonists, IL-2 receptor antagonists, cytotoxic agents, immunomodulators, antibiotics, T-cell joint Irritant blockers, disease modifying antirheumatic agents, B cell scavengers, immunosuppressants, antilymphocyte antibodies, alkylating agents, anti-metabolites, plant alkaloids, terpenoids, topoisomerase inhibitors, antitumor antibiotics, monoclonal antibodies, hormone therapy (Eg, aromatase inhibitors), or a combination thereof.

In some embodiments, the second agent is niacin, bezapiperate; Ciprobiplate; Clofibrate; Gemfibrozil; Fenofibrate; DF4 (Ac-D-W-F-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-NH2); DF5; RVX-208 from Resverlogix; Abashimib; Factimid sulfate (CS-505); CI-1011 (2,6-diisopropylphenyl [(2,4,6-triisopropylphenyl) acetyl] sulfamate); CI-976 (2,2-dimethyl-N- (2,4,6-trimethoxyphenyl) dodecanamide); VULM1457 (1- (2,6-diisopropyl-phenyl) -3- [4- (4'-nitrophenylthio) phenyl] urea); CI-976 (2,2-dimethyl-N- (2,4,6-trimethoxyphenyl) dodecanamide); E-5324 (n-butyl-N '-(2- (3- (5-ethyl-4-phenyl-1H-imidazol-1-yl) propoxy) -6-methylphenyl) urea); HL-004 (N- (2,6-diisopropylphenyl) tetradecylthioacetamide); KY-455 (N- (4,6-dimethyl-1-pentylindolin-7-yl) -2,2-dimethylpropanamide); FY-087 (N- [2- [N'-pentyl- (6,6-dimethyl-2,4-peptadiinyl) amino] ethyl]-(2-methyl-1-naphthyl-thio) acetamide) ; MCC-147 from Mitsubishi Pharma; F 12511 ((S) -2 ', 3', 5'-trimethyl-4'-hydroxy-alpha-dodecylthioacetanilide); SMP-500 from Sumitomo Pharmaceuticals; CL 277082 (2,4-difluoro-phenyl-N [[4- (2,2-dimethylpropyl) phenyl] methyl] -N- (heptyl) urea); F-1394 ((1s, 2s) -2- [3- (2,2-dimethylpropyl) -3-nonylureido] aminocyclohexan-1-yl 3- [N- (2,2,5,5 -Tetramethyl-1,3-dioxane-4-carbonyl) amino] propionate); CP-113818 (N- (2,4-bis (methylthio) -6-methylpyridin-3-yl) -2- (hexylthio) decanoic acid amide); YM-750; Torcetrapib; Anacetrapide; JTT-705 from Japan Tobacco / Roche; Absiximab; Eptipibatide; Tyropiban; Roxypiban; Variabiline; XV 459 (N (3)-(2- (3- (4-formamidinophenyl) isoxazolin-5-yl) acetyl) -N (2)-(1-butyloxycarbonyl) -2,3 Diaminopropionate); SR 121566A (3- [N- {4- [4- (aminoiminomethyl) phenyl] -1,3-thiazol-2-yl} -N- (1-carboxymethylpiperid-4-yl) aminol Propionic acid, trihydrochloride); FK419 ((S) -2-acetylamino-3-[(R)-[1- [3- (piperidin-4-yl) propionyl] piperidin-3-ylcarbonyl] amino] propionic acid 3 Hydrate); Clopidogrel; Prasugrel; Cancrylol; AZD6140 from AstraZeneca; MRS 2395 (2,2-Dimethyl-propionic acid 3- (2-chloro-6-methylaminopurin-9-yl) -2- (2,2-dimethyl-propionyloxymethyl) -propyl ester); BX 667 from Berlex Biosciences; BX 048 from Berlex Biosciences; Daraplapip (SB 480848); SB-435495 from Glaxo Smith Kline; SB-222657 from Glaxo Smith Kline; SB-253514 from Glaxo Smith Kline; Alefacept, epalizumab, methotrexate, acitretin, isotretinoin, hydroxyurea, mycophenolate mofetil, sulfasalazine, 6-thioguanine, walknex, taclonex, betamethasone, tazarotene, hydroxychloroquine, Sulfasalazine, etanercept, adalimumab, infliximab, avataravacept, rituximab, trastuzumab, anti-CD45 monoclonal antibody AHN-12 (NCI), iodine-131 anti-B1 antibody (Corixa Corp. ), Anti-CD66 monoclonal antibody BW 250/183 (NCI, Southampton General Hospital), anti-CD45 monoclonal antibody (NCI, Baylor College of Medicine), antibody antianb3 integrin (NCI), BIW-8962 (BioWa Inc.). ), Antibody BC8 (NCI), antibody muJ591 (NCI), indium In 111 monoclonal antibody MN-14 (NCI), yttrium Y 90 monoclonal antibody MN-14 (NCI), F105 monoclonal antibody (NIAID) , Monoclonal antibody RAV12 (Raven Biotechnologies), CAT-192 (human anti-TGF-beta1 monoclonal antibody, Genzyme), antibody 3F8 (NCI), 177Lu-J591 (Weill Medical College of Cornell University), TB-403 (Biolnvent International AB), Anakinra, Azathioprine, Cyclophosphamide, Cyclosporin A, Leflunomide, d-Penisilamine, Amitriptyline, or Notriptyline, chlorambucil, nitrogen mustard, prasterone, LJP 394 (sodium abetatimus), LJP 1082 (La Jolla Pharmaceutical), eculizumab, belimumab, rhuCD40L (NIAID), epratuzumab, silolimus, Tacrolimus, pimecrolimus, thalidomide, anti-thymocyte globulin-horse (Atgam, Pharmacia Upjohn), anti-thymocyte globulin-rabbit (Thymoglobulin, Genzyme), Muromoab-CD3 (FDA Office of Orphan Products Development), basiliximab, daclizumab, rilusol, cladribine, natalizumab, interferon beta-1b, interferon beta-1a, tizanidine, baclofen, mesalazine, asacol, pentasa , Mesalamine, valsalazide, olsalazine, 6-mercaptopurine, AIN457 (anti IL-17 monoclone Antibody, Novartis), theophylline, D2E7 (human anti-TNF mAb, Knoll Pharmaceuticals), mepolizumab (anti-IL-5 antibody, SB 240563), canakinumab (anti-IL-1 beta antibody, NIAMS), anti-IL-2 Receptor Antibody (Daclixumab, NHLBI), CNTO 328 (Anti IL-6 Monoclonal Antibody, Centocor), ACZ885 (Full Human Anti-Interleukin-1beta Monoclonal Antibody, Novartis), CNTO 1275 (Fully Human Anti-IL- 12 monoclonal antibody, Centocor), (3S) -N-hydroxy-4-({4-[(4-hydroxy-2-butynyl) oxy] phenyl} sulfonyl) -2,2-dimethyl- 3-thiomorpholine carboxamide (apratastat), golimumab (CNTO 148), oneercept, BG9924 (Biogen Idee), certolizumab pegol (CDP870, UCB Pharma), AZD9056 (AstraZeneca), AZD5069 (AstraZeneca), AZD9668 (AstraZeneca), AZD7928 (AstraZeneca), AZD2914 (AstraZeneca), AZD6067 (AstraZeneca), AZD3342 (AstraZeneca), AZD8309 (AstraZeneca), [(1R) -3-methyl-1-(((2S) -3-phenyl -2-[(pyrazin-2-ylcarbonyl) amino] propanoyl} amino) butyl] boronic acid (Bortezomib), AMG-714, ( IL 15 human monoclonal antibody, Amgen), ABT-874 (anti-IL-12 monoclonal antibody, Abbott Labs), MRA (Tocilizumab, anti-IL-6 receptor monoclonal antibody, Chugai Pharmaceutical), CAT-354 ( Human antiinterleukin-13 monoclonal antibody, Cambridge Antibody Technology, Medlmmune), aspirin, salicylic acid, gentisic acid, choline magnesium salicylate, choline salicylate, choline magnesium salicylate, choline salicylate, magnesium salicylate Latex, sodium salicylate, diflunisal, carpropene, phenopropene, phenopropene calcium, fluroprofen, ibuprofen, kentoprofen, nabutone, ketololac, ketorolac tromethamine , Naproxen, oxaprozin, diclofenac, etodolak, indomethacin, sulindac, tolmethine, meclofenamate, meclofenamate sodium, mefenamic acid, pyrocampam, meloxycamp, celecoxib, lofecoxib, Valdecockship, Parekcockship, Etoricockship, Lumina Cockship, CS-502 (Sankyo), JTE-522 (Japan Tobacco Inc.), L-745,337 (Almirall), NS398 (Sigma), Betamethasone (Celestone), Prednisone (Deltasone), Alcromethasone, Aldosterone, Amcinonini De, beclomethasone, betamethasone, budesonide, ciclesonide, clobetasol, clobetason, clocortolone, clofredol, cortisone, cortibazole, deplazacoat, deoxycorticosterone, desonide , Desoxymethasone, desoxy corton, dexamethasone, diploson, diflucortolone, diflufredate, fluchlorone, fludrocortisone, fluoxycortide, flumethasone, flunisolid, Nolon acetonide, fluorosinide, fluorocortin, fluorocortolone, fluorometholone, fluperolone, fluprednidene, fluticasone, formocoltal, formoterol, halcinolone, halomethasone, hydrocortisone Hydrocortisone acenate, hy Locortisone Buterate, Hydrocortisone Butyrate, Loteprednol, Medridone, Mefrednison, Methylprednisolone, Methylprednisolone Acynate, Mometasone Furoate, Paramethasone, Prednicabate, Prednisone, Limexolone, Thixocor Tol, triamcinolone, ulabetasol; Cisplatin; Carboplatin; Oxaliplatin; Mechlorethamine; Cyclophosphamide; Chlorambucil; Vincristine; Vinblastine; Vinorelbine; Bindeseo; Azathioprine; Mercaptopurine; Fludarabine; Pentostatin; Cladribine; 5-fluorouracil (5FU); Phloxuridine (FUDR); Cytosine arabinoside; Methotrexate; Trimetapririm; Pyrimethamine; Pemetrexed; Paclitaxel; Docetaxel; Etoposide; Tenifocide; Irinotecan; Topotecan; Amsacrine; Etoposide; Etoposide phosphate; Tenifocide; Dactinomycin; Doxorubicin; Daunorubicin; Varubicin; Idarubicin; Epirubicin; Bleomycin; Plicamycin; Mitomycin; Trastuzumab; Cetuximab; Rituximab; Bevacizumab; Finasteride; Goserelin; Aminoglutetimides; Anastrozole; Letrozole; Borosol; Exemestane; 4-Androsten-3,6,17-trione ("6-OXO"; 1,4,6-androstatriene-3,17-dione (ATD); formmestan; testosterone; padro Sol, milatuzumab, milatuzumab conjugated to doxorubicin, or a combination thereof.

Gene therapy

In some embodiments, disclosed herein is (a) an agent disclosed herein; And (b) a combination of gene therapies. In some embodiments, disclosed herein is (a) an agent disclosed herein; And (b) co-administering a combination of gene therapies.

In some embodiments, gene therapy comprises adjusting the concentration of lipids and / or lipoproteins (eg, HDL) in the blood of an individual in need of adjustment of lipid and / or lipoprotein (eg, HDL) concentrations in the blood. Include. In some embodiments, adjusting the concentration of lipids and / or lipoproteins (eg, HDL) in the blood comprises transfecting the DNA into an individual in need thereof. In some embodiments, the DNA encodes an Apo A1 gene, LCAT gene, LDL gene, IL-4 gene, IL-10 gene, IL-1ra gene, galectin-3 gene, or a combination thereof. In some embodiments, the DNA transfects liver cells.

In some embodiments, the DNA is transfected into liver cells through the use of ultrasound. See US Pat. No. 7,211,248 for the disclosure of techniques related to Apo A1 DNA transfection using ultrasound (these patents are incorporated herein by reference for the disclosure).

In some embodiments, the individual is administered a vector (eg, a "gene vector") engineered to carry human genes. See US Pat. No. 6,784,162 for the disclosure of techniques for generating LDL gene vectors (these patents are incorporated herein by reference for such disclosure). In some embodiments, the genetic vector is a retrovirus. In some embodiments, the genetic vector is not a retrovirus (eg, adenovirus; lentivirus; or a polymer delivery system, eg, METAFECTENE, SUPERFECT®, EFFECTENE®, or MIRUS TRANSIT). In certain cases, retroviruses, adenoviruses, or lentiviruses may be mutated to render the virus incapable.

In some embodiments, the vector is in vivo (ie, the vector is injected directly into an individual, eg, into liver cells), in vitro (ie, cells from the individual are grown in vitro and transduced into a gene vector). Introduced, inserted into a carrier and then implanted into a subject), or a combination thereof.

In certain cases, after administration of the gene vector, the gene vector infects the cells at the site of administration (eg, liver). In certain instances, the gene sequence (eg, when the gene vector is a retrovirus) is introduced into an individual's genome. In certain cases, the therapy (eg, if the gene vector is not a retrovirus) will require periodic re-administration. In some embodiments, the therapy is re-administered annually. In some embodiments, the therapy is re-administered every half year. In some embodiments, the therapy is re-administered when the individual's HDL level decreases below about 60 mg / dL. In some embodiments, the therapy is re-administered when the individual's HDL level decreases below about 50 mg / dL. In some embodiments, the therapy is re-administered when the individual's HDL level decreases below about 45 mg / dL. In some embodiments, the therapy is re-administered when the individual's HDL level decreases below about 40 mg / dL. In some embodiments, the therapy is re-administered when the individual's HDL level decreases below about 35 mg / dL. In some embodiments, the therapy is re-administered when the individual's HDL level decreases below about 30 mg / dL.

RNAi  therapy

In some embodiments, disclosed herein is (a) an agent disclosed herein; And (b) a combination of RNAi molecules designed to silence the expression of genes ("target genes") that participate in the onset and / or progression of MIF mediated disease. . In some embodiments, disclosed herein is (a) an agent disclosed herein; And (b) administering a combination of RNAi molecules designed to silence the expression of genes ("target genes") that participate in the onset and / or progression of MIF mediated disease. It starts. In some embodiments, the target gene is Apo B: Apolipoprotein B, Heat Shock Protein 110 (Hsp 110), Proprotein Convertase Subtilisin Kexin 9 (Pcsk9). , CyD1, TNFα, IL-1β, Natriuretic Peptide Receptor A (NPRA), GATA-3, Syk, VEGF, MIP-2, FasL, DDR-1, C5aR, AP-1, or these Is a combination.

In some embodiments, the target gene is silenced by RNA interference (RNAi). In some embodiments, RNAi therapy comprises the use of siRNA molecules. In some embodiments, a double stranded RNA (dsRNA) molecule having a sequence complementary to the mRNA sequence of a gene (eg, Apo B, Hsp 110 and Pcsk9) to be silenced (eg, by PCR ). Create In some embodiments, a 20-25 bp siRNA molecule is generated having a sequence complementary to the mRNA of the gene to be silenced. In some embodiments, a 20-25 bp siRNA molecule has 2-5 bp overhangs, 5 'phosphate ends, and 3' hydroxyl ends on the 3 'end of each strand. In some embodiments, 20-25 bp siRNA molecules have blunt ends. Techniques for generating RNA sequences are described in Molecular Cloning: A Laboratory Manual, second edition (Sambrook et al., 1989) and Molecular Cloning: A Laboratory Manual, third edition (Sambrook and Russel, 2001). (Current Protocols in Molecular Biology (FM Ausubel et al., Eds., 1987, including supplements through 2001)); [Current Protocols in Nucleic Acid Chemistry John Wiley & Sons, Inc., New York, 2000))), which are incorporated herein by reference in their entirety for this disclosure.

In some embodiments, siRNA molecules are “completely complementary” (ie, 100% complementary) to a target gene. In some embodiments, the antisense molecule is “almost complementary” to the target gene (eg, 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90 %, 85%, 80%, 75%, or 70% complementary). In some embodiments, there is a 1 bp mismatch, 2 bp mismatch, 3 bp mismatch, 4 bp mismatch, or 5 bp mismatch.

In certain cases, following administration of a dsRNA or siRNA molecule, cells at the site of administration (eg, liver and / or small intestine cells) are transformed with dsRNA or siRNA molecules. In certain cases, after transformation, the dsRNA molecule is cleaved into a plurality of fragments of about 20-25 bp to produce siRNA molecules. In certain cases, the fragments have about 2 bp overhangs on the 3 'end of each strand.

In certain cases, siRNA molecules are divided into two strands by RNA-induced Silencing Complex (RISC) (guide strand and antiguide strand). In certain cases, the guide strand is introduced into the catalytic component of the RISC (ie, argonaute). In certain cases, the guide strand specifically binds to the complementary RB1 mRNA sequence. In certain cases, RISC cleaves the mRNA sequence of the gene to be silenced. In certain cases, the expression of the gene to be silenced is down regulated.

In some embodiments, a sequence complementary to the mRNA sequence of the target gene is introduced into the vector. In some embodiments, the sequence is located between two promoters. In some embodiments, the promoters are oriented in opposite directions. In some embodiments, the vector is contacted with a cell. In certain cases, the cells are transformed with the vector. In certain cases, after transformation, sense and antisense strands of the sequence are generated. In certain cases, dsRNA molecules are produced in which the sense and antisense strands hybridize and are cleaved into siRNA molecules. In certain cases, the strands hybridize to produce siRNA molecules. In some embodiments, the vector is a plasmid (eg, pSUPER; pSUPER.neo; pSUPER.neo + gfp).

In some embodiments, siRNA molecules are administered in vivo (ie, vectors are injected directly into an individual, eg, into liver cells or small intestine cells, or into the bloodstream).

In some embodiments, siRNA molecules are delivery vehicles (eg, liposomes, biodegradable polymers, cyclodextrins, PLGA microspheres, PLCA microspheres, biodegradable nanocapsules, bioadhesive microspheres, or proteinaceous vectors), carriers And diluents, and pharmaceutically acceptable excipients. Methods for formulating and administering nucleic acid molecules to individuals in need thereof are described in Akhtar et al., 1992, Trends Cell Bio., 2, 139; Delivery Strategies for Antisense Oligonucleotide Therapeutics, ed.Akhtar, 1995. Maurer et al., 1999, Mol. Membr. Biol., 16, 129-140; Hofland and Huang, 1999, Handb.Exp. Pharmacol., 137, 165-192; Lee et al., 2000, ACS Symp. Ser., 752, 184-192; US Pat. No. 6,395,713 (Beigelman et al.); PCT WO 94/02595 (Sullivan et al.); Gonzalez et al., 1999, Bioconjugate Chem., 10, 1068-1074; International PCT Publication Nos. WO 03/47518 and WO 03/46185 to Wang et al .; US Pat. No. 6,447,796; US Patent Application Publication No. US 2002130430; International PCT Publication No. WO 00/53722 (O 'Hare and Normand) and US Patent Application Publication No. 20030077829; US Provisional Patent Publication No. 60 / 678,531, all of which are incorporated herein by reference for the above disclosure.

In some embodiments, siRNA molecules described herein are administered to the liver in any suitable manner (eg, see Wen et al., 2004, World J Gastroenterol., 10, 244-9; Murao et al., 2002, Pharm Res., 19, 1808-14; Liu et al., 2003, Gene Ther., 10, 180-7; Hong et al., 2003, J Pharm Pharmacol., 54 , 51-8; Herrmann et al., 2004, Arch Virol., 149, 161 1-7; and Matsuno et al., 2003, Gene Ther., 10, 1559-66).

In some embodiments, siRNA molecules described herein are ionically administered to, for example, certain organs or compartments (eg, liver or small intestine). Non-limiting examples of iontophoretic delivery include, for example, , WO 03/043689 and WO 03/030989, which documents are incorporated herein by reference for the above disclosure.

In some embodiments, siRNA molecules described herein are administered systemically (ie, distributed systemically after systemic absorption or accumulation in vivo of siRNA molecules in the bloodstream). Routes of administration contemplated for systemic administration include, but are not limited to, intravenous, subcutaneous, portal vein, intraperitoneal and intramuscular. Each of these routes of administration exposes the siRNA of the invention to accessible diseased tissue (eg, liver).

In certain cases, the therapy will require periodic re-administration. In some embodiments, the therapy is re-administered annually. In some embodiments, the therapy is re-administered every half year. In some embodiments, the therapy is administered monthly. In some embodiments, the therapy is administered weekly. In some embodiments, the therapy is re-administered when the individual's HDL level decreases below about 60 mg / dL. In some embodiments, the therapy is re-administered when the individual's HDL level decreases below about 50 mg / dL. In some embodiments, the therapy is re-administered when the individual's HDL level decreases below about 45 mg / dL. In some embodiments, the therapy is re-administered when the individual's HDL level decreases below about 40 mg / dL. In some embodiments, the therapy is re-administered when the individual's HDL level decreases below about 35 mg / dL. In some embodiments, the therapy is re-administered when the individual's HDL level decreases below about 30 mg / dL.

See US Publication No. 2007/0293451, which is incorporated herein by reference for this disclosure, for disclosures of techniques related to silencing the expression of Apo B and / or Hsp110. For disclosures of techniques related to Pcsk9 expression silencing, see US Publication No. 2007/0173473, which is incorporated herein by reference for the disclosure.

Antisense  therapy

In some embodiments, disclosed herein is (a) an agent disclosed herein; And (b) a combination of antisense molecules designed to inhibit the expression and / or activity of DNA or RNA sequences ("target sequences") that participate in the development and / or progression of MIF mediated diseases. A composition is disclosed. In some embodiments, disclosed herein is (a) an agent disclosed herein; And (b) co-administering an antisense molecule designed to inhibit the expression and / or activity of a DNA or RNA sequence ("target sequence") that participates in the onset and / or progression of a MIF mediated disease. It discloses a method of adjusting. In some embodiments, inhibition of expression and / or activity of the target sequence includes the use of antisense molecules complementary to the target sequence. In some embodiments, the target sequence is microRNA-122 (miRNA-122 or mRNA-122), secretory phospholipase A2 (sPLA2: secretory phospholipase A2), intracellular adhesion molecule 1 (ICAM-1: intracellular adhesion molecule- 1), GATA-3, NF-κB, Syk, or a combination thereof. In certain cases, inhibiting the expression and / or activity of miRNA-122 (partially or completely) reduces the concentration of cholesterol and / or lipids in the blood.

In some embodiments, an antisense molecule complementary to a target sequence (eg, by PCR). Create. In some embodiments, the antisense molecule is about 15 to about 30 nucleotides. In some embodiments, the antisense molecule is about 17 to about 28 nucleotides. In some embodiments, the antisense molecule is about 19 to about 26 nucleotides. In some embodiments, the antisense molecule is about 21 to about 24 nucleotides. Techniques for generating RNA sequences are described in Molecular Cloning: A Laboratory Manual, second edition (Sambrook et al., 1989) and Molecular Cloning: A Laboratory Manual, third edition (Sambrook and Russel, 2001). Referred to herein as "Sambrook"); (Current Protocols in Molecular Biology (FM Ausubel et al., Eds., 1987, including supplements through 2001); [Current Protocols in Nucleic Acid Chemistry John Wiley & Sons, Inc. , New York, 2000)), which are incorporated herein by reference in their entirety for this disclosure.

In some embodiments, the antisense molecule is single stranded, double stranded, cyclic or hairpin. In some embodiments, the antisense molecule contains structural elements (eg, inner or terminal bulges or loops).

In some embodiments, the antisense molecule is “completely complementary” to the target sequence (ie, 100% complementary). In some embodiments, the antisense molecule is “almost complementary” to the target RNA sequence (eg, 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, 85%, 80%, 75%, or 70% complementary). In some embodiments, there is a 1 bp mismatch, 2 bp mismatch, 3 bp mismatch, 4 bp mismatch, or 5 bp mismatch.

In some embodiments, the antisense molecule is hybridized to the target sequence. As used herein, “hybridized” means that the nucleotides of the antisense molecule are paired with the corresponding nucleotides of the target sequence. In certain cases, hybridization involves forming one or more hydrogen bonds between paired nucleotides (eg, Watson-Crick, Hugstin, or reverse Hugstin hydrogen bonds).

In certain cases, hybridization occurs (partially or completely) to degrade, cleave and / or isolate RNA sequences.

In some embodiments, siRNA molecules are delivery vehicles (eg, liposomes, biodegradable polymers, cyclodextrins, PLGA microspheres, PLCA microspheres, biodegradable nanocapsules, bioadhesive microspheres, or proteinaceous vectors), carriers And diluents, and pharmaceutically acceptable excipients. Methods for formulating and administering nucleic acid molecules to individuals in need thereof are described in Akhtar et al., 1992, Trends Cell Bio., 2, 139; Delivery Strategies for Antisense Oligonucleotide Therapeutics, ed.Akhtar, 1995. Maurer et al., 1999, Mol. Membr. Biol., 16, 129-140; Hofland and Huang, 1999, Handb.Exp. Pharmacol., 137, 165-192; Lee et al., 2000, ACS Symp. Ser., 752, 184-192; US Pat. No. 6,395,713 (Beigelman et al.); PCT WO 94/02595 (Sullivan et al.); Gonzalez et al., 1999, Bioconjugate Chem., 10, 1068-1074; International PCT Publication Nos. WO 03/47518 and WO 03/46185 to Wang et al .; US Pat. No. 6,447,796; US Patent Application Publication No. US 2002130430; International PCT Publication No. WO 00/53722 (O 'Hare and Normand) and US Patent Application Publication No. 20030077829; US Provisional Patent Publication No. 60 / 678,531, all of which are incorporated herein by reference for the above disclosure.

In some embodiments, siRNA molecules described herein are administered to the liver in any suitable manner (eg, see Wen et al., 2004, World J Gastroenterol., 10, 244-9; Murao et al., 2002, Pharm Res., 19, 1808-14; Liu et al., 2003, Gene Ther., 10, 180-7; Hong et al., 2003, J Pharm Pharmacol., 54 , 51-8; Herrmann et al., 2004, Arch Virol., 149, 161 1-7; and Matsuno et al., 2003, Gene Ther., 10, 1559-66).

In some embodiments, siRNA molecules described herein are ionically administered to, for example, certain organs or compartments (eg, liver or small intestine). Non-limiting examples of iontophoretic delivery include, for example, , WO 03/043689 and WO 03/030989, which documents are incorporated herein by reference for the above disclosure.

In some embodiments, siRNA molecules described herein are administered systemically (ie, distributed systemically after systemic absorption or accumulation in vivo of siRNA molecules in the bloodstream). Routes of administration contemplated for systemic administration include, but are not limited to, intravenous, subcutaneous, portal vein, intraperitoneal and intramuscular. Each of these routes of administration exposes the siRNA of the invention to accessible diseased tissue (eg, liver).

In certain cases, the therapy will require periodic re-administration. In some embodiments, the therapy is re-administered annually. In some embodiments, the therapy is re-administered every half year. In some embodiments, the therapy is administered monthly. In some embodiments, the therapy is administered weekly. In some embodiments, the therapy is re-administered when the individual's HDL level decreases below about 60 mg / dL. In some embodiments, the therapy is re-administered when the individual's HDL level decreases below about 50 mg / dL. In some embodiments, the therapy is re-administered when the individual's HDL level decreases below about 45 mg / dL. In some embodiments, the therapy is re-administered when the individual's HDL level decreases below about 40 mg / dL. In some embodiments, the therapy is re-administered when the individual's HDL level decreases below about 35 mg / dL. In some embodiments, the therapy is re-administered when the individual's HDL level decreases below about 30 mg / dL.

See WO 07 / 027775A2, which is incorporated herein by reference for this disclosure, for disclosures of techniques related to silencing expression of miRNA-122.

Device paratherapy

In some embodiments, the device mediated strategy comprises removing lipids from HDL molecules in an individual in need thereof (degreasing), removing LDL molecules from blood or plasma of the individual in need thereof (degreasing), or Combinations thereof. For disclosures of techniques for lipid removal from HDL molecules and removal of LDL molecules from blood or plasma of individuals in need thereof, see US Publication No. 2008/0230465, which is incorporated herein by reference for said disclosure. ).

In certain cases, the therapy will require periodic re-administration. In some embodiments, the therapy is re-administered annually. In some embodiments, the therapy is re-administered every half year. In some embodiments, the therapy is administered monthly. In some embodiments, the therapy is administered weekly. In some embodiments, the therapy is re-administered when the individual's HDL level decreases below about 60 mg / dL. In some embodiments, the therapy is re-administered when the individual's HDL level decreases below about 50 mg / dL. In some embodiments, the therapy is re-administered when the individual's HDL level decreases below about 45 mg / dL. In some embodiments, the therapy is re-administered when the individual's HDL level decreases below about 40 mg / dL. In some embodiments, the therapy is re-administered when the individual's HDL level decreases below about 35 mg / dL. In some embodiments, the therapy is re-administered when the individual's HDL level decreases below about 30 mg / dL.

Pharmaceutical composition

In some embodiments, disclosed herein is a pharmaceutical composition for treating an inflammatory disease, disease, condition, or condition comprising a therapeutically effective amount of a formulation disclosed herein.

The pharmaceutical compositions herein are formulated using one or more physiologically acceptable carriers, including excipients and auxiliaries that facilitate processing of the formulation into a pharmaceutically used formulation. Proper formulation is dependent upon the route of administration chosen. A summary of pharmaceutical compositions can be found, for example, in Remington: The Science and Practice of Pharmacy, Nineteenth Ed (Easton, Pa .: Mack Publishing Company, 1995); Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liman, HA and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, NY, 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Ed (Lippincott Williams & Wilkins, 1999)].

In some embodiments, the pharmaceutical composition for modulating cardiovascular disease further comprises a pharmaceutically acceptable diluent (s), excipient (s) or carrier (s). In some embodiments, the pharmaceutical composition comprises other pharmaceutical or pharmaceutical agents, carriers, adjuvants, such as preservatives, stabilizers, wetting agents or emulsifiers, solution promoters, osmotic pressure regulating salts and / or buffers. In addition, the pharmaceutical compositions may also include other therapeutically valuable substances.

 The pharmaceutical formulations described herein may optionally contain multiple, oral, parenteral (eg, intravenous, subcutaneous, intramuscular), nasal, buccal, topical, rectal or transdermal routes of administration. Administration to the subject is via the route of administration. The pharmaceutical formulations described herein include aqueous liquid dispersions, self-emulsifying dispersions, solid solutions, liposome dispersions, aerosols, solid formulations, powders, immediate release formulations, controlled release formulations, rapid melt formulations, tablets, capsules, pills, delays Release formulations, long-release formulations, pulsatile release formulations, multiparticulate formulations, and mixed immediate and controlled release formulations.

The pharmaceutical compositions described herein can be used in oral dispersions, solutions, gels, syrups, elixirs, slurries, suspensions and the like, solid oral formulations, aerosols, controlled release formulations, rapid melt formulations, for oral ingestion of the subjects to be treated, Effervescent formulations, lyophilized formulations, tablets, powders, pills, dragees, capsules, modified release formulations, delayed release formulations, long-term release formulations, pulsatile release formulations, multiparticulate formulations, and mixed immediate and controlled release formulations. It is formulated in any suitable formulation, including but not limited to.

In some embodiments, the pharmaceutical compositions described herein are formulated into multiparticulate formulations. In some embodiments, the pharmaceutical compositions described herein comprise a first population of particles and a second population of particles. In some embodiments, the first group comprises an agent. In some embodiments, the second group comprises an agent. In some embodiments, the dose of the agent in the first group is the same as the dose of the agent in the second group. In some embodiments, the dose of the agent in the first group is not equal to (eg, more or less) the dose of the agent in the second group.

In some embodiments, the formulation of the first group is released before the formulation of the second group. In some embodiments, the second particle population comprises a modified release (eg, delayed release, controlled release or prolonged release) coating. In some embodiments, the second particle population comprises a modified release (eg, delayed release, controlled release or long term release) matrix.

Coating materials for use with the pharmaceutical compositions described herein include polymeric coating materials (eg, cellulose acetate phthalate, cellulose acetate trimaleate, hydroxy propyl methylcellulose phthalate, polyvinyl acetate phthalate); Ammonio methacrylate copolymers (eg, Eudragit® RS and RL); Polyacrylic acid and polyacrylate and methacrylate copolymers (eg, Eudragite S and L, polyvinyl acetaldiethylamino acetate, hydroxypropyl methylcellulose acetate succinate, cellac); Hydrogels and gel formers (e.g., carboxyvinyl polymers, sodium alginates, sodium carmelloses, calcium carmelloses, sodium carboxymethyl starch, polyvinyl alcohol, hydroxyethyl cellulose, methyl cellulose, gelatin, starch, hydroxy Propyl cellulose, hydroxypropyl methylcellulose, polyvinylpyrrolidone, crosslinked starch, microcrystalline cellulose, chitin, aminoacryl-methacrylate copolymer, pullulan, collagen, casein, agar, arabic gum, sodium carboxymethyl cellulose, (Swellable hydrophilic polymer) poly (hydroxyalkyl methacrylate) (m. Wt. To 5 k-5,000 k), polyvinylpyrrolidone (m. Wt. To 10 k-360 k), anionic and cationic Hydrogel, swellable mixture of polyvinyl alcohol, agar and carboxymethyl cellulose with low acetate residues, copolymers of maleic anhydride and sterin, ethylene, propylene or Isobutylene, pectin (m. Wt. ˜30 k-300 k), polysaccharides, for example agar, acacia, karaya, tragacanth, algin and guar, polyacrylamide, polyox® polyethylene oxide (m. wt. ˜100 k-5,000 k), AquaKeep® acrylate polymers, diesters of polyglucans, crosslinked polyvinyl alcohols and poly N-vinyl-2-pyrrolidone, sodium starch; hydrophilic polymers (E.g., polysaccharides, methyl cellulose, sodium or calcium carboxymethyl cellulose, hydroxypropyl methyl cellulose, hydroxypropyl cellulose, hydroxyethyl cellulose, nitro cellulose, carboxymethyl cellulose, cellulose ether, polyethylene oxide, methyl ethyl cellulose , Ethylhydroxy ethylcellulose, cellulose acetate, cellulose butyrate, cellulose propionate, gelatin, collagen, starch, maltodextrin, pullulan, polyvinyl pyrrolidone, Polyvinyl alcohol, polyvinyl acetate, glycerol fatty acid esters, polyacrylamides, polyacrylic acids, copolymers of methacrylic acid or methacrylic acid, other acrylic acid derivatives, sorbitan esters, natural gums, lecithin, pectin, alginates, ammonia Alginate, sodium, calcium, potassium alginate, propylene glycol alginate, agar, gum arabic, karaya gum, locust bean gum, tragacanth gum, carrageen gum, guar gum, xanthan gum, screroglucan sword); Or combinations thereof. In some embodiments, the coating material includes a plasticizer, a lubricant, a solvent, or a combination thereof. Suitable plasticizers include acetylated monoglycerides; Butylphthalyl butyl glycolate; Dibutyl tartrate; Diethyl phthalate; Dimethyl phthalate; Ethyl phthalyl ethyl glycolate; glycerin; Propylene glycol; Triacetin; Citrate; Tripropioin; Diacetin; Dibutyl phthalate; Acetyl monoglycerides; Polyethylene glycol; Castor oil; Triethyl citrate; Polyhydric alcohols, glycerol, acetate esters, glycerol triacetate, acetyl triethyl citrate, dibenzyl phthalate, dihexyl phthalate, butyl octyl phthalate, diisononyl phthalate, butyl octyl phthalate, dioctyl azelate, epoxidized talate, Triisooctyl trimellitate, diethylhexyl phthalate, di-i-octyl phthalate, di-i-decyl phthalate, di-n-undecyl phthalate, di-n-tridecyl phthalate, tri-2-ethylhexyl trimelli Tate, di-2-ethylhexyl adipate, di-2-ethylhexyl sebacate, di-2-ethylhexyl azelate, dibutyl sebacate.

In some embodiments, the second particle population comprises a modified release matrix material. Materials for use in the pharmaceutical compositions described herein include microcrystalline cellulose, sodium carboxymethylcellulose, hydroxyalkylcelluloses (e.g., hydroxypropylmethylcellulose and hydroxypropylcellulose), polyethylene oxide, alkylcelluloses ( Methylcellulose and ethylcellulose), polyethylene glycol, polyvinylpyrrolidone, cellulose acetate, cellulose acetate butyrate, cellulose acetate phthalate, cellulose acetate trimellitate, polyvinylacetate phthalate, polyalkylmethacrylate, polyvinyl Acetate, or combinations thereof.

In some embodiments, the first particle population comprises an agent for cardiovascular disease. In some embodiments, the second particle population comprises (1) a modulator of MIF; (2) modulators of interaction between RANTES and platelet factor 4; Or (3) combinations thereof. In some embodiments, the first particle population comprises (1) a modulator of MIF; (2) modulators of interaction between RANTES and platelet factor 4; Or (3) combinations thereof. In some embodiments, the second particle population comprises an agent for cardiovascular disease.

Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions are generally used, optionally with gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol, and / or titanium dioxide, lacquer liquor, and suitable organic solvents or solvents. It contains a mixture. Optionally, dyes or pigments are added to the tablets or dragee coatings to identify or characterize different combinations of formulation doses.

In some embodiments, the solid formulations disclosed herein comprise tablets (suspension tablets, quick melt tablets, occlusal disintegrating tablets, rapid disintegrating tablets, effervescent tablets or caplets), pills, powders (sterile packaged powders, dispersible powders, or boiling). Ingredients), capsules (both soft or hard capsules, for example, capsules made of animal gelatin or vegetable HPMC, or "sprinkle capsules"), solid dispersions, solid solutions, biocorrosive formulations, In the form of controlled release formulations, pulsatile release formulations, multiparticulate formulations, pellets, granules or aerosols. In other embodiments, the pharmaceutical formulation is in powder form. In another embodiment, the pharmaceutical formulation is in the form of a tablet, including but not limited to rapid melt tablets. In addition, the pharmaceutical formulations disclosed herein are optionally administered in a single capsule or multiple capsule formulation. In some embodiments, the pharmaceutical formulation is administered in two, or three, or four capsules or tablets.

In another aspect, the formulation comprises a microencapsulated formulation. In some embodiments, one or more other compatible materials are present as microencapsulation materials. Exemplary materials include pH modifiers, corrosion promoters, defoamers, antioxidants, flavoring agents, and carrier materials, such as binders, suspending agents, disintegrating agents, fillers, surfactants, solubilizers, stabilizers, lubricants, wetting agents And diluents.

Exemplary microencapsulation materials useful for delaying the release of agents, including MIF receptor inhibitors, include hydroxypropyl cellulose ethers (HPC), such as Klucel® or Nisso HPC, Low-substituted hydroxypropyl cellulose ethers (L-HPC), hydroxypropyl methyl cellulose ethers (HPMC), for example Sepppifilm-LC, Pharmacote Pharmacoat®, Metolose SR, Methocel®-E, Opadry YS, PrimaFlo, Benecel MP824, and Venecel MP843, methylcellulose polymers such as methocel®-A, hydroxypropylmethylcellulose acetate stearate Aqoat (HF-LS, HF-LG, HF-MS) and metholose®, ethylcellulose ( EC) and mixtures thereof such as E461, Ethocel®, Aqualon®-EC, Surelease®, Polyvinyl alcohol (PVA), for example Opadry AMB, hydroxyethylcellulose, for example Natrosol ®, salts of carboxymethylcellulose and carboxymethylcellulose (CMC), for example Aquaron®-CMC, polyvinyl alcohol and polyethylene glycol copolymers, such as Kollicoat IR®, mono Glycerides (Myverol), triglycerides (KLX), polyethylene glycols, modified food starch, acrylic polymers and mixtures of acrylic polymers and cellulose ethers, for example Eudragit® EPO, Eudragit ® L30D-55, Eudragit® FS 30D Eudragit® L100-55, Eudragit® L100, Eudragit® S 100, Eudragit® RD100, Eudragit® E100, Eudragit® L12. 5, Eudragit® S 12.5, Eudragit® NE30D, and Eudragit® NE 40D, Cellulose Acetate Phthalates, sepfifilms such as mixtures of HPMC and stearic acid, cyclodextrins, and mixtures of the foregoing materials.

Liquid formulation formulations for oral administration optionally include an aqueous suspension selected from the group including, but not limited to, aqueous oral dispersions, emulsions, solutions, elixirs, gels, and syrups. See, eg, Singh et al., Encyclopedia of Pharmaceutical Technology, 2nd Ed., Pp. 754-757 (2002). In addition to MIF receptor inhibitors, liquid formulations may optionally include additives such as (a) disintegrants; (b) dispersants; (c) wetting agents; (d) at least one preservative, (e) a viscosity enhancer, (f) at least one sweetener, and (g) at least one flavoring agent. In some embodiments, the aqueous dispersions further comprise a crystal formation inhibitor.

In some embodiments, the pharmaceutical formulations described herein are self-emulsifying drug delivery systems (SEDDS). Emulsions are usually dispersions in the form of droplets, an immiscible phase in another phase. In general, emulsions are produced by strong mechanical dispersion. Unlike emulsions or microemulsions, SEDDS spontaneously forms emulsions upon addition of excess water without any external mechanical dispersion or agitation. The advantage of SEDDS is that only mild mixing is required to distribute the droplets throughout the solution. In addition, water or an aqueous layer is optionally added immediately before administration to ensure the stability of the labile or hydrophobic active ingredient. Thus, SEDDS provides an efficient delivery system for oral and parenteral delivery of hydrophobic active ingredients. In some embodiments, SEDDS provides an improvement in the bioavailability of the hydrophobic active ingredient. Methods of making self-emulsifying formulations include, but are not limited to, for example, US Pat. Nos. 5,858,401, 6,667,048, and 6,960,563.

Suitable nasal preparations include, for example, those described in US Pat. Nos. 4,476,116, 5,116,817, and 6,391,452. Nasal formulations generally contain large amounts of water in addition to the active ingredient. Small amounts of other components, such as pH adjusters, emulsifiers or dispersants, preservatives, surfactants, gelling agents, or buffers and other stabilizers and solubilizers, are optionally present.

For inhalation administration, the pharmaceutical compositions disclosed herein are optionally in aerosol, mist or powder form. The pharmaceutical compositions described herein present aerosol sprays from a pressurized pack or inhaler using a suitable propellant such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, or other suitable gas. It is easily delivered in form. In the case of a pressurized aerosol, the unit dose is determined by providing a valve to deliver a metered amount. For example, as a mere example of a powder mix and suitably used in an inhaler or blower containing a powder base such as lactose or starch, cartridges and capsules of gelatin are formulated.

Buccal formulations include, but are not limited to, US Pat. Nos. 4,229,447, 4,596,795, 4,755,386, and 5,739,136. In addition, the buccal formulations described herein optionally comprise a biocorrosive (hydrolyzable) polymer carrier that also serves to attach the formulation to the buccal mucosa. The buccal formulation is made to gradually erode over a period of time. Buccal drug delivery can avoid the disadvantages encountered with oral drug administration, eg, slow absorption, degradation of the formulation by fluids present in the gastrointestinal tract and / or rapid passage inactivation in the liver. Bioerodible (hydrolyzable) polymer carriers generally include hydrophilic (water soluble and water expandable) polymers that adhere to the wet surface of the buccal mucosa. Examples of polymeric carriers useful herein include acrylic acid polymers and copolymers, such as those known as “carbomers” (Carbopol® obtained from B.F. Goodrich is one such polymer). Other components introduced into the buccal formulations described herein also include, but are not limited to, disintegrants, diluents, binders, lubricants, flavors, colorants, preservatives, and the like. For buccal or sublingual administration, the compositions may optionally take the form of tablets, rosin or gels formulated in a conventional manner.

Transdermal formulations of the pharmaceutical compositions disclosed herein are described, for example, in US Pat. Nos. 3,598,122, 3,598,123, 3,710,795, 3,731,683, 3,742,951, 3,814,097, 3,921,636, 3,972,995, 3,993,072, 3,993,073, 3,996,934, 4,031,894, 4,060,084, 4,069,307, 4,077,407, 4,201,211, 4,230,105, 4,292,299, 4,292,303,78,78,78,78 , 5,837,280, 5,869,090, 6,923,983, 6,929,801 and 6,946,144.

The transdermal formulations described herein comprise three or more components: (1) formulations; (2) penetration enhancers; And (3) aqueous adjuvants. In addition, transdermal formulations include ingredients including, but not limited to, gelling agents, creams, ointment bases, and the like, for example. In some embodiments, the transdermal formulation further comprises a woven or nonwoven support material to enhance absorption and prevent the removal of the transdermal formulation from the skin. In other embodiments, the transdermal formulations described herein remain saturated or supersaturated to facilitate diffusion into the skin.

In some embodiments, suitable formulations for transdermal administration are transdermal delivery devices and transdermal delivery patches, are lipophilic emulsions or buffered aqueous solutions, and are dissolved and / or dispersed in a polymer or adhesive. Such patches are optionally made continuously, pulsating, or as required for the delivery of the pharmaceutical formulation. In addition, transdermal delivery is optionally performed via iontophoretic patches and the like. In addition, transdermal patches are provided in controlled delivery. The rate of absorption is slowed down, optionally through the use of rate controlled membranes or by entrapping the formulation in a polymer matrix or gel. In contrast, absorption enhancers are used to increase absorption. Absorption enhancers or carriers include absorbable pharmaceutically acceptable solvents to aid in the passage of skin. For example, the transdermal device may be a support member, optionally a reservoir containing a carrier and the agent, a rate-controlled barrier for delivering the agent to the skin of the host, optionally at a controlled rate over a prolonged period of time, and for maintaining the device on the skin. It is in the form of a band containing means for.

Formulations suitable for intramuscular, subcutaneous or intravenous injection include physiologically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injections or dispersions. Examples of suitable aqueous and non-aqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols (propylene glycol, polyethylene glycol, glycerol, cremophors, etc.), suitable mixtures thereof, vegetable oils (eg olive oil) and injections. Possible organic esters such as ethyl oleate. Proper fluidity is maintained, for example, through the use of coatings such as lecithin, in the case of dispersions, through the maintenance of the required particle size, and through the use of surfactants. Formulations suitable for subcutaneous injection also include any additives such as preservatives, wetting agents, emulsifiers and dispersants.

For intravenous injection, the preparation is optionally formulated in an aqueous solution, preferably physiologically compatible, in a buffer such as Hank's solution, Ringer's solution or physiological saline buffer. For transmucosal administration, penetrants suitable for the barrier to be permeated are used in the formulation. For other parenteral injections, suitable formulations preferably include aqueous or non-aqueous solutions with physiological compatibility with buffers or excipients.

Parenteral injections optionally include bolus injection or continuous infusion. Injectable preparations are optionally present in unit dosage forms, such as ampoules or multiple dose containers, with an added preservative. In some embodiments, the pharmaceutical compositions described herein are in a form suitable for parenteral injection as an sterile suspension, solution or emulsion in an oily or aqueous vehicle, containing formulated preparations such as suspending, stabilizing and / or dispersing agents. do. Pharmaceutical formulations for parenteral administration include aqueous solutions of the formulations in water-soluble form. In addition, suspensions are optionally prepared as suitable oily suspensions for injection.

In some embodiments, the formulations disclosed herein are topically administered and formulated into a variety of topically administrable compositions, for example, in solutions, suspensions, lotions, gels, pastes, medical sticks, balms, creams, or ointments. do. Such pharmaceutical compositions optionally include solubilizers, stabilizers, isotonicity enhancers, buffers and preservatives.

The formulations disclosed herein are also optionally formulated with a rectal composition such as an enema, rectal gel, rectal foaming agent, rectal aerosol, suppository, jelly suppository, or retention type enema, which is a conventional suppository base, for example Synthetic polymers such as polyvinylpyrrolidone, PEG, and the like, including cocoa butter or other glycerides. In the form of suppositories, low melting waxes, such as, but not limited to, a mixture of fatty acid glycerides, optionally in combination with cocoa butter, are first melted.

The formulations disclosed herein are used in the manufacture of a medicament for the prophylactic and / or therapeutic treatment of inflammatory diseases, diseases, conditions and symptoms or conditions, optionally where at least partially improvement may be beneficial. In addition, a method of treating any of the diseases or conditions described herein in an individual in need thereof may comprise an agent described herein, or a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable N oxide, a pharmaceutical A pharmaceutical composition comprising an active metabolite, a pharmaceutical prodrug, a pharmaceutically acceptable solvate, is administered to the subject in a therapeutically effective amount.

If the condition of the subject has not improved at the physician's discretion, administration of the formulations disclosed herein is chronically ie, life of the subject, optionally to ameliorate, or otherwise control or limit the symptoms of the disorder or condition of the subject. It is administered long term throughout this duration.

At the physician's discretion, when the condition of the subject improves, the formulations described herein are optionally administered continuously; Alternatively, the dose of drug is temporarily reduced or temporarily suspended for a certain period of time (ie, a “drug holiday”). The drug holiday period can optionally vary from 2 days to 1 year, with only one example being 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 10 days, 12 days, 15 days, 20 days, 28, 35, 50, 70, 100, 120, 150, 180, 200, 250, 280, 300, 320, 350, or 365 days. Drug reduction rates during the drug holiday include 10% -100%, for example only 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60 %, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%.

Once the subject's condition improves, maintenance doses are administered as needed. The dose or frequency of administration, or both, is then reduced to a level where symptoms, conditions, or conditions that improve with symptoms are maintained. In some embodiments, the individual optionally requires intermittent treatment for a long time if symptoms recur.

In some embodiments, the pharmaceutical compositions described herein are unit dosage forms suitable for single administration of precise dosages. In unit dosage form, the formulation is divided into unit dosage forms containing appropriate amounts of the formulations disclosed herein. In some embodiments, the unit dosage form is in the form of a package containing discrete amounts of the formulation. Non-limiting examples are packaged tablets or capsules, and powders in vials or ampoules. In some embodiments, the aqueous suspension composition is packaged into a single dose nonreclosable container. Alternatively, a reclosed container for multiple dosing is used, which typically includes a preservative in the composition. By way of example only, parenteral preparations are present in unit dosage forms, including but not limited to ampoules, or in multi-dose containers, with a preservative added thereto.

Suitable daily dosages for the formulations disclosed herein are about 0.01 to 3 mg / kg body weight. The indicated daily dosage in large mammals, including but not limited to humans, ranges from about 0.5 mg to about 100 mg, including but not limited to divided doses up to four times per day or for a long time It may be conveniently administered in a released form. Suitable unit dosage forms for oral administration comprise from about 1 to 50 mg of active ingredient. The range is only for suggestion and the number of variables associated with the individual treatment regimen may be large, so there is a general deviation from these recommendations. Such doses include, but are not limited to, the activity of the MIF receptor inhibitor optionally used, the disease or condition to be treated, the mode of administration, the individual requirement, the severity of the disease or condition to be treated, and the judgment of the attending physician. It can be changed according to the variable of.

Toxicity and therapeutic efficacy of the treatment regimen are measured in cell culture or experimental animals, including, but not limited to, LD50 (fatal dose to 50% population) and ED50 (therapeutically effective dose at 50% population). It is not limited. The dose ratio between toxic and therapeutic effects is the therapeutic index and is expressed as the ratio between LD50 and ED50. Preferred are the formulations disclosed herein having a high therapeutic index. Data obtained from cell culture assays and animal studies are used to formulate a range of dosages for use in humans. Dosages of the formulations disclosed herein are preferably in the range of circulating concentrations that include ED50 of minimal toxicity. Dosages may vary within this range depending upon the dosage form employed and the route of administration utilized.

[Example]

The following specific examples are illustrative only and are not intended to limit the disclosure or claims.

Example  One

Cell Lines and Reagents

Human aorta (Schober, A., et al., (2004) Circulation 109, 380-385) and umbilical vein (Weber, KS, et al., (1999) Eur . J. Immunol . 29, 700-712) endothelial cells (PromoCell), MonoMac6 cells (Weber, C, et al., (1993) Eur . J. Immunol . 23, 852-859) and Chinese hamster ovary (CHO : Chinese hamster ovary) ICAM-1-transfectants (Ostermann, G., et al., (2002) Nat. Immunol . 3, 151-158) were used as described. Jurkat cells and RAW264.7 macrophages were transfected with pcDNA3-CXCR2. HL-60 cells were transfected with pcDNA3.1 / V5-HisTOPO-TA-CD74 or vector control (Nucleofector Kit V, Amaxa). L1.2 cells were transferred to pcDNA3-CXCR or pcDNA-CCR5 (UMR cDNA Resource Center) for analysis on simian virus-40-transformed mouse microvascular endothelial cells (SVECs). Transfection. Peripheral vascular mononuclear cells are subjected to monocytes, mithohamagglutinin / interleukin-2 (Biosource) stimulation and / or immunomagnetic screening (antibodies against CD3 / M-450 Dynabeads) through smoke, adherence or immunomagnetic separation (Miltenyi). Primary T cells by, and neutrophils by Ficoll gradient centrifugation. Human embryonic kidney-CXCR2 transfectants (HEK293-CXCR2) were as described above (Ben-Baruch, A., et al., (1997) Cytokine 9, 37-45).

Recombinant MIF (Bernhagen, J., et al., (1993) Nature 365, 756-759). Chemokines were obtained from PeproTech. Human VCAM-1.Fc Chimeric Blocking Antibodies to CXCR1 (42705, 5A12), Human VCAM-1.Fc Chimeric Blocking Antibodies to CXCR2 (48311), Human VCAM-1.Fc Chimeric Blocking Antibodies to CXCR4 (44708, FABSP2 Cocktail, R & D), human VCAM-1.Fc chimeric blocking antibody (NIHIII.D.9) against human MIF and mouse MIF (Lan, HY, et al., (1997) J. Exp . Med . 185, 1455-1465]), human VCAM-1.Fc chimeric blocking antibody to CD74 (M-B741, Pharmingen), human VCAM-1.Fc chimeric blocking antibody to TS ₂ integrin (TS1 / 18), α ₄ integrin Human VCAM-1.Fc chimeric blocking antibody (HP2 / 1) for (Weber, C., et al., (1996) J. Cell Biol . 134, 1063-1073]) and human VCAM-1.Fc chimeric blocking antibody (RII115) against CXCR2, and human VCAM-1.Fc chimeric blocking antibody (327C) against α _L integrin (Shamri, R., et al., (2005) Nat . Immunol . 6, 497-506). PTX and B-oligomers were obtained from Merck.

Method used in the examples

Adhesion  Analysis method

Monocytes, T cells and L1.2 transfectants labeled with calcein-AM (Molecular Probes) were quantified for their suspension by flowing in a parallel-wall chamber (1.5 dynes / cm 2, 5 min) (Schober , A., et al., (2004) Circulation 109, 380-385; Ostermann, G., et al., (2002) Nat . Immunol . 3, 151-158; Weber, C, et al. , (1996) J. Cell Biol . 134, 1063-1073]). Confluent endothelial cells, CHO-ICAM-1 cells, VCAM-1.Fc-coated plates and leukocytes were pretreated with MIF, chemokines or antibodies. CHO-ICAM-1 cells incubated with MIF (2 hours) were subjected to antibodies against MIF Ka565 (Leng, L., et al., (2003) J. Exp . Med . 197, 1467-1476) and Staining with FITC conjugated antibody.

Chemotaxis method

Using a transwell chamber (Costar), we quantified primary leukocyte mobility towards MIF or chemokines using fluorescence microscopy or calcein-AM labeling and FluoroBlok filters (Falcon). Cells were pretreated with PTX / B-oligomer, Ly294002, MIF (for desensitization), antibodies to CXCR or CD74, or isotype IgG. Pore sizes and time intervals were 5 μm and 3 h (monocytes), 3 μm and 1.5 h (T cells), and 3 mm and 1 h (neutrophils).

Q- PCR  And ELISA

RNA was reverse transcribed using oligo-dT primers. RTPCR was performed using a QuantiTect kit (Qiagen) using SYBR Green, specific primers and MJ Opticon2 (Biozym). CXCL8 was quantified using Quanticine ELISA (R & D).

alpha _L β ₂ Integreen  Activation Method

MIF or Mg ^{2 +} / EGTA secure the mononuclear cells were stimulated with the (positive control), it was reacted with FITC-conjugated antibodies to mouse IgG and preparation 327C. LFA-1 activation, analyzed by flow cytometry, was recorded as an increase in mean fluorescent intensity (MFI) or as a relative increase over a positive control (Shamri, R., et al., (2005) Nat . Immunol . 6, 497-506].

Calcium transfer

Neutrophils or L1.2 CXCR2 transfectants were labeled with Fluo-4 AM (Molecular Probes). After addition of a primary or subsequent stimulant (MIF, CXCL8 or CXCL7), MFA was monitored by measuring cytosolic Ca ⁺² concentration for 120 s using BD FACSSAria (BD FACSAria). The L1.2 control group showed negligibly small calcium influx.

Receptor binding assay

Since iodinated MIF is inactive (Leng, L., et al., (2003) J. Exp . Med. 197, 1467-1476); Kleemann, R., et al., (2002) J. Interferon Cytokine Res. 22, 351-363])), radioiodized tracer (Amersham): [I ¹²⁵ ] CXCL8 (reconstituted at 4 nM (80 μCi / ml) to a final concentration of 40 pM); [I ¹²⁵ ] competitive receptor binding using CXCL12 (reconstituted at 5 nM (100 μCi / ml) to a final concentration of 50 pM) (Hayashi, S., et al., (1995) J. Immunol . 154, 814 -824]). Tracers and cold MIFs and / or CXCLs were loaded with HEK293-CXCR2 or CXCR4- for either MIF and [I ¹²⁵ ] CXCL8 competition for CXCR2 binding or MIF and [I ¹²⁵ ] CXCL12 competition for CXCR4 binding in an equilibrium binding assay. Added to cells. This assay was performed via liquid scintillation counting. To calculate EC ₅₀ and K _d values, a one-site receptor-ligand binding model was taken and the Cheng / Prusoff-equation and GraphPad Prism were used.

For pulldown of the biotin-MIF CXCR complex, HEK293-CXCR2 transfectants or controls were incubated with biotin-labeled MIF (Kleemann, R., et al. (2002) J. Interferone Cytokine Res . 22, 351-363), and washed with coimmunoprecipitation (CoIP) buffer and dissolved. The complex was isolated from the clear lysate by magnetic beads (M280, Dynal) coated with streptavidin and analyzed by western blotting using antibodies against CXCR2 or streptavidin-peroxidase. For flow cytometry, HEK293-CXCR2 transfectants or Jurkat cells pretreated with AMD3465 and / or 20-fold unlabeled MIF were incubated with fluorescein-labeled MIF and BD FACSCalibur. Analyze using.

CXCR  Internalization Method

HEK293-CXCR2 or Jurkat cells were treated with CXCL8 or CXCL12, respectively, treated with MIF, washed with acidic glycine buffer, stained with antibodies to CXCR2 or CXCR4, and analyzed by flow cytometry. Internalization was calculated as relative to surface expression of isotype control staining (0% control) and cells treated with buffer (100% control): geometric MFI [experimental] -MFI [0% control] / MFI [100% Control] -MFI [0% control] x 100.

CXCR2  And CD74 Co Localization

RAW264.7-CXCR2 transfectants were co-stained with rat antibody (In-1, Pharmingen) against CXCR2 and mouse CD74, followed by FITC conjugated antibody against rat IgG and Cy3 conjugated antibody against mouse IgG And confocal laser scanning microscope (Zeiss).

CXCR2  And CD74 of Coimmunoprecipitation Method

HEK293-CXCR2 cells transiently transfected with pcDNA3.1 / V5-HisTOPO-TA-CD74 were lysed with undenatured CoIP buffer. Supernatants were incubated with CXCR2 antibody RII115 or an isotype control and blocked overnight with protein G-Sepharose. Proteins were analyzed via Western blot using the preparation for His-tag (Santa Cruz). Similarly, CoIP and immunoblot were performed using antibodies against His-tag and CXCR2, respectively. Immunoprecipitation with antibodies against CXCR2 was performed against L1.2-CXCR2 cells, and immunoblotting with antibodies against mouse CD74.

Carotid In vitro  Perfusion and Biomarkers Microscopy

MIF ^/- (Fingerle-Rowson, G., et al., (2003) Proc . Natl . Acad . Sci . USA 100, 9354-9359) and Ldlr ^-/- mice (Charles River) hybridized The resulting MIF ^/ -Ldlr ^-/- mice and Mif ^{+ / +} Ldlr ^-/- littermates controls, and Apoe ^-/- mice provided an atheromatous diet (21% fat; altromine) for 6 weeks. All single knockout variants were backcrossed 10 times in the C57BL / 6 background. Mif ^{+ / +} and MIF ^{/ −} mice were treated with TNFα (intraperitoneal (ip), 4 h). The explanted artery was transferred onto a stage of a fluorescence microscope and calcein-AM-labeled monoMac6 cells treated with an antibody against CD74 or CXCR2, isotype control IgG at 4 μl / min, or left untreated. Perfusion (Huo, Y., et al., (2001) J. Clin . Invest . 108, 1307-1314). Monocyte cells left untreated were blocked with antibody to MIF for 30 min and then perfused. For in vivo microscopy, Rhodamine-G (Molecular Probes) was administered intravenously (iv) and the carotid arteries of anesthetized mice were exposed. Stopping of labeled leukocytes (> 30 s) was analyzed by fluorescence microscopy (Zeiss Axiotech, 20x water immersion). All studies were approved by local authorities (Bezirksregierung Koeln) and in accordance with German Animal Protection Act: 50.203.2-AC 36, 19/05.

Atherosclerotic  Mouse model of disease progression

Apoe ^-/- mice receiving a 12-week atherogenic diet had an antibody against MIF (NIHIIID.9), an antibody against CXCL12 (79014) or an antibody against CXCL1 (124014, R & D) for an additional 4 weeks (n = 6-10 mice) were injected (three injections per week, 50 μg each). The aortic root was fixed via in situ perfusion and quantified by staining the cross section with Oil Red O (Oil-Red-O). Relative macrophage and T cell contents were determined by staining with an antibody against MOMA-2 (MCA519, Serotec) or an antibody against CD3 (PC3 / 188A, Dako) and an FITC conjugated antibody. In MIF ^{− / −} Ldlr ^{− / −} mice and Mif ^{+ / +} Ldlr ^{− / −} mice that were fed for 30 weeks, the presence of lesion macrophages and luminal monocytes in the aortic root (see Verschuren, L., et. al., (2005) Arterioscler . Thromb . Vase . Biol . 25, 161-167).

Testicular muscle  Microcirculation Model

Human MIF (1 μg) was injected intrascrotum, and testicular muscle muscles were extracted out of the body in mice pretreated with antibodies to CXCR2 (100 μg, ip). After 4 h, biomicroscopy (Zeiss Axioplan; 20x) was performed in the posterior capillary lavage veins (Gregory, JL, et al., (2004) Arthritis Rheum . 50, 3023-3034; Keane, MP, et al., (2004) J. Immunol . 172, 2853-2860]). Adhesion was measured as white blood cells stopped for more than 30 s and release was measured as the number of extravascular white blood cells per field of view.

Bone marrow transplantation

The femur and tibia were aseptically isolated from donor Il8rb ^-/- (Jackson Laboratories) or BALB / c mice. Cells oozed from the bone marrow at 24 h after excisional systemic irradiation were administered iv into Mif ^{+ / +} or MIF ^{− / −} mice (Zernecke, A., et al., (2005) Circ . Res . 96 , 784-791].

Acute Peritonitis Model

Mice (200 ng) were injected ip with mice reproliferated with Il8rb ^{+ / +} or Il8rb ^{− / −} bone marrow. After 4 h, peritoneal lavage was performed and Gr-1 ⁺ CD115 ^- F4 / 80 ^- neutrophils were quantified by flow cytometry using the associated conjugated antibody.

Statistical analysis

Statistical analyzes were performed using Welch's (GraphPad Prism) analysis using one-way analysis of variance (ANOVA) and Newman-Keuls post test or unpaired Student's t test. .

Example  2

CXCR2 Surface through Combined MIF  Inducible monocyte arrest

Monoclonal antibodies and pertussis toxin (PTX) were used to study whether MIF-induced monocyte arrest is dependent on the G _αi coupled activity of CXCR2. Human aortic endothelial cells pretreated with recombinant MIF for 2 h substantially increased the arrest of primary human monocytes under flow conditions, an effect blocked by antibodies to MIF (FIG. 1A). Note that monocyte arrest, which was induced by MIF but was not spontaneous, was eliminated by antibody or PTX to CXCR2, which means that the G _αi coupled CXCR2 is involved. The ability of MIF to induce monocyte arrest through CXCR2 was confirmed using monocyte monoMac6 cells and this activity was associated with immobilization of MIF on aortic endothelial cells (FIG. 1B). These data suggest that MIF is present on endothelial cell surfaces and exerts chemokine-like stop function as a cognate CXCR2 ligand. Classical CXCR2 agonist (CXCL1 / CXCL8) blockade did not interfere with this effect of MIF (FIG. 1A).

Chinese hamster ovary (CHO) transfectants expressing the β ₂ integrin ligand, ICAM-1 (intracellular adhesion molecule 1), were used to analyze in detail the mechanism by which MIF promotes integrin dependent arrest. Quantification under flow conditions revealed the exposure of CHO transfectants to MIF for 2 h, which was presented on the surface (FIG. 1B), and similarly, exposure of the phenotype to CXCL8 increased monocyte cell arrest (FIG. 1C). ). This effect was completely sensitive to antibodies to PTX and β ₂ integrins (FIG. 1C), which confirmed the role of G _αi in β ₂ integrin mediated arrest induced by MIF. Primary monocytes and monoMac6 cells express both CXCR1 and CXCR2 (Weber, KS, et al., (1999) Eur . J. Immunol . 29, 700-712). While CXCR1 blockade was ineffective, CXCR2 blockade substantially damaged MIF induced and CXCL8 induced monocyte cell arrest, but not completely. Adding antibodies against both CXCR1 and CXCR2 completely inhibited the stopping function of MIF or CXCL8 (FIGS. 1D & 8). The use of an antibody against CD74 resulted in association of this protein with CXCR2 at MIF inducible arrest (FIG. 1D). Spontaneous stop was not affected (FIG. 8). Thus, CXCR2 aided by CD74 mediates MIF inducible arrest.

CXCR4 Through MIF  Inducible T cell arrest

MIF or CXCL12 immobilized on aortic endothelial cells induced quiescence of primary human effector T cells (FIG. 1E). T cell arrest induced by MIF but not spontaneous is sensitive to PTX and inhibited by antibodies to CXCR4 (FIG. 1E). Although less pronounced than in monocytes expressing CXCR2, the presentation of MIF (or CXCL12) on CHO transfectants expressing ICAM-1 induced α _L β ₂ dependent arrest of Jurkat T cells, which effect was CXCR4. Mediated by (FIG. 1F).

The extra testicular expression of CXCR2 in Jurkat T cells increased MIF induced arrest (FIG. 1G) and confirmed that CXCR2 confers responsiveness to MIF in leukocytes. Controls using L1.2 pre-B lymphoma transfectants expressing CXCR1, CXCR2 or CXCR3, and cells expressing only endogenous CXCR4 were used in the presence of CXCR4 antagonist AMD3465. MIF induced quiescence of CXCR2 transfectants and CXCR4-bearing controls on endothelial cells with efficacy similar to regular ligands CXCL8 and CXCL12, while CXCR1 and CXCR3 transfectants were reactive to CXCL8 and CXCL10, respectively, but not to MIF. (FIG. 1H). This data demonstrated that CXCR2 and CXCR4, but not CXCR1 or CXCR3, support MIF inducible arrest.

Example  3

CXCR2 With / 4 activation MIF  Inducible leukocyte chemotaxis

Chemokines are defined by their names as inducers of chemotaxis (Baggiolini, M., et al., (1994) Adv . Immunol . 55, 97-179; Weber, C, et al., (2004) Arterioscler . Thromb . Vase . Biol . 24, 1997-2008]). Paradoxically, MIF was initially considered to inhibit the "random" movement (lit. [Calandra, T., et al. , (2003) Nat. Rev. Immunol. 3, 791-800]). While this may be due to the active repulsion or desensitization of the designated effluent, the specific mechanisms caused by MIF to control migration still remain unclear. Based on our results showing that MIF induced the G _αi mediated functions of CXCR2 and CXCR4, we quickly tested whether MIF directly induced leukocyte chemotaxis through these receptors.

Using the Transwell system, the premigration effects of MIF and CXCL8 were compared on monocytes derived from primary human peripheral blood mononuclear cells. CCL2 was also used as circular chemokine for monocytes. Similar to CXCL8 and CCL2, migration was induced by the addition of MIF to the lower chamber, resulting in a bell-shaped dose response curve typical for chemokines, although at low peak shift indices, the optimal value was 25-. 50 ng / ml (FIG. 2A). Neutralizing antibodies to heat treatment or MIF eliminated MIF inducible transition. In contrast, isotype matched immunoglobulin (IgG) was ineffective (FIG. 2B). When added to the upper chamber, the MIF desensitized the movement towards the MIF in the lower chamber in a dose dependent manner (FIG. 2C), but did not induce movement when present only in the upper chamber, which caused the MIF to be chemotactic. Rather, it produces true chemotaxis. Consistent with G _ai dependent signaling via phosphinositide-3-kinase, MIF induced monocyte chemotaxis was PTX sensitive and was removed by Ly294002 (FIG. 2D). Both CXCR2 and CD74 specifically contributed to MIF induced monocyte chemotaxis (FIG. 2E). The role for CXCR2 has been shown to show MIF mediated cross-sensitization of CXCL8 inducible chemotaxis in L1.2 cells transfected with CXCR2. Chemotactic activity of MIF was demonstrated in RAW264.7 macrophages (FIG. 8) and THP 1 monocytes. These data demonstrate that MIF induces monocyte chemotaxis through CXCR2.

To demonstrate functional MIF CXCR4 interaction, the transition of primary CD3 ⁺ T lymphocytes without CXCR1 and CXCR2 was evaluated. Similar to CXCL12, known CXCR4 ligands and T cell chemotactic factors, the way MIF depends on the dose-dependent transition, a chemotactic, process transformed via CXCR4, as shown through cross-sensitization and antibody blocking of CXCL12. Induced (FIG. 2F & FIG. 8). Thus, MIF causes directed T cell migration through CXCR4. In primary human neutrophils, a major cell type bearing CXCR2, MIF exerted CXCR2-mediated chemotactic activity, not CXCR1-mediated, and exhibited bell-shaped dose response curves and cross-sensitized CXCL8 (FIG. 2G, h). . Its extraordinary expression in CD74 ^- promyelocytic HL-60 cells increased MIF inducible migration (FIG. 8), suggesting that moderate chemotactic activity of neutrophils towards MIF is related to the absence of CD74 on neutrophils. Similar to other CXCR2 ligands, MIF functions as a stationary chemokine, but the data revealed that MIF also has significant chemotactic properties on monocytes and neutrophils.

Example  4

MIF Swift Integreen  Cause activation and calcium influx.

Although the stop function of MIF may reflect direct MIF / CXCR signaling, it cannot be completely ruled out that MIF induces other stop chemokines for the time required for MIF immobilization. To confirm that MIF directly induces leukocyte arrest (FIG. 1), real-time PCR and ELISA were performed, and long-term preincubation of human aortic (or venous) endothelial cells with MIF for 2 h resulted in CXCR2 involvement. Upregulation of a typical stop chemokine known to have been shown to fail (FIG. 3A).

Short-term exposure to chemokines that are immobilized in parallel with integrin ligands (eg, vascular cell adhesion molecule (VCAM) -1) or present in solution can rapidly upregulate integrin activity, which is white blood cells. Mediates arrest (Laudanna, C, et al., (2006) Thromb . Haemost . 95, 5-11). This is done by clustering (eg α ₄ β ₁ ) or conformational changes (eg α _L β ₂ ) just before ligand binding. Stimulation of monocyte cells with MIF (or CXCL8) for 1-5 min induced α _L β ₂ dependent arrest on CHO / ICAM-1 cells (FIG. 3B). To obtain evidence for direct stimulation of monocyte integrins, the reporter antibody 327C, which recognizes the extended high affinity conformation of α _L β ₂ , was used (Shamri, R., et al., (2005) Nat . Immunol . 6, 497-506]. These analyzes revealed that α _L β ₂ activation in monoMac6 cells (FIG. 3C) and human blood monocytes (FIG. 3D) occurs prematurely and lasts 30 minutes after exposure to MIF. To determine whether the MIF effect is limited to α _L β ₂ , we tested α ₄ β ₁ dependent monocyte cell arrest on VCAM-1. Exposure to MIF for 1-5 min resulted in a significant stop, which was mediated by CXCR2, CD74 and α ₄ β ₁ (FIG. 3E). Similar to the effect of CXCL12, stimulation of Jurkat T cells with MIF for 1-5 min resulted in CXCR4 dependent adhesion on VCAM-1 (FIG. 8).

Since CXCR2 can mediate cytosolic calcium increase induced by CXCL8 (Jones, SA, et al., (1997) J. Biol . Chem . 272, 16166-16169), it stimulates calcium influx and CXCL8 The ability of the MIF to desensitize the signal was tested. Indeed, similar to CXCL8, MIF induced calcium influx in primary human neutrophils and desensitized transient changes in calcium in response to CXCL8 or MIFM (FIG. 3F), thereby allowing MIF to deliver GPCR / G _αi signaling. It was confirmed that it activates. Partial desensitization of CXCL8 signaling by MIF observed in neutrophils was similar to that observed for other CXCR2 ligands (Jones, SA, et al., (1997) J. Biol . Chem . 272, 16166-16169 ]), Reflects the presence of CXCR1. In L1.2 transfectants expressing CXCR2, MIF completely desensitized CXCL8 inducible calcium influx, and in neutrophils, MIF desensitized transient changes induced by the selective CXCR2 ligand CXCL7 (and CXCL7 is MIF Transient changes induced by desensitization) (FIG. 3F). In CXCR2 transfectants, MIF induced calcium influx in a dose dependent manner and showed slightly less potency and effect than CXCL8 or CXCL7 (FIG. 3G). In conclusion, MIF acts on CXCR2 and CXCR4 to induce rapid integrin activation and calcium influx.

Example 5

MIF The CXCR2  And CXCR4 Interact with

To test the physical interaction of MIF with CXCR2 and CXCR4, we conducted receptor-binding competition and internalization studies. In HEK293 cells expressing CXCR2 out of testis, MIF competed strongly with CXCL8 labeled ¹²⁵ I for CXCR2 binding under equilibrium conditions. Binding of the CXCL8 tracer to CXCR2 was inhibited by MIF and the effector concentration (EC ₅₀ ) for half maximal response was 1.5 nM (FIG. 4A). The affinity of CXCR2 for MIF ( K _d = 1.4 nM) was close to the affinity for CXCL8 ( K _d = 0.7 nM) and was contained within the MIF concentration range (2-4 nM) leading to optimal chemotaxis. . To confirm binding to CXCR2, a receptor internalization assay was used to record specific receptor-ligand interactions. FACS analysis of surface CXCR2 on stable HEK293 transfectants showed that MIF induced CXCR2 internalization in a dose response similar to CXCL8 (FIG. 4B). Similar data were obtained from RAW264.7 macrophages transfected with CXCR2 (insertion of FIG. 4B).

To demonstrate the interaction of MIF with CXCR4, it was performed on Jurkat T cells expressing CXCR4 endogenously. MIF competed with CXCL12 labeled ¹²⁵ I for CXCR4 binding ( K _d = 1.5 nM for CXCL12; EC ₅₀ = 19.9 nM, K _d = 19.8 nM for MIF) (FIG. 4C). The K _d was based on the MIF concentration that induces T cell chemotaxis. Consistently, MIF induced CXCR4 internalization in a dose dependent manner, such as CXCL12 (FIG. 4D). Unlike cognate ligand CCL5, MIF failed to induce CCR5 internalization in L1.2 CCR5 transfectants, so MIF inducible internalization of CXCR2 and CXCR4 was specific for these receptors.

To confirm the interaction with CXCR, MIF was labeled with biotin or fluorescein to allow direct receptor-binding assays, unlike iodinated MIF. As demonstrated by flow cytometry (FIG. 4E), pull down using streptavidin beads (insertion of FIG. 4E), and fluorescence microscopy, CXCR2 transfectants supported direct binding of labeled MIF unlike vector controls. . In addition, specific binding of fluorescein-MIF to CXCR4-bearing Jurkat cells was inhibited by CXCR4 antagonist AMD3465.

CXCR2  And CD74  Complex formation between

Our data suggest that the functional MIF receptor complex includes both GPCR and CD74. Thus, co-localization of endogenous CD74 and CXCR2 was visualized using confocal fluorescence microscopy in RAW264.7 macrophages expressing human CXCR2. Using this technique, pronounced co-localization with a polarization pattern was observed in ˜50% of cells (FIG. 4F).

In addition, coimmunoprecipitation analysis revealed that CXCR2 physically interacts with CD74. CXCR2 / CD74 complex was detected in HEK293 cells stably overexpressing CXCR2 and transiently expressing His-tagged CD74. These complexes were observed through precipitation with antibodies to CXCR2 and coprecipitated CD74 detection using Western blots for His-tags. Coprecipitation was also observed when the sequence of antibodies used was reversed (FIG. 4G). The complex was also detected using CD74 in L1.2 transfectants stably expressing human CXCR2, as assessed by coimmunoprecipitation with an antibody against CXCR2. In contrast, no complexes were observed when using the L1.2 control or the isotype control (FIG. 4H). These data are consistent with the model that CD74 forms a signaling complex with CXCR2 to mediate MIF function.

Example 6

CXCR2 In the artery MIF  Mediates inducible monocyte arrest.

MIF promotes the formation of complex plaques with abundant cell proliferation, macrophage infiltration and lipid deposition (Weber, C, et al., (2004) Arterioscler . Thromb . Vase . Biol. 24, 1997-2008) ;... [Morand, EF , et al, (2006) Nat Rev Drug Discov . 5, 399-410]). This is associated with the induction of endothelial MIF by oxLDL, which causes monocyte arrest (Schober, A., et al., (2004) Circulation 109, 380-385). CXCR2 ligand CXCL1 may also induce α ₄ β ₁ dependent monocyte accumulation in the ex vivo perfused carotid arteries of mice with early atherosclerotic endothelial (Huo, Y., et al., (2001) J. Clin . Invest . 108, 1307-1314]. This system was used to test whether MIF acted to induce mobilization through CXCR2. Intracarotid monocyte arrest in Apoe ^-/- mice received on a high ^- fat diet was inhibited by antibodies to CXCR2, CD74 or MIF (FIGS. 5A & 9), which caused MIF to recruit atherogenicity via CXCR2 and CD74. Suggests that After blocking of MIF, CXCR2 and CD74 for 24 h, a similar pattern was observed for monocyte arrest in the arteries of wild-type mice treated with tumor necrosis factor (TNF), resembling acute vascular inflammation (FIG. 5B). . In the arteries of MIF ^{− / −} mice treated with TNFα, the inhibitory effect on CD74 was attenuated and there was no MIF blocking effect, whereas there was residual CXCR2 inhibition, suggesting the involvement of other inducible ligands (FIG. 5C). Compared with the ^mice as compared to the MIF deficiency effect observed with TNFα stimulation, monocyte accumulation MIF ^{- / -} Ldlr ^{- / -} in the artery of the mouse was clearly damage than by MIF deficiency (atherogenic Mif ^{+ / +} Ldlr ^{- /} When; Fig. 5d, e). In the absence of MIF, CXCR2 clearly did not make any contribution. In addition, MIF blockade was ineffective (FIG. 5D, e). The inhibitory effect of CXCR2 blockade was restored by loading of exogenous MIF (FIG. 5F).

To further demonstrate the opinion that CXCR2 is required for in vivo MIF mediated monocyte recruitment, in vivo microscopy of carotid arteries of chimeric wild type Mif ^{+ / +} and MIF ^{− / −} mice reconstituted with wild type or Il8rb ^{− / −} bone marrow (Il8rb encodes CXCR2; FIG. 5G, h). After treatment with TNFα for 4 h, accumulation of rhodamine G labeled leukocytes was attenuated in MIF ^{− / −} mice reconstituted with wild type bone marrow compared to that in wild type mice reconstituted with wild type bone marrow. Leukocyte accumulation reduction due to bone marrow CXCR2 deficiency was more pronounced in chimeric wild type mice than in chimeric MIF ^{− / −} mice (FIG. 5G, h).

Example  7

In vivo MIF  Inductive inflammation CXCR2 Depends on

The importance of CXCR2 for MIF mediated leukocyte recruitment under atheromatous or inflammatory diseases, diseases, conditions and symptoms was verified in vivo. Adherence of monocytes to the lumen surface of the aortic root was reduced in MIF ^{− / −} Ldlr ^{− / −} mice compared with Mif ^{+ / +} Ldlr ^{− / −} mice suffering from primary atherosclerosis, which showed a marked increase in the macrophage content of the lesion. Reflected by the decrease (FIG. 6A). In vivo microscopic examination of intramuscular microcirculation of testicular muscle muscles resulted in a significant increase in leukocyte adhesion and excretion in postcapillary vasculature (usually CD68) and inhibited by antibodies to CXCR2. It was found to be (Fig. 6b, c). The count of circulating monocytes was not affected.

The MIF induced peritonitis model was then used in chimeric mice reconstituted with wild type or Il8rb ^{− / −} bone marrow. Intraperitoneal injection of MIF induced neutrophil recruitment after 4 h in mice with wild-type bone marrow and disappeared in mice with Il8rb ^{− / −} bone marrow (FIG. 6D). Overall, these results demonstrate that MIF induces leukocyte recruitment under CXCR2 under in vivo atheromatous and inflammatory diseases, diseases, conditions and symptoms.

MIF Targeting Atherosclerosis  Regress.

As described herein, MIF acts through both CXCR2 and CXCR4. Given the role of MIF and CXCR2 in the development of atherosclerotic lesions, rather than CXCL1 or CXCL12, targeting of MIF was investigated as a method for modifying progressive lesions and their CXCR2 ⁺ monocytes and CXCR4 ⁺ T cell content. Apoe ^{− / −} mice receiving high fat diet for 12 weeks and developing severe atherosclerotic lesions were treated with neutralizing antibodies against MIF, CXCL1 or CXCL12 for 4 weeks. Immunoblotting and adhesion assays were used to demonstrate the specificity of the MIF antibody. These analyzes confirmed that MIF blocked MIF inducible arrest but not CXCL1 or CXCL8 inducible arrest (FIG. 10).

Blocking of MIF, but not CXCL1 or CXCL12, reduced plaque area near the aortic root at week 16, and at week 12 there was a significant (P <0.05) plaque degeneration compared to baseline (FIGS. 6E, f). In addition, blockade of MIF, but not CXCL1 or CXCL12, was associated with a decrease in inflammatory plaque phenotype at week 16, as evidenced by a lower content of both macrophages and CD3 ⁺ T cells (FIG. 6g, h). Thus, targeting MIF and inhibiting the activation of CXCR2 and CXCR4 resulted in the therapeutic regression and stabilization of advanced atherosclerotic lesions. In some embodiments, the present invention provides a subject in need of (i) Binding of MIF to CXCR2 and / or CXCR4 and / or (ii) MIF activation of CXCR2 and / or CXCR4; Or (iii) reducing the plaque area in an individual in need thereof by administering one or more agents that inhibit any combination of (i) and (ii).

Example 8

CXCR4  Interference Atherosclerosis  Worsen.

To investigate the role of CXCR4 in atherosclerosis, Apoe ^-/- mice receiving an atheromatous diet were continuously treated with CXCR4 antagonist AMD3465 or vehicle (control) through an osmotic minipump, and after 12 weeks atherosclerotic plaque The formation was analyzed. Compared with the control group, AMD3465 treatment significantly aggravated lesion formation in the oil red O-stained aortic root section (FIG. 9A) and the anteriorly prepared thoracoabdominal aorta (FIG. 9B). In addition, successive treatment of Apoe ^{− / −} mice with AMD3465 induced significant peripheral blood leukocytosis (which persisted for the entire study period) within 2 days, leading to a relative increase in the number of circulating neutrophils. It increased further during this process (FIG. 9C).

Example 9

In a Mouse Model of Multiple Sclerosis Th -17 block occurrence

C57BL / 6 mice aged 8 to 12 weeks (obtained from The Jackson Laboratory, Bar Harbor, Maine) were treated with 5 mg / kg control antibody (group 1), antagonistic anti-mouse MIF antibody (group 2), MIF binding And / or antibodies to CXCR2 that block activation of CXCR2 (group 3), MIF binding and / or antibodies to CXCR4 that block activation of CXCR4 (group 4), or MIF binding and / or to block activation of CXCR4. Antibodies against CXCR4, and antibodies against CXCR2 (group 5) that block MIF binding and / or activation of CXCR2 (group 5) were pretreated -1 days and thereafter weekly. The following day, mice (n = 30 per group) were immunized with a total of 200 μl of an emulsion of MOG35-55 peptide (MEVGWYRSPFSRVVHLYRNGK; Bachem AG, Bobendorf, Switzerland) in CFA twice subcutaneously. Final concentrations of peptide and M. tuberculosis were 150 μg / mouse and 1 mg / mouse, respectively. PTX (400 ng; LIST Biological Laboratories Inc., Campbell, Calif.) Was administered intraperitoneally on days 0 and 2. This disease was monitored by measuring paralysis on a 0-6 scale daily as described above. Mean maximum disease scores were compared between groups using one-way ANOVA.

Paralysis results were compared with group 2 and group 1 mice to determine the efficacy of antagonistic anti-MIF antibodies for the treatment or prevention of EAE. Group 5 mice were compared to group 1 mice to determine the efficacy of agents that block MIF binding and / or activation of CXCR2 and CXCR4 for the treatment or prevention of EAE. Group 5 mice were compared to groups 3 & 4 to determine the effect of blocking MIF binding and / or activation of both CXCR2 and CXCR4 for the effect of blocking CXCR2 or CXCR4 individually.

Mixed T cells were prepared by draining lymph nodes and spleen 7-11 days after immunization. Surviving cells (3.75 × 10 ⁶ / mL) were cultured in complete medium (restimulation) with or without varying concentrations of MOG peptides (amino acids 35-55). Supernatants from activated cells were recovered after 72 h and TNF, IFN-γ, IL-23 & IL-17 were measured by ELISA (BD Pharmingen). High levels of IL-17 and IL-23 suggest the development of Th-17 cells and Th-17 mediated disease phenotype. Treatment of mouse or cell cultures with MIF blocking antibodies (group 2), or inhibition of these cytokines by blocking MIF binding and / or activation of both CXCR2 and CXCR4 (group 5), resulted in the inhibition of Th-17 cells. An important regulatory role of MIF in the development and progression of Th-17 mediated inflammatory diseases (ie, multiple sclerosis) has been described.

For intracellular cytokine staining, spleen and lymph nodes from immunized mice are stimulated with peptide antigens for 24 h and GolgiPlug (BD Pharmingen) is added at the last 5 h, or Golgi Plug + 500 ng / ml of ionomycin and 50 ng / ml of phorbol 12-myristate 13-acetate (PMA; Sigma-Aldrich) were added for 5 h. For cell staining, cells were made permeable using Cytofix / Cytoperm Plus Kit (BD Pharmingen) according to the manufacturer's protocol. Gated CD4-positive T-cells were analyzed for the presence of intracellular IL-17, IL-23 or cell surface IL23 receptor (IL23R) by flow cytometry. The presence of CD4 +, IL-17 + double positive T-cells suggests the development of Th-17 phenotypes that drive disease progression. In addition, up-regulation of IL-23R on CD4 +, IL-17 double positive cells provides evidence supporting the Th-17 phenotype. The presence of high levels of intracellular IL-23 in CD4 +, IL-17 double positive cells or any leukocytes provides additional supportive evidence for Th-17 cell proliferation and / or maintenance driven by IL-23. By treating mice with MIF blocking agents (group 2 mice) or agents that block MIF binding and / or activation of CXCR2 and CXCR4 (group 5 mice), IL-17, IL-23R or As measured through low levels of IL-23, inhibition of Th-17 cell development has demonstrated a predominant role of MIF in driving the progression of Th-17 mediated autoimmune disease. Inhibition of Th-17 cell development and suppression of EAE progression in mice by MIF blockade have been described in (i) CXCR2 and / or CXCR4 for the treatment and / or prevention of Th-17 mediated autoimmune diseases such as multiple sclerosis. Binding of MIF and / or (ii) MIF activation of CXCR2 and / or CXCR4; Or (iii) confirming the valuable utility of agents that inhibit any combination of (i) and (ii).

Example 10

MIF  Identification of agents that disrupt signaling

Peptide libraries were generated covering the extracellular N-terminal motifs / domains of CXCR2. The peptide ranged in size from about 12 amino acids to about 15 amino acids.

The peptide library was screened for inhibition of MIF mediated signaling through CXCR2 using the HTS GPCR screening technique.

Peptides that inhibit MIF mediated signaling from IL-8 and / or SDF-1 mediated signaling inhibition on CXCR2 were then screened.

For further investigation, peptides were selected that inhibit MIF signaling through CXCR2 but allow SDF-1 and IL-8 mediated signaling through CXCR2.

Example  11

MIF Trimerization  Identification of destructive agents

Polypeptides comprising amino acid residues 38-44 (beta-2 strand) of MIFM were generated.

The polypeptide was screened for inhibition of MIF mediated signaling through CXCR2 using the HTS GPCR screening technique.

The polypeptides that inhibit MIF mediated signaling were then screened for IL-8 and / or SDF-1 mediated signaling inhibition on CXCR2.

For further investigation, peptides were selected that inhibit MIF signaling through CXCR2 but allow SDF-1 and IL-8 mediated signaling through CXCR2.

Example  12

Human clinical trial

Experimental Objective (s): The primary objective of this study was the efficacy of (P1; LMAFGGSSEP) (P1; 20 mg, 40 mg, 80 mg) in individuals suffering from homozygous familial hypercholesterolemia (HoFH). Was to evaluate.

Way

Study Design: This study was a multicenter, open-label, single-group forced titration study conducted on fixed combination P2 in ≥ 18 years old male and female subjects with HoFH. After the initial screening, eligible individuals entered a four-week screening period, consisting of two visits (weeks 4 and -1), during which all lipid lowering drugs were stopped (bile acid sequestrants and cholesterol absorption). Counseling changes in therapeutic lifestyle according to the National Cholesterol Education Program (NCEP) Adult Treatment Panel-III (ATP-III) Clinical Guidelines or equivalents (TLC: therapeutic lifestyle change counseling). Subjects already undergoing constitutive treatment continued their treatment regimen while maintaining a consistent state and interval during the study. At Visit 3 (Week 0), baseline efficacy / safety values were measured and subjects started treatment with the initial dose of P2 (20 mg) once daily (QD) for 6 weeks. At 6 weeks (4 visits), the dose was titrated with P2 40 mg QD for 6 weeks, and if the subject showed resistance to the previous dose, then again at 12 weeks (5 visits) with P2 80 mg QD for 6 weeks. . The final visit (sixth visit) occurred at week 18. Study visits were timed to the subjecting procedure performed immediately before the visit procedure, if applicable. If the interval between chemotherapy did not match the study drug treatment period, the subjects were kept on the same drug treatment until the next planned chemotherapy and until the interval returned to the original time length. Efficacy was measured at least 2 weeks after the previous collection and immediately before the planned collection procedure with study visits.

Number of Participants: 30 to 50 individuals

Key criteria for diagnosis and inclusion: There is clear evidence of familial hypercholesterolemia (FH) homozygotes according to the guidelines of the World Health Organization, and serum fasting triglycerides (TG)> Women and men over the age of 18, 400 mg / dL (4.52 mmol / L) and 200 mg / dL (2.26 mmol / L) in 18-20 year old subjects were screened for study participation.

Experimental Treatment: During three 6-week open label treatment periods, subjects took 1 tablet QD, with food, immediately after breakfast. No down titration was allowed. If the subject could not tolerate an increase in dose, this experiment was discontinued.

Efficacy Assessment: The primary endpoint was the mean rate of change of HDL-C and LDL-C from baseline to the end of each treatment period (ie, weeks 6, 12 and 18 weeks). Lipid profiles, including HDL-C and LDL-C, were obtained at each study visit.

Safety Assessment: Safety was assessed using conventional clinical laboratory assessments (hemistological and urinalysis panels at weeks 4, 0 and 18, and chemical tests at weeks 6 and 12). Vital signs were monitored at each visit and physical examination and electrocardiogram (ECG) were performed at weeks 0 and 18. Urine pregnancy tests were performed at each visit except week -1. Subjects were monitored from week 0 to week 18 for adverse events (AEs). At 18 weeks, the safety assessment was completed at the early termination of adverse events.

Statistical Methods: The primary efficacy endpoint was the rate of change of HDL-C and LDL-C from baseline to the end of each treatment period (ie, weeks 6, 12 and 18 weeks). The primary efficacy analysis population is a full analysis set (FAS) that includes all individuals who receive study drug at one or more doses and have both baseline and at least one valid baseline measurement at each analysis period. .

The primary efficacy endpoint was analyzed by computerizing the mean rate of change (or nominal mean change), 95% confidence interval (CI), 1-sample t-test statistics, and corresponding p-values of the samples. Incremental treatment deviations between different dose levels were also predicted and 95% CIs were obtained. The hypothesis test is bilateral with an overall familywise type I error rate of 5% (ie p = 0.05 significance level). Hochberg's treatment procedure was used to control foot-to-foot error rates for multiple comparisons.

Example  13

Abdominal aortic aneurysm ( AAA Animal models for treatment

Animal models were prepared as follows. Adult, male rats were infused with elastase for 2 hours. Histological analysis was performed 12-24 hours after injection to confirm the presence of fragmented and de-organized elastin. Ultrasound was performed daily to identify and monitor the aortic dilatation area.

Two weeks after elastase administration, rats received peptide 2 (P1; LMAFGGSSEP). Initial administration of P2 was injected into the subject at a rate of 0.5 mg / hr. In the absence of infusion toxicity, the infusion rate was increased up to 2.0 mg / hr with a 0.5 mg / hr increase every 30 minutes. Each week thereafter, P2 was injected at a rate of 1.0 mg / hr. In the absence of infusion toxicity, the rate was increased up to 4.0 mg / hr in increments of 1.0 mg / hr in 30 minute intervals.

Efficacy Assessment: The primary endpoint was the average rate of change in AAA size (ie, aortic diameter) from week 3, week 6, and week 12 from baseline.

Example  14

Abdominal aortic aneurysm ( AAA A) human clinical trial for treatment

Study Objective (s): The primary objective of this study was to assess the efficacy of peptide 1 (P1; LMAFGGSSEP) in individuals with early AAA.

Way

Study Design: This study was a multicenter, open-label, single-group study in which early AAA was performed on P2 in male and female subjects ≥18 years old. The presence of early AAA was confirmed by a series of cross-sectional imaging. At week 0, baseline efficacy / safety values were measured and subjects started treatment with an initial dose of P2. Subjects received P2 once weekly for 12 weeks.

Number of Participants: 30-50 individuals.

Study Treatment: Initial administration of P2 was injected into the subject at a rate of 50 mg / hr. In the absence of infusion toxicity, the infusion rate was increased up to 400 mg / hr with a 50 mg / hr increase every 30 minutes. Then each week, P2 was injected at a rate of 100 mg / hr. In the absence of infusion toxicity, the rate was increased up to 400 mg / hr in increments of 100 mg / hr in 30 minute intervals.

Efficacy Assessment: The primary endpoint was the average rate of change in AAA size (ie, aortic diameter) from week 3, week 6, and week 12 from baseline.

Example  15: inflammatory bowel disease ( IBD A) human clinical trial for treatment

Study Objective (s): The primary objective of this study was to evaluate the efficacy of peptide 2 (P2; cyclic CNVPRASVPDGC) in individuals with rheumatoid arthritis (RA).

Study type:

Arbitration.

Study design:

Allocation: non-random.

Control group: non-control group.

Endpoint Classification: Safety Study.

Intervention Model: Single Military Assignment.

Shielding: Open Sign.

Primary purpose: treatment.

Primary outcome measure:

Number of subjects improved based on American College of Rheumatology (ACR) criteria consisting of 20%, 50%, and 70% (ACR20 / 50/70 responders, respectively).

20%, 50%, and 70% (ACR20 / 50/70, respectively) reduction in tenderness or swelling joint count (tender joint count (TJC) or swollen joint count (SJC), respectively) Physician's global assessment of disease activity (PGA), 2) subject's assessment of disease activity, 3) subject's assessment of pain, and 4) health assessment questionnaire (DI-HAQ) Subject's assessment of the disorder, 5) Improved response based on ACR criteria, consisting of 20%, 50%, and 70% improvement in C-reactive protein (CRP) at each visit, respectively. embroidery.

Secondary Outcome Measures:

Average number of changes from baseline of tender joint count (TJC, max = 68), which is a component of the American Society for Rheumatology (ACR) at visit.

Average number of changes from baseline of swelling joint number (SJC, max = 66), a component of the American Society for Rheumatology (ACR) at visit.

At visit, the average number of changes from baseline in the physician's overall assessment of disease activity (PGA), which is a component of the ACR criteria.

The mean number of changes from baseline in the subject's assessment of disease activity using the Visual Analog Scale, a component of the ACR criteria, at visit.

Average number of changes from baseline in the subject's assessment of pain at the time of visit, using the visual pain thought rating component, which is a component of the ACR criteria.

At visit, the average number of changes from baseline in the Disability Index of the Health Assessment Questionaire (DI-HAQ), a component of the American Society for Rheumatology (ACR).

Average number of changes from baseline of C-reactive protein (CRP), a component of the American Society for Rheumatology (ACR), at visit.

Whether morning stiffness exists.

Average number of changes from baseline of morning stiffness duration, in visits.

The presence of Rheumatoid Factor (RF).

Mean number of changes from baseline of rheumatoid factor (RF) (IU / ML) at visit.

Arm

Peptide 2 was administered in a single oral dose (10 mg / Kg) once daily.

Placebo was administered via oral administration once daily.

Eligibility Criteria

20 years old or older.

Male and female.

Inclusion Criteria:

Participation and completion of pre-adalimumab dose range study at week 24.

Women should be postmenopausal for at least one year, infertility surgically, or have birth control restrictions throughout the study and for 90 days after study completion.

For women of childbearing potential, female subjects with a negative test result in the pregnancy test (serum test) at week 24 of the prior adalimumab study.

Exclusion Criteria:

Subjects who experienced any of the following during the pre-study period:

Diabetes, which is not progressive or limited

Joint Surgery (joint evaluated in this study).

Subjects are prescribed to not take medication during the pre-study period.

If you have a history of:

Clinically significant drug or alcohol abuse,

Intravenous (iv) drug abuse,

Active infections, including listeria or tuberculosis (TB),

Lymphoma, leukemia, and

Any malignant tumor except nonmetastatic basal cell carcinoma of the skin that has been successfully treated.

Subjects who received the live vaccine during the prior study, or subjects who were scheduled to complete the live vaccine administration during the study period.

Example  16: acute respiratory distress syndrome ARDS : Acute Respiratory Distress  Syndrome) Human Clinical Trial for Treatment

Study Objective (s): The primary objective of this study was to evaluate the efficacy of peptide 2 (P3; LMAFGGSS) in individuals with ARDS.

Study type:

Arbitration.

Study design:

Assignment: Random.

Control: Placebo Control.

Endpoint Classification: Safety / Efficacy Study.

Arbitration Model: Parallel Assignment.

Shielding: double blind (subject, caregiver, researcher, outcome assessor).

Primary purpose: treatment.

Primary outcome measure:

Safety profile of the study drug.

Number of days not to use ventilator until day 28.

Secondary Outcome Measures:

Mortality at 28 days.

Hospitalization period until the 28th.

Length of stay in ICU until day 28.

Days that did not show malfunction of organs other than the lungs by day 28.

Changes in physiological parameters of pulmonary disorders.

Change in disease severity and lung disorder scores.

Effect of the study drug and the etiology of the disease (ie pulmonary or extrapulmonary organs).

Pharmacokinetic and Pharmacodynamic Properties.

Immunogenicity.

Arm

Peptide 3 was administered via intravenous infusion over a 15 minute period at a single dose (0.06 mg / Kg).

Placebo was administered via intravenous infusion over 15 minutes.

Eligibility Criteria.

18 years old or older.

Male and female

Healthy volunteers allowed: Not allowed.

Inclusion Criteria:

If you suspect an infection or have proven infected.

Hypoxemia: Pa02 / Fi02 ≦ 300 mm Hg.

Bilateral infiltration consistent with pulmonary edema.

Mechanical ventilation of positive pressure through the endotracheal tube.

There is no clinical evidence of left atrial hypertension that may explain bilateral infiltration.

Three or more of the SIRS criteria exist. If only two are proven, one should be either temperature or WBC.

Criteria 2 and 3 should occur within a 24 hour time interval. The 48 hour registration time window begins when the criteria 2, 3, and 4 are met.

Exclusion Criteria:

<18 years old.

If you do not get a consent form.

The patient, agent, or doctor cannot concentrate on overall support.

The death to death is perceived to be imminent.

Morbid obesity.

If you have a malignant tumor or other irreversible disease or condition that is predicted to have> 50% mortality to die within 6 months.

The terminal process is known and known to be HIV positive.

Overall neurological function recovery has not been demonstrated if there was a prior cardiac arrest requiring CPR; Or New York Heart Association Class IV.

If you are pregnant or nursing.

ALI / ARDS is mechanically or chemically induced (including burns, trauma, and drowning).

If elapsed time is> 48 hours after all inclusion criteria are met.

You have a neuromuscular disorder that impairs your ability to ventilate without assistance.

Severe chronic respiratory disease, severe pulmonary hypertension, or relying on a ventilator.

Severe muscle limitation, erythrocytosis, or dependence on the respiratory system due to chest wall deformation.

Have a history of organ transplants (including bone marrow transplants).

Have severe chronic liver disease, measured as a Child-Pugh Score> 10.

When hemoglobin is constantly maintained at <8.0 g / dL.

Platelet count <50,000 / dl.

Long term INR> 3.

If you have hemorrhagic disease even without corrective surgery.

If internal bleeding is active.

Drug Major surgery within 48 hours prior to drug infusion. Or if bleeding is proven to be active postoperatively, or if you plan to undergo any major surgery within 3 days of study drug infusion.

Diffuse alveolar hemorrhage resulting from vasculitis.

Known for bleeding constitution.

Drugs If you had an epidural catheter or lumbar puncture within 48 hours prior to drug infusion, or if you are expected to receive an epidural catheter or lumbar puncture within 48 hours after study drug infusion.

Had a stroke within 3 months prior to study entry.

Have trauma with increased risk of life-threatening bleeding.

You have a history of severe head trauma that requires hospitalization or undergoes intracranial surgery, and is within 2 months prior to study entry.

Have any history of intravenous arteriovenous malformation, cerebral aneurysm, or central nervous system mass lesion.

Limited use of certain medications or treatment regimens, such as chemotherapy, unfractionated heparin, low molecular weight heparin, warfarin, antithrombin III, acetylsalicylic acid, glycoprotein IIb / IIIa antagonists, thrombolytic therapy, and activated protein C If it is.

If you have participated in another experimental drug study within 30 days prior to study entry except for the Nutritional Testing of ARDSNet Pharmacological Nutritional Supplement (OMEGA).

Example  17: Human clinical trial for the treatment of glomerulonephritis

Study Objective (s): The primary objective of this study was to evaluate the efficacy of peptide 2 (P4; VHVVPDLLMA) in individuals with glomerulonephritis.

Study type :

Arbitration.

Study design:

Assignment: Random.

Control group: active control group.

Endpoint Classification: Efficacy Study.

Arbitration Model: Parallel Assignment.

Shielding: Open Sign.

Primary purpose: treatment.

Primary outcome measure:

Acute dialysis onset or twice the serum creatinine level [time frame: 3 months] [designated as a safety issue: yes].

Secondary Outcome Measures:

ESRD (defined as requiring long-term dialysis).

details

This study was a randomized, open marker, comparative study.

Group A was treated with methylprednisolone (15 mg / kg / day or up to 1 g / day from day 1 to day 3) at a pulsed dose by standard intravenous administration for 3 months, and then prednisolone 0.5 by oral administration. Treatment was at -1.0 mg / kg / day (day 4 to day 30).

Group B was treated with peptide 4 (0.33-0.66 mg / kg / h, day 1 to day 7) by the same corticosteroid therapy and intravenous administration followed by 400-800 mg / peptide 4 by oral administration. Treatment was done with day (day 8 to day 90). The dose of pentoxifylline administered intravenously was determined by GFR estimates: patients with a GFR of 30-59 ml / min / 1.73 m 2 received 0.66 mg / kg / h and patients less than 30 ml / min / 1.73 m 2 Received 0.33 mg / kg / h. The dose of peptide 4 administered orally was also determined by GFR prediction. Patients with GFR estimates between 30-59 ml / min / 1.73 m 2 received 800 mg / day and patients under 30 ml / min / 1.73 m 2 received 400 mg / day.

Serum and single urination samples were collected at the hospital before the start of therapy (day 0) and on days 8, 15, 30, and 90 after the start of therapy. Kidney function was calculated by the Cockcroft-Gault formula and simplified to the MDRE formula. Serum and urine samples were measured for inflammatory mediators such as TNFalpha, IL-1beta, IL-6, MCP-1, CX3CL1 (frectalcaine), IL-8 using commercially available ELISA kits.

Eligibility Criteria

20 to 80 years old.

Male and female.

Inclusion Criteria:

Crescent glomerulonephritis with rapid progressive renal failure has been demonstrated by biopsy.

Exclusion Criteria:

If you have anti-GBM disease.

Rely on dialysis or have bleeding from the lungs.

If the woman is a nursing mother or pregnant woman.

Suffer from congestive heart failure.

Have experienced unstable angina, myocardial infarction, coronary artery bypass surgery, or percutaneous coronary intervention within the last 6 months before signing informed consent.

Have had a cerebral hemorrhage within the last 6 months before signing the informed consent.

You have had retinal bleeding within the last 6 months before you signed the informed consent.

Known or suspected of having secondary hypertension.

If you have uncontrolled hypertension or diabetes.

Suffer from cirrhosis of the liver or liver dysfunction, which is defined as ALT or AST greater than 2 times the upper limit of normal range (> 2 times).

Suffer from obstructive biliary tract disease.

If you have an active tumor or infection.

Example  18: inflammatory bowel disease ( IBD A) human clinical trial for treatment

Study Objective (s): The primary objective of this study was to evaluate the efficacy of peptide 2 (P5; QLMAFGGSSE) in individuals with IBD.

This study was a diagnostic, open marker, non-control, multicenter, one arm study.

A total of 24 patients received 35 mg P5 tablets once daily for 12 weeks. First, patients went through a one-week screening period and follow-up visits earlier than four weeks after study drug discontinuation or if recurrence occurred during subsequent periods. Total study duration was up to 17 weeks.

Eight study visits were made: one screening visit, six visits during the treatment period, and one follow-up visit. On the sixth and tenth weeks, two telephone visits were made.

The overall study duration (the period from the first patient's entry to the last patient's exit) was estimated to be about 13 months.

Primary outcome measure:

Number of patients who showed complete or partial remission after 12 weeks of advanced therapy with P5 at a daily dose of 35 mg in patients with Crohn's Disease (CD) or Ulcerative Colitis (UC). Efficacy of P5 as measured by.

Secondary Outcome Measures:

The secondary objective of this study was to assess the safety and tolerability of P5 and to investigate its plasma levels (lowest) when P5 was administered once daily at a dose of 35 mg in CD or UC.

Eligibility Criteria

18 to 70 years old.

Male and female.

Inclusion Criteria:

Crohn's disease related criteria:

Have been diagnosed with CD clearly by standard criteria (eg endoscopy, ultrasound, X-ray).

Patients should show clinical remission for steroid therapy progressed for at least 2 weeks (Crohn's disease activity index [CDAI] <150 points).

If the CD is found to be dependent on steroids: Patients who do not completely reduce steroids within 3 months after steroid initiation, even if there is no recurrent active disease, or relapses within 2 months after steroid discontinuation.

Individual threshold * dose of previous relapse should be a steroid dose equal to, or less than, or equivalent to 20 mg / day prednisolone.

The patient has received a stable glycocorticosteroid therapy using a dose between 20 and 40 mg / day prednisolone or equivalent steroid doses over the past week.

Ulcerative Colitis Related Criteria:

A clear diagnosis of UC, as determined by standard criteria (eg endoscopy, ultrasound, X-ray).

Patients should show clinical remission for steroid therapy progressed for at least 2 weeks (clinical activity index [CAI] <4 points).

If UC is found to be dependent on steroids: Patients who do not completely reduce steroids within 3 months after steroid initiation, even if there are no recurrent active diseases, or relapse within 2 months after steroid discontinuation.

Individual threshold * dose of previous relapse should be a steroid dose equal to, or less than, or equivalent to 20 mg / day prednisolone.

The patient has received a stable glycocorticosteroid therapy using a dose between 20 and 40 mg / day prednisolone or equivalent steroid doses over the past week.

Exclusion Criteria:

If you have short bowel syndrome.

If you have had an ileostomy, colonic intestinal tract, or rectal sac.

If relapse occurs during screening.

Example  19: Human clinical trial for the treatment of sepsis

Study Objective (s): The primary objective of this study was to evaluate the efficacy of peptide 2 (P6; NVPRASVPDG) in individuals with sepsis.

Study type:

Arbitration.

Study design:

Assignment: Random.

Control: Placebo Control.

Endpoint Classification: Safety / Efficacy Study.

Intervention Model: Single Military Assignment.

Shielding: double blind.

Primary purpose: treatment.

Arm

P7 0.5 (twice daily, for 1 week, by IV infusion).

P7 1 mg (twice daily, for 1 week, by IV infusion).

Placebo (2 times daily for 1 week, by IV infusion).

Eligibility Criteria

18 to 85 years old.

Male and female.

Inclusion Criteria:

You are currently in the ICU, or are about to be taken back to the ICU.

Within hGG analysis within 24 hours prior to drug administration, women of childbearing potential should have negative serum (or urine).

All races.

Suffer from severe sepsis (newly developed respiratory failure, refractory shock, renal dysfunction, liver dysfunction, or metabolic acidosis and SIRS (three or more of the signs for systematic inflammatory response syndrome)).

If there are objective signs of an infection that may be caused by a bacterial or fungal pathogen.

Patients should receive study medication within 8-12 hours after recognizing organ failure associated with early sepsis.

The death risk predicted score by APACHE is between 20% and 80%.

Your doctor and family are willing to actively treat you during the 28-day study.

Exclusion Criteria:

Suffer from cardiac or low blood shock.

Acute third degree burn is> 20% of body surface.

If your visa has received a tissue-type organ transplant within the past year.

If you are pregnant.

In chronic vegetative conditions.

Have severe, uncontrolled bleeding (having 0.2 units of blood / platelet in the previous 24 hours). If the bleeding has stopped, the patient can be considered as a subject to enroll, otherwise the patient is still eligible.

You are unwilling or unable to get a full evaluation at every subsequent visit.

Patients classified as "do not revive" or "do not cure".

Patients develop severe sepsis after trauma or within 36 hours of surgery. If the patient has met other inclusion criteria, the patient may be considered subject to enrollment after trauma or after more than 36 hours post surgery.

Patients with a predicted death risk score of <20% or> 80% after a limited organ malfunction.

Patients with a predicted death risk score of <51% for patients with Xigris® use plan.

Example  20: Lupus  Human Clinical Trial for Treatment

Study Objective (s): The primary objective of this study was to evaluate the efficacy of peptide 2 (P7; NVPRASVPD) in individuals with lupus.

Study type:

Arbitration.

Study design:

Assignment: Random.

Endpoint Classification: Safety / Efficacy Study

Arbitration Model: Parallel Assignment.

Shielding: double blind (subject, researcher, outcome assessor).

Primary purpose: treatment.

Primary result measurement :

Changes in safety, tolerability, swelling and tender joint count.

Arm

0.5 mg P7 once daily for 12 weeks.

P7 1 mg (oral, once daily for 12 weeks).

Placebo (oral, once daily for 12 weeks).

Eligibility criteria .

18 to 75 years old.

Male and female.

Inclusion Criteria:

Subjects diagnosed with SLE.

Have 4 or more tenderness and 4 or more swelling joints,

Subjects with active lupus arthritis, which prove to be active synovialitis ≧ 1 joint (some loss of functional range of motion).

Exclusion Criteria:

Subjects with severe renal dysfunction or who are on dialysis.

Suffer from severe instability and / or progressive CNS lupus.

Subjects with clinically significant or unstable medical or surgical conditions.

Pregnant or nursing mothers during the study, or women who are willing to be the subjects.

If you are a woman of childbearing potential who does not implement acceptable birth control measures.

Example  21: Human clinical trial for the treatment of asthma

Study Objective (s): The primary objective of this study was to evaluate the efficacy of peptide 2 (P8; cyclic CLMAFGGSSEPCALC) in individuals with asthma.

Study type:

Arbitration.

Study design:

Assignment: Random.

Control: Placebo Control.

Endpoint Classification: Safety / Efficacy Study.

Arbitration Model: Parallel Assignment.

Shielding: double blind (subject, caregiver, researcher, outcome assessor)

Primary purpose: treatment.

Primary outcome measure:

Airway reactivity was measured using the methacholine challenge test, which is conducted according to the following ATS guidelines.

Secondary Outcome Measures:

Pulmonary function measured by FEV1 and FVC in accordance with ATS guidelines.

Asthma symptoms and management were objectively monitored using the Juniper Questionnaire, the Asthma Quality of Life Questionnaire, and the St. George Respiratory Questionnaire.

details:

Participants in this study were randomly assigned to P8 or placebo (inactive pill). The participants were given daily study medication for 12 weeks (3 months).

Participants completed a number of asthma questions and lung function tests. Participants underwent physical examination and electrocardiogram and collected blood for leptin, adiponectin, inflammatory markers, blood counts, blood glucose levels, BNP hormone levels, and liver function measurements.

Follow-up visits were made at weeks 2, 6, and 12 weeks after the study drug was initiated to continue monitoring participants during the study. At this visit, a number of pulmonary function tests and questionnaires were repeated.

Eligibility Criteria

18 to 60 years old.

Male and female.

Inclusion Criteria:

You have been diagnosed with asthma by your doctor at least one year before enrolling in the study.

Inadequate asthma control at study enrollment.

Non-smokers (if you started smoking at least a year ago) and have a very large smoking history throughout your life.

If the body mass index is 30-60.

Responds to PC20 <16 mg / ml for the methacholine challenge test.

You received inhaled corticosteroids at a stable dose for at least 4 weeks prior to study entry.

When FEV1> 60% (predicted).

If you can weigh your weight every week at home.

Exclusion Criteria:

Systemic steroids within the last 4 weeks.

If you have pathological abnormalities in your lungs other than asthma.

Concomitant diseases other than other significant pulmonary diseases, such as coronary artery disease, peripheral vascular disease, cerebrovascular disease, ejection rate <50%, congestive heart failure, liver disease or elevated liver enzymes at baseline, malignant tumors (non Melanoma skin cancer), AIDS, heart failure with serum creatinine> 3.0, or a disease requiring steroid treatment, such as vasculitis, lupus, rheumatoid arthritis.

B-type natriuretic peptide (BNP)> 400 pg / ml

If you are pregnant or nursing.

Currently taking a vector blocker, CYP2C8 inhibitor or inducer such as gemfibrozil or rifampin, TZD (thiazolidinedione), or are allergic to TZD.

Taking antioxidants (when taking multivitamins prior to enrollment should be a stable therapy).

Use of illegal drugs within the past year.

For current / active upper respiratory infections (if active URI, wait for 1 week until symptoms disappear until enrollment).

If your asthma has worsened within the last 4 weeks (including ER, urgent care, or hospital visits due to asthma that has increased asthma medication),

If you have had a sleep apnea assessment or plan to start sleep apnea treatment (patients who have been reliably treated for sleep apnea for the last three months may participate).

A clinically significant abnormality appears to be present on the 12-guided electrocardiogram.

Women of childbearing potential who use oral contraceptives during the study are not willing to use a second method.

Example  22: Human clinical trial for the treatment of colon cancer

Study Objective (s): The primary objective of this study was to evaluate the efficacy of peptide 1 (P1; LMAFGGSSEP) in individuals with colon cancer.

Study type:

Arbitration.

Study design:

Allocation: non-random.

Control group: non-control group.

Endpoint Classification: Efficacy Study.

Intervention Model: Single Military Assignment.

Shielding: Open Sign.

Primary purpose: treatment.

Arm

Oral Peptide 1 (15 mg / kg) (once daily for four months).

Oral placebo (once daily for four months).

Eligibility Criteria

18 years old or older.

Male and female.

Inclusion Criteria:

You have had a complete resection of the colon for stage I or II adenocarcinoma for healing purposes within the past year.

Have had preoperative or postoperative colonoscopy of the cecum (or small intestine anastomosis) using the appropriate bowel formulation within the last 180 days.

The distal border location of the tumor is ≥ 12 cm from the anal dermal gland.

Not typical familial adenomatous polyposis, attenuated familial adenomatous polyposis (ie ≥ 20 adenomas, simultaneous or at other times), or hereditary nonpolypical colorectal cancer (Lynch syndrome).

Patient features:

ECOG Performance Status 0-1.

Serum creatinine is less than or equal to 1.5 times the upper limit of normal (ULN) (≦ 1.5 times).

AST and / or ALT is less than or equal to 3.01 times ULN (≦ 3.0 times).

Total bilirubin is less than or equal to 1.5 times ULN (≤ 1.5 times).

Not pregnant or lactating mothers.

Voice in pregnancy test.

Fertility patients should use effective contraception during study treatment and for a period of at least 3 months (≧ 3 months) after termination of study treatment.

Oral medications may be swallowed.

It does not suffer from malabsorption syndrome, ulcerative colitis, inflammatory bowel disease, gastric or small bowel resection, or other disease that can have a significant effect on gastrointestinal (GI) function.

There is no documented history for upper GI bleeding or upper GI ulcer disease.

It is not hyperlipidemia clinically indicated as statin therapy (measured lipid levels at fasting should follow current dyslipidemia management guidelines).

Researchers have confirmed that they have not been improperly treated for hypothyroidism.

There is no history of myopathy or rhabdomyolysis.

Except for in situ skin cancer or basal cell or squamous cell carcinoma of the skin, no other malignant tumor has been experienced within the last five years.

It is not anaphylactic or intolerant to statins.

There is no other nonmalignant systemic disease that can interfere with rosuvastatin administration or long term subsequent treatment.

Pre-Concurrent Therapy:

Disease feature observation

More than 30 days elapsed since prestatin.

More than 30 days since the elapsed time of the preliminary trial preparation.

There was no prior total or total colon resection.

There is no concurrent chronic use of NSAIDs.

Cyclosporin, coumarin anticoagulants, gemfibrozil, other lipid lowering therapies (eg fibrates or niacin), lopinavir / ritonavir, or drugs that lower the level or activity of steroid hormones (eg ketoconazole There is no concurrent chronic drug therapy using spironolactone, or cimetidine).

While preferred embodiments of the invention have been presented and described herein, it is apparent to those skilled in the art that these embodiments are provided by way of example only. Without departing from the present invention, one of ordinary skill in the art can add many modifications, changes and substitutions. It should be understood that various alternatives to the embodiments of the invention described herein may be used to practice the invention. The following claims define the scope of the invention and are intended to include methods and structures and equivalents thereof that fall within the scope of the claims.

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Met Pro Met Phe Val Val Asn Thr Asn Val Pro Arg Ala Ser Val Pro 1 5 10 15 Asp Gly Phe Leu Ser Glu Leu Thr Gln Gln Leu Val Gln Ala Met Gly             20 25 30 Lys Pro Ala Gln Tyr Ile Ala Val His Val Val Pro Asp Gln Leu Met         35 40 45 Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu Cys Ser Leu His Ser     50 55 60 Ile Gly Lys Ile Gly Gly Ala Gln Asn Arg Ser Tyr Ser Lys Leu Leu 65 70 75 80 Cys Gly Leu Leu Ala Glu Arg Leu Arg Ile Ser Pro Asp Arg Ile Tyr                 85 90 95 Ile Asn Tyr Tyr Asp Met Asn Ala Ala Asn Val Gly Trp Asn Gly Ser             100 105 110 Thr Phe Ala Leu         115 <210> 3 <211> 116 <212> PRT <213> Bos sp. <400> 3 Met Pro Met Phe Val Val Asn Thr Asn Val Pro Arg Ala Ser Val Pro 1 5 10 15 Asp Gly Leu Leu Ser Glu Leu Thr Gln Gln Leu Ala Gln Ala Thr Gly             20 25 30 Lys Pro Ala Gln Tyr Ile Ala Val His Val Val Pro Asp Gln Leu Met         35 40 45 Thr Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu Cys Ser Leu His Ser     50 55 60 Ile Gly Lys Ile Gly Gly Ala Gln Asn Arg Ser Tyr Ser Lys Leu Leu 65 70 75 80 Cys Gly Leu Leu Thr Glu Arg Leu Arg Ile Ser Pro Asp Arg Ile Tyr                 85 90 95 Ile Asn Phe Cys Asp Met Asn Ala Ala Asn Val Gly Trp Asn Gly Ser             100 105 110 Thr Phe Ala Leu         115 <210> 4 <211> 116 <212> PRT <213> Mus sp. <400> 4 Met Pro Met Phe Ile Val Asn Thr Asn Val Pro Arg Ala Ser Val Pro 1 5 10 15 Glu Gly Phe Leu Ser Glu Leu Thr Gln Gln Leu Ala Gln Ala Thr Gly             20 25 30 Lys Pro Pro Ala Tyr Ile Ala Val His Val Val Pro Asp Gln Leu Met         35 40 45 Thr Phe Ser Gly Thr Asn Asp Pro Cys Ala Leu Cys Ser Leu His Ser     50 55 60 Ile Gly Lys Ile Gly Gly Ala Gln Asn Arg Asn Tyr Ser Lys Leu Leu 65 70 75 80 Cys Gly Leu Leu Ser Asp Arg Leu His Ile Ser Pro Asp Arg Val Tyr                 85 90 95 Ile Asn Tyr Tyr Asp Met Asn Ala Ala Asn Val Gly Trp Gly Asn Ser             100 105 110 Thr Phe Ala Leu         115 <210> 5 <211> 116 <212> PRT <213> Rattus sp. <400> 5 Met Pro Met Phe Ile Val Asn Thr Asn Val Pro Arg Ala Ser Val Pro 1 5 10 15 Glu Gly Phe Leu Ser Glu Leu Thr Gln Gln Leu Ala Gln Ala Thr Gly             20 25 30 Lys Pro Pro Ala Tyr Ile Ala Val His Val Val Pro Asp Gln Leu Met         35 40 45 Thr Phe Ser Gly Thr Ser Asp Pro Cys Ala Leu Cys Ser Leu His Ser     50 55 60 Ile Gly Lys Ile Gly Gly Ala Gln Asn Arg Asn Tyr Ser Lys Leu Leu 65 70 75 80 Cys Gly Leu Leu Ser Asp Arg Leu His Ile Ser Pro Asp Arg Val Tyr                 85 90 95 Ile Asn Tyr Tyr Asp Met Asn Ala Ala Asn Val Gly Trp Gly Asn Ser             100 105 110 Thr Phe Ala Leu         115 <210> 6 <211> 21 <212> PRT <213> Homo sapiens <400> 6 Ser Glu Ala Asp Asp Arg Tyr Ile Cys Asp Arg Phe Tyr Pro Asn Asp 1 5 10 15 Leu Trp Val Val Val             20 <210> 7 <211> 15 <212> PRT <213> Homo sapiens <400> 7 Asp Asp Arg Tyr Ile Cys Asp Arg Phe Tyr Pro Asn Asp Leu Trp 1 5 10 15 <210> 8 <211> 30 <212> PRT <213> Unknown <220> <223> Description of Unknown: MIF peptide sequence <400> 8 Ile Gly Lys Ile Gly Gly Ala Gln Asn Arg Ser Tyr Ser Lys Leu Leu 1 5 10 15 Cys Gly Leu Leu Ala Glu Arg Leu Arg Ile Ser Pro Asp Arg             20 25 30 <210> 9 <211> 16 <212> PRT <213> Homo sapiens <400> 9 Leu Cys Gly Leu Leu Ala Glu Arg Leu Arg Ile Ser Pro Asp Arg Val 1 5 10 15 <210> 10 <211> 43 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       polypeptide <400> 10 Val Asn Thr Asn Val Pro Pro Arg Ala Ser Val Pro Asp Gly Phe Leu 1 5 10 15 Ser Glu Leu Thr Gln Gln Leu Ala Gln Ala Thr Gly Lys Pro Pro Gln             20 25 30 Tyr Ile Ala Val His Val Val Pro Asp Gln Leu         35 40 <210> 11 <211> 19 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 11 Pro Asp Gln Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu 1 5 10 15 Cys ser leu              <210> 12 <211> 58 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       polypeptide <400> 12 Val Asn Thr Asn Val Pro Pro Arg Ala Ser Val Pro Asp Gly Phe Leu 1 5 10 15 Ser Glu Leu Thr Gln Gln Leu Ala Gln Ala Thr Gly Lys Pro Pro Gln             20 25 30 Tyr Ile Ala Val His Val Val Pro Asp Gln Leu Met Ala Phe Gly Gly         35 40 45 Ser Ser Glu Pro Cys Ala Leu Cys Ser Leu     50 55 <210> 13 <211> 22 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 13 Pro Asp Gln Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu 1 5 10 15 Cys Ser Leu His Ser Ile             20 <210> 14 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 14 Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu Cys 1 5 10 <210> 15 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 15 Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu 1 5 10 <210> 16 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 16 Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala 1 5 10 <210> 17 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 17 Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys 1 5 10 <210> 18 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 18 Leu Met Ala Phe Gly Gly Ser Ser Glu Pro 1 5 10 <210> 19 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 19 Leu Met Ala Phe Gly Gly Ser Ser Glu 1 5 <210> 20 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 20 Leu Met Ala Phe Gly Gly Ser Ser 1 5 <210> 21 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 21 Leu Met Ala Phe Gly Gly Ser 1 5 <210> 22 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 22 Leu Met Ala Phe Gly Gly 1 5 <210> 23 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 23 Met Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu Cys 1 5 10 <210> 24 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 24 Met Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu 1 5 10 <210> 25 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 25 Met Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala 1 5 10 <210> 26 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 26 Met Ala Phe Gly Gly Ser Ser Glu Pro Cys 1 5 10 <210> 27 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 27 Met Ala Phe Gly Gly Ser Ser Glu Pro 1 5 <210> 28 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 28 Met Ala Phe Gly Gly Ser Ser Glu 1 5 <210> 29 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 29 Met Ala Phe Gly Gly Ser Ser 1 5 <210> 30 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 30 Met Ala Phe Gly Gly Ser 1 5 <210> 31 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 31 Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu Cys 1 5 10 <210> 32 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 32 Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu 1 5 10 <210> 33 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 33 Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala 1 5 10 <210> 34 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 34 Ala Phe Gly Gly Ser Ser Glu Pro Cys 1 5 <210> 35 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 35 Ala Phe Gly Gly Ser Ser Glu Pro 1 5 <210> 36 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 36 Ala Phe Gly Gly Ser Ser Glu 1 5 <210> 37 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 37 Ala Phe Gly Gly Ser Ser 1 5 <210> 38 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 38 Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu Cys 1 5 10 <210> 39 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 39 Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu 1 5 10 <210> 40 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 40 Phe Gly Gly Ser Ser Glu Pro Cys Ala 1 5 <210> 41 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 41 Phe Gly Gly Ser Ser Glu Pro Cys 1 5 <210> 42 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 42 Phe Gly Gly Ser Ser Glu Pro 1 5 <210> 43 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 43 Phe Gly Gly Ser Ser Glu 1 5 <210> 44 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 44 Gly Gly Ser Ser Glu Pro Cys Ala Leu Cys 1 5 10 <210> 45 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 45 Gly Gly Ser Ser Glu Pro Cys Ala Leu 1 5 <210> 46 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 46 Gly Gly Ser Ser Glu Pro Cys Ala 1 5 <210> 47 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 47 Gly Gly Ser Ser Glu Pro Cys 1 5 <210> 48 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 48 Gly Gly Ser Ser Glu Pro 1 5 <210> 49 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 49 Gly Ser Ser Glu Pro Cys Ala Leu Cys 1 5 <210> 50 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 50 Gly Ser Ser Glu Pro Cys Ala Leu 1 5 <210> 51 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 51 Gly Ser Ser Glu Pro Cys Ala 1 5 <210> 52 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 52 Gly Ser Ser Glu Pro Cys 1 5 <210> 53 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 53 Ser Ser Glu Pro Cys Ala Leu Cys 1 5 <210> 54 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 54 Gly Ser Ser Glu Pro Cys Ala Leu Cys 1 5 <210> 55 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 55 Gly Ser Ser Glu Pro Cys Ala Leu 1 5 <210> 56 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 56 Gly Ser Ser Glu Pro Cys Ala 1 5 <210> 57 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 57 Gly Ser Ser Glu Pro Cys 1 5 <210> 58 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 58 Ser Ser Glu Pro Cys Ala Leu Cys 1 5 <210> 59 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 59 Ser Ser Glu Pro Cys Ala Leu 1 5 <210> 60 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 60 Ser Ser Glu Pro Cys Ala 1 5 <210> 61 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 61 Ser Glu Pro Cys Ala Leu Cys 1 5 <210> 62 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 62 Ser Glu Pro Cys Ala Leu 1 5 <210> 63 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 63 Glu Pro Cys Ala Leu Cys 1 5 <210> 64 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 64 Gln Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu Cys 1 5 10 15 <210> 65 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 65 Gln Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu 1 5 10 <210> 66 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 66 Gln Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala 1 5 10 <210> 67 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 67 Gln Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys 1 5 10 <210> 68 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 68 Gln Leu Met Ala Phe Gly Gly Ser Ser Glu Pro 1 5 10 <210> 69 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 69 Gln Leu Met Ala Phe Gly Gly Ser Ser Glu 1 5 10 <210> 70 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 70 Gln Leu Met Ala Phe Gly Gly Ser Ser 1 5 <210> 71 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 71 Gln Leu Met Ala Phe Gly Gly Ser 1 5 <210> 72 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 72 Gln Leu Met Ala Phe Gly Gly 1 5 <210> 73 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 73 Gln Leu Met Ala Phe Gly 1 5 <210> 74 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 74 Cys Ser Ser Glu Pro Cys Ala Leu Cys 1 5 <210> 75 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 75 Cys Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu Cys 1 5 10 <210> 76 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 76 Cys Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu Cys 1 5 10 15 <210> 77 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 77 Cys Ala Phe Gly Gly Ser Ser Cys 1 5 <210> 78 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 78 Cys Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys Cys 1 5 10 <210> 79 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 79 Cys Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala Cys 1 5 10 <210> 80 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 80 Cys Met Ala Phe Gly Gly Ser Ser Glu Pro Cys 1 5 10 <210> 81 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 81 Cys Gly Gly Ser Ser Glu Pro Cys Ala Cys 1 5 10 <210> 82 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 82 Asn Val Pro Arg Ala Ser Val Pro Asp 1 5 <210> 83 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 83 Val Pro Asp Gly Phe Leu Ser Glu Leu 1 5 <210> 84 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 84 Cys Phe Gly Gly Ser Ser Glu Pro Cys 1 5 <210> 85 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 85 Ile Ala Val His Val Val Pro Asp Gln Leu Met Ala Phe Gly Gly Ser 1 5 10 15 Ser Glu Pro Cys             20 <210> 86 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 86 Cys Leu His Ser Ile Gly Lys Ile Gly Gly Ala Gln Asn Arg Ser Tyr 1 5 10 15 Ser Lys Leu Leu             20 <210> 87 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 87 Pro Cys Ala Leu Leu Cys Ser Leu His Ser Ile Gly Lys Ile Gly 1 5 10 15 <210> 88 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 88 Cys Ser Leu His Ser Ile Gly Lys Ile Gly Gly Ala Gln Asn Arg 1 5 10 15 <210> 89 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 89 Ile Gly Lys Ile Gly Gly Ala Gln Asn Arg Ser Tyr Ser Lys Leu 1 5 10 15 <210> 90 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 90 Gly Ala Gln Asn Arg Ser Tyr Ser Lys Leu Leu Cys Gly Leu Leu Ala 1 5 10 15 <210> 91 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 91 Cys Gly Leu Leu Ala Glu Arg Leu Arg Ile Ser Pro Asp Arg Val 1 5 10 15 <210> 92 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 92 Glu Arg Leu Arg Ile Ser Pro Asp Arg Val Tyr Ile Asn Tyr Tyr 1 5 10 15 <210> 93 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 93 Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu Cys 1 5 10 <210> 94 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 94 Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu 1 5 10 <210> 95 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 95 Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala 1 5 10 <210> 96 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 96 Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys 1 5 10 <210> 97 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 97 Leu Met Ala Phe Gly Gly Ser Ser Glu Pro 1 5 10 <210> 98 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 98 Leu Met Ala Phe Gly Gly Ser Ser Glu 1 5 <210> 99 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 99 Leu Met Ala Phe Gly Gly Ser Ser 1 5 <210> 100 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 100 Leu Met Ala Phe Gly Gly Ser 1 5 <210> 101 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 101 Leu Met Ala Phe Gly Gly 1 5 <210> 102 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 102 Met Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu Cys 1 5 10 <210> 103 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 103 Met Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu 1 5 10 <210> 104 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 104 Met Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala 1 5 10 <210> 105 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 105 Met Ala Phe Gly Gly Ser Ser Glu Pro Cys 1 5 10 <210> 106 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 106 Met Ala Phe Gly Gly Ser Ser Glu Pro 1 5 <210> 107 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 107 Met Ala Phe Gly Gly Ser Ser Glu 1 5 <210> 108 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 108 Met Ala Phe Gly Gly Ser Ser 1 5 <210> 109 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 109 Met Ala Phe Gly Gly Ser 1 5 <210> 110 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 110 Gly Ser Ser Glu Pro Cys Ala Leu Cys 1 5 <210> 111 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 111 Gly Ser Ser Glu Pro Cys Ala Leu 1 5 <210> 112 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 112 Gly Ser Ser Glu Pro Cys Ala 1 5 <210> 113 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 113 Gly Ser Ser Glu Pro Cys 1 5 <210> 114 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 114 Ser Ser Glu Pro Cys Ala Leu Cys 1 5 <210> 115 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 115 Ser Ser Glu Pro Cys Ala Leu 1 5 <210> 116 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 116 Ser Ser Glu Pro Cys Ala 1 5 <210> 117 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 117 Ser Glu Pro Cys Ala Leu Cys 1 5 <210> 118 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 118 Ser Glu Pro Cys Ala Leu 1 5 <210> 119 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 119 Glu Pro Cys Ala Leu Cys 1 5 <210> 120 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 120 Gln Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu Cys 1 5 10 15 <210> 121 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 121 Gln Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu 1 5 10 <210> 122 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 122 Gln Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala 1 5 10 <210> 123 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 123 Gln Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys 1 5 10 <210> 124 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 124 Gln Leu Met Ala Phe Gly Gly Ser Ser Glu Pro 1 5 10 <210> 125 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 125 Gln Leu Met Ala Phe Gly Gly Ser Ser Glu 1 5 10 <210> 126 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 126 Gln Leu Met Ala Phe Gly Gly Ser Ser 1 5 <210> 127 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 127 Gln Leu Met Ala Phe Gly Gly Ser 1 5 <210> 128 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 128 Gln Leu Met Ala Phe Gly Gly 1 5 <210> 129 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 129 Gln Leu Met Ala Phe Gly 1 5 <210> 130 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 130 Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu Cys 1 5 10 <210> 131 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 131 Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu 1 5 10 <210> 132 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 132 Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala 1 5 10 <210> 133 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <133> 133 Ala Phe Gly Gly Ser Ser Glu Pro Cys 1 5 <210> 134 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 134 Ala Phe Gly Gly Ser Ser Glu Pro 1 5 <210> 135 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 135 Ala Phe Gly Gly Ser Ser Glu 1 5 <210> 136 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 136 Ala Phe Gly Gly Ser Ser 1 5 <210> 137 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 137 Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu Cys 1 5 10 <210> 138 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 138 Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu 1 5 10 <210> 139 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 139 Phe Gly Gly Ser Ser Glu Pro Cys Ala 1 5 <210> 140 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 140 Phe Gly Gly Ser Ser Glu Pro Cys 1 5 <210> 141 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 141 Phe Gly Gly Ser Ser Glu Pro 1 5 <210> 142 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 142 Phe Gly Gly Ser Ser Glu 1 5 <210> 143 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 143 Gly Gly Ser Ser Glu Pro Cys Ala Leu Cys 1 5 10 <210> 144 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 144 Gly Gly Ser Ser Glu Pro Cys Ala Leu 1 5 <210> 145 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 145 Gly Gly Ser Ser Glu Pro Cys Ala 1 5 <210> 146 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 146 Gly Gly Ser Ser Glu Pro Cys 1 5 <210> 147 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 147 Gly Gly Ser Ser Glu Pro 1 5 <210> 148 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 148 Cys Ser Ser Glu Pro Cys Ala Leu Cys 1 5 <210> 149 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 149 Cys Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu Cys 1 5 10 <210> 150 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 150 Cys Phe Gly Gly Ser Ser Glu Pro Cys Cys 1 5 10 <210> 151 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 151 Cys Phe Gly Gly Ser Ser Glu Pro Cys 1 5 <210> 152 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 152 Cys Gly Ser Ser Glu Pro Cys Ala Leu Cys 1 5 10 <210> 153 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 153 Cys Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala Cys 1 5 10 <210> 154 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 154 Cys Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu Cys 1 5 10 15 <210> 155 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 155 Cys Ala Phe Gly Gly Ser Ser Cys 1 5 <210> 156 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 156 Cys Thr Asn Val Pro Arg Ala Ser Val Pro Asp Gly Cys 1 5 10 <210> 157 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 157 Cys Val Pro Arg Ala Ser Cys 1 5 <210> 158 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 158 Val Asn Thr Asn Val Pro Arg Ala Ser Val Pro Asp Gly Phe Leu Ser 1 5 10 15 Glu leu          <210> 159 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 159 Asn Thr Asn Val Pro Arg Ala Ser Val Pro Asp Gly Phe Leu Ser Glu 1 5 10 15 Leu      <210> 160 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 160 Thr Asn Val Pro Arg Ala Ser Val Pro Asp Gly Phe Leu Ser Glu Leu 1 5 10 15 <210> 161 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 161 Asn Val Pro Arg Ala Ser Val Pro Asp Gly Phe Leu Ser Glu Leu 1 5 10 15 <210> 162 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 162 Val Pro Arg Ala Ser Val Pro Asp Gly Phe Leu Ser Glu Leu 1 5 10 <210> 163 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 163 Pro Arg Ala Ser Val Pro Asp Gly Phe Leu Ser Glu Leu 1 5 10 <210> 164 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 164 Arg Ala Ser Val Pro Asp Gly Phe Leu Ser Glu Leu 1 5 10 <210> 165 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 165 Ala Ser Val Pro Asp Gly Phe Leu Ser Glu Leu 1 5 10 <210> 166 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 166 Ser Val Pro Asp Gly Phe Leu Ser Glu Leu 1 5 10 <210> 167 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 167 Val Pro Asp Gly Phe Leu Ser Glu Leu 1 5 <210> 168 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 168 Asn Val Pro Arg Ala Ser Val Pro Asp Gly Phe Leu Ser Glu 1 5 10 <210> 169 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 169 Asn Val Pro Arg Ala Ser Val Pro Asp Gly Phe Leu Ser 1 5 10 <210> 170 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 170 Asn Val Pro Arg Ala Ser Val Pro Asp Gly Phe Leu 1 5 10 <210> 171 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 171 Asn Val Pro Arg Ala Ser Val Pro Asp Gly Phe 1 5 10 <210> 172 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 172 Asn Val Pro Arg Ala Ser Val Pro Asp Gly 1 5 10 <210> 173 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 173 Asn Val Pro Arg Ala Ser Val Pro Asp 1 5 <210> 174 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 174 Asn Val Pro Arg Ala Ser Val Pro 1 5 <210> 175 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 175 Val Pro Arg Ala Ser Val Pro 1 5 <210> 176 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 176 Pro Arg Ala Ser Val Pro 1 5 <210> 177 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 177 Val Pro Arg Ala Ser Val Pro Asp Gly Phe Leu 1 5 10 <210> 178 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 178 Val Pro Arg Ala Ser Val Pro Asp Gly Phe 1 5 10 <210> 179 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 179 Val Pro Arg Ala Ser Val Pro Asp Gly 1 5 <210> 180 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 180 Val Pro Arg Ala Ser Val Pro Asp 1 5 <210> 181 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 181 Val Pro Arg Ala Ser Val Pro 1 5 <210> 182 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 182 Val Pro Arg Ala Ser 1 5 <210> 183 <211> 23 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 183 Met Pro Met Phe Ile Val Asn Thr Asn Val Pro Arg Ala Ser Val Pro 1 5 10 15 Asp Gly Phe Leu Ser Glu Cys             20 <210> 184 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 184 Met Pro Met Phe Ile Val Asn Thr Asn Val Pro Arg Ala Ser Val 1 5 10 15 <210> 185 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 185 Phe Ile Val Asn Thr Asn Val Pro Arg Ala Ser Val Pro Asp Gly 1 5 10 15 <210> 186 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 186 Asn Thr Asn Val Pro Arg Ala Ser Val Pro Asp Gly Phe Leu Ser 1 5 10 15 <210> 187 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 187 Val Pro Arg Ala Ser Val Pro Asp Gly Phe Leu Ser Glu Leu Thr 1 5 10 15 <210> 188 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 188 Cys Ser Leu His Ser Ile Gly Lys Ile Gly Gly Ala Gln Asn Arg 1 5 10 15 <210> 189 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 189 Ile Gly Lys Ile Gly Gly Ala Gln Asn Arg Ser Tyr Ser Lys Leu 1 5 10 15 <210> 190 <211> 22 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 190 His Ser Ile Gly Lys Ile Gly Gly Ala Gln Asn Arg Ser Tyr Ser Lys 1 5 10 15 Leu Leu Cys Gly Leu Leu             20 <210> 191 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 191 His Ser Ile Gly Lys Ile Gly Gly Ala Gln Asn Arg Ser Tyr Ser Lys 1 5 10 15 Leu Leu Cys Gly             20 <210> 192 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 192 His Ser Ile Gly Lys Ile Gly Gly Ala Gln Asn Arg Ser Tyr Ser Lys 1 5 10 15 Leu Leu          <210> 193 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 193 His Ser Ile Gly Lys Ile Gly Gly Ala Gln Asn Arg Ser Tyr Ser Lys 1 5 10 15 <210> 194 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 194 His Ser Ile Gly Lys Ile Gly Gly Ala Gln Asn Arg Ser Tyr Ser 1 5 10 15 <210> 195 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 195 Ile Gly Lys Ile Gly Gly Ala Gln Asn Arg Ser Tyr Ser Lys Leu Leu 1 5 10 15 Cys      <210> 196 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 196 Lys Ile Gly Gly Ala Gln Asn Arg Ser Tyr Ser Lys Leu Leu Cys 1 5 10 15 <210> 197 <211> 22 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 197 Gly Gly Ala Gln Asn Arg Ser Tyr Ser Lys Leu Leu Cys Gly Leu Leu 1 5 10 15 Ala Glu Arg Leu Arg Ile             20 <210> 198 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 198 Ala Gln Asn Arg Ser Tyr Ser Lys Leu Leu Cys Gly Leu Leu Ala Glu 1 5 10 15 Arg Leu Arg Ile             20 <210> 199 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 199 Asn Arg Ser Tyr Ser Lys Leu Leu Cys Gly Leu Leu Ala Glu Arg Leu 1 5 10 15 Arg Ile          <210> 200 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 200 Ser Tyr Ser Lys Leu Leu Cys Gly Leu Leu Ala Glu Arg Leu Arg Ile 1 5 10 15 <210> 201 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 201 Tyr Ser Lys Leu Leu Cys Gly Leu Leu Ala Glu Arg Leu Arg Ile 1 5 10 15 <210> 202 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 202 Gly Ala Gln Asn Arg Ser Tyr Ser Lys Leu Leu Cys Gly Leu Leu Ala 1 5 10 15 Glu      <210> 203 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 203 Gly Ala Gln Asn Arg Ser Tyr Ser Lys Leu Leu Cys Gly Leu Leu 1 5 10 15 <210> 204 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 204 Gln Asn Arg Ser Tyr Ser Lys Leu Leu Cys Gly Leu Leu Ala Glu 1 5 10 15 <210> 205 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 205 His Ser Ile Gly Lys Ile Gly Gly Ala Gln Asn Arg Ser Tyr 1 5 10 <210> 206 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 206 His Ser Ile Gly Lys Ile Gly Gly Ala Gln Asn Arg 1 5 10 <210> 207 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 207 His Ser Ile Gly Lys Ile Gly Gly Ala Gln Asn Arg Ser Tyr Ser Lys 1 5 10 15 <210> 208 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 208 Ile Gly Lys Ile Gly Gly Ala Gln Asn Arg Ser Tyr Ser Lys Leu Leu 1 5 10 15 Cys      <210> 209 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 209 Lys Ile Gly Gly Ala Gln Asn Arg Ser Tyr Ser Lys Leu Leu Cys 1 5 10 15 <210> 210 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 210 Lys Ile Gly Gly Ala Gln Asn Arg Ser Tyr Ser 1 5 10 <210> 211 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 211 Gly Ala Gln Asn Arg Ser Tyr Ser Lys Leu Leu Cys Gly Leu Leu Ala 1 5 10 15 Glu      <210> 212 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 212 Gly Ala Gln Asn Arg Ser Tyr Ser Lys Leu Leu Cys Gly Leu Leu 1 5 10 15 <210> 213 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 213 Gly Ala Gln Asn Arg Ser Tyr Ser Lys Leu Leu Cys Gly 1 5 10 <210> 214 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 214 Gly Ala Gln Asn Arg Ser Tyr Ser Lys Leu Leu 1 5 10 <210> 215 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 215 Gln Asn Arg Ser Tyr Ser Lys Leu Leu Cys Gly Leu Leu Ala Glu 1 5 10 15 <210> 216 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 216 Arg Ser Tyr Ser Lys Leu Leu Cys Gly Leu Leu Ala Glu 1 5 10 <210> 217 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 217 Tyr Ser Lys Leu Leu Cys Gly Leu Leu Ala Glu 1 5 10 <210> 218 <211> 43 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       polypeptide <400> 218 Ile Ala Val His Val Val Pro Asp Gln Leu Met Ala Phe Gly Gly Ser 1 5 10 15 Ser Glu Pro Cys Ala Leu Cys Ser Leu His Ser Ile Gly Lys Ile Gly             20 25 30 Gly Ala Gln Asn Arg Ser Tyr Ser Lys Leu Leu         35 40 <210> 219 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       polypeptide <400> 219 Ile Ala Val His Val Val Pro Asp Gln Leu Met Ala Phe Gly Gly Ser 1 5 10 15 Ser Glu Pro Cys Ala Leu Cys Ser Leu His Ser Ile Gly Lys Ile Gly             20 25 30 Gly Ala Gln Asn Arg Ser Tyr         35 <210> 220 <211> 35 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       polypeptide <400> 220 Ile Ala Val His Val Val Pro Asp Gln Leu Met Ala Phe Gly Gly Ser 1 5 10 15 Ser Glu Pro Cys Ala Leu Cys Ser Leu His Ser Ile Gly Lys Ile Gly             20 25 30 Gly ala gln         35 <210> 221 <211> 31 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       polypeptide <400> 221 Ile Ala Val His Val Val Pro Asp Gln Leu Met Ala Phe Gly Gly Ser 1 5 10 15 Ser Glu Pro Cys Ala Leu Cys Ser Leu His Ser Ile Gly Lys Ile             20 25 30 <210> 222 <211> 27 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 222 Ile Ala Val His Val Val Pro Asp Gln Leu Met Ala Phe Gly Gly Ser 1 5 10 15 Ser Glu Pro Cys Ala Leu Cys Ser Leu His Ser             20 25 <210> 223 <211> 23 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 223 Ile Ala Val His Val Val Pro Asp Gln Leu Met Ala Phe Gly Gly Ser 1 5 10 15 Ser Glu Pro Cys Ala Leu Cys             20 <210> 224 <211> 19 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 224 Ile Ala Val His Val Val Pro Asp Gln Leu Met Ala Phe Gly Gly Ser 1 5 10 15 Ser Glu Pro              <210> 225 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 225 Ile Ala Val His Val Val Pro Asp Gln Leu Met Ala Phe Gly Gly 1 5 10 15 <210> 226 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 226 Ile Ala Val His Val Val Pro Asp Gln Leu Met 1 5 10 <210> 227 <211> 43 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       polypeptide <400> 227 Ile Ala Val His Val Val Pro Asp Gln Leu Met Ala Phe Gly Gly Ser 1 5 10 15 Ser Glu Pro Cys Ala Leu Cys Ser Leu His Ser Ile Gly Lys Ile Gly             20 25 30 Gly Ala Gln Asn Arg Ser Tyr Ser Lys Leu Leu         35 40 <210> 228 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       polypeptide <400> 228 Val Val Pro Asp Gln Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys 1 5 10 15 Ala Leu Cys Ser Leu His Ser Ile Gly Lys Ile Gly Gly Ala Gln Asn             20 25 30 Arg Ser Tyr Ser Lys Leu Leu         35 <210> 229 <211> 35 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       polypeptide <400> 229 Gln Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu Cys Ser 1 5 10 15 Leu His Ser Ile Gly Lys Ile Gly Gly Ala Gln Asn Arg Ser Tyr Ser             20 25 30 Lys Leu Leu         35 <210> 230 <211> 31 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       polypeptide <400> 230 Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu Cys Ser Leu His Ser Ile 1 5 10 15 Gly Lys Ile Gly Gly Ala Gln Asn Arg Ser Tyr Ser Lys Leu Leu             20 25 30 <210> 231 <211> 27 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 231 Ser Glu Pro Cys Ala Leu Cys Ser Leu His Ser Ile Gly Lys Ile Gly 1 5 10 15 Gly Ala Gln Asn Arg Ser Tyr Ser Lys Leu Leu             20 25 <210> 232 <211> 23 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 232 Ala Leu Cys Ser Leu His Ser Ile Gly Lys Ile Gly Gly Ala Gln Asn 1 5 10 15 Arg Ser Tyr Ser Lys Leu Leu             20 <210> 233 <211> 19 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 233 Leu His Ser Ile Gly Lys Ile Gly Gly Ala Gln Asn Arg Ser Tyr Ser 1 5 10 15 Lys Leu Leu              <210> 234 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 234 Gly Lys Ile Gly Gly Ala Gln Asn Arg Ser Tyr Ser Lys Leu Leu 1 5 10 15 <210> 235 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 235 Ile Gly Gly Ala Gln Asn Arg Ser Tyr Ser Lys Leu Leu 1 5 10 <210> 236 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 236 Gln Asn Arg Ser Tyr Ser Lys Leu Leu 1 5 <210> 237 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 237 Ile Gly Lys Ile Gly Gly Ala Gln Asn Arg Ser Tyr Ser Lys Leu 1 5 10 15 <210> 238 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 238 Ile Gly Lys Ile Gly Gly Ala Gln 1 5 <210> 239 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 239 Cys Ile Gly Lys Ile Gly Gly Ala Gln Cys 1 5 10 <210> 240 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 240 Cys Ile Gly Lys Ile Gly Gly Ala Gln Cys 1 5 10 <210> 241 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 241 Arg Ser Tyr Ser Lys Leu Leu Cys Gly Leu Leu Ala Glu 1 5 10 <210> 242 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 242 Cys Arg Ser Tyr Ser Lys Leu Leu Cys Gly Leu Leu Ala Glu Cys 1 5 10 15 <210> 243 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 243 Cys Arg Ser Tyr Ser Lys Leu Leu Cys Gly Leu Leu Ala Glu Cys 1 5 10 15 <210> 244 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 244 Cys Gly Leu Leu Ala Glu Arg Leu Arg Ile Ser Pro Asp Arg 1 5 10 <210> 245 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 245 Cys Gly Leu Leu Ala Glu Arg Leu Arg Ile Ser Pro Asp Arg Cys 1 5 10 15 <210> 246 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 246 Cys Gly Leu Leu Ala Glu Arg Leu Arg Ile Ser Pro Asp Arg Cys 1 5 10 15 <210> 247 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 247 Ala Tyr Phe Leu Tyr Gln Gln Gln 1 5 <210> 248 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 248 Gln Gln Gln Gly Arg Leu Asp Lys Leu Thr Val Thr Gly Arg Leu 1 5 10 15 <210> 249 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 249 Gly Arg Leu Asp Lys Leu Thr Val Thr Ser Gln Asn Leu Gln Leu 1 5 10 15 <210> 250 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 250 Ser Gln Asn Leu Gln Leu Glu Asn Leu Arg Met 1 5 10 <210> 251 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 251 Thr Val Thr Gly Arg Leu Asp Lys Leu Thr Val Thr Ser Gln Asn 1 5 10 15 <210> 252 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 252 Thr Val Thr Ser Gln Asn Leu Gln Leu Glu Asn Leu Arg Met 1 5 10 <210> 253 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 253 Leu Glu Asn Leu Arg Met Lys Leu Pro Lys Pro Pro Lys Pro Val 1 5 10 15 <210> 254 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 254 Lys Leu Pro Lys Pro Pro Lys Pro Val Ser Lys Met Arg Met Ala 1 5 10 15 <210> 255 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 255 Ser Lys Met Arg Met Ala Thr Pro Leu 1 5 <210> 256 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 256 Leu Met Gln Ala Leu Pro Met Gly Ala Leu Pro Gln Gly Pro Met 1 5 10 15 <210> 257 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 257 Leu Pro Gln Gly Pro Met Gln Asn Ala Thr Lys Tyr Gly Asn Met 1 5 10 15 <210> 258 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 258 Thr Lys Tyr Gly Asn Met Thr Glu Asp His Val Met His Leu Leu 1 5 10 15 <210> 259 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 259 His Val Met His Leu Leu Gln Asn Ala Asp Pro Leu Lys Val Tyr 1 5 10 15 <210> 260 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 260 Asp Pro Leu Lys Val Tyr Pro Pro Leu Lys Gly Ser Phe Pro Glu 1 5 10 15 <210> 261 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 261 Lys Gly Ser Phe Pro Glu Asn Leu Arg His Leu Lys Asn Thr Met 1 5 10 15 <210> 262 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 262 His Leu Lys Asn Thr Met Glu Thr Ile Asp Trp Lys Val Phe Glu 1 5 10 15 <210> 263 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 263 Asp Trp Lys Val Phe Glu Ser Trp Met His His Trp Leu Leu Phe 1 5 10 15 <210> 264 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 264 His His Trp Leu Leu Phe Glu Met Ser Arg His Ser Leu Glu Gln 1 5 10 15 <210> 265 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 265 Arg His Ser Leu Glu Gln Lys Pro Thr Asp Ala Pro Pro Lys Glu 1 5 10 15 <210> 266 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 266 Asp Ala Pro Pro Lys Glu Ser Leu Glu Leu Glu Asp Pro Ser Ser 1 5 10 15 <210> 267 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 267 Leu Glu Asp Pro Ser Ser Gly Leu Gly Val Thr Lys Gln Asp Leu 1 5 10 15 <210> 268 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 268 Val Thr Lys Gln Asp Leu Gly Pro Val Pro Met 1 5 10 <210> 269 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 269 Met Asp Asp Gln Arg Asp Leu Ile Ser Asn Asn Glu Gln Leu Pro 1 5 10 15 <210> 270 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 270 Leu Pro Met Leu Gly Arg Arg Pro Gly Ala Pro Glu Ser Lys Cys 1 5 10 15 <210> 271 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 271 Cys Ser Arg Gly Ala Leu Tyr Thr Gly Phe Ser Ile Leu Val Thr 1 5 10 15 <210> 272 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 272 Phe Ser Ile Leu Val Thr Leu Leu Leu Ala Gly Gln Ala Thr Thr 1 5 10 15 <210> 273 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 273 Ala Tyr Phe Leu Tyr Gln Gln Gln Gly Arg Leu Asp Lys Leu Thr 1 5 10 15 <210> 274 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 274 Gly Arg Leu Asp Lys Leu Thr Val Thr Ser Gln Asn Leu Gln Leu 1 5 10 15 <210> 275 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 275 Ser Gln Asn Leu Gln Leu Glu Asn Leu Arg Met Lys Leu Pro Lys 1 5 10 15 <210> 276 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 276 Lys Leu Pro Lys Pro Pro Lys Pro Val Ser Lys Met Arg Met Ala 1 5 10 15 <210> 277 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 277 Ser Lys Met Arg Met Ala Thr Pro Leu Leu Met Gln Ala Leu Pro 1 5 10 15 <210> 278 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 278 Leu Met Gln Ala Leu Pro Met Gly Ala Leu Pro Gln Gly Pro Met 1 5 10 15 <210> 279 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 279 Leu Pro Gln Gly Pro Met Gln Asn Ala Thr Lys Tyr Gly Asn Met 1 5 10 15 <210> 280 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 280 Thr Lys Tyr Gly Asn Met Thr Glu Asp His Val Met His Leu Leu 1 5 10 15 <210> 281 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 281 His Val Met His Leu Leu Gln Asn Ala Asp Pro Leu Lys Val Tyr 1 5 10 15 <210> 282 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 282 Asp Pro Leu Lys Val Tyr Pro Pro Leu Lys Gly Ser Phe Pro Glu 1 5 10 15 <210> 283 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 283 Lys Gly Ser Phe Pro Glu Asn Leu Arg His Leu Lys Asn Thr Met 1 5 10 15 <210> 284 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 284 His Leu Lys Asn Thr Met Glu Thr Ile Asp Trp Lys Val Phe Glu 1 5 10 15 <210> 285 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 285 Asp Trp Lys Val Phe Glu Ser Trp Met His His Trp Leu Leu Phe 1 5 10 15 <210> 286 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 286 His His Trp Leu Leu Phe Glu Met Ser Arg His Ser Leu Glu Gln 1 5 10 15 <210> 287 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 287 Arg His Ser Leu Glu Gln Lys Pro Thr Asp Ala Pro Pro Lys Glu 1 5 10 15 <210> 288 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 288 Asp Ala Pro Pro Lys Glu Ser Leu Glu Leu Glu Asp Pro Ser Ser 1 5 10 15 <210> 289 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 289 Leu Glu Asp Pro Ser Ser Gly Leu Gly Val Thr Lys Gln Asp Leu 1 5 10 15 <210> 290 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 290 Ser Gly Leu Gly Val Thr Lys Gln Asp Leu Gly Pro Val Pro Met 1 5 10 15 <210> 291 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 291 Met Asp Asp Gln Arg Asp Leu Ile Ser Asn His Glu Gln Leu Pro 1 5 10 15 <210> 292 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 292 Leu Pro Ile Leu Gly Asn Arg Pro Arg Glu Pro Glu Arg Cys Ser 1 5 10 15 <210> 293 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 293 Cys Ser Arg Gly Ala Leu Tyr Thr Gly Val Ser Val Leu Val Ala 1 5 10 15 <210> 294 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 294 Val Ser Val Leu Val Ala Leu Leu Leu Ala Gly Gln Ala Thr Thr 1 5 10 15 <210> 295 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 295 Ala Tyr Phe Leu Tyr Gln Gln Gln Gly Arg Leu Asp Lys Leu Thr 1 5 10 15 <210> 296 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 296 Leu Thr Ile Thr Ser Gln Asn Leu Gln Leu Glu Ser Leu Arg Met 1 5 10 15 <210> 297 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 297 Arg Met Lys Leu Pro Lys Ser Ala Lys Pro Val Ser Gln Met Arg 1 5 10 15 <210> 298 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 298 Met Arg Met Ala Thr Pro Leu Leu Met Arg Pro Met Ser Met Asp 1 5 10 15 <210> 299 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 299 Met Asp Asn Met Leu Leu Gly Pro Val Lys Asn Val Thr Lys Tyr 1 5 10 15 <210> 300 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 300 Lys Tyr Gly Asn Met Thr Gln Asp His Val Met His Leu Leu Thr 1 5 10 15 <210> 301 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 301 Arg Ser Gly Pro Leu Glu Tyr Pro Gln Leu Lys Gly Thr Phe Pro 1 5 10 15 <210> 302 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 302 Phe Pro Glu Asn Leu Lys His Leu Lys Asn Ser Met Asp Gly Val 1 5 10 15 <210> 303 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 303 Gly Val Asn Trp Lys Ile Phe Glu Ser Trp Met Lys Gln Trp Leu 1 5 10 15 <210> 304 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 304 Trp Leu Leu Phe Glu Met Ser Lys Asn Ser Leu Glu Glu Lys Lys 1 5 10 15 <210> 305 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 305 Glu Lys Lys Pro Thr Glu Ala Pro Pro Lys Val Leu Thr Lys Cys 1 5 10 15 <210> 306 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 306 Cys Gln Glu Glu Val Ser His Ile Pro Ala Val Tyr Pro Gly Ala 1 5 10 15 <210> 307 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 307 Gly Ala Phe Arg Pro Lys Cys Asp Glu Asn Gly Asn Tyr Leu Pro 1 5 10 15 <210> 308 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 308 Leu Pro Leu Gln Cys His Gly Ser Thr Gly Tyr Cys Trp Cys Val 1 5 10 15 <210> 309 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 309 Cys Val Phe Pro Asn Gly Thr Glu Val Pro His Thr Lys Ser Arg 1 5 10 15 <210> 310 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 310 Ser Arg Gly Arg His Asn Cys Ser Glu Pro Leu Asp Met Glu Asp 1 5 10 15 <210> 311 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 311 Glu Asp Leu Ser Ser Gly Leu Gly Val Thr Arg Gln Glu Leu Gly 1 5 10 15 <210> 312 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 312 Ser Gly Leu Gly Val Thr Arg Gln Glu Leu Gly Gln Val Thr Leu 1 5 10 15 <210> 313 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 313 Asp Leu Ser Asn Tyr Ser Tyr Ser Ser Thr Leu Pro Pro Phe Leu 1 5 10 15 <210> 314 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 314 Asp Leu Ser Asn Tyr Ser Tyr Ser Ser Thr Leu Pro Pro 1 5 10 <210> 315 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 315 Asp Leu Ser Asn Tyr Ser Tyr Ser Ser Thr Leu 1 5 10 <210> 316 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 316 Asp Leu Ser Asn Tyr Ser Tyr Ser Ser 1 5 <210> 317 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 317 Asp Leu Ser Asn Tyr Ser Tyr 1 5 <210> 318 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 318 Asp Leu Ser Asn Tyr 1 5 <210> 319 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 319 Lys Val Asn Gly Trp Ile Phe Gly Thr Phe Leu 1 5 10 <210> 320 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 320 Lys Val Asn Gly Trp Ile Phe Gly Thr 1 5 <210> 321 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 321 Lys Val Asn Gly Trp Ile Phe 1 5 <210> 322 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 322 Lys Val Asn Gly Trp 1 5 <210> 323 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 323 Arg Arg Thr Val Tyr Ser Ser Asn Val Ser Pro Ala Cys 1 5 10 <210> 324 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 324 Arg Arg Thr Val Tyr Ser Ser Asn Val Ser Pro 1 5 10 <210> 325 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 325 Arg Arg Thr Val Tyr Ser Ser Asn Val 1 5 <210> 326 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 326 Arg Arg Thr Val Tyr Ser Ser 1 5 <210> 327 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 327 Arg Arg Thr Val Tyr 1 5 <210> 328 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 328 Glu Asp Met Gly Asn Asn Thr Ala Asn Trp Arg Met Leu 1 5 10 <210> 329 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 329 Glu Asp Met Gly Asn Asn Thr Ala Asn Trp Arg 1 5 10 <210> 330 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 330 Glu Asp Met Gly Asn Asn Thr Ala Asn 1 5 <210> 331 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 331 Glu Asp Met Gly Asn Asn Thr 1 5 <210> 332 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 332 Glu Asp Met Gly Asn 1 5 <210> 333 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 333 Met Arg Thr Gln Val Ile Gln Glu Thr Cys Glu Arg Arg 1 5 10 <210> 334 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 334 Met Arg Thr Gln Val Ile Gln Glu Thr Cys Glu 1 5 10 <210> 335 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 335 Met Arg Thr Gln Val Ile Gln Glu Thr 1 5 <210> 336 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 336 Met Arg Thr Gln Val Ile Gln 1 5 <210> 337 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 337 Met Arg Thr Gln Val 1 5 <210> 338 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 338 Cys Glu Arg Arg Asn His Ile Asp Arg Ala Leu Asp Ala 1 5 10 <210> 339 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 339 Cys Glu Arg Arg Asn His Ile Asp Arg Ala Leu 1 5 10 <210> 340 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 340 Cys Glu Arg Arg Asn His Ile Asp Arg 1 5 <210> 341 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 341 Cys Glu Arg Arg Asn His Ile 1 5 <210> 342 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 342 Cys Glu Arg Arg Asn 1 5 <210> 343 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 343 Asp Arg Tyr Ile Cys Asp Arg Phe Tyr Pro Asn Asp Leu 1 5 10 <210> 344 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 344 Asp Arg Tyr Ile Cys Asp Arg Phe Tyr Pro Asn 1 5 10 <210> 345 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 345 Asp Arg Tyr Ile Cys Asp Arg Phe Tyr 1 5 <210> 346 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 346 Asp Arg Tyr Ile Cys Asp Arg 1 5 <210> 347 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 347 Asp Arg Tyr Ile Cys 1 5 <210> 348 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 348 Ile Cys Asp Arg Phe Tyr Pro Asn Asp Leu Trp Val Val 1 5 10 <210> 349 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 349 Ile Cys Asp Arg Phe Tyr Pro 1 5 <210> 350 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 350 Ile Cys Asp Arg Phe 1 5 <210> 351 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 351 Arg Phe Tyr Pro Asn Asp Leu Trp Val Val Val Phe Gln 1 5 10 <210> 352 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <352> 352 Arg Phe Tyr Pro Asn Asp Leu Trp Val Val Val 1 5 10 <210> 353 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 353 Arg Phe Tyr Pro Asn Asp Leu Trp Val 1 5 <210> 354 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 354 Arg Phe Tyr Pro Asn Asp Leu 1 5 <210> 355 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 355 Arg Phe Tyr Pro Asn 1 5 <210> 356 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 356 Ile Cys Asp Arg Phe Tyr Pro Asn Asp Leu Trp 1 5 10 <210> 357 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 357 Ile Cys Asp Arg Phe Tyr Pro Asn Asp 1 5 <210> 358 <211> 361 <212> PRT <213> Homo sapiens <400> 358 Met Asp Lys Phe Trp Trp His Ala Ala Trp Gly Leu Cys Leu Val Pro 1 5 10 15 Leu Ser Leu Ala Gln Ile Asp Leu Asn Ile Thr Cys Arg Phe Ala Gly             20 25 30 Val Phe His Val Glu Lys Asn Gly Arg Tyr Ser Ile Ser Arg Thr Glu         35 40 45 Ala Ala Asp Leu Cys Lys Ala Phe Asn Ser Thr Leu Pro Thr Met Ala     50 55 60 Gln Met Glu Lys Ala Leu Ser Ile Gly Phe Glu Thr Cys Arg Tyr Gly 65 70 75 80 Phe Ile Glu Gly His Val Val Ile Pro Arg Ile His Pro Asn Ser Ile                 85 90 95 Cys Ala Ala Asn Asn Thr Gly Val Tyr Ile Leu Thr Ser Asn Thr Ser             100 105 110 Gln Tyr Asp Thr Tyr Cys Phe Asn Ala Ser Ala Pro Pro Glu Glu Asp         115 120 125 Cys Thr Ser Val Thr Asp Leu Pro Asn Ala Phe Asp Gly Pro Ile Thr     130 135 140 Ile Thr Ile Val Asn Arg Asp Gly Thr Arg Tyr Val Gln Lys Gly Glu 145 150 155 160 Tyr Arg Thr Asn Pro Glu Asp Ile Tyr Pro Ser Asn Pro Thr Asp Asp                 165 170 175 Asp Val Ser Ser Gly Ser Ser Ser Glu Arg Ser Ser Thr Ser Gly Gly             180 185 190 Tyr Ile Phe Tyr Thr Phe Ser Thr Val His Pro Ile Pro Asp Glu Asp         195 200 205 Ser Pro Trp Ile Thr Asp Ser Thr Asp Arg Ile Pro Ala Thr Arg Asp     210 215 220 Gln Asp Thr Phe His Pro Ser Gly Gly Ser His Thr Thr His Gly Ser 225 230 235 240 Glu Ser Asp Gly His Ser His Gly Ser Gln Glu Gly Gly Ala Asn Thr                 245 250 255 Thr Ser Gly Pro Ile Arg Thr Pro Gln Ile Pro Glu Trp Leu Ile Ile             260 265 270 Leu Ala Ser Leu Leu Ala Leu Ala Leu Ile Leu Ala Val Cys Ile Ala         275 280 285 Val Asn Ser Arg Arg Arg Cys Gly Gln Lys Lys Lys Leu Val Ile Asn     290 295 300 Ser Gly Asn Gly Ala Val Glu Asp Arg Lys Pro Ser Gly Leu Asn Gly 305 310 315 320 Glu Ala Ser Lys Ser Gln Glu Met Val His Leu Val Asn Lys Glu Ser                 325 330 335 Ser Glu Thr Pro Asp Gln Phe Met Thr Ala Asp Glu Thr Arg Asn Leu             340 345 350 Gln Asn Val Asp Met Lys Ile Gly Val         355 360 <210> 359 <211> 365 <212> PRT <213> Mus sp. <400> 359 Met Asp Lys Phe Trp Trp His Thr Ala Trp Gly Leu Cys Leu Leu Gln 1 5 10 15 Leu Ser Leu Ala His Pro His Gln Gln Ile Asp Leu Asn Val Thr Cys             20 25 30 Arg Tyr Ala Gly Val Phe His Val Glu Lys Asn Gly Arg Tyr Ser Ile         35 40 45 Ser Arg Thr Glu Ala Ala Asp Leu Cys Gln Ala Phe Asn Ser Thr Leu     50 55 60 Pro Thr Met Asp Gln Met Lys Leu Ala Leu Ser Lys Gly Phe Glu Thr 65 70 75 80 Cys Arg Tyr Gly Phe Ile Glu Gly Asn Val Val Ile Pro Arg Ile His                 85 90 95 Pro Asn Ala Ile Cys Ala Ala Asn His Thr Gly Val Tyr Ile Leu Val             100 105 110 Thr Ser Asn Thr Ser His Tyr Asp Thr Tyr Cys Phe Asn Ala Ser Ala         115 120 125 Pro Pro Glu Glu Asp Cys Thr Ser Val Thr Asp Leu Pro Asn Ser Phe     130 135 140 Asp Gly Pro Val Thr Ile Thr Ile Val Asn Arg Asp Gly Thr Arg Tyr 145 150 155 160 Ser Lys Lys Gly Glu Tyr Arg Thr His Gln Glu Asp Ile Asp Ala Ser                 165 170 175 Asn Ile Ile Asp Asp Asp Val Ser Ser Gly Ser Thr Ile Glu Lys Ser             180 185 190 Thr Pro Glu Ser Tyr Ile Leu His Thr Tyr Leu Pro Thr Glu Gln Pro         195 200 205 Thr Gly Asp Gln Asp Asp Ser Phe Phe Ile Arg Ser Thr Leu Ala Thr     210 215 220 Arg Asp Arg Asp Ser Ser Lys Asp Ser Arg Gly Ser Ser Arg Thr Val 225 230 235 240 Thr His Gly Ser Glu Leu Ala Gly His Ser Ser Ala Asn Gln Asp Ser                 245 250 255 Gly Val Thr Thr Thr Ser Gly Pro Met Arg Arg Pro Gln Ile Pro Glu             260 265 270 Trp Leu Ile Ile Leu Ala Ser Leu Leu Ala Leu Ala Leu Ile Leu Ala         275 280 285 Val Cys Ile Ala Val Asn Ser Arg Arg Arg Cys Gly Gln Lys Lys Lys     290 295 300 Leu Val Ile Asn Gly Gly Asn Gly Thr Val Glu Asp Arg Lys Pro Ser 305 310 315 320 Glu Leu Asn Gly Glu Ala Ser Lys Ser Gln Glu Met Val His Leu Val                 325 330 335 Asn Lys Glu Pro Ser Glu Thr Pro Asp Gln Cys Met Thr Ala Asp Glu             340 345 350 Thr Arg Asn Leu Gln Ser Val Asp Met Lys Ile Gly Val         355 360 365 <210> 360 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 360 Met Asp Lys Phe Trp Trp His Ala Ala Trp Gly Leu Cys Leu Val 1 5 10 15 <210> 361 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 361 Leu Val Pro Leu Ser Leu Ala Gln Ile Asp Leu Asn Ile Thr Cys 1 5 10 15 <210> 362 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 362 Cys Arg Phe Ala Gly Val Phe His Val Glu Lys Asn Gly Arg Tyr 1 5 10 15 <210> 363 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 363 Arg Tyr Ser Ile Ser Arg Thr Glu Ala Ala Asp Leu Cys Lys Ala 1 5 10 15 <210> 364 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 364 Lys Ala Ala Phe Asn Ser Thr Leu Pro Thr Met Ala Gln Met Glu 1 5 10 15 <210> 365 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 365 Lys Ala Leu Ser Ile Gly Phe Glu Thr Cys Arg Tyr Gly Phe Ile 1 5 10 15 <210> 366 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 366 Phe Ile Glu Gly His Val Val Ile Pro Arg Ile His Pro Asn Ser 1 5 10 15 <210> 367 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 367 Asn Ser Ile Cys Ala Ala Asn Asn Thr Gly Val Tyr Ile Leu Thr 1 5 10 15 <210> 368 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 368 Leu Thr Ser Asn Thr Ser Gln Tyr Asp Thr Tyr Cys Phe Asn Ala 1 5 10 15 <210> 369 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 369 Asn Ala Ser Ala Pro Pro Glu Glu Asp Cys Thr Ser Val Thr Asp 1 5 10 15 <210> 370 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 370 Thr Asp Leu Pro Asn Ala Phe Asp Gly Pro Ile Thr Ile Thr Ile 1 5 10 15 <210> 371 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 371 Thr Ile Val Asn Arg Asp Gly Thr Arg Tyr Val Gln Lys Gly Glu 1 5 10 15 <210> 372 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 372 Gly Glu Tyr Arg Thr Asn Pro Glu Asp Ile Tyr Pro Ser Asn Pro 1 5 10 15 <210> 373 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 373 Asn Pro Thr Asp Asp Asp Val Ser Ser Gly Ser Ser Ser Glu Arg 1 5 10 15 <210> 374 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 374 Glu Arg Ser Ser Thr Ser Gly Gly Tyr Ile Phe Tyr Thr Phe Ser 1 5 10 15 <210> 375 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 375 Phe Ser Thr Val His Pro Ile Pro Asp Glu Asp Ser Pro Trp Ile 1 5 10 15 <210> 376 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 376 Trp Ile Thr Asp Ser Thr Asp Arg Ile Pro Ala Thr Arg Asp Gln 1 5 10 15 <210> 377 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 377 Asp Gln Asp Thr Phe His Pro Ser Gly Gly Ser His Thr Thr His 1 5 10 15 <210> 378 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 378 Thr His Gly Ser Glu Ser Asp Gly His Ser His Gly Ser Gln Glu 1 5 10 15 <210> 379 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 379 Gln Glu Gly Gly Ala Asn Thr Thr Ser Ser Gly Pro Ile Arg Thr Pro 1 5 10 15 <210> 380 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 380 Thr Pro Gln Ile Pro Glu Trp Leu Ile Ile Leu Ala Ser Leu Leu 1 5 10 15 <210> 381 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 381 Leu Leu Ala Leu Ala Leu Ile Leu Ala Val Cys Ile Ala Val Asn 1 5 10 15 <210> 382 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 382 Val Asn Ser Arg Arg Arg Cys Gly Gln Lys Lys Lys Leu Val Ile 1 5 10 15 <210> 383 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 383 Val Ile Asn Ser Gly Asn Gly Ala Val Glu Asp Arg Lys Pro Ser 1 5 10 15 <210> 384 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 384 Pro Ser Gly Leu Asn Gly Glu Ala Ser Lys Ser Gln Glu Met Val 1 5 10 15 <210> 385 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 385 Met Val His Leu Val Asn Lys Glu Ser Ser Glu Thr Pro Asp Gln 1 5 10 15 <210> 386 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 386 Asp Gln Phe Met Thr Ala Asp Glu Thr Arg Asn Leu Gln Asn Val 1 5 10 15 <210> 387 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 387 Asp Glu Thr Arg Asn Leu Gln Asn Val Asp Met Lys Ile Gly Val 1 5 10 15 <210> 388 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 388 Met Asp Lys Phe Trp Trp His Thr Ala Trp Gly Leu Cys 1 5 10 <210> 389 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 389 Leu Leu Gln Leu Ser Leu Ala His Pro His Gln Gln Ile 1 5 10 <210> 390 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 390 Gln Ile Asp Leu Asn Val Thr Cys Arg Tyr Ala Gly Val 1 5 10 <210> 391 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 391 Gly Val Phe His Val Glu Lys Asn Gly Arg Tyr Ser Ile 1 5 10 <210> 392 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 392 Ser Ile Ser Arg Thr Glu Ala Ala Asp Leu Cys Gln Ala 1 5 10 <210> 393 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 393 Gln Ala Phe Asn Ser Thr Leu Pro Thr Met Asp Gln Met 1 5 10 <210> 394 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 394 Gln Met Lys Leu Ala Leu Ser Lys Gly Phe Glu Thr Cys 1 5 10 <210> 395 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 395 Cys Arg Tyr Gly Phe Ile Glu Gly Asn Val Val Ile Pro 1 5 10 <210> 396 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 396 Ile Pro Arg Ile His Pro Asn Ala Ile Cys Ala Ala Asn 1 5 10 <210> 397 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 397 Ala Asn His Thr Gly Val Tyr Ile Leu Val Thr Ser Asn 1 5 10 <210> 398 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 398 Ser Asn Thr Ser His Tyr Asp Thr Tyr Cys Phe Asn Ala 1 5 10 <210> 399 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 399 Asn Ala Ser Ala Pro Pro Glu Glu Asp Cys Thr Ser Val 1 5 10 <210> 400 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 400 Ser Val Thr Asp Leu Pro Asn Ser Phe Asp Gly Pro Val 1 5 10 <210> 401 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 401 Pro Val Thr Ile Thr Ile Val Asn Arg Asp Gly Thr Arg 1 5 10 <210> 402 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 402 Thr Arg Tyr Ser Lys Lys Gly Glu Tyr Arg Thr His Gln 1 5 10 <210> 403 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 403 His Gln Glu Asp Ile Asp Ala Ser Asn Ile Ile Asp Asp 1 5 10 <210> 404 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 404 Asp Asp Val Ser Ser Gly Ser Thr Ile Glu Lys Ser Thr 1 5 10 <210> 405 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 405 Ser Thr Pro Glu Ser Tyr Ile Leu His Thr Tyr Leu Pro 1 5 10 <210> 406 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 406 Leu Pro Thr Glu Gln Pro Thr Gly Asp Gln Asp Asp Ser 1 5 10 <210> 407 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 407 Asp Ser Phe Phe Ile Arg Ser Thr Leu Ala Thr Arg Asp 1 5 10 <210> 408 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 408 Arg Asp Arg Asp Ser Ser Lys Asp Ser Arg Gly Ser Ser 1 5 10 <210> 409 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 409 Ser Ser Arg Thr Val Thr His Gly Ser Glu Leu Ala Gly 1 5 10 <210> 410 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 410 Ala Gly His Ser Ser Ala Asn Gln Asp Ser Gly Val Thr 1 5 10 <210> 411 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 411 Thr Thr Ser Gly Pro Met Arg Arg Pro Gln Ile Pro Glu 1 5 10 <210> 412 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 412 Pro Glu Trp Leu Ile Ile Leu Ala Ser Leu Leu Ala Leu 1 5 10 <210> 413 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 413 Ala Leu Ala Leu Ile Leu Ala Val Cys Ile Ala Val Asn 1 5 10 <210> 414 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 414 Val Asn Ser Arg Arg Arg Cys Gly Gln Lys Lys Lys Leu 1 5 10 <210> 415 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 415 Lys Leu Val Ile Asn Gly Gly Asn Gly Thr Val Glu Asp 1 5 10 <210> 416 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 416 Glu Asp Arg Lys Pro Ser Glu Leu Asn Gly Glu Ala Ser 1 5 10 <210> 417 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 417 Ala Ser Lys Ser Gln Glu Met Val His Leu Val Asn Lys 1 5 10 <210> 418 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 418 Asn Lys Glu Pro Ser Glu Thr Pro Asp Gln Cys Met Thr 1 5 10 <210> 419 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 419 Met Thr Ala Asp Glu Thr Arg Asn Leu Gln Ser Val Asp 1 5 10 <210> 420 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 420 Thr Arg Asn Leu Gln Ser Val Asp Met Lys Ile Gly Val 1 5 10 <210> 421 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 421 Val His Val Val Pro Asp Gln Leu Met Ala 1 5 10 <210> 422 <211> 362 <212> DNA <213> Homo sapiens <400> 422 ggtaccggat cccccatgtt catcgtgaac accaacgtgc ccagagccag cgtgcccgac 60 gcttcctgag cgagctgaca cagcagctgg cccaggccac cggcaagccc cctcagtata 120 tcgccgtgca cgtggtgccc gaccagctga tggccttcgg cggcagcagc gagccttgcg 180 ccctgtgtag cctgcacagc atcggcaaga tcggcggagc ccagaacaga agctacagca 240 agctgctgtg cggcctgctg gccgagagac tgagaatcag ccccgacaga gtgtacatca 300 actactacga catgaacgcc gccaacgtgg gctggaacaa cagcaccttc gccctcgagc 360 tc 362 <210> 423 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 423 Cys Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu Cys 1 5 10 15 <210> 424 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 424 Cys Gly Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Gly Cys 1 5 10 <210> 425 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 425 Cys Gly Gly Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Gly Gly Cys 1 5 10 15 <210> 426 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 426 Cys Gly Gly Ser Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Ser Gly 1 5 10 15 Gly cys          <210> 427 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 427 Cys Gly Gly Ser Gly Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Gly 1 5 10 15 Ser Gly Gly Cys             20 <210> 428 <211> 22 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 428 Cys Gly Gly Ser Gly Gly Leu Met Ala Phe Gly Gly Ser Ser Glu Pro 1 5 10 15 Gly Gly Ser Gly Gly Cys             20 <210> 429 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <220> <221> MOD_RES (222) (12) .. (12) <223> Abu <400> 429 Cys Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Xaa Ala Leu Cys 1 5 10 15 <210> 430 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <221> MOD_RES <222> (4) (4) <223> NLe <220> <221> MOD_RES (222) (14) .. (14) <223> NLe <400> 430 Leu Met Ala Leu Ala Phe Gly Gly Ser Ser Glu Pro Cys Leu 1 5 10 <210> 431 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <221> MOD_RES <222> (4) (4) L-cyclohexylalanine <220> <221> MOD_RES (222) (14) .. (14) L-cyclohexylalanine <400> 431 Leu Met Ala Ala Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala 1 5 10 <210> 432 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <221> MOD_RES <222> (4) (4) <223> D-cyclohexylalanine <220> <221> MOD_RES (222) (14) .. (14) <223> D-cyclohexylalanine <400> 432 Leu Met Ala Ala Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala 1 5 10 <210> 433 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <221> MOD_RES <222> (4) (4) <223> D-Phe <220> <221> MOD_RES (222) (14) .. (14) <223> D-Phe <400> 433 Leu Met Ala Phe Ala Phe Gly Gly Ser Ser Glu Pro Cys Phe 1 5 10 <210> 434 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <221> MOD_RES (222) (1) .. (1) <223> D-Met <400> 434 Met Ala Phe Gly Gly Ser Ser Glu Pro Cys 1 5 10 <210> 435 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <221> MOD_RES (222) (1) .. (1) <223> D-Cys <400> 435 Cys Pro Glu Ser Ser Gly Gly Phe Ala Met Leu 1 5 10 <210> 436 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 436 Cys Pro Glu Ser Ser Gly Gly Phe Ala Met Leu 1 5 10 <210> 437 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 437 Val Asn Thr Asn Val Pro Arg Ala Ser Val Pro Asp Gly 1 5 10 <210> 438 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 438 Cys Pro Arg Ala Ser Val Pro Asp Gly Cys 1 5 10 <210> 439 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 439 Cys Gly Gly Ser Gly Gly Pro Arg Ala Ser Val Pro Asp Gly Gly Gly 1 5 10 15 Ser Gly Gly Cys             20 <210> 440 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 440 Cys Asn Val Pro Arg Ala Ser Val Pro Asp Gly Cys 1 5 10 <210> 441 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 441 Cys Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu Cys 1 5 10 <210> 442 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 442 Cys Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu Cys 1 5 10 15 <210> 443 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 443 Cys Ser Ser Glu Pro Cys Ala Leu Cys 1 5 <210> 444 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 444 Cys Phe Gly Gly Ser Ser Glu Pro Cys Cys 1 5 10 <210> 445 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 445 Cys Phe Gly Gly Ser Ser Glu Pro Cys 1 5 <210> 446 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 446 Cys Gly Ser Ser Glu Pro Cys Ala Leu Cys Cys 1 5 10 <210> 447 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 447 Cys Ala Phe Gly Gly Ser Ser Cys 1 5 <210> 448 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 448 Cys Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala Cys 1 5 10 <210> 449 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 449 Cys Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala Cys 1 5 10 <210> 450 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 450 Cys Ala Phe Gly Gly Ser Ser Cys 1 5 <210> 451 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 451 Cys Val His Val Val Pro Asp Gln Leu Met Ala Cys 1 5 10 <210> 452 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 452 Cys Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala Cys Gly 1 5 10 15 <210> 453 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 453 Cys Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys Cys Gly Gly 1 5 10 15 <210> 454 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 454 Cys Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys Gly Gly Gly 1 5 10 15 <210> 455 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 455 Cys Leu Met Ala Phe Gly Gly Ser Ser Glu Cys Gly Gly Gly Gly 1 5 10 15 <210> 456 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 456 Cys Leu Met Ala Phe Gly Gly Ser Ser Cys Gly Gly Gly Gly Gly 1 5 10 15 <210> 457 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 457 Cys Leu Met Ala Phe Gly Gly Ser Cys Gly Gly Gly Gly Gly 1 5 10 <210> 458 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 458 Cys Leu Met Ala Phe Gly Gly Cys Gly Gly Gly Gly Gly Gly Gly 1 5 10 15 <210> 459 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 459 Cys Leu Met Ala Phe Gly Cys Gly Gly Gly Gly Gly Gly Gly 1 5 10 15 <210> 460 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 460 Cys Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys Ala Leu Gly 1 5 10 15 <210> 461 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 461 Val Val Pro Asp Gln Leu Met Ala Phe Gly 1 5 10 <210> 462 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 462 Asp Gln Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys 1 5 10 <210> 463 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 463 Cys Leu Met Ala Phe Gly Gly Ser Ser Glu Cys 1 5 10 <210> 464 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 464 Cys Leu Met Ala Phe Gly Gly Ser Ser Glu Cys 1 5 10 <210> 465 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 465 Val His Val Val Pro Asp Gln Leu Met Ala 1 5 10 <210> 466 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 466 Val Val Pro Asp Gln Leu Met Ala Phe Gly 1 5 10 <210> 467 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 467 Asp Gln Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys 1 5 10 <210> 468 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 468 Gln Leu Met Ala Phe Gly Gly Ser Ser Glu 1 5 10 <210> 469 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 469 Cys Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Ser Ala Leu Cys 1 5 10 15 <210> 470 <211> 14 <212> PRT <213> Unknown <220> <223> Description of Unknown: MIF N-terminal Loop motif / domain       sequence <400> 470 Pro Asp Gln Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys 1 5 10 <210> 471 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> Cyclic peptide <400> 471 Cys Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys 1 5 10 <210> 472 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 472 Pro Arg Ala Ser Val Pro Asp Gly 1 5 <210> 473 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 473 Asn Val Pro Arg Ala 1 5 <210> 474 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 474 Ser Val Pro Asp Gly 1 5 <210> 475 <211> 4 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 475 Leu Gln Asp Pro One <210> 476 <211> 4 <212> PRT <213> Unknown <220> <223> Description of Unknown: MIF Loop peptide sequence <400> 476 Ser Pro Asp Arg One <210> 477 <211> 42 <212> DNA <213> Homo sapiens <220> <221> CDS (222) (1) .. (42) <400> 477 ttc ctg agc gag ctg aca cag cag ctg gcc cag gcc acc ggc 42 Phe Leu Ser Glu Leu Thr Gln Gln Leu Ala Gln Ala Thr Gly 1 5 10 <210> 478 <211> 21 <212> DNA <213> Homo sapiens <220> <221> CDS (222) (1) .. (21) <400> 478 ccc atg ttc atc gtg aac acc 21 Pro Met Phe Ile Val Asn Thr 1 5 <210> 479 <211> 30 <212> DNA <213> Homo sapiens <220> <221> CDS (222) (1) .. (30) <400> 479 aac gtg ccc aga gcc agc gtg ccc gac ggc 30 Asn Val Pro Arg Ala Ser Val Pro Asp Gly 1 5 10 <210> 480 <211> 21 <212> DNA <213> Homo sapiens <220> <221> CDS (222) (1) .. (21) &Lt; 400 > 480 aag ccc cct cag tat atc gcc 21 Lys Pro Pro Gln Tyr Ile Ala 1 5 <210> 481 <211> 42 <212> DNA <213> Homo sapiens <220> <221> CDS (222) (1) .. (42) <400> 481 ccc gac cag ctg atg gcc ttc ggc ggc agc agc gag cct tgc 42 Pro Asp Gln Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys 1 5 10 <210> 482 <211> 24 <212> DNA <213> Homo sapiens <220> <221> CDS (222) (1) .. (24) <400> 482 atc ggc aag atc ggc gga gcc cag 24 Ile Gly Lys Ile Gly Gly Ala Gln 1 5 <210> 483 <211> 54 <212> DNA <213> Homo sapiens <220> <221> CDS (222) (1) .. (54) <400> 483 aac aga agc tac agc aag ctg ctg tgc ggc ctg ctg gcc gag aga ctg 48 Asn Arg Ser Tyr Ser Lys Leu Leu Cys Gly Leu Leu Ala Glu Arg Leu 1 5 10 15 aga atc 54 Arg Ile                                                                            <210> 484 <211> 33 <212> DNA <213> Homo sapiens <220> <221> CDS (222) (1) .. (33) <400> 484 agc ccc gac aga gtg tac atc aac tac tac gac 33 Ser Pro Asp Arg Val Tyr Ile Asn Tyr Tyr Asp 1 5 10 <210> 485 <211> 42 <212> DNA <213> Homo sapiens <220> <221> CDS (222) (1) .. (42) <400> 485 atg aac gcc gcc aac gtg ggc tgg aac aac agc acc ttc gcc 42 Met Asn Ala Ala Asn Val Gly Trp Asn Asn Ser Thr Phe Ala 1 5 10 <210> 486 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <220> <223> N-term Ac <220> <223> C-term NH2 <400> 486 Asp Trp Phe Lys Ala Phe Tyr Asp Lys Val Ala Glu Lys Phe Lys Glu 1 5 10 15 Ala Phe          <210> 487 <211> 21 <212> PRT <213> Unknown <220> <223> Description of Unknown: MOG35-55 peptide sequence <400> 487 Met Glu Val Gly Trp Tyr Arg Ser Pro Phe Ser Arg Val Val His Leu 1 5 10 15 Tyr Arg Asn Gly Lys             20 <210> 488 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic       peptide <400> 488 Val His Val Val Pro Asp Leu Leu Met Ala 1 5 10 <210> 489 <211> 7 <212> PRT <213> Homo sapiens <400> 489 Pro Met Phe Ile Val Asn Thr 1 5 <210> 490 <211> 10 <212> PRT <213> Homo sapiens <400> 490 Asn Val Pro Arg Ala Ser Val Pro Asp Gly 1 5 10 <210> 491 <211> 14 <212> PRT <213> Homo sapiens <400> 491 Phe Leu Ser Glu Leu Thr Gln Gln Leu Ala Gln Ala Thr Gly 1 5 10 <210> 492 <211> 7 <212> PRT <213> Homo sapiens <400> 492 Lys Pro Pro Gln Tyr Ile Ala 1 5 <210> 493 <211> 14 <212> PRT <213> Homo sapiens <400> 493 Pro Asp Gln Leu Met Ala Phe Gly Gly Ser Ser Glu Pro Cys 1 5 10 <210> 494 <211> 8 <212> PRT <213> Homo sapiens <400> 494 Ile Gly Lys Ile Gly Gly Ala Gln 1 5 <210> 495 <211> 18 <212> PRT <213> Homo sapiens <400> 495 Asn Arg Ser Tyr Ser Lys Leu Leu Cys Gly Leu Leu Ala Glu Arg Leu 1 5 10 15 Arg Ile          <210> 496 <211> 11 <212> PRT <213> Homo sapiens <400> 496 Ser Pro Asp Arg Val Tyr Ile Asn Tyr Tyr Asp 1 5 10 <210> 497 <211> 14 <212> PRT <213> Homo sapiens <400> 497 Met Asn Ala Ala Asn Val Gly Trp Asn Asn Ser Thr Phe Ala 1 5 10

Claims (44)

The following domains of MIF: N-terminal / pseudo ELR motif / domain, alpha-helix # 1 motif / domain, MIF N-loop motif / domain, loop-barrel-loop motif / domain, C-terminal motif / A peptide that competitively binds to a binding partner of one of the domains, or a combination thereof. The peptide of claim 1, wherein the peptide has the following domains: N-terminal tail, pseudo ELR-loop, alpha-helix # 1 motif / domain, PPQ-loop, PDQ-loop, IGK-loop, NRS-helix, SPDR-loop A peptide that competitively binds to a binding partner of one of C, terminal tail, or a combination thereof. The peptide of claim 1, wherein the peptide binds competitively with a binding partner of the N-loop domain. The peptide of claim 2, wherein the peptide comprises an amino acid that competitively binds with a binding partner of MIF leu47. The peptide of claim 1, wherein the peptide binds competitively with a binding partner of the pseudo ELR domain. The composition of claim 1, wherein the peptide is LMAFGGSSEP (SEQ ID NO: 18); LMAFGGSS (SEQ ID NO: 20); Cyclic CLMAFGGSSEPCALC (SEQ ID NO: 423); VHVVPDQLMA (SEQ ID NO: 465); QLMAFGGSSE (SEQ ID NO: 468); VNTNVPRASVPDG (SEQ ID NO: 437); NVPRASVPDG (SEQ ID NO: 172); Cyclic CNVPRASVPDGC (SEQ ID NO: 440); NVPRASVPD (SEQ ID NO: 82); Cyclic CLMAFGGSSEP [Abu] ALC (SEQ ID NO: 429), wherein Abu is isosteric L-amino acid, alpha-aminobutyric acid; Or a cyclic CLMAFGGSSEPSALC (SEQ ID NO: 469). Peptide that competitively binds to the binding partner of one motif / domain of CXCR2. The peptide of claim 7 wherein the peptide competitively binds to a binding partner of one of the following domains: CXCR2 extracellular loop 1, CXCR2 extracellular loop 2, CXCR2 extracellular loop 3, or CXCR2 N-terminal / domain. . Peptide that competitively binds to the binding partner of one motif / domain of CXCR4. The method of claim 9, wherein the peptide is selected from SEADDRYICDRFYPNDLWVVV; Or competitively binds to a binding partner of DDRYICDRFYPNDLW. Peptide that competitively binds to the binding partner of one motif / domain of CD44. Peptide that competitively binds to the binding partner of one motif / domain of CD74. (a) a first peptide that competitively binds to the binding partner of the N-loop motif of MIF; And (b) a second peptide that competitively binds to the binding partner of the pseudo ELR motif of MIF; Wherein the first peptide and the second peptide maintain their activity in the fusion peptide. The method of claim 13, wherein the peptide comprises (a) a first peptide that competitively binds to a binding partner of the N-loop motif of MIF; (b) a second peptide that competitively binds to the binding partner of the pseudo ELR motif of MIF; And (c) a third peptide that competitively binds to the binding partner of the pseudo ELR motif of MIF; Wherein the first peptide and the second peptide maintain their activity in the fusion peptide. The method of claim 13 or 14, wherein the fusion peptide is LMAFGGSSEP (SEQ ID NO: 18); LMAFGGSS (SEQ ID NO: 20); Cyclic CLMAFGGSSEPCALC (SEQ ID NO: 423); VHVVPDQLMA (SEQ ID NO: 465); QLMAFGGSSE (SEQ ID NO: 468); VNTNVPRASVPDG (SEQ ID NO: 437); NVPRASVPDG (SEQ ID NO: 172); Cyclic CNVPRASVPDGC (SEQ ID NO: 440); NVPRASVPD (SEQ ID NO: 82); Cyclic CLMAFGGSSEP [Abu] ALC (SEQ ID NO: 429), wherein Abu is isosteric L-amino acid, alpha-aminobutyric acid; Or a peptide selected from cyclic CLMAFGGSSEPSALC (SEQ ID NO: 469). The fusion peptide according to any one of claims 13 to 15, wherein the fusion peptide is represented by formula (IV):

. The fusion peptide of claim 13, wherein the fusion peptide is represented by formula (V):

. 18. The ringer of claim 16 or 17, wherein the linker is alkyl, heteroalkyl, alkylene, alkenylene, alkynylene, heteroalkylene, carbocycle, heterocycle, aromatic ring, nonaromatic ring, substituted ring, monocy. A fusion peptide comprising a click ring, a polycyclic ring, or a combination thereof. (a) an antibody, and (b) a peptide disclosed herein; Wherein the peptide and antibody maintain their activity in the peptibody. The peptibody of claim 19, wherein the peptide is indirectly bound to the antibody. The peptibody of claim 19, wherein the peptide is directly bound to the antibody. The peptibody of claim 19, wherein the peptide is covalently bound to the antibody. The peptibody of claim 19, wherein the peptide is bound to the Fab region of the antibody. The peptibody of claim 19, wherein the peptide is bound to the antigen binding site of the antibody. The peptibody of claim 19, wherein the peptide is bound to the antibody via a reactive side chain. The peptibody of claim 19, wherein the peptide is indirectly bound to the antibody via a linker. 20. The ringer of claim 19 wherein the linker is alkyl, heteroalkyl, alkylene, alkenylene, alkynylene, heteroalkylene, carbocycle, heterocycle, aromatic ring, non-aromatic ring, substituted ring, monocyclic ring, polish A peptibody comprising a click ring, or a combination thereof. The peptibody of claim 19, wherein the antibody is IgA, IgD, IgE, IgG, or IgM, and in some embodiments, the antibody is a humanized antibody. The peptibody of claim 19, wherein the peptibody is CovX ™ body. Use of a substance composition of any one of claims 1 to 29 for the treatment of an inflammatory disease, disease or condition. 31. The method of claim 30, wherein the inflammatory disease, disease or condition is atherosclerosis; Abdominal aortic aneurysm; Acute disseminated encephalomyelitis; Moyamoya disease; Takayasu's disease; Acute coronary syndrome; Cardiac allograft angiopathy; Lung inflammation; Acute respiratory distress syndrome; Pulmonary fibrosis; Acute disseminated encephalomyelitis; Addison's disease; Ankylosing spondylitis; Antiphospholipid antibody syndrome; Autoimmune hemolytic anemia; Autoimmune hepatitis; Autoimmune inner ear disease; Bullous whey; Shaga's disease; Chronic obstructive pulmonary disease; Celiac disease; Dermatitis; Type 1 diabetes; Type 2 diabetes; Endometriosis; Goodpasture syndrome; Graves' disease; Guillain-Barré syndrome; Hashimoto's disease; Idiopathic thrombocytopenic purpura; Interstitial cystitis; Systemic lupus erythematosus (SLE); Metabolic syndrome; Multiple sclerosis; Myasthenia gravis; myocarditis; Narcolepsy; obesity; Vulgaris vulgaris; Pernicious anemia; Polymyositis; Primary biliary cirrhosis; Rheumatoid arthritis; Schizophrenia; Scleroderma; Sjogren's syndrome; Vasculitis; Vitiligo; Wegener's granulomatosis; Allergic rhinitis; Prostate cancer; Non-small cell lung carcinoma; Ovarian cancer; Breast cancer; Melanoma; Stomach cancer; Colon cancer; Brain cancer; Metastatic bone disease; Pancreatic cancer; Bladder cancer; Hepatocellular carcinoma; Liver cancer; Lung adenocarcinoma; Esophageal squamous cell carcinoma; CNS tumors; Blood tumors; Lymphoma; Nasal polyps; Gastrointestinal cancer; Ulcerative colitis; Crohn's disease; Collagen colitis; Lymphocytic colitis; Ischemic colitis; Converting colitis; Behcet's syndrome; Infectious colitis; Indeterminate colitis; Inflammatory liver disease; Endotoxic shock; Septic shock; Rheumatoid spondylitis; Ankylosing spondylitis; Gout arthritis; Rheumatic polymyalgia; Alzheimer's disease; Parkinson's disease; epilepsy; AIDS dementia; asthma; Adult respiratory distress syndrome; bronchitis; Cystic fibrosis; Acute leukocyte-mediated lung injury; Distal proctitis; Wegener's granulomatosis; Fibromyalgia; bronchitis; Uveitis; conjunctivitis; psoriasis; eczema; dermatitis; Smooth muscle hyperplasia disease; meningitis; Shingles; encephalitis; Nephritis; Tuberculosis; Retinitis; Atopic dermatitis; Pancreatitis; Periodontal gingivitis; Coagulation necrosis; Liquefied necrosis; Fibrotic necrosis; Neointimal hyperplasia; Myocardial infarction; stroke; Organ transplant rejection; Influenza, or a combination thereof. 31. The use according to claim 30, wherein the inflammatory disease, disease or condition is cancer. 31. The method of claim 30, wherein the inflammatory disease, disease or condition is prostate cancer; Non-small cell lung carcinoma; Ovarian cancer; Breast cancer; Melanoma; Stomach cancer; Colon cancer; Brain cancer; Metastatic bone disease; Pancreatic cancer; Bladder cancer; Hepatocellular carcinoma; Liver cancer; Lung adenocarcinoma; Esophageal squamous cell carcinoma; CNS tumors; Blood tumors; Lymphoma; Or combinations thereof. 31. The use of claim 30, wherein the inflammatory disease, disease or condition is rheumatoid arthritis. 31. The use according to claim 30, wherein the inflammatory disease, disease or condition is acute respiratory distress syndrome. 31. The use according to claim 30, wherein the inflammatory disease, disease or condition is glomerulonephritis. The use of claim 30, wherein the inflammatory disease, disease or condition is an inflammatory bowel disease. 31. The use of claim 30, wherein the inflammatory disease, disease or condition is an abdominal aortic aneurysm disease. 31. The use of claim 30, wherein the inflammatory disease, disease or condition is a chronic obstructive pulmonary disease. 31. The use according to claim 30, wherein the inflammatory disease, disease or condition is asthma. 31. The use of claim 30, wherein the inflammatory disease, disease or condition is lupus. 31. The use according to claim 30, wherein the inflammatory disease, disease or condition is sepsis. Use of the substance composition of any one of claims 1 to 29 for treating, preventing or reducing angiogenesis. A pharmaceutical composition for treating an inflammatory disease, disease, condition or condition in an individual in need thereof, comprising the substance composition of any one of claims 1 to 29.

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