Antibiotic peptides

Abstract

The invention relates to a peptide or peptide derivative having the general formula: Sub 1 -X 1 -D 2 K 3 -P 4 -P 5 -Y 6 -L 7 -P 8 -R 9 -P 10 -X 2 -P 12 - P 13 -R 14 -X 3 -I 16 -P 17 /Y 17 -N 18 -N 19 -X 4 -Sub 2 , wherein X 1 is a non-polar, hydrophobic group or a positively charged group, D 2 is asparagine or glutamine, K 3 , X 2 , and X 4 are positively charged groups, X 3 is a positively charged group, proline, or a proline derivative: L 7 and I 16 are non-polar, hydrophobic groups, Y 6 and Y 17 are tyrosine, R 9 and R 14 are arginine, N 18 and N 19 are asparagine or glutamine, P 4 , P 5 , P 8 , P 10 , P 12 , P 13 and P 17 are proline, hydroxyproline, or derivatives thereof, wherein possibly one or two of the groups selected from D 2 , P 4 , P 5 , P 8 , P 10 , P 12 , P 13 , P 17 and Y 17 are replaced by an arbitrary group and/or P 13 and R 14 are exchanged. Sub 1 is the free or modified N-terminus, and Sub 2 is the free or modified C-terminus. The invention further relates to the use of the peptides and peptide derivatives in medicine, as an antibiotic, in a disinfectant or cleaning agent, as a preservative or in a packaging material, in pharmaceutical research, or in a screening method.

1 . A peptide or peptide derivative, which contains one of the following sequences:
 Sub 1 -X 1 -D 2 -K 3 -P 4 -P 5 -Y 6 -L 7 -P 8 -R 9 -P 10 -X 2 -P 12 -P 13 -R 14 -X 3 -I 16 -P 17 /Y 17 -N 18 -N 19 -X 4 -Sub 2      X 1  is a residue with a nonpolar, hydrophobic side chain or with a positive net charge or a side chain that is positively charged under physiological conditions;   D 2  is an aspartic acid or glutamic acid residue,   K 3  is a residue with a positive net charge or a side chain that is positively charged under physiological conditions, preferably lysine or arginine,   X 2  and X 4  are selected independently of one another from residueswith a positive net charge or a side chain that is positively charged under physiological conditions;   X 3  is a residue with a positive net charge or a side chain that is positively charged under physiological conditions or proline or a proline derivative;   L 7  is selected from residues with a nonpolar, hydrophobic side chain, preferably leucine, isoleucine and valine,   I 16  is selected from residues with a nonpolar, hydrophobic side chain, preferably leucine, isoleucine, tert-butylglycine and valine,   Y 6  and Y 17  are in each case tyrosine, R 9  and R 14  are in each case arginine, N 18  is asparagine or glutamine, N 19  is asparagine or glutamine or is absent, P 4 , P 5 , P 8 , P 10 , P 12 , P 13  and P 17  are selected independently of one another from proline and proline derivatives or hydroxyproline and hydroxyproline derivatives,   Sub 1  is the free N-terminus of the amino acid X 1  or a modified N-terminal amino group;   Sub 1  is the free C-terminal carboxyl group of the C-terminal amino acid (—COOH) or a modified C-terminal carboxyl group.   
     
     
         2 . The peptide or peptide derivative as claimed in  claim 1 , wherein P 13  and R 14  are exchanged, and/or one or two of the residues selected from D 2 , P 4 , P 5 , P 8 , P 10 , P 12 , P 13 , P 17  and Y 17  are replaced with any residue. 
     
     
         3 . The peptide or peptide derivative as claimed in  claim 1  or  2 , wherein the N-terminus is linked to another peptide directly or via a linker. 
     
     
         4 . The peptide or peptide derivative as claimed in any one of  claims 1  to  3  containing at least one additional residue X 5  and/or X 6  in Sub 2 , wherein X 5  is selected from the group comprising proline, a proline derivative and a building block, which has a positive net charge or bears a side chain that is positively charged under physiological conditions and in that X 6  is selected from proline, a proline derivative, a polar building block or a hydrophobic building block. 
     
     
         5 . The peptide or peptide derivative as claimed in any one of  claims 1  to  4 , characterized in that the residue X 1  is selected from arginine, lysine, δ-hydroxylysine, homoarginine, 2,4-diaminobutyric acid, β-homoarginine, D-arginine, arginal, 2-amino-3-guanidinopropionic acid, nitroarginine, N-methylarginine, ε-N-methyllysine, allo-hydroxylysine, 2,3-diaminopropionic acid, 2,2′-diaminopimelic acid, ornithine, sym-dimethylarginine, asym-dimethylarginine, 2,6-diaminohexinic acid, p-aminobenzoic acid, 3-aminotyrosine, valine, isoleucine, leucine and methionine, alanine, phenylalanine, N-methylleucine, tert-butylglycine, cyclohexylalanine, β-alanine, 1-aminocyclohexylcarboxylic acid, N-methylisoleucine, norleucine, norvaline and N-methylvaline. 
     
     
         6 . the peptide or peptide derivative as claimed in any one of  claims 1  to  5 , characterized in that the residue X 2  and/or the residue X 4  are selected independently of one another from arginine, lysine, δ-hydroxylysine, homoarginine, β-homoarginine, D-arginine, arginal, 2,4-diaminobutyric acid, 2-amino-3-guanidinopropionic acid, nitroarginine, nitrosoarginine, N-methyl-arginine, ε-N-methyllysine, allo-hydroxylsine, 2,3-diaminopropionic acid, 2,2′-diaminopimelic acid, ornithine, sym-dimethylarginine, asym-dimethylarginine, 2,6-diaminohexinic acid, p-aminobenzoic acid and 3-aminotyrosine. 
     
     
         7 . The peptide or peptide derivative as claimed in any one of  claims 1  to  6 , characterized in that the residue X3 is selected from arginine, lysine, δ-hydroxylysine, homoarginine, β-homoarginine, D-arginine, arginal, 2,4-diaminobutyric acid, β-homoarginine, 2-amino-3-quanidinopropionic acid, nitroarginine, nitrosoarginine, N-methyl-arginine, ε-N-methyllysine, allo-hydroxylysine, 2,3-diaminopropionic acid, 2,2′-diaminopimelic acid, ornithine, sym-dimethylarginine, asym-dimethylarginine, 2,6-diaminohexinic acid, p-aminobenzoic acid, 3-aminotyrosine, proline, cis-4-hydroxyproline, trans-4-hydroxyproline, cis-3-hydroxyproline, trans- 3 -hydroxyproline, β-cyclohexylalanine, 3,4-cis-methanoproline, 3,4-dehydroproline, homoproline and pseudoproline. 
     
     
         8 . The peptide or peptide derivative as claimed in any one of  claims 1  to  7 , selected from the group of sequences comprising SEQ ID No. 5 to 9, 14 to 26, 29, 30, 32, 33, 36, 38, 40, 41, 44 to 46, 49, 50, 532 to 59, 61 to 85, 93, 94, 101, 102 and 107 to 112. 
     
     
         9 . A peptide or peptide derivative containing an antibacterial peptide and a cell-penetrating peptide. 
     
     
         10 . The peptide or peptide derivative as claimed in  claim 9 , wherein said antibacterial peptide is selected from the group comprising apidaecin, drosocin, formaecin 1, pyrrhocoricin and metalnikowin 1. 
     
     
         11 . The peptide or peptide derivative as claimed in  claim 9 , wherein said antibacterial peptide comprises a peptide or peptide derivative as claimed in any one of  claims 1  to  8 . 
     
     
         12 . The peptide or peptide derivative as claimed in any one of  claims 9  to  11 , wherein said cell-penetrating peptide is selected from the group comprising penetratin, tat peptides, model amphipathic peptides, transportan, SynB and cis-γ-amino-1-proline-containing peptides. 
     
     
         13 . The peptide or peptide derivative as claimed in any one of  claims 1  to  12  selected from the group comprising SEQ ID NO. 95 to 102 and 106. 
     
     
         14 . The peptide or peptide derivative as claimed in any one of  claims 1  to  13 , characterized in that at least one of the peptide bonds of the peptide backbone is chemically modified. 
     
     
         15 . The peptide or peptide derivative as claimed in  claim 14 , characterized in that the bond between X 6 -X 7  is a chemical bond that cannot be cleaved under physiological conditions. 
     
     
         16 . The peptide or peptide derivative as claimed in  claim 14  or  15 , characterized in that the chemically modified bond is selected from a reduced amide bond, an alkylated amide bond or a thioamide bond. 
     
     
         17 . The peptide or peptide derivative as claimed in any one of  claims 1  to  16 , which is joined to a protein or coupled to a polymer or bound to a carrier. 
     
     
         18 . A multimer, in which at least two peptides or peptide derivatives are joined together, characterized in that at least one of the peptides or peptide derivatives is a peptide or peptide derivative as claimed in any one of  claims 1  to  17 . 
     
     
         19 . A pharmaceutical composition, characterized in that it contains at least one peptide, peptide derivative or multimer as claimed in any one of  claims 1  to  10 . 
     
     
         20 . A method of production of a peptide or peptide derivative or multimer as claimed in any one of  claims 1  to  18 , characterized in that the peptide or peptide derivative or multimer is produced by chemical synthesis or by means of recombinant methods. 
     
     
         21 . The use of a peptide, peptide derivative or multimer as claimed in any one of  claims 1  to  18  for the production of a medical product. 
     
     
         22 . The use of a peptide, peptide derivative or multimer as claimed in any one of  claims 1  to  18  as disinfectant and/or cleaning agent, as preservative and/or in a packaging material. 
     
     
         23 . The peptide, peptide derivative and/or multimer as claimed in any one of  claims 1  to  18  for use in the treatment of microbial, bacterial or fungel infections. 
     
     
         24 . The use of a peptide, peptide derivative and/or multimer as claimed in any one of  claims 1  to  18  for removing contamination by microbes, bacteria or fungi. 
     
     
         25 . The use of a peptide, peptide derivative or multimer as claimed in any one of  claims 1  to  18  in pharmaceutical research or in a screening process. 
     
     
         26 . A method for identifying a substance with antibacterial or antimycotic action, comprising the following steps:
 (i) carrying out a competitive assay with:   (a) a microorganism that is sensitive to a peptide, peptide derivative or multimer as claimed in any one of  claims 1  to  18 ;   (b) a peptide, peptide derivative or multimer as claimed in any one of  claims 1  to  18  and   (c) at least one substance to be tested by bringing (a) in contact with (b) and (c); and   (ii) selecting a test substance, which competitively displaces the binding of the peptide, peptide derivative or multimer on the microorganism.   
     
     
         27 . The method as claimed in  claim 26 , wherein the microorganism is a species selected from the group of genera comprising  Escherichia coli, Enterobacter cloacae, Erwinia amylovora, Klebsiella pneumoniae, Morganella morganii, Salmonella typhimurium, Salmonella typhi, Shigella dysenteriae, Yersinia enterocolitica, Acinetobacter calcoaceticus, Acinetobacter baumannii, Agrobacterium tumefaciens, Francisella tularensis, Legionella pneumophila, Pseudomonas syringae, Rhizobium meliloti, Pseudomonas aeruginosa, Proteus vulgaris, Proteus mirabilis, Stenotrophomonas maltophilia and Haemophilus influenzae.    
     
     
         28 . A nucleic acid coding for a peptide or multimer as claimed in any one of  claims 1  to  18 . 
     
     
         29 . A host cell, comprising one or more nucleic acids as claimed in  claim 28 .