Cell-Permeable Peptide Inhibitors of the JNK Signal Transduction Pathway

Abstract

Protein kinase inhibitors and more specifically inhibitors of the protein kinase c-Jun amino terminal kinase are herein described. Additionally, JNK inhibitor sequences, chimeric peptides, nucleic acids encoding same as well as pharmaceutical compositions for treating pathophysiologies associated with JNK signaling are herein provided.

1 . JNK inhibitor sequence comprising less than 150 amino acids in length, wherein the inhibitor sequence comprises or consists of at least one amino acid sequence according to SEQ ID NOs: 1, 2, 3 or 4, or a fragment, derivative or variant thereof. 
     
     
         2 . The JNK inhibitor sequence according to  claim 1 , wherein the JNK inhibitor sequence comprises a range of 5 to 150 amino acid residues, more preferably 10 to 100 amino acid residues, even more preferably 10 to 75 amino acid residues and most preferably a range of 15 to 50 amino acid residues. 
     
     
         3 . The JNK inhibitor sequence of any of  claims 1  to  2 , wherein the JNK inhibitor sequence binds c-jun amino terminal kinase (JNK). 
     
     
         4 . The JNK inhibitor sequence of any of  claims 1  to  3 , wherein the JNK inhibitor sequence inhibits the activation of at least one JNK targeted transcription factor when the JNK inhibitor sequence is present in a JNK expressing cell. 
     
     
         5 . The JNK inhibitor sequence of any of  claims 1  to  4 , wherein the JNK targeted transcription factor is selected from the group consisting of c-Jun, ATF2, and Elkl. 
     
     
         6 . The JNK inhibitor sequence of any of  claims 1  to  5 , wherein the JNK inhibitor sequence alters a JNK effect when the peptide is present in a JNK expressing cell. 
     
     
         7 . A chimeric peptide comprising at least one first domain and at least one second domain linked by a covalent bond, the first domain comprising a trafficking sequence, and the second domain comprising a JNK inhibitor sequence. 
     
     
         8 . The peptide of  claim 7 , wherein the trafficking sequence comprises the amino acid sequence of a human immunodeficiency virus TAT polypeptide. 
     
     
         9 . The peptide of any of  claims 7  to  8 , wherein the trafficking sequence comprises the amino acid sequence of SEQ ID NO: 5, 6, 7 or 8. 
     
     
         10 . The peptide of any of  claims 7  to  9 , wherein the trafficking sequences augments cellular uptake of the peptide. 
     
     
         11 . The peptide of any of  claims 7  to  9 , wherein the trafficking sequence directs nuclear localization of the peptide. 
     
     
         12 . The peptide of any of  claims 7  to  11 , wherein the JNK inhibitor sequence comprises a JNK inhibitor sequence according to any of  claims 1  to  6 . 
     
     
         13 . The peptide of any of  claims 7  to  12 , wherein the peptide comprises the amino acid sequence of any of SEQ ID NOs: 9, 10, 11 or 12, or a fragment, or variant thereof. 
     
     
         14 . An isolated nucleic acid encoding a JNK inhibitor sequence according to any of  claims 1  to  6  or a chimeric peptide according to any of  claims 7  to  13 . 
     
     
         15 . A vector comprising the nucleic acid of  claim 14 . 
     
     
         16 . A cell comprising the vector of  claim 15 . 
     
     
         17 . An antibody which binds immunospecifically to a JNK inhibitor sequence according to any of  claims 1  to  6  or to a chimeric peptide according to any of  claims 7  to  13 . 
     
     
         18 . A pharmaceutical composition comprising a JNK inhibitor sequence according to any of  claims 1  to  6  or a chimeric peptide according to any of  claims 7  to  13  or a nucleic acid according to  claim 14 , and a pharmaceutically acceptable carrier. 
     
     
         19 . Use of a JNK inhibitor sequence according to any of  claims 1  to  6  or a chimeric peptide according to any of  claims 9  to  13  for the preparation of a pharmaceutical composition for treating a pathophysiology associated with activation of JNK in a subject. 
     
     
         20 . Use according to  claim 19 , wherein the pathophysiology is selected from malignancies of lung, breast, lymphoid, gastrointestinal, and genito-urinary tract as well as adenocarcinomas, including malignancies such as colon cancers, renal-cell carcinoma, prostate cancer, non-small cell carcinoma of the lung, cancer of the small intestine and cancer of the esophagus, as well as leukemia, disorders or pathophysiologies associated with oncogenic transformation as well as cancers with Bcr-Abl oncogenic transformations, non-malignant or immunological-related cell proliferative diseases selected from psoriasis, pemphigus vulgaris, Behcet's syndrome, acute respiratory distress syndrome (ARDS), ischemic heart disease, post-dialysis syndrome, rheumatoid arthritis, acquired immune deficiency syndrome, vasculitis, septic shock, pathophysiologies associated with activation of JNK in a cell selected from restenosis, hearing loss, ear trauma, ischemia, stroke and/or disorders or pathophysiologies associated with maturation and differentiation of immune cells, reperfusion injuries, hypoxia, apoptosis-related, response to stressful stimuli, and with secondary effects due to treatment with proinflammatory cytokines, effects associated with diabetes or with cellular shear stress, selected from pathological states induced by arterial hypertension, including heart and cardiac hypertrophy and arteriosclerotic lesions, and at bifurcations of blood vessels, by ionizing radiation as used in radiotherapy and ultraviolet light (UV lights), by free radicals, DNA damaging agents, including chemotherapeutic drugs, by ischemia/reperfusion injuries, by hypoxia, by hypo- and hyperthermia, or to inhibit inflammatory, autoinflammatory, immune and autoimmune diseases, degenerative diseases, myopathies, cardiomyopathies, and graft rejection. 
     
     
         21 . Use according to  claim 19  or  20 , wherein the pharmaceutical composition is to be administered by an administration route selected from the group consisting of intraperitoneal, nasal, intravenous, oral and patch delivery. 
     
     
         22 . A method for preparing a chimeric peptide according to any of  claims 7  to  13 , the method comprising: (a) culturing a cell containing a nucleic acid according to  claim 14  under conditions that provide for expression of the chimeric peptide; and (b) recovering the expressed peptide. 
     
     
         23 . Kit comprising a JNK inhibitor sequence according to any of  claims 1  to  6  and/or a chimeric peptide according to any of  claims 7  to  13  and/or a nucleic acid according to  claim 14  and/or a vector according to  claim 15  and/or a cell according to  claim 16  and/or an antibody according to  claim 17 .