Cationic antimicrobial peptides

Abstract

The present invention relates to a peptide comprising an amino acid sequence of SEQ ID NO: 1 having antimicrobial activity, a polynucleotide encoding the peptide of the invention, a composition comprising the peptide of the invention and methods using the peptide or composition of the invention.

Description

[0001] New PCT Patent Application

[0002] Applicant: Space Pharmaceuticals AG

[0003] Vossius Ref.: AJ4188 PCT BS

[0004] Cationic antimicrobial peptides

[0005] The present invention relates to the field of antimicrobial peptides. Particularly, the invention relates to a peptide comprising an amino acid sequence of SEQ ID NO: 1 having antimicrobial activity, a polynucleotide encoding the peptide of the invention, a composition comprising the peptide of the invention and methods using the peptide or composition of the invention.

[0006] Related Art

[0007] In recent years, the extensive use and overuse of antibiotics have led to the emergence of drug-resistant and multidrug-resistant bacteria, posing significant challenges to clinical treatment. To combat antimicrobial resistance, the development of novel antimicrobial agents has become an urgent priority. Among these, cationic antimicrobial peptides (AMPs) have emerged as a promising new class of potent antimicrobials. Found naturally in various organisms, AMPs serve as valuable templates for next-generation antibiotics. They exhibit bactericidal activity against both Gram -positive and Gram-negative bacteria, are effective against drug-resistant strains, and significantly reduce the likelihood of resistance development.

[0008] AMPs demonstrate broad-spectrum antimicrobial activities, including antibacterial, antiviral, antitumor, anti-inflammatory, and immunoregulatory effects, and also play roles in signal transduction. Unlike traditional antibiotics, which target specific bacterial pathways and structures, making them susceptible to resistance through bacterial mutations, AMPs primarily act by physically disrupting bacterial cell membranes. This mechanism causes leakage of intracellular contents and is less prone to resistance due to the highly conserved nature of bacterial membrane structures. Consequently, AMPs hold tremendous potential as alternatives to traditional antibiotics, offering a viable solution to the escalating issue of bacterial resistance.

[0009] However, AMPs known in the prior art face notable limitations, such as challenges in separation and purification, weak antibacterial activity, high hemolytic activity andcytotoxicity, and poor stability, which impede their large-scale production and clinical application. To overcome these challenges, the structural optimization of AMPs is critical. Designing peptides with smaller molecular weights, stronger antibacterial activity, improved stability, reduced cytotoxicity, and scalability for mass production is essential for developing clinically viable alternatives to traditional antibiotics and addressing the growing global antimicrobial resistance crisis.

[0010] In light of the above, there is still a need for compounds and compositions having broad-spectrum antimicrobial activity.

[0011] Summary of the invention

[0012] This present invention provides a peptide, which exhibits broad-spectrum activity against microorganisms, i.e., an antimicrobial activity, including antibacterial effects against both Gram-positive and Gram-negative bacteria, as well as antifungal properties. The peptide of the invention has a high stability, is easy to produce in large quantities, and effectively inhibits bacterial and fungal growth and biofilm formation in a dose-dependent manner while demonstrating low hemolytic activity and cytotoxicity. These attributes render the peptide of the invention suitable for use as a novel therapeutic agent for treating bacterial and fungal infections and for use in an antimicrobial composition.

[0013] Accordingly, the invention relates to, inter alia, the following aspects and embodiments:

[0014] 1. A peptide comprising an amino acid sequence of SEQ ID NO: 1 :

[0015] X1 -X2-X3-X4-X5-X6-X7-X8-X9-X10-X11 ,

[0016] wherein X1 to X7 are, independently of each other, a D- or L-amino acid selected from the group consisting of K (lysine), R (arginine), W (tryptophan), L (leucine), an amino acid variant of K, R, W, or L, and a conservative substitute amino acid of K, R, W or L; and

[0017] wherein X8 to X11 are, independently of each other, a deletion or a D- or L- amino acid selected from the group consisting of K, W, R, L, an amino acidvariant of K, R, W or L, and a conservative substitute amino acid of K, R, W or L.

[0018] 2. The peptide of embodiment 1, wherein the peptide comprising the amino acid sequence of SEQ ID NO: 1 , wherein X1 to X7 are, independently of each other, a D- or L-amino acid selected from the group consisting of K, R, W and L; and wherein X8 to X11 are, independently of each other, a D- or L-amino acid selected from the group consisting of K, R, W and L.

[0019] 3. The peptide of any one of the preceding embodiments, wherein X1 is a D- or L- amino acid selected from the group consisting of K, an amino acid variant of K, and a conservative substitute amino acid of K.

[0020] 4. The peptide of any one of the preceding embodiments, wherein X2 is a D- or L- amino acid selected from the group consisting of R, an amino acid variant of R, and a conservative substitute amino acid of R.

[0021] 5. The peptide of any one of the preceding embodiments, wherein X3 is a D- or L- amino acid selected from the group consisting of W, an amino acid variant of W, and a conservative substitute amino acid of W.

[0022] 6. The peptide of any one of the preceding embodiments, wherein X4 is a D- or L- amino acid selected from the group consisting of: W, K and R, an amino acid variant of W, K and R, and a conservative substitute amino acid of W, K and R.

[0023] 7. The peptide of any one of the preceding embodiments, wherein X5 and X6 are independently of each other a D- or L-amino acid selected from the group consisting of: W, K, L, and R, an amino acid variant of W, K, L and R, and a conservative substitute amino acid of W, K, L, and R.

[0024] 8. The peptide of any one of the preceding embodiments, wherein X7 and X8 are independently of each other a D- or L-amino acid selected from the group consisting of: W, L, and R, an amino acid variant of W, L and R, and a conservative substitute amino acid of W, L, and R.The peptide of any one of the preceding embodiments, wherein the C terminal amino acid, preferably the last C terminal amino acid is a D- or L-amino acid selected from the group consisting of: W, L, and R, an amino acid variant of W, L and R, and a conservative substitute amino acid of W, L, and R;

[0025] more preferably, the last C terminal amino acid is a D- or L-amino acid, preferably an L-amino acid, selected from the group consisting of: L, R, an amino acid variant of L, an amino acid variant of R, a conservative substitute amino acid of L, and a conservative substitute amino acid of R;

[0026] again more preferably, the last C terminal amino acid is a D- or L-amino acid, preferably an L-amino acid, selected from the group consisting of: L, an amino acid variant of L, and a conservative substitute amino acid of L;

[0027] again more preferably, the last C terminal amino acid is a D- or L-amino acid of leucine, preferably an L-amino acid of leucine.

[0028] The peptide of any one of the preceding embodiments, wherein the C terminus of the peptide consists of one or more, preferably one, two or three D- or L-, preferably L-amino acids selected from the group consisting of: L, an amino acid variant of L, or a conservative substitute amino acid of L; or

[0029] the C terminus of the peptide consists of one or two or three D- or L-, preferably L-amino acids selected from the group consisting of: L, an amino acid variant of L, or a conservative substitute amino acid of L, or

[0030] the C terminus of the peptide consists of one or two L-amino acids selected from the group consisting of: L, an amino acid variant of L, or a conservative substitute amino acid of L; or

[0031] the C terminus of the peptide consists of one, two or three D- or L-, preferably L-amino acids of leucine;

[0032] the C terminus of the peptide consists of one or two D- or L-, preferably L-amino acids of leucine; orthe C terminus of the peptide consists of one or two L-amino acids of leucine.

[0033] The peptide of any one of the preceding embodiments comprising at least one alpha helical domain.

[0034] The peptide of any one of the preceding embodiments, wherein the peptide is a cationic peptide, preferably the cationic peptide has a net positive charge, more preferably the net positive charge ranges from +2 to +12.

[0035] The peptide of any one of the preceding embodiments, wherein the peptide has a length of 5 to 1000 amino acids, preferably 5 to 500 amino acids, more preferably 5 to 100 amino acids, or 5 to 50 amino acids, or 5 to 30 amino acids, or 5 to 20 amino acids, or 5 to 15 amino acids, or 7 to 11 amino acids.

[0036] The peptide of any one of the preceding embodiments, wherein the peptide comprises one or more sequences selected from the group consisting of: SEQ ID NO: 2 to 154, and a derivative of SEQ ID NO: 2 to 154;

[0037] wherein preferably the peptide comprises one or more sequences selected from the group consisting of: SEQ ID NO: 2, 4, 5, 7 to 154, and a derivative of SEQ ID NO: 2, 4, 5, 7 to 154;

[0038] wherein the derivative of SEQ ID NO: 2 to 154 or SEQ ID NO: 2, 4, 5, 7 to 154 comprises a chemical modification of one or more amino acids or a chemical modification of the peptide of SEQ ID NO: 2 to 154, preferably the chemical modification is selected from the group consisting of phosphorylation, acetylation, oxidation, ubiquitylation, amidation, deamidation, esterification, alkylation, preferably methylation, nitration, hydroxylation, formylation, sulfation, covalent binding of a lipidic moiety, sulphonation, glycation, acylation, myristoylation, PEGylation, palmitoylation, halogenation, chemical modifications of part or all peptide bonds, and / or post-translational amino acid modification; and / or

[0039] wherein in the derivative of SEQ ID NO: 2 to 154 or SEQ ID NO: 2, 4, 5, 7 to 154, one, two, three or four amino acids are each exchanged by an amino acidvariant, preferably by an analogue amino acid or a conservative substitute amino acid; and / or

[0040] wherein in the derivative of SEQ ID NO: 2 to 154 or SEQ ID NO: 2, 4, 5, 7 to 154, one, two, three or four amino acids are deleted, wherein the derivative of SEQ ID NO: 2 to 151 or SEQ ID NO: 2, 4, 5, 7 to 154, has a length of at least 7 amino acids, and / or

[0041] wherein in the derivative of SEQ ID NO: 2 to 154 or SEQ ID NO: 2, 4, 5, 7 to 154, one, two, three or four single, double, or triple amino acids are added to SEQ ID NO: 2 to 154 or SEQ ID NO: 2, 4, 5, 7 to 151 , wherein the added amino acids are D- or L-amino acids selected from the group consisting of K, R, W and L.

[0042] The peptide of any one of the preceding embodiments, wherein the peptide comprises one or more sequences selected from the group consisting of: SEQ ID NO: 2 to 151 , and a derivative of SEQ ID NO: 2 to 151 ;

[0043] wherein preferably the peptide comprises one or more sequences selected from the group consisting of: SEQ ID NO: 2, 4, 5, 7 to 151, and a derivative of SEQ ID NO: 2, 4, 5, 7 to 151;

[0044] wherein the derivative of SEQ ID NO: 2 to 151 or SEQ ID NO: 2, 4, 5, 7 to 151 comprises a chemical modification of one or more amino acids or a chemical modification of the peptide of SEQ ID NO: 2 to 151, preferably the chemical modification is selected from the group consisting of phosphorylation, acetylation, oxidation, ubiquitylation, amidation, deamidation, esterification, alkylation, preferably methylation, nitration, hydroxylation, formylation, sulfation, covalent binding of a lipidic moiety, sulphonation, glycation, acylation, myristoylation, PEGylation, palmitoylation, halogenation, chemical modifications of part or all peptide bonds, and / or post-translational amino acid modification; and / or

[0045] wherein in the derivative of SEQ ID NO: 2 to 151 or SEQ ID NO: 2, 4, 5, 7 to151 , one, two, three or four amino acids are each exchanged by an amino acid variant, preferably by an analogue amino acid or a conservative substitute amino acid; and / or

[0046] wherein in the derivative of SEQ ID NO: 2 to 151 or SEQ ID NO: 2, 4, 5, 7 to 151, one, two, three or four amino acids are deleted, wherein the derivative of SEQ ID NO: 2 to 151 or SEQ ID NO: 2, 4, 5, 7 to 151 , has a length of at least 7 amino acids, and / or

[0047] wherein in the derivative of SEQ ID NO: 2 to 151 or SEQ ID NO: 2, 4, 5, 7 to 151, one, two, three or four single, double, or triple amino acids are added to SEQ ID NO: 2 to 151 or SEQ ID NO: 2, 4, 5, 7 to 151 , wherein the added amino acids are D- or L-amino acids selected from the group consisting of K, R, W and L.

[0048] The peptide of any one of the preceding embodiments, wherein the peptide comprises one or more sequences selected from the group consisting of SEQ ID NO: 2, 4, 5, and 7, or selected from the group consisting of a derivative of SEQ ID NO: 2, 4, 5, and 7;

[0049] wherein the derivative of SEQ ID NO: 2, 4, 5, and 7, comprises a chemical modification of one or more amino acids or a chemical modification of the peptide of SEQ ID NO: 2, 4, 5, and 7, preferably the chemical modification is selected from the group consisting of phosphorylation, acetylation, oxidation, ubiquitylation, amidation, deamidation, esterification, alkylation, preferably methylation, nitration, hydroxylation, formylation, sulfation, covalent binding of a lipidic moiety, sulphonation, glycation, acylation, myristoylation, PEGylation, palmitoylation, halogenation, chemical modifications of part or all peptide bonds, and / or post-translational amino acid modification; and / or

[0050] wherein in the derivative of SEQ ID NO: 2, 4, 5, and 7, one, two, three or four amino acids are each exchanged by an amino acid variant, preferably by an analogue amino acid or a conservative substitute amino acid; and / orwherein in the derivative of SEQ ID NO: 2, 4, 5, and 7, one, two, three or four amino acids are deleted, wherein the derivative of SEQ ID NO: 2, 4, 5, and 7, has a length of at least 7 amino acids, and / or

[0051] wherein in the derivative of SEQ ID NO: 2, 4, 5, and 7, one, two, three or four single, double, or triple amino acids are added to SEQ ID NO: 2, 4, 5, and 7, wherein the added amino acids are D- or L-, preferably L-amino acids selected from the group consisting of K, R, W and L.

[0052] The peptide of any one of the preceding embodiments, wherein the chemical modification is an amidation and / or acetylation, more preferably a C terminal amidation and / or N terminal acetylation.

[0053] The peptide of any one of the preceding embodiments, wherein

[0054] the amino acid variant of K (lysine) is selected from the group consisting of: diaminobutyric acid, 2,3-diaminopropanoic acid, (2s)-2,8-diaminooctanoic acid, ornithine, thialysine, histidine, arginine, 5-methyl-arginine, lysine, c-gamma-hydroxy arginine, citrulline, 2-amino-4-guanidinobutryric acid, 2-amino-3-guanidinopropionic acid, canavanine, homoarginine, thio-citrulline, histidine, 2-fluoro-l-histidine, 2-fluoro-l-histidine, 2-fluoro-l-histidine, (2-furyl)-alanine, 3-(1-pyrazolyl)-alanine, 3-(2-tetrazolyl)-alanine, 3-(3-thienyl)-alanine, 3-(2-thienyl)-alanine, 3-(1 ,2,4-triazol-1 -yl)-alanine, and (4-thiazolyl)-alanine;

[0055] preferably the amino acid variant of K (lysine) is selected from the group consisting of: diaminobutyric acid, 2,3-Diaminopropanoic acid, (2s)-2,8-diaminooctanoic acid, ornithine, thialysine, histidine, arginine, 2-fluoro-l-histidine, 2-fluoro-l-histidine, 2-fluoro-l-histidine, (2-furyl)-alanine, 3-(1-Pyrazolyl)-alanine, 3-(2-Tetrazolyl)-alanine, 3-(3-thienyl)-alanine, 3-(2-thienyl)-alanine, 3-(1 ,2,4-Triazol-1-yl)-alanine, and (4-thiazolyl)-alanine;

[0056] more preferably the amino acid variant of K (lysine) is selected from the group consisting of: diaminobutyric acid, 2,3-Diaminopropanoic acid, (2s)-2,8-diaminooctanoic acid, ornithine, thialysine, histidine and arginine;the amino acid variant of R (arginine) is selected from the group consisting of: diaminobutyric acid, 2,3-diaminopropanoic acid, (2s)-2,8-diaminooctanoic acid, ornithine, thialysine, histidine, lysine, 5-methyl-arginine, c-gamma-hydroxy arginine, citrulline, 2-amino-4-guanidinobutryric acid, 2-amino-3-guanidinopropionic acid, canavanine, homoarginine, thio-citrulline, histidine, 2-fluoro-l-histidine, 2-fluoro-l-histidine, 2-fluoro-l-histidine, (2-furyl)-alanine, 3-(1-pyrazolyl)-alanine, 3-(2-tetrazolyl)-alanine, 3-(3-thienyl)-alanine, 3-(2-thienyl)-alanine, 3-(1 ,2,4-triazol-1 -yl)-alanine, and (4-thiazolyl)-alanine;

[0057] preferably the amino acid variant of R (arginine) is selected from the group consisting of: 5-methyl-arginine, lysine, histidine, c-gamma-hydroxy arginine, citrulline, 2-amino-4-guanidinobutryric acid, 2-amino-3-guanidinopropionic acid, canavanine, homoarginine, thio-citrulline, 2-fluoro-l-histidine, 2-fluoro-l-histidine, 2-fluoro-l-histidine, (2-furyl)-alanine, 3-(1-Pyrazolyl)-alanine, 3-(2-Tetrazolyl)-alanine, 3-(3-thienyl)-alanine, 3-(2-thienyl)-alanine, 3-(1 ,2,4-Triazol-1-yl)-alanine, and (4-thiazolyl)-alanine;

[0058] more preferably the amino acid variant of R (arginine) is selected from the group consisting of 5-methyl-arginine, c-gamma-hydroxy arginine, citrulline, 2-amino- 4-guanidinobutryric acid, 2-amino-3-guanidinopropionic acid, canavanine, homoarginine, thio-citrulline, lysine, and histidine;

[0059] the amino acid variant of W (tryptophan) is selected from the group consisting of: 7-hydroxy-l-tryptophan, 3-(4H-thieno[3,2-b]pyrrol-6-yl)-L-alanine, 4-fluoro-tryptophan, 4-hydroxy-tryptophan , 4-amino-l-tryptophan, 6-chloro-l-tryptophan, 3-(3-benzothienyl)-alanine, 6-bromo-tryptophan, 7-chloro-tryptophan, 6-fluoro-l-tryptophan, 5-Fluoro-tryptophan, 5-Hydroxy-tryptophan, beta-hydroxy-tryptophane, 5-Methoxy-tryptophan, 5-Methyl-tryptophan, 6-Methyl-tryptophan, 2-hydroxy-tryptophan, 6-hydroxy-tryptophan, 6-amino-7-hydroxy-l-tryptophan, leucine, isoleucine, valine, proline, phenylalanine, methionine, histidine, and tyrosine

[0060] preferably the amino acid variant of W (tryptophan) is selected from the group consisting of: 7-hydroxy-l-tryptophan, 3-(4H-thieno[3,2-b]pyrrol-6-yl)-L-alanine,4-fluoro-tryptophan, 4-hydroxy-tryptophan, 4-amino-l-tryptophan, 6-chloro-l-tryptophan, 3-(3-benzothienyl)-alanine, 6-bromo-tryptophan, 7-chloro-tryptophan, 6-fluoro-l-tryptophan, 5-Fluoro-tryptophan, 5-Hydroxy-tryptophan, beta-hydroxy-tryptophane, 5-Methoxy-tryptophan, 5-Methyl-tryptophan, 6-Methyl-tryptophan, 2-hydroxy-tryptophan, 6-hydroxy-tryptophan, 6-amino-7-hydroxy-l-tryptophan, phenylalanine, histidine, and tyrosine;

[0061] the amino acid variant of L (leucine) is selected from the group consisting of: alpha-amino-2-indanacetic acid, isoleucine, tert-leucine, allo-isoleucine, norleucine, 5-bromo-L-isoleucine, homoleucine, 5,5,5-trifluoro-leucine, 3-methyl-L-alloisoleucine, 4-hydroxy-L-isoleucine, 4,5-dihydroxy-isoleucine, betahydroxyleucine, 6-hydroxy-L-norleucine, (4r)-5-oxo-l-leucine, 5-oxo-l-norleucine, (4s)-5-fluoro-l-leucine, 2-aminobutyric acid, 2-aminoheptanoic acid, diethylalanine, norvaline, (2s)-2-amino-4,4-difluorobutanoic acid, hydroxynorvaline, valine, 3-fluoro-valine, 3-hydroxy-l-valine, 4-oxo-l-valine, 3-hydroxy-l-valine, alanine, proline, phenylalanine, histidine, methionine, tryptophan, serine, threonine, and tyrosine;

[0062] preferably the amino acid variant of L (leucine) is selected from the group consisting of: alpha-amino-2-indanacetic acid, isoleucine, tert-leucine, allo-isoleucine, norleucine, 5-bromo-L-isoleucine, homoleucine, 5,5,5-trifluoro-leucine, 3-methyl-L-alloisoleucine, 4-hydroxy-L-isoleucine, 4,5-dihydroxy-isoleucine, beta-hydroxyleucine, 6-hydroxy-L-norleucine, (4r)-5-oxo-l-leucine, 5-oxo-l-norleucine, (4s)-5-fluoro-l-leucine, 2-aminobutyric acid, 2-aminoheptanoic acid, diethylalanine, norvaline, (2s)-2-amino-4,4-difluorobutanoic acid, hydroxynorvaline, valine, methionine, 3-fluoro-valine, 3-hydroxy-l-valine, 4-oxo-l-valine, and 3-hydroxy-l-valine.

[0063] The peptide of any one of the preceding embodiments, wherein the amino acid variant is the conservative substitute amino acid, wherein

[0064] the conservative substitute amino acid of K (lysine) is D- or L-histidine or D- or L-arginine, preferably arginine;the conservative substitute amino acid of R (arginine) is D- or L-lysine or D- or L-histidine, preferably D- or L-lysine;

[0065] the conservative substitute am ino acid of W (tryptophan) is a D- or L- amino acid selected from the group consisting of: alanine, leucine, isoleucine, valine, proline, phenylalanine, methionine, histidine, and tyrosine; preferably the conservative substitute amino acid W (tryptophan) is a D- or L- amino acid selected from the group consisting of phenylalanine, histidine, and tyrosine;

[0066] the conservative substitute amino acid of L (leucine) is a D- or L- amino acid selected from the group consisting of: alanine, isoleucine, norleucine, valine, proline, phenylalanine, tryptophan, serine, threonine, and methionine; preferably the conservative substitute amino acid of L (leucine) is a D- or L-amino acid selected from the group consisting of isoleucine, valine, methionine, and norleucine.

[0067] The peptide of any one of the preceding embodiments, wherein

[0068] the conservative substitute amino acid of K (lysine) is D- or L-arginine;

[0069] the conservative substitute amino acid of R (arginine) is D- or L-lysine;

[0070] the conservative substitute am ino acid of W (tryptophan) is a D- or L- amino acid selected from the group consisting of phenylalanine, histidine, and tyrosine;

[0071] the conservative substitute amino acid of L (leucine) is a D- or L- amino acid selected from the group consisting of isoleucine, valine, methionine, and norleucine.

[0072] The peptide of any one of the preceding embodiments, wherein the peptide comprises an amino acid sequence selected of the group consisting of SEQ ID NO: 1-151, or a derivative thereof, wherein up to 200 amino acids, preferably up to 100 amino acids, more preferably up to 50 amino acids are added to the N terminus and / or C terminus of a sequence of SEQ ID NO: 1-151.

[0073] The peptide of any one of the preceding embodiments, wherein the peptidecomprises more than one sequence of SEQ ID NO: 1-151, preferably an n-mer, wherein n is an integer between 2 and 20, preferably 2 and 10.

[0074] The peptide of any one of the preceding embodiments, wherein the peptide is a diastereomer comprising at least one D-amino acid.

[0075] The peptide of any one of the preceding embodiments, wherein the amino acids of the peptide comprise or consist of a racemic mixture of D- and L-amino acids.

[0076] The peptide of any one of the preceding embodiments, wherein the amino acids ofX1 toX11 of the sequence of SEQ ID NO: 1 comprises or consist of a racemic mixture of D- and L-amino acids.

[0077] A polynucleotide encoding the peptide of any one of embodiments 1-22.

[0078] A composition comprising the peptide of any one of embodiments 1 -22.

[0079] The composition of any one of embodiments 24 further comprising a pharmaceutically acceptable carrier.

[0080] The composition of embodiment 24 or 25 further comprising a compound selected from the group consisting of an antibiotic, lysozyme, adjuvant, vaccines, immunostimulant and combinations thereof.

[0081] The peptide of any one of embodiments 1 -22, or the composition of any one of embodiments 24-26, for use in treating and / or preventing an infection and / or inflammation in a subject or on a subject.

[0082] A method for treating or preventing a microbial infection and / or inflammation in or on a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the peptide of any one of embodiments 1-22, or the composition of any one of embodiments 23-26.

[0083] The peptide or the composition for use according to embodiment 27, or the method of embodiment 28, further comprising administering to the subject at least one antibiotic agent, wherein the antibiotic is administered simultaneouslyor sequentially with the disclosed peptide.

[0084] The peptide or the composition for use according to embodiment 27 or 29, or the method of embodiment 28 or 29, wherein the infection and / or inflammation is caused by one or more microorganisms.

[0085] A non-in vivo, preferably an in vitro or ex vivo, method of inhibiting growth of one or more microorganisms, the method comprises contacting the microorganisms with an effective amount of the peptide of any one of the preceding embodiments 1-22, or the composition of any one of embodiments 24-26.

[0086] A non-in vivo , preferably an in vitro or ex vivo, method of inhibiting biofilm formation and / or dissolution of a biofilm, the method comprises contacting the microorganisms with an effective amount of the peptide of any one of the preceding embodiments 1-22, or the composition of any one of embodiments 24-26.

[0087] The method of embodiments 31 or 32, wherein the step of contacting the microorganisms with an effective amount of the peptide or the composition comprises applying the peptide or the composition to an article or a surface of an article.

[0088] The peptide or the composition for use according to embodiment 30, or the method of embodiment 30, or the methods of any one of embodiments 31-33, wherein the one or more microorganisms are selected from the group consisting of bacteria, fungi, viruses, archaea, protozoa, and combinations of two or more thereof.

[0089] The peptide or the composition for use according to embodiment 30, or the method of embodiment 30, or the methods of any one of embodiments 31-33, wherein the one or more microorganisms are selected from the group consisting of bacteria, fungi, and a combination thereof.

[0090] The peptide or the composition for use according to embodiment 30, or themethod of embodiment 30, or the methods of any one of embodiments 31-33, wherein the bacteria are selected from the group consisting of Gram-positive bacteria, Gram-negative bacteria, and a combination thereof.

[0091] The peptide or the composition for use according to embodiment 30, or the method of embodiment 30, or the methods of any one of embodiments 31-33, wherein the bacteria are from a genus selected from the group consisting of Pseudomonas, Escherichia, Staphylococcus, Bacillus, Gardnerella, Neisseria, and a combination thereof;

[0092] preferably the bacteria are from a species selected from the group consisting of Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis, Bacillus subtilis, Gardnerella vaginalis and Neisseria gonorrhoeae and a combination thereof;

[0093] more preferably the bacteria are from the species Staphylococcus aureus and / or Bacillus subtilis.

[0094] The peptide or the composition for use according to embodiment 30, or the method of embodiment 30, or the methods of any one of embodiments 31-33, wherein the fungus / fungi is / are from the genus Candida and / or Aspergillus, preferably the fungus / fungi is / are is of the species Candida albicans and / or Aspergillus niger.

[0095] A method for identifying a compound, preferably the compound is a peptide, having antimicrobial activity, the method comprises the steps of:

[0096] mixing a bacterial suspension with the compound to be tested;

[0097] incubating the mixture comprising the bacterial suspension and the compound to be tested, wherein incubation time and incubation temperature are adjusted to the bacteria included in the bacterial suspension;

[0098] quantifying bacterial growth by defining a minimum inhibitory concentration (MIC), wherein MIC is defined as the lowest concentration of the compound orpeptide that inhibits bacterial growth to a non-detectable level, wherein preferably bacterial growth is quantified by measuring density or metabolic activity of the bacteria.

[0099] Detailed Description of the Invention

[0100] In a first aspect, the invention relates to a peptide comprising an amino acid sequence of SEQ ID NO: 1 : X1 -X2-X3-X4-X5-X6-X7-X8-X9-X10-X11 ,

[0101] wherein X1 to X7 are, independently of each other, a D- or L-amino acid selected from the group consisting of K (lysine), R (arginine), W (tryptophan), L (leucine), an analogue amino acid of K, R, W, or L, and a conservative substitute amino acid of K, R, W or L; and wherein X8 to X11 are, independently of each other, a deletion or a D-or L-amino acid selected from the group consisting of K, W, R, L, an analogue amino acid of K, R, W or L, and a conservative substitute amino acid of K, R, W or L.

[0102] Preferably, the amino acids in SEQ ID NO: 1 are contiguous amino acids. In certain embodiments, the invention discloses an isolated and / or purified peptide comprising a contiguous amino acid sequence of SEQ ID NO: 1. The term “isolated”, as used herein in reference to a peptide, refers to a peptide substantially free of proteins, lipids, nucleic acids, for example, with which it might be naturally associated. The term "isolated" is not meant to exclude artificial or synthetic mixtures with other compounds or materials, or the presence of impurities that do not interfere with the fundamental activity, and that may be present, for example, due to incomplete purification, or the addition of stabilizers.

[0103] X1 is preferably a D- or L-amino acid, preferably an L-amino acid, selected from the group consisting of K, an analogue amino acid of K, and a conservative amino acid substitute of K. X2 is preferably a D- or L-amino acid, preferably an L-amino acid, selected from the group consisting of R, an analogue amino acid of R, and a conservative amino acid substitute of R. X3 is preferably a D- or L-amino acid, preferably an L-amino acid, selected from the group consisting of W, an analogue amino acid of W, and a conservative amino acid substitute of W. X4 is a D- or L-amino acid, preferably an L-amino acid, selected from the group consisting of: W, K and R,an analogue amino acids of W, K and R, and a conservative amino acid substitutes of W, K and R. X5 is a D- or L-amino acid, preferably an L-amino acid, selected from the group consisting of: W, K and R, and L, analogue amino acids of W, K, R, and L, and conservative amino acid substitutes of W, K, R, and L. X6 is a D- or L-amino acid, preferably an L-amino acid, selected from the group consisting of: W, K and R, and L, analogue amino acids of W, K, R, and L, and conservative amino acid substitutes of W, K, R, and L. X7 and X8 are independently a D- or L-amino acid, preferably an L-amino acid, selected from the group consisting: of W, R, and L, analogue amino acids of W, R, and L, and conservative amino acid substitutes of W, R, and L. Preferably, the C terminus of the peptide comprises or preferably consists of one, two or three D-or L-leucine, preferably one, two or three L-leucine. More preferably, the C terminus of the peptide comprises or preferably consists of one or two D- or L-leucine, preferably one, or two L-leucine.

[0104] In certain embodiments, X1 is a D- or L-amino acid selected from the group consisting of K, an analogue amino acid of K and a conservative amino acid substitute of K, and X2 is a D- or L-amino acid selected from the group consisting of R, an analogue amino acid of R, and a conservative amino acid substitute of R, in the peptide of the invention. In certain embodiments, X1 is a D- or L-amino acid, preferably an L-amino acid, selected from the group consisting of K, an analogue amino acid of K and a conservative amino acid substitute of K, and X2 is a D- or L-amino acid, preferably an L-amino acid, selected from the group consisting of R, an analogue amino acid of R, and a conservative amino acid substitute of R, in the peptide of the invention, and the C terminus of the peptide comprises or preferably consists of one, two or three D- or L-leucine, preferably one, two or three L-leucine.

[0105] In certain embodiments, X1 is a D- or L-amino acid selected from the group consisting of K, an analogue amino acid of K, and a conservative amino acid substitute of K, and X3 is a D- or L-amino acid selected from the group consisting of W, an analogue amino acid of W, and a conservative amino acid substitute of W, in the peptide of the invention. In certain embodiments, X1 is a D- or L-amino acid, preferably an L-amino acid, selected from the group consisting of K, an analogue amino acid of K, and a conservative amino acid substitute of K, and X3 is a D- or L-amino acid selected fromthe group consisting of W, an analogue amino acid of W, and a conservative amino acid substitute of W, in the peptide of the invention, and the C terminus of the peptide comprises or preferably consists of one, two or three D- or L-leucine, preferably one, two or three L-leucine.

[0106] In certain embodiments, X2 is a D- or L-amino acid selected from the group consisting of R, an analogue amino acid of R, and a conservative amino acid substitute of R, and X3 is a D- or L-amino acid selected from the group consisting of W, an analogue amino acid of W, and a conservative amino acid substitutes of W, in the peptide of the invention. In certain embodiments, X2 is a D- or L-amino acid, preferably an L-amino acid, selected from the group consisting of R, an analogue amino acid of R, and a conservative amino acid substitute of R, and X3 is a D- or L-amino acid selected from the group consisting of W, an analogue amino acid of W, and a conservative amino acid substitutes of W, in the peptide of the invention, and the C terminus of the peptide comprises or preferably consists of one, two or three D- or L-leucine, preferably one, two or three L-leucine.

[0107] In certain embodiments, X1 is a D- or L-amino acid selected from the group consisting of K, an analogue amino acid of K, and a conservative amino acid substitute of K, and X2 is a D- or L-amino acid selected from the group consisting of R, an analogue amino acid of R, and a conservative amino acid substitute of R, and X3 is a D- or L-amino acid selected from the group consisting of W, an analogue amino acid of W, and a conservative amino acid substitute of W, in the peptide of the invention. In certain embodiments, X1 is a D- or L-amino acid, preferably an L-amino acid, selected from the group consisting of K, an analogue amino acid of K, and a conservative amino acid substitute of K; X2 is a D- or L-amino acid, preferably an L-amino acid, selected from the group consisting of R, an analogue amino acid of R, and a conservative amino acid substitute of R; X3 is a D- or L-amino acid, preferably an L-amino acid, selected from the group consisting of W, an analogue amino acid of W, and a conservative amino acid substitute of W, in the peptide of the invention, and the C terminus of the peptide comprises, preferably consists of one, two or three D- or L-leucine, preferably L-leucine.In certain embodiments, X1 is a D- or L-amino acid selected from the group consisting of K, an analogue amino acid of K, and a conservative amino acid substitute of K, and X2 is a D- or L-amino acid selected from the group consisting of R, an analogue amino acid of R, and a conservative amino acid substitute of R, X3 is a D- or L-amino acid selected from the group consisting of W, an analogue amino acid of W, and a conservative amino acid substitute of W, and X4 is a D- or L-amino acid selected from the group consisting of W, Kand R, an analogue amino acid, and a conservative amino acid substitute thereof, in the peptide of the invention. In certain embodiments, X1 is a D- or L-amino acid, preferably an L-amino acid, selected from the group consisting of K, an analogue amino acid of K, and a conservative amino acid substitute of K, and X2 is a D- or L-amino acid, preferably an L-amino acid, selected from the group consisting of R, an analogue amino acid of R, and a conservative amino acid substitute of R, X3 is a D- or L-amino acid, preferably an L-amino acid, selected from the group consisting of W, an analogue amino acid of W, and a conservative amino acid substitute of W, and X4 is a D- or L-amino acid, preferably an L-amino acid, selected from the group consisting of W, K and R, an analogue amino acid, and a conservative amino acid substitute thereof, in the peptide of the invention, and the C terminus of the peptide comprises or preferably consists of one, two or three D- or L-leucine, preferably one, two or three L-leucine.

[0108] In certain embodiments, X1 is K, and X2 is R, X3 is W, and X4 is a D- or L-amino acid selected from the group consisting of W, K and R, in the peptide of the invention. In certain embodiments, X1 is a D- or L-amino acid of K, and X2 is a D- or L-amino acid of R, X3 is a D- or L-amino acid of W, and X4 is a D- or L-amino acid of W, in the peptide of the invention. In certain embodiments, X1 is a D- or L-amino acid of K, and X2 is a D- or L-amino acid of R, X3 is a D- or L-amino acid of W, and X4 is a D- or L-amino acid of K, in the peptide of the invention. In certain embodiments, X1 is a D- or L-amino acid of K, and X2 is a D- or L-amino acid of R, X3 is a D- or L-amino acid of W, and X4 is a D- or L-amino acid of R, in the peptide of the invention. In certain embodiments, X1 is K, and X2 is R, X3 is W, and X4 is a D- or L-amino acid, preferably an L-amino acid, selected from the group consisting of W, K and R, in the peptide of the invention, and the C terminus of the peptide comprises or preferably consists ofone, two or three D- or L-leucine, preferably one, two or three L-leucine. In certain embodiments, X1 is a D- or L-amino acid, preferably an L-amino acid, of K, and X2 is a D- or L-amino acid, preferably an L-amino acid, of R, X3 is a D- or L-amino acid, preferably an L-amino acid, of W, and X4 is a D- or L-amino acid, preferably an L-amino acid, of W, in the peptide of the invention, and the C terminus of the peptide comprises or preferably consists of one, two or three D- or L-leucine, preferably one, two or three L-leucine. In certain embodiments, X1 is a D- or L-amino acid, preferably an L-amino acid, of K, and X2 is a D- or L-amino acid, preferably an L-amino acid, of R, X3 is a D- or L-amino acid, preferably an L-amino acid, of W, and X4 is a D- or L-amino acid, preferably an L-amino acid, of K, in the peptide of the invention, and the C terminus of the peptide comprises or preferably consists of one, two or three D- or L-leucine, preferably one, two or three L-leucine. In certain embodiments, X1 is a D- or L-amino acid, preferably an L-amino acid, of K, and X2 is a D- or L-amino acid, preferably an L-amino acid, of R, X3 is a D- or L-amino acid, preferably an L-amino acid, of W, and X4 is a D- or L-amino acid, preferably an L-amino acid, of R, in the peptide of the invention, and the C terminus of the peptide comprises or preferably consists of one, two or three D- or L-leucine, preferably one, two or three L-leucine.

[0109] The inventors demonstrated that one or more amino acids can be deleted from the N-terminus or C-terminus without substantial loss of biological function. Accordingly, the present invention provides polypeptides having one or more residues deleted from the amino terminus. Especially, the inventors provide biologically functional peptides with C-terminal deletions. Accordingly, the present invention provides polypeptides having one or more residues deleted from the carboxy terminus. Thus, X8 to X11 are, independently of each other, a deletion or a D- or L-amino acid, preferably an L-amino acid, selected from the group consisting of K, W, R, L, an amino acid variant of K, R, W or L, and a conservative substitute amino acid of K, R, W or L. The invention also provides polypeptides having one or more amino acids deleted from both the amino and the carboxyl termini.

[0110] Preferably, the peptide of the invention has antimicrobial, preferably an antibacterial and / or anti-fungal effect. The terms "antimicrobial" as used herein, means to inhibit, prevent, or stop proliferation of microbes or to destroy microbes, such as bacteria,fungi, viruses, parasites or the like. , fungicidal activity and / or virucidal activity. The term "antibacterial" or “bactericidal activity” as used herein, means to inhibit, prevent, or stop proliferation of bacteria or to destroy bacteria. The term "antifungal", as used herein, means to inhibit, prevent, or stop proliferation of fungi or to destroy fungi.

[0111] The peptide of any one of the preceding claims comprising at least one alpha helical domain. The term “alpha helix” (or a-helix), as used herein, is a secondary structure in a peptide sequence, wherein the amino acid sequence is twisted into a coil (a helix). The term “alpha helix” includes right- and left-handed helices, preferably right-handed helices.

[0112] The term “domain”, as used herein, refers to a section or portion of a peptide entity. A “domain” can be associated with a particular structural and / or functional feature of the peptide. A domain can be a section of a polypeptide. Preferably, a domain is characterized by a particular structural element (e.g., a particular amino acid sequence or sequence motif, alpha-helix character, b-sheet character, coiled-coil character, random coil character, etc.).

[0113] As used herein, an alpha helical domain is a section comprised in the peptide of the invention which has an alpha-helix character. In a certain embodiment, the complete peptide of the invention has an alpha-helix character.

[0114] The peptide of the invention is a cationic peptide. Preferably the cationic peptide has a net positive charge, more preferably the net positive charge ranges from +2 to +12. The step of determining most common net charge of the anti-microbial peptides within the set involves calculating the charge of each peptide in the database, then dividing it by the peptide length. These charges are plotted to assess distribution and then averaged to determine a most common charge per unit length for the set.

[0115] The peptide of the invention can comprise more than one sequence of SEQ ID NO: 1-151. The more than one sequences have either the same or different sequences (homo- or heteromer). The peptide of the invention can comprise n different or identical sequences of SEQ ID NO: 1-151, wherein n is an integer between 2 and 20, preferably 2 and 10. The polypeptides of the present invention can be produced as multimersincluding dimers, trimers, and tetramers. Multimerization can be facilitated by linkers, introduction of cysteines to permit creation of interchain disulfide bonds, or recombinantly though heterologous polypeptides, such as Fc regions.

[0116] The peptide of the invention can be a fusion protein, i.e. , a type of protein that has an amino acid sequence that results from the linkage of the amino acid sequences of two or more normally separate polypeptides.

[0117] Derivatives of the peptide of the invention

[0118] The invention further includes variants and derivatives of the peptides of the invention and the amino acids included therein, including amino acid mutations, such as deletions, insertions, inversions, repeats, and substitutions selected according to general rules known in the art so as to have little effect on activity.

[0119] A derivative of the peptide of the invention can comprise a chemical modification of the peptide of the invention and the amino acids included therein; one, two, three or four amino acids exchanges, preferably conservative amino acid exchanges; one, two, three or four amino acid deletions; and / or one, two, three or four additions of single, double, or triple amino acids.

[0120] Preferably, the chemical modification is selected from the group consisting of phosphorylation, acetylation, oxidation, ubiquitylation, amidation, deamidation, esterification, alkylation, preferably methylation, nitration, hydroxylation, formylation, sulfation, covalent binding of a lipidic moiety, sulphonation, glycation, acylation, myristoylation, PEGylation, palmitoylation, halogenation, chemical modifications of part or all peptide bonds, and / or post-translational amino acid modification.

[0121] The one, two, three or four exchanged amino acids are each preferably exchanged by a corresponding number of amino acid variant. The term “amino acid variant” preferably comprises and relates to analogue amino acids and conservative substitute amino acids.

[0122] The analogue amino acid of K (lysine) is preferably selected from the group consisting of: diaminobutyric acid, 2,3-diaminopropanoic acid, (2s)-2,8-diaminooctanoic acid,ornithine, thialysine, 5-methyl-arginine, lysine, c-gamma-hydroxy arginine, citrulline, 2-amino-4-guanidinobutryric acid, 2-amino-3-guanidinopropionic acid, canavanine, homoarginine, thio-citrulline, histidine, 2-fluoro-l-histidine, 2-fluoro-l-histidine, 2-fluoro-l-histidine, (2-furyl)-alanine, 3-(1-pyrazolyl)-alanine, 3-(2-tetrazolyl)-alanine, 3-(3-thienyl)-alanine, 3-(2-thienyl)-alanine, 3-(1 ,2,4-triazol-1 -yl)-alanine, and (4-thiazolyl)-alanine.

[0123] The analogue amino acid of K (lysine) is more preferably selected from the group consisting of: diaminobutyric acid, 2,3-Diaminopropanoic acid, (2s)-2,8-diaminooctanoic acid, ornithine, thialysine, 2-fluoro-l-histidine, 2-fluoro-l-histidine, 2-fluoro-l-histidine, (2-furyl)-alanine, 3-(1-Pyrazolyl)-alanine, 3-(2-Tetrazolyl)-alanine, 3-(3-thienyl)-alanine, 3-(2-thienyl)-alanine, 3-(1 ,2,4-Triazol-1-yl)-alanine, and (4-thiazolyl)-alanine.

[0124] The analogue amino acid of K (lysine) is again more preferably selected from the group consisting of: diaminobutyric acid, 2,3-Diaminopropanoic acid, (2s)-2,8-diaminooctanoic acid, ornithine, and thialysine.

[0125] The analogue amino acid of R (arginine) is preferably selected from the group consisting of: diaminobutyric acid, 2,3-diaminopropanoic acid, (2s)-2,8-diaminooctanoic acid, ornithine, thialysine, 5-methyl-arginine, c-gamma-hydroxy arginine, citrulline, 2-amino-4-guanidinobutryric acid, 2-amino-3-guanidinopropionic acid, canavanine, homoarginine, thio-citrulline, histidine, 2-fluoro-l-histidine, 2-fluoro-l-histidine, 2-fluoro-l-histidine, (2-furyl)-alanine, 3-(1-pyrazolyl)-alanine, 3-(2-tetrazolyl)-alanine, 3-(3-thienyl)-alanine, 3-(2-thienyl)-alanine, 3-(1 ,2,4-triazol-1-yl)-alanine, and (4-thiazolyl)-alanine;

[0126] The analogue amino acid of R (arginine) is more preferably selected from the group consisting of: 5-methyl-arginine, c-gamma -hydroxy arginine, citrulline, 2-amino-4-guanidinobutryric acid, 2-amino-3-guanidinopropionic acid, canavanine, homoarginine, thio-citrulline, 2-fluoro-l-histidine, 2-fluoro-l-histidine, 2-fluoro-l-histidine, (2-furyl)-alanine, 3-(1-Pyrazolyl)-alanine, 3-(2-Tetrazolyl)-alanine, 3-(3-thienyl)-alanine, 3-(2-thienyl)-alanine, 3-(1 ,2,4-Triazol-1-yl)-alanine, and (4-thiazolyl)-alanine.The analogue amino acid of R (arginine) is again more preferably selected from the group consisting of 5-methyl-arginine, c-gamma-hydroxy arginine, citrulline, 2-amino- 4-guanidinobutryric acid, 2-amino-3-guanidinopropionic acid, canavanine, homoarginine, and thio-citrulline.

[0127] The analogue amino acid of W (tryptophan) is preferably selected from the group consisting of: 7-hydroxy-l-tryptophan, 3-(4H-thieno[3,2-b]pyrrol-6-yl)-L-alanine, 4-fluoro-tryptophan, 4-hydroxy-tryptophan, 4-amino-l-tryptophan, 6-chloro-l-tryptophan, 3-(3-benzothienyl)-alanine, 6-bromo-tryptophan, 7-chloro-tryptophan, 6-fluoro-l-tryptophan, 5-Fluoro-tryptophan, 5-Hydroxy-tryptophan, beta-hydroxy-tryptophane, 5-Methoxy-tryptophan, 5-Methyl-tryptophan, 6-Methyl-tryptophan, 2-hydroxy-tryptophan, 6-hydroxy-tryptophan, and 6-amino-7-hydroxy-l-tryptophan.

[0128] The analogue amino acid of W (tryptophan) is more preferably selected from the group consisting of: 7-hydroxy-l-tryptophan, 3-(4H-thieno[3,2-b]pyrrol-6-yl)-L-alanine, 4-fluoro-tryptophan, 4-hydroxy-tryptophan, 4-amino-l-tryptophan, 6-chloro-l-tryptophan, 3-(3-benzothienyl)-alanine, 6-bromo-tryptophan, 7-chloro-tryptophan, 6-fluoro-l-tryptophan, 5-Fluoro-tryptophan, 5-Hydroxy-tryptophan, beta-hydroxy-tryptophane, 5-Methoxy-tryptophan, 5-Methyl-tryptophan, 6-Methyl-tryptophan, 2-hydroxy-tryptophan, 6-hydroxy-tryptophan, and 6-amino-7-hydroxy-l-tryptophan;

[0129] The analogue amino acid of L (leucine) is preferably selected from the group consisting of: alpha-amino-2-indanaceticacid, isoleucine, tert-leucine, allo-isoleucine, norleucine, 5-bromo-L-isoleucine, homoleucine, 5,5,5-trifluoro-leucine, 3-methyl-L-alloisoleucine, 4-hydroxy-L-isoleucine, 4,5-dihydroxy-isoleucine, beta-hydroxyleucine, 6-hydroxy-L-norleucine, (4r)-5-oxo-l-leucine, 5-oxo-l-norleucine, (4s)-5-fluoro-l-leucine, 2-aminobutyric acid, 2-aminoheptanoic acid, diethylalanine, norvaline, (2s)-2-amino-4,4-difluorobutanoic acid, hydroxynorvaline, 3-fluoro-valine, 3-hydroxy-l-valine, and 4-oxo-l-valine, 3-hydroxy-l-valine.

[0130] The analogue amino acid of L (leucine) is more preferably selected from the group consisting of: alpha-amino-2-indanacetic acid, isoleucine, tert-leucine, allo-isoleucine, norleucine, 5-bromo-L-isoleucine, homoleucine, 5,5,5-trifluoro-leucine, 3-methyl-L-alloisoleucine, 4-hydroxy-L-isoleucine, 4,5-dihydroxy-isoleucine, beta-hydroxyleucine,6-hydroxy-L-norleucine, (4r)-5-oxo-l-leucine, 5-oxo-l-norleucine, (4s)-5-fluoro-l-leucine, 2-aminobutyric acid, 2-aminoheptanoic acid, diethylalanine, norvaline, (2s)-2-amino-4,4-difluorobutanoic acid, hydroxynorvaline, 3-fluoro-valine, 3-hydroxy-l-valine, 4-oxo-l-valine, and 3-hydroxy-l-valine.

[0131] In certain embodiments, the invention discloses a peptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1-151, preferably SEQ ID NO: 2-151 , again more preferably SEQ ID NO: 2, 4, 5, and 7, with one or more amino acid substitutions, modifications, insertions, or deletions. Preferably, the peptide has antimicrobial, preferably an antibacterial and / or anti-fungal effect.

[0132] In other embodiment, the invention discloses a peptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1-151, preferably SEQ ID NO: 2-151, again more preferably SEQ ID NO: 2, 4, 5, and 7, with one or more conservative amino acid substitutions. Preferably, the peptide has antimicrobial, preferably an antibacterial and / or anti-fungal effect.

[0133] The peptide may have internal peptide deletions and substitutions as well as deletions and substitutions at the N-terminus and C-terminus based on SEQ ID NO: 1-151, preferably SEQ ID NO: 2-151 again more preferably SEQ ID NO: 2, 4, 5, and 7. In some embodiments, the peptide has about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, or 20 or more amino acid substitutions, modifications, insertions, or deletions.

[0134] In some embodiments, the peptide of the invention has at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 2-151 or SEQ ID NO: 2, 4, 5, and 7.

[0135] In some embodiment, any peptide of the invention comprises at least one D-amino acid.

[0136] Polynucleotide encoding the peptide of the invention

[0137] In a further aspect, the invention relates to a polynucleotide encoding the peptide ofthe invention. The term “polynucleotide", “nucleic acid” or “nucleic acid molecule”, as used interchangeably herein, is understood as a polymeric or oligomeric macromolecule made from nucleotide monomers. Nucleic acid molecules comprise DNA, RNA, mixtures thereof such as RNA-DNA hybrids, single-stranded, doublestranded, or more highly aggregated hybridization motifs, and any chemical modifications thereof.

[0138] In a further aspect, the invention relates to a vector comprising the polynucleotide encoding the peptide of the invention. As used herein, the term "vector" refers to a protein or a polynucleotide or a mixture thereof which is capable of being introduced or of introducing a peptide and / or nucleic acid, preferably a peptide and / or nucleic acid of the invention or disclosed therein into a cell. In the context of the present invention, it is preferred that the polynucleotide of the invention is expressed within the cell upon introduction of the vector or vectors. Examples of suitable vectors include but are not limited to plasmids, cosmids, phages, viruses, and artificial chromosomes.

[0139] Composition comprising the peptide of the invention

[0140] In a further aspect, the invention relates to a composition comprising the peptide of the invention.

[0141] The composition can further comprise a pharmaceutically acceptable carrier. The characteristics of the carrier are dependent on the route of administration. One route of administration is topical administration. For example, for topical administrations, a preferred carrier is an emulsified cream comprising the active peptide, but other common carriers such as certain petrolatum / mineral-based and vegetable-based ointments can be used, as well as polymer gels, liquid crystalline phases and microemulsions.

[0142] Alternatively, or additionally, the composition further comprises a compound selected from the group consisting of an antibiotic, adjuvant, vaccines, immunostimulant, lysozyme, and combinations thereof. Classes of antibiotics that can be used with the peptides of the invention include, but are not limited to, aminoglycoside, penicillin, cephalosporin, fluoroquinolone, carbapenem, tetracycline and macrolide. Thepharmaceutical compositions of the invention may be administered alone or in combination with other therapeutic agents, such as antibiotic or antiseptic agents such as anti-bacterial agents, anti-fuingicides, anti-viral agents, and anti-parasitic agents. These agents may be incorporated as part of the same pharmaceutical composition or may be administered separately.

[0143] The antimicrobial / pharmaceutical compositions may comprise one or more peptides of the invention, such as 1 , 2, 3 or 4 different peptides in the antimicrobial / pharmaceutical compositions. By using a combination of different peptides, the antimicrobial effect may be increased as well as decrease of the possibility that the microorganism to combat might be resistant and / or tolerant against the antimicrobial agent.

[0144] The antimicrobial / pharmaceutical compositions may also be used to in cleansing solutions, such as lens disinfectants and storage solutions or used to prevent bacterial infection in association with urinary catheter use or use of central venous catheters.

[0145] Additionally, the antimicrobial compositions may be used for prevention of infection post-surgery in plasters, adhesives, sutures, or be incorporated in wound dressings. The antimicrobial peptides may also be used in polymers, textiles or the like to create antibacterial surfaces or Cosmetics, and personal care products (soap, shampoos, toothpaste, anti-acne, suncreams, tampons, diapers, etc.) may be supplemented with the antimicrobial / pharmaceutical compositions.

[0146] Medical treatment

[0147] In a further aspect, the peptide of the invention, or the composition of the invention, is used for treating and / or preventing an infection and / or inflammation.

[0148] Infections that can be reduced or prevented using a compound of formula (I) are caused by a microorganism.

[0149] The term “microorganism” or “microbe”, as used herein, refers broadly to bacteria, fungi, viruses, archaea, protozoa, and combinations of two or more thereof. The bacteria, fungi, viruses, and protozoa as known in the art are generally encompassed.The term “bacteria” includes gram-positive or gram-negative bacteria. The microbe can be a gram-negative bacterium, such as Pseudomonas spp, Escherichia spp, Neisseria spp, or Salmonella spp. The microbe can be a gram-positive bacterium, such as Staphylococcus spp, Staphylococcus spp, Bacillus spp, Gardnerella spp, or Enterococcus spp. Alternatively, the microorganism can be a fungus, such as a yeast, e.g., Candida spp and Aspergillus spp.

[0150] Further bacteria that are known to cause infections include Streptococci such as Group A beta-hemolytic Streptococci, Group B Streptococci, Streptococcus pneumoniae, Viridans Streptococci, Enterococci, Pneumococci and Streptococcus Bovis, Staphylococci including Coagulase negative Staphylococci (CoNS), Coagulase positive Staphylococci, Staphylococcus aureus, Staphylococcus epidermidis and Staphylococcus capitus, Haemophilus influenzae, Moraxella catarrhalis, Pseudomonas aeruginosa, E. coli, Klebsiella including Klebsiella pneumoniae, enterobacteria, Proteus including Proteus mirabilis, Serratia, Corynebacteria, Enterococcus faecalis, Bacterioides spp., Gardenerella spp., Corynebacterium spp., Enterococci, coccobacilli including gram-negative coccobacilli, Providencia stuartii, M. Morganii, Propionibacterium acne, Acinetobacter calcoaceticus, Enterobacter aerogenes, Lactobacilli including anaerobic lactobacilli, lactobacillus plantarum, and Micrococcus.

[0151] In a preferred embodiment, the bacterium is drug resistant, preferably antibiotic resistant, more preferably penicillin resistant. Drug resistant bacteria are known in the art. Preferred examples of drug-resistant bacteria are methicillin-resistant Staphylococcus aureus and multiple-resistant Staphylococcus aureus (MRSA).

[0152] Non-bacterial microbes that are known to cause infections arising from medical devices include Candida spp., for example Candida Albicans, and Aspergillus spp.

[0153] Use for inhibiting growth of microorganisms

[0154] In a further aspect, the invention relates to use of the peptide of the invention or the composition of the invention as biocide. The biocide preferably exhibits bactericidal activity, fungicidal activity and / or virucidal activity. In an embodiment, the inventionprovides a disinfecting solution comprising at least one peptide of the invention and optionally an acceptable carrier.

[0155] In a further aspect, the invention relates to use of the peptide of the invention or the composition of the invention for inhibiting growth of microorganisms.

[0156] In a further aspect, the invention relates to use of the peptide of the invention or the composition of the invention for inhibiting biofilm formation and / or dissolution of a biofilm.

[0157] In a further aspect, the invention relates to a method of inhibiting the growth of microorganisms, such as bacteria and / or fungi, the method comprises contacting the bacteria and / or fungi with an effective amount of the peptide or the composition of the invention. In a further aspect, the invention relates to a method of inhibiting biofilm formation and / or dissolution of a biofilm, the method comprises contacting the bacteria and / or fungi with an effective amount of the peptide or the composition of the invention.

[0158] The term “contacting” refers to exposing the microorganisms to the peptide or composition, so that the peptide can effectively inhibit, kill, or lyse bacteria, bind endotoxin (LPS), or permeabilize Gram negative bacterial outer membranes. Contacting may be in vitro, for example by adding the peptide to a bacterial culture to test for susceptibility of the bacteria to the peptide. Contacting may be in vivo, for example administering the peptide to a subject with a microorganism disorder, such as septic shock or infection. Contacting may further involve coating an object (e.g., medical device) such as a catheter to inhibit microorganisms with which it comes into contact, thus preventing it from becoming colonized with the bacteria. “Inhibiting” or “inhibiting effective amount” refers to the amount of peptide that is required to cause a biostatic or biocidal effect. The method of inhibiting the growth of bacteria may further include the addition of antibiotics for combination or synergistic therapy. The peptides and / or analogs or derivatives thereof may be administered to any host, including a human or non-human animal, in an amount effective to inhibit growth of a bacterium or fungus. The peptides and / or analogs or derivatives thereof may be administered to any in vitro sample, device, object or surface, in an amount effective to inhibit growth of a bacterium or fungus.The invention also relates to a method of disinfecting a surface of an article, the method comprising applying to the surface of the article a composition comprising at least one of the peptides of the invention. Such article may be clothes (e.g., clothes of healthcare workers or military personnel), walls (e.g., walls in hospital rooms), or particles intended for release into the air to kill or reduce pathogens.

[0159] In another embodiment, the present invention may be used as a food preservative or in treating food products to control, reduce, or eliminate potential pathogens or contaminants. A peptide of the invention may be used as a disinfectant, for use in or with any product that must remain microbial free or be within certain tolerances. In an embodiment, treatment with a peptide provides at least partial regulation of infection or contamination.

[0160] In an embodiment it is also possible to incorporate or distribute the peptides within materials, on devices, or on objects (e.g., on an accessible surface), where microbial growth or viable presence is undesirable, as a method of microbicidal or microbistatic inhibition of microbial growth by administering to the devices or objects a microbicidal or microbistatic effective amount of peptide. In an embodiment, such devices or objects include, but are not limited to, linens, cloth, plastics, latex fabrics, natural rubbers, implantable devices, surfaces, or storage containers.

[0161] The invention further provides a method of protecting objects from bacterial colonization. The invention further relates to a medical device or implant comprising at least one peptide of the invention. Preferably, at least one peptide of the invention is covalently attached to the surface of the medical device or implant.

[0162] The peptides of the invention remain active when conjugated to solid surfaces. Thus, the peptides may be used for protecting objects such as medical devices from colonization with pathogenic bacteria by chemically conjugating, or coating by any other means, at least one peptide of the invention to the surface of the medical device. Such medical devices include indwelling catheters, and the like. The invention also relates to a disinfecting solution comprising at least one peptide of the invention and an acceptable carrier.In a further aspect, the invention provides a method for identifying a compound, preferably a peptide, having antimicrobial activity, the method comprises the steps of: mixing a bacterial suspension with the compound to be tested;

[0163] incubating the mixture, wherein incubation time and incubation temperature are adjusted to the bacteria included in the suspension;

[0164] quantifying bacterial growth in the mixture by defining a minimum inhibitory concentration (MIC), preferably bacterial growth is quantified by measuring density or metabolic activity of the bacteria in the mixture,

[0165] wherein the minimum inhibitory concentration (MIC) is defined as the lowest concentration of the compound or peptide that inhibited bacterial growth to a non-measurable or non-detectable level.

[0166] Bacterial density can be measured by measuring turbidity of the mixture, e.g., via spectrophotometry. Metabolic activity of the bacteria can be measured by a colorimetric assay, such as an assay using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). With the MTT assay, the minimum inhibitory concentration (MIC) is defined as the lowest concentration of the peptide that completely inhibited visible bacterial growth, as assessed after addition of MTT to the mixture.

[0167] For E. coli, P. aeruginosa, S. aureus, S. epidermidis, B. subtilis, G. vaginalis, N. gonorrhoeae, C. albicans, and A. niger, incubation temperature was at 35°C ± 2°C. Conventional aerobic bacteria require about 16-20 hours incubation, but incubation times can be adjusted as required.

[0168] The method for identifying a compound or peptide having antimicrobial activity can comprise the step of comparing the MIC measured for the compound or peptide to be tested with the MIC of a reference compound.

[0169] Definitions

[0170] As used herein, the term “amino acid” is used in its broadest sense and may refer toan amino acid in its many different chemical forms including a single administration amino acid, its physiologically active salts or esters, its combinations with its various salts, its tautomeric, polymeric and / or isomeric forms, its analogue forms, its derivative forms, its products of biosynthesis, and / or its decarboxylation products. Compounds containing both a carboxyl group and an amino group are typically known as amino acids. Amino acids typically have the basic formula X-R, wherein X is NH2-CH-COOH. Amino acids typically differ from one another with respect to the structure of the R group. It is the structure of the R group that typically determines the individuality and character of each amino acid. Amino acids are referred to herein by either their commonly known three letter symbols or by the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission. The term “amino acid” comprises naturally occurring amino acids, and synthetic amino acids (e.g., non-naturally occurring, non-encoded amino acids). Naturally occurring amino acids are those encoded by the genetic code and include the proteinogenic amino acids. Naturally occurring amino acids also include those modified after translation in cells. Synthetic amino acids include non-canonical amino acids such as selenocysteine and pyrrolysine. Typically, synthetic amino acids are non-proteinogenic amino acids. The term “amino acid” as used herein comprises D- and L-amino acids. Preferably, and unless indicated to the contrary, the amino acids mentioned herein are L-amino acids.

[0171] Amino acids used in the peptide of the invention and the peptides of the invention may include at least one chemical modification, such as phosphorylation, acetylation, oxidation, ubiquitylation, amidation, esterification, deamidation, alkylation, preferably methylation, nitration, hydroxylation, formylation, sulfation, covalent binding of a lipidic moiety, sulphonation, glycation, acylation, myristoylation, PEGylation, palmitoylation, halogenation, chemical modifications of part or all peptide bonds, and / or post-translational amino acid modification, and the like. The peptides of the invention may have at least one modification selected from modification at the C-terminal part by amidation or esterification and modification at the N-terminal part by acylation, acetylation, myristoylation, PEGylation, alkylation, palmitoylation, and the like.

[0172] Additionally the peptides of the invention may be operably linked to other known antimicrobial peptides or other substances, such other peptides, proteins,oligosaccharides, polysaccharides, other organic compounds, or inorganic substances. For example, the antimicrobial peptides may be coupled to a substance which protect the antimicrobial peptides from being degraded within a mammal prior to the antimicrobial peptides has inhibited, prevented or destroyed the life of the microorganism.

[0173] Further, any amino acid residue of the peptides of the invention may comprise an amino acid having at least one protecting group. The protecting groups may comprise Boc, But, Fmoc, be or any other protecting group. A protected amino acid is an amino acid with one or more of its reactive groups modified with an inert molecule, to reduce and / or prevent chemical reactions of the reactive group.

[0174] The terms “peptide”, "protein", and "polypeptide" are used interchangeably herein and refer to any peptide-bond-linked chain(s) of amino acids, regardless of length or post-translational modification. Proteins usable in the present invention (including protein derivatives, protein variants, protein fragments, protein segments, protein epitopes and protein domains) can be further modified by chemical modification. This means such a chemically modified polypeptide comprises other chemical groups than the 20 naturally occurring amino acids. Examples of such other chemical groups include without limitation glycosylated amino acids and phosphorylated amino acids. Chemical modifications of a polypeptide may provide advantageous properties as compared to the parent polypeptide, e.g., one or more of enhanced stability, increased biological half-life, or increased water solubility. In the context of present invention, the primary structure of a protein or polypeptide is the sequence of amino acids in the polypeptide chain. The secondary structure in a protein is the general three-dimensional form of local segments of the protein, such as the alpha helical structure. In proteins, the secondary structure is defined by patterns of hydrogen bonds between backbone amide and carboxyl groups. The tertiary structure of a protein is the three-dimensional structure of the protein determined by the atomic coordinates. The quaternary structure is the arrangement of multiple folded or coiled protein or polypeptide molecules in a multi-subunit complex. The terms “amino acid chain” and “polypeptide chain” are used synonymously in the context of present invention.The peptide of the invention may include chemical modifications. Examples of chemical peptide modifications include those corresponding to post-translational modifications, for example phosphorylation, acetylation, oxidation, ubiquitylation, and deamidation (Engelhard et al., Curr Opin Immunol. 2006 Feb;18(1):92-7; Berlett and Stadtman, J Biol Chem, 1997. 272(33):20313-16; Kerscheret al., Annu. Rev. Cell Dev. Biol. 200622(1): 159-180). Chemical modifications may not correspond to those that may be present in vivo. For example, the N or C terminal ends of the peptide may be modified to improve the stability, bioavailability and or affinity of the peptides (see for example, Brinckerhoff et al Int J Cancer. 1999 Oct 29;83(3):326-34). Further examples of non-natural modifications include incorporation of photoreactive cross-linking amino acids, N-methylated amino acids, backbone reduction, retro-inversion by using D-amino acids, fluorescent amino acids, N-terminal methylation and C-terminal amidation, and pegylation. Examples of non-natural modifications include incorporation of non-encoded a-amino acids and [3-amino acids.

[0175] In certain embodiments, the invention discloses mutations of the peptides of SEQ ID NO: 1 or SEQ ID NO: 2-151. The term “mutation”, as used herein, refers preferably to a sequence modification, i.e., an addition, deletion, or substitution of at least one amino acid at a particular position in an amino acid sequence. In one embodiment, the addition or deletion of an amino acid refers to the addition or deletion of at least 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 50, or 100 amino acids at a particular position in an amino acid sequence.

[0176] The term “amino acid substitution” as used herein refers to the exchange of an amino acid at a particular position in an amino acid sequence by one or more other amino acids. The term “amino acid substitution” encompasses both conservative and nonconservative amino acid substitutions, wherein conserved amino acid substitutions are preferred. A conservative amino acid substitution is an amino acid exchange by a functionally similar amino acid. A conservative substitution can be an exchange within the same amino acid class. The peptide of the present disclosure, especially the peptide of SEQ ID NO: 1 or the peptides of SEQ ID NO: 2-151, preferably comprise one or more of the following conservative amino acid substitutions:Replacement of a neutral / uncharged amino acids including glycine, serine, threonine, cysteine, tyrosine, asparagine, and glutamine, with another neutral amino acid.

[0177] Replacement of a nonpolar (hydrophobic) amino acid, including alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophan, and methionine, with another nonpolar amino acid.

[0178] Replacement of an aliphatic amino acid, including alanine, valine, leucine, and isoleucine, with another aliphatic amino acid.

[0179] Replacement of an aliphatic amino acid, including alanine, serine, threonine, valine, leucine, and isoleucine, with another aliphatic amino acid.

[0180] Replacement of serine with threonine or vice versus.

[0181] Replacement of cysteine with methionine or vice versus

[0182] Replacement of an acidic residue, including aspartic acid and glutamic acid, with another acidic residue.

[0183] Replacement of a residue bearing an amide group, including asparagine and glutamine, with another residue bearing an amide group.

[0184] Replacement of a basic residue, including histidine, lysine, and arginine, preferably lysine and arginine, with another basic residue.

[0185] Replacement of an aromatic residue, including phenylalanine, tryptophane, histidine and tyrosine, with another aromatic residue.

[0186] The following groups are particularly preferred conservative substitutions for one another:

[0187] 1) Alanine (A), Serine (S), Threonine (T), Valine (V), Glycine (G);

[0188] 2) Aspartic acid (D), Glutamic acid (E);3) Asparagine (N), Glutamine (Q);

[0189] 4) Arginine (R), Lysine (K);

[0190] 5) Isoleucine (I), Leucine (L), Methionine (M), Valine (V), Norleucine;

[0191] 6) Phenylalanine (F), Tyrosine (Y), Tryptophan (W), Histidine (H); and

[0192] 7) Methionine (M), and Serine (S).

[0193] Amino acid analogues refers to agents that have the same basic chemical structure as a naturally occurring amino acid, i.e., an a carbon that is bound to a hydrogen, a carboxyl group, an amino group, and an R group, such as, homoserine, norleucine, methionine sulfoxide, methionine methyl sulfonium. Such analogues have modified R groups (such as, norleucine) or modified peptide backbones, but retain the same basic chemical structure as a naturally occurring amino acid.

[0194] The term “N terminus” as used herein refers preferably to amino acid residues that are located towards the amino terminus having a free amino group. The term “C terminus” as used herein refers to amino acid residues that are located towards the carboxy end of the amino acid sequence having a free carboxyl group. In one embodiment, the N and C terminus refers to the amino acid residues located in the terminal 10% or 5% of the respective amino acid sequence.

[0195] The terms “identical” or percent “identity”, in the context of two or more nucleic acids or polypeptide sequences, refers to two or more sequences or sub-sequences that are the same or have a specified percentage of amino acid residues or nucleotides that are the same, when compared and aligned for maximum correspondence over a comparison window or designated region, as measured using for example a BLAST sequence comparison algorithms with default parameters described below, or by manual alignment and visual inspection. The term also includes sequences that have deletions and / or additions, as well as those that have substitutions. As described below, the preferred algorithms can account for gaps and the like. Preferably, identity exists over a region that is at least about 25 amino acids or nucleotides in length, or more preferably over a region that is 50-100 amino acids or nucleotides in length.The term “a corresponding position” as used herein refers to an analogous position between two or more sequences. Amino acid exchanges or substitutions relate to the corresponding amino acid position. In one embodiment, the corresponding position is determined through a sequence alignment of at least a first and a second sequence. The corresponding position in the at least first and second sequences is aligned in the output from the sequence alignment and hence can be identified. In one embodiment, the sequence alignment is a pairwise or a multiple sequence alignment. In one embodiment, the sequence alignment is performed by an algorithm. In one embodiment, the algorithm is BLAST or EMBOSS Needle as discussed above.

[0196] The terms "effective amount", "therapeutically effective amount" or "pharmaceutically effective amount" as used herein, refer to an amount of the peptide of the invention that is sufficient to treat a disorder. In some embodiments, the result is a reduction in and / or alleviation of the signs, symptoms, or causes of a disorder, or the reduction on the number of microorganisms, preferably bacteria and fungi, or any other desired alteration of a biological system. “Ameliorate” refers to a lessening of the detrimental effect of the disease state or disorder in the recipient of the therapy.

[0197] The term "pharmaceutically acceptable carrier" as used herein, refers to a material, (e.g., a carrier, excipient, and / or diluent), which does not abrogate the biological activity or properties of the peptide of the invention described herein, and is preferably nontoxic (i.e. , the material is administered to an individual without causing undesirable biological effects or interacting in a deleterious manner with any of the components of the composition in which it is contained). Suitable carriers can be found in Remington: The Science and Practice of Pharmacy, 20th Edition, Lippincott Williams & Wilkins, Philadelphia, Pa., 2000.

[0198] As used herein, the terms “methods of treating or preventing” mean amelioration, prevention or relief from the symptoms and / or effects associated with a microbial infection. The term “preventing” as used herein refers to administering a medicament beforehand to forestall or obtund an acute episode. The person of ordinary skill in the medical art (to which the present method claims are directed) recognizes that the term“prevent” is not an absolute term. In the medical art it is understood to refer to the prophylactic administration of a drug to substantially diminish the likelihood or seriousness of a condition, and this is the sense intended in applicants' claims. As used herein, reference to “treatment” of a subject is intended to include prophylaxis.

[0199] As used herein, the terms "subject", "individual" and "patient" are used interchangeably. None of the terms are to be interpreted as requiring the supervision of a medical professional (e.g., a doctor, nurse, physician’s assistant, orderly, hospice worker). As used herein, the subject can be any animal, including mammals (e.g., a human or non-human animal) and non-mammals. In one embodiment of the methods and compositions provided herein, the mammal is a human.

[0200] The term "disease" and "disorder" are used interchangeably herein, referring to an abnormal condition, especially an abnormal medical condition such as an illness or injury, wherein cells, a tissue, an organ, or an individual is not able to efficiently fulfil its function anymore. Typically, but not necessarily, a disease is associated with specific symptoms or signs indicating the presence of such disease. The presence of such symptoms or signs may thus, be indicative for cells, a tissue, an organ, or an individual suffering from a disease.

[0201] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.

[0202] The methods and techniques of the present invention are generally performed according to conventional methods well known in the art and as described in various general and more specific references that are cited and discussed throughout the present specification unless otherwise indicated. See, e.g., Sambrook et al., Molecular Cloning: A Laboratory Manual, 2d ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1989) and Ausubel et al., Current Protocols in Molecular Biology,Greene Publishing Associates (1992), and Harlow and Lane Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1990).

[0203] While the invention is illustrated and described in detail in the drawings and description, such illustration and description are to be considered illustrative or exemplary and not restrictive. It will be understood that changes and modifications may be made by those of ordinary skill within the scope and spirit of the following claims. In particular, the present invention covers further embodiments with any combination of features from different embodiments described above and below.

[0204] The invention also covers all further features shown in the figures individually, although they may not have been described in the previous or following description. Also, single alternatives of the embodiments described in the figures and the description and single alternatives of features thereof can be disclaimed from the subject matter of the other aspect of the invention.

[0205] Throughout this specification and the claims, unless the context requires otherwise, the word "comprise" or “include”, and variations such as "comprises / includes" and "comprising / including" are to be understood to imply the inclusion of an element, stated integer, step or a group thereof but not the exclusion of any other element, stated integer, step or a group thereof.

[0206] As used in this specification and the appended claims, the singular forms "a", "an", and "the" include plural referents, unless the content clearly dictates otherwise.

[0207] A single unit may fulfill the functions of several features recited in the claims.

[0208] The term “essentially”, “about” or “approximately” and the like when used in connection with an attribute or numerical value particularly defines exactly the attribute or value and additionally encompass, unless the context indicates otherwise, numerical values within a range having a lower limit that is 0-10% smaller than the indicated numerical value and having an upper limit that is 0-10% larger than the indicated numerical value. The term “about” or “approximately” means preferably ±10%, more preferably ±5%, again more preferably ±3% or most preferably ±0% (referring to the given numericvalue, respectively). In each of the invention embodiments, „about” can be deleted. All ranges of values disclosed herein, should refer and include to any and all values falling within said range including the values defining the range.

[0209] EXAMPLES EXAMPLE 1 - Solid Solid-Phase Peptide Synthesis of SP-CAKT 1-150

[0210] Protected Amino Acids Used: Fmoc-Leu-OH, Fmoc-Arg(Pbf)-OH, Fmoc-Trp(Boc)-OH, Fmoc-Lys(Boc)-OH, Fmoc-Gly-OH, Fmoc-Val-OH, Fmoc-Phe-OH, Fmoc-Asn(Trt)-OH, Fmoc-lle-OH, Fmoc-D-Phe-OH, Fmoc-D-Val-OH, Fmoc-D-Lys(Boc)-OH, Fmoc-Asn(Trt)-OH, Fmoc-D-lle-OH, Fmoc-D-Leu-OH, Fmoc-D-Ser(tBu)-OH, Fmoc-D-Gln(Trt)-OH, Fmoc-D-Arg(Pbf)-OH, Fmoc-D-Pro-OH, Fmoc-D-Met-OH, Fmoc-D-Thr(tBu)-OH.

[0211] 1) The solid-phase synthesis method was employed, using 2-CTC resin as the solid support.

[0212] 2) After swelling the 2-CTC resin in DCM, 1.5 equivalents (eq) of Fmoc-AA-OH and 7.5 eq of DIEA were added, with DCM as the solvent. The reaction was carried out at room temperature for 30-60 minutes. The reaction solution was removed by suction, and the resin was washed. To block remaining active sites on the resin, a reaction solution of MeOH: DIEA: DCM = 2:1 :17 was added, and the reaction was conducted at room temperature for 10-30 minutes, followed by washing.

[0213] 3) The swollen resin was treated with 10 times the volume (V / m) of 20% piperidine / DMF solution and reacted at room temperature for 5-10 minutes. The solution was removed, and the resin was washed three times each with 10* V / m of DMF and DCM. After drying, the resin with exposed amino groups was tested with ninhydrin and showed a positive result. Subsequently, 3 eq of the protected amino acid (Fmoc-AA-OH), 3 eq of coupling agents HoBt / DIC or TBTU / DIEA, and 10x V / m of DMF as the solvent were added. Coupling was carried out at room temperature for 30-120minutes. After the reaction, the resin was washed with 10* V / m of DMF, and a ninhydrin test showed a negative result.

[0214] 4) Sequential coupling at step 3) was repeated to sequentially couple the subsequent Fmoc-AA-OH amino acids according to the peptide sequence. The Fmoc protecting group of the last amino acid at the resin’s terminal was removed. The resin was then washed, dried, and the peptide resin was collected and dried to constant weight.

[0215] 5) The peptide chain was cleaved from the resin and deprotected using a cleavage reagent mixture (TFA:TIS:DTT:Mesph:Meoph:H2O = 80:1:5:5:5:4). The resin was filtered, and the residue was washed with TFA (1.2 eq of V / m). The filtrate was combined, and TFA was partially removed by rotary evaporation.

[0216] 6) Methyl tert-butyl ether was added to the filtrate from step 5) at a volume ratio of filtrate: methyl tert-butyl ether = 1:5. The mixture was centrifuged, and the resulting precipitate was washed with methyl tert-butyl ether and centrifuged three times. The target polypeptide was obtained after vacuum drying.

[0217] 7) The crude polypeptide was dissolved, filtered through a 0.22 pm microporous filter, and purified by preparative liquid chromatography using gradient elution. The mobile phase A was 0.1% TFA in water, and mobile phase B was acetonitrile. The purified polypeptide was collected and lyophilized to yield the final pure product.

[0218] EXAMPLE 2 - Minimum Inhibitory Concentration (MIC) determination of antimicrobial peptides

[0219] The standard strains used in the following examples of the present invention, Pseudomonas aeruginosa CMCC(B)26069, Escherichia coli 8099, Staphylococcus aureus CMCC(B)26003, Staphylococcus epidermidis CMCC(B)26069, Bacillus subtilis CMCC(B)63501, Candida albicans CMCC(F)98001 , Aspergillus niger CMCC(F)98003.

[0220] The results of MIC of antibacterial peptide SP-CAKT340 (KKRLKKIFKKPMVIGVTIPF), SP-CAKT341 (VFQFLGKIIKKVGNFVKGFSKVF), SP-CAKT343(VFQFLGKIIKKVGNFVKGFSKVF consisting of a racemic mixture of the indicated D-and L-amino acids at each position with a randomized stereochemistry), and SP-CAKT345 (KRWWKWWRRLL) against Gram-positive and Gram-negative bacteria, and fungi were determined using the broth dilution method. Stock solutions of the samples were prepared at a concentration of 8 mg mL"1in H2O and diluted to initial concentrations of 32, 64, 128, or 256 pg mL-1in 300 pL of MH medium. These solutions were added to the first well of a 96-well microtiter plate (TPP, untreated) and serially diluted by a factor of .

[0221] A bacterial colony was cultured overnight in MH medium at 37°C. The bacterial concentration was determined by measuring the absorbance at 600 nm (OD600) and diluted to an OD600of 0.022 in MH medium. Sample solutions (150 pL) were then mixed with 4 pL of the diluted bacterial suspension, resulting in a final inoculation concentration of approximately 5 x 105CFU. The plates were incubated at 37°C for approximately 18 hours, allowing satisfactory bacterial growth.

[0222] For each test, two columns of the plate were designated as controls: one for sterility control (broth only) and the other for growth control (broth with bacterial inoculum, no antibiotics). The minimum inhibitory concentration (MIC) was defined as the lowest concentration of the peptide dendrimer that completely inhibited visible bacterial growth, as assessed after treatment with MTT.

[0223] Table 1. Antimicrobial activity of peptides SP-CAKT340, 341, 343, and 345.

[0224]

[0225] 8099

[0226] (B)26069 (B)10104 (B)26003 (B)63501 (F)98001 (F)98003

[0227] polymyxin B 8 <0.25 8 <0.25 2 >8 >8

[0228] ST-CAK345 4 4 <2 4 8 16 64ST-CAK340 16 8 8 4 16 32 >64

[0229] ST-CAK341 >64 >64 8 >64 8 >64 >64

[0230] ST-CAK343 >64 >64 <2 >64 <2 >64 >64

[0231] As shown in Table 1, the antimicrobial peptide ST-CAK345 demonstrates strong activity against Gram-negative bacteria (P. aeruginosa and E. coli), Gram-positive bacteria (S. aureus, S. epidermidis, B. subtilis), and fungi (C. albicans). ST-CAK340 displays similar antimicrobial activity to ST-CAK345 but with reduced efficacy against S. aureus. ST-CAK341 shows moderate activity, specifically against Gram-positive bacteria S. aureus and B. subtilis. In contrast, ST-CAK343 exhibits strong activity exclusively against the two tested Gram-positive bacteria S. aureus and B. subtilis. For preparation of peptide ST-CAK343, the inventors randomized the stereochemistry of the amino acids at each position of the peptide sequence. For each amino acid coupling, L and D-stereoisomers of the respective amino acid were mixed in a 1 : 1 ratio.

[0232] Table 2. Antimicrobial activity of SP-CAKT345 against challenging pathogens

[0233] Neisseria

[0234] Gardnerella

[0235] gonorrhoeae

[0236] vaginalis

[0237] (Zopf) Trevisan

[0238] BNCC354890 BNCC270346

[0239] polymyxin B 16 64

[0240] Gastrodia elata >256 >256

[0241] ST-CAK345 16 16

[0242] As shown in Table 2, ST-CAK345 was selected for further testing due to its broad-spectrum antimicrobial activity. It was evaluated against challenging pathogens,including Gardnerella vaginalis and Neisseria gonorrhoeae (Zopf). ST-CAK345 demonstrated comparable activity to polymyxin B against G. vaginalis but exhibited superior activity against N. gonorrhoeae.

[0243] Table 3. Exemplary peptides of the invention: K (lysine), R (arginine), W (tryptophan), L (leucine)

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[0245]

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[0247]

[0248]

[0249]

[0250] Example 3 - Minimum Inhibitory Concentration (MIC) Assay - Microdilution Method

[0251] A microdilution method was applied in order to determine the minimum inhibitory concentration of antimicrobial agents which allows evaluating their antibacterial activity and bacteriostatic efficacy.

[0252] Methods

[0253] Mueller-Hinton broth (MHB) was prepared and sterilized. Test strains were adjusted to a 0.5 McFarland standard turbidity suspension for use. Drug solutions were added at varying concentrations to a 96-well plate with CAMHB to complete serial dilutions. Subsequently, the diluted bacterial suspension was added to all wells, achieving a final volume of 200 pL per well. The final inoculum contains 5 x 104CFU per well, with drug concentrations ranging from 64 to 0.25 pg / mL. Simultaneously, positive control wells (containing bacteria but no drug) and negative control wells (bacteria-free, drug-free) were established. The plate was incubated at 35°C ± 2°C in a constant-temperature incubator. Conventional aerobic bacteria required 16-20 hours incubation. Incubationtimes was extended as required for fastidious bacteria or bacteria targeted by specific drugs. The minimum inhibitory concentration (MIC) was defined as the lowest concentration of the peptide that completely inhibited visible bacterial growth, as assessed after treatment with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT).

[0254] Results

[0255] Table 4. Minimum inhibitory concentrations (MICs) of CAKT1-6

[0256]

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[0262]

[0263] ‘Minimum inhibitory concentration (MIC, pg / mL) was determined on E. coli 8099, P. aeruginosa CMCC(B)10104, S. aureus CMCC(B)26003, S. epidermidis CMCC(B)26069, B. subtilis CMCC(B)63501 , in Muller-Hinton medium, after incubation for 16-20 h at 37 °C. Minimum inhibitory concentration (MIC, pg / mL) was determined on C. Albicans CMCC(F)98001 and A. niger CMCC(F)98003 in Sabouraud Dextrose Broth media. Minimum inhibitory concentration (MIC, pg / mL) was determined on G.Vaginalis BNCC354890 and N. gonorrhoeae BNCC270346 in brain heart infusion broth.

[0264] Minimum inhibitory concentrations (MICs) of the CAKT analogues (CAKT1-6) were determined against nine representative microbial strains: Escherichia coli 8099, Pseudomonas aeruginosa CMCC(B)10104, Staphylococcus aureus CMCC(B)26003, Staphylococcus epidermidis CMCC(B)26069, Bacillus subtilis CMCC(B)63501 , Candida albicans CMCC(F)98001, Aspergillus niger CMCC(F)98003, Gardnerella vaginalis BNCC354890, and Neisseria gonorrhoeae BNCC270346 (Table 4). The reference compound CAKT7 and polymyxin B were included as comparators.

[0265] Among the analogues, CAK345 (a synonymous term of CAKT1) retained the antimicrobial activity of the reference compound against E. coli, S. aureus, and B. subtilis. Notably, it exhibited an 8-fold improvement in activity against P. aeruginosa and S. epidermidis, with MIC values of 8 pg / mL for both strains.

[0266] In addition, CAK345 preserved the activity of the reference compound against G. vaginalis (MIC 2-4 pg / mL) and demonstrated strong activity against N. gonorrhoeae (MIC 8 pg / mL). Importantly, both the reference compound and polymyxin B were inactive against N. gonorrhoeae.

[0267] CAK345 was the most active compound tested against G. vaginalis and N. gonorrhoeae, whereas polymyxin B and the reference compound displayed reduced activity against G. vaginalis and were completely inactive against N. gonorrhoeae. It is noteworthy that N. gonorrhoeae has been classified by the World Health Organization as a high-priority pathogen due to its increasing antibiotic resistance and the growing difficulty of treatment (Multi-drug resistant gonorrhoea, https: / / www.who.int / news-room / fact-sheets / detail / multi-drug-resistant-gonorrhoea (accessed 2026-02-13).

Claims

CLAIMS1. A peptide comprising an amino acid sequence of SEQ ID NO: 1 :X1 -X2-X3-X4-X5-X6-X7-X8-X9-X10-X11 ,wherein X1 to X7 are, independently of each other, a D- or L-amino acid selected from the group consisting of K (lysine), R (arginine), W (tryptophan), L (leucine), an amino acid variant of K, R, W, or L, and a conservative substitute amino acid of K, R, W or L; andwherein X8 to X11 are, independently of each other, a deletion or a D- or L- amino acid selected from the group consisting of K, R, W, L, an amino acid variant of K, R, W or L, and a conservative substitute amino acid of K, R, W or L.

2. The peptide of claim 1 , wherein the peptide comprising the amino acid sequence of SEQ ID NO: 1 , wherein X1 to X7 are, independently of each other, a D- or L- amino acid selected from the group consisting of K, R, W and L; and wherein X8 to X11 are, independently of each other, a D- or L-amino acid selected from the group consisting of K, R, W and L.

3. The peptide of any one of the preceding claims, wherein X1 is a D- or L-, preferably L-amino acid selected from the group consisting of K, an amino acid variant of K, and a conservative substitute amino acid of K.

4. The peptide of any one of the preceding claims, wherein X2 is a D- or L-, preferably L-amino acid selected from the group consisting of R, an amino acid variant of R, and a conservative substitute amino acid of R.

5. The peptide of any one of the preceding claims, wherein X3 is a D- or L-, preferably L-amino acid selected from the group consisting of W, an amino acid variant of W, and a conservative substitute amino acid of W.

6. The peptide of any one of the preceding claims, wherein X4 is a D- or L-,preferably L-amino acid selected from the group consisting of: W, K and R, an amino acid variant of W, K and R, and a conservative substitute amino acid of W, K and R.

7. The peptide of any one of the preceding claims, wherein X5 and X6 are independently of each other a D- or L-, preferably L-amino acid selected from the group consisting of: W, K, L, and R, an amino acid variant of W, K, L and R, and a conservative substitute amino acid of W, K, L, and R.

8. The peptide of any one of the preceding claims, wherein X7 and X8 are independently of each other a D- or L-, preferably L-amino acid selected from the group consisting of: W, L, and R, an amino acid variant of W, L and R, and a conservative substitute amino acid of W, L, and R.

9. The peptide of any one of the preceding claims, wherein the C terminal amino acid, preferably the last C terminal amino acid is a D- or L-amino acid, preferably an L-amino acid, selected from the group consisting of: W, L, and R, an amino acid variant of W, L and R, and a conservative substitute amino acid of W, L, and R;preferably, the last C terminal amino acid is a D- or L-amino acid, preferably an L-amino acid, selected from the group consisting of: L, R, an amino acid variant of L, an amino acid variant of R, a conservative substitute amino acid of L, and a conservative substitute amino acid of R;more preferably, the last C terminal amino acid is a D- or L-amino acid, preferably an L-amino acid, selected from the group consisting of: L, an amino acid variant of L, and a conservative substitute amino acid of L;again more preferably, the last C terminal amino acid is a D- or L-amino acid of leucine, preferably an L-amino acid of leucine.

10. The peptide of any one of the preceding claims comprising at least one alpha helical domain.

11. The peptide of any one of the preceding claims, wherein the peptide is a cationic peptide, preferably the cationic peptide has a net positive charge, more preferably the net positive charge ranges from +2 to +12.

12. The peptide of any one of the preceding claims, wherein the peptide has a length of 5 to 1000 amino acids, preferably 5 to 500 amino acids, more preferably 5 to 100 amino acids, or 5 to 50 amino acids, or 5 to 30 amino acids, or 5 to 20 amino acids, or 5 to 15 amino acids, or 7 to 11 amino acids.

13. The peptide of any one of the preceding claims, wherein the peptide comprises one or more sequences selected from the group consisting of: SEQ ID NO: 2 to 151 , and a derivative of SEQ ID NO: 2 to 151 ;wherein the derivative of SEQ ID NO: 2 to 151 comprises a chemical modification of one or more amino acids or a chemical modification of the peptide of SEQ ID NO: 2 to 151, preferably the chemical modification is selected from the group consisting of phosphorylation, acetylation, oxidation, ubiquitylation, amidation, deamidation, esterification, alkylation, preferably methylation, nitration, hydroxylation, formylation, sulfation, covalent binding of a lipidic moiety, sulphonation, glycation, acylation, myristoylation, PEGylation, palmitoylation, halogenation, chemical modifications of part or all peptide bonds, and / or post-translational amino acid modification; and / orwherein in the derivative of SEQ ID NO: 2 to 151 , one, two, three or four amino acids are each exchanged by an amino acid variant, preferably by an analogue amino acid or a conservative substitute amino acid; and / orwherein in the derivative of SEQ ID NO: 2 to 151 , one, two, three or four amino acids are deleted, wherein the derivative of SEQ ID NO: 2 to 151 has a length of at least 7 amino acids, and / orwherein in the derivative of SEQ ID NO: 2 to 151 , one, two, three or four single, double, or triple amino acids are added to SEQ ID NO: 2 to 151, wherein the added amino acids are D- or L-amino acids selected from the group consistingof K, R, Wand L.

14. The peptide of any one of the preceding claims, wherein the chemical modification is an amidation and / or acetylation, more preferably a C terminal amidation and / or N terminal acetylation.

15. The peptide of any one of the preceding claims, whereinthe amino acid variant of K (lysine) is selected from the group consisting of: diaminobutyric acid, 2,3-diaminopropanoic acid, (2s)-2,8-diaminooctanoic acid, ornithine, thialysine, histidine, arginine, 5-methyl-arginine, lysine, c-gamma- hydroxy arginine, citrulline, 2-amino-4-guanidinobutryric acid, 2-amino-3- guanidinopropionic acid, canavanine, homoarginine, thio-citrulline, histidine, 2- fluoro-l-histidine, 2-fluoro-l-histidine, 2-fluoro-l-histidine, (2-furyl)-alanine, 3-(1-pyrazolyl)-alanine, 3-(2-tetrazolyl)-alanine, 3-(3-thienyl)-alanine, 3-(2- thienyl)-alanine, 3-(1 ,2,4-triazol-1 -yl)-alanine, and (4-thiazolyl)-alanine; preferably the amino acid variant of K (lysine) is selected from the group consisting of: diaminobutyric acid, 2,3-Diaminopropanoic acid, (2s)-2,8- diaminooctanoic acid, ornithine, thialysine, histidine, arginine, 2-fluoro-l- histidine, 2-fluoro-l-histidine, 2-fluoro-l-histidine, (2-furyl)-alanine, 3-(1- Pyrazolyl)-alanine, 3-(2-Tetrazolyl)-alanine, 3-(3-thienyl)-alanine, 3-(2-thienyl)- alanine, 3-(1 ,2,4-Triazol-1-yl)-alanine, and (4-thiazolyl)-alanine;more preferably the amino acid variant of K (lysine) is selected from the group consisting of: diaminobutyric acid, 2,3-Diaminopropanoic acid, (2s)-2,8- diaminooctanoic acid, ornithine, thialysine, histidine and arginine;the amino acid variant of R (arginine) is selected from the group consisting of: diaminobutyric acid, 2,3-diaminopropanoic acid, (2s)-2,8-diaminooctanoic acid, ornithine, thialysine, histidine, lysine, 5-methyl-arginine, c-gamma-hydroxy arginine, citrulline, 2-amino-4-guanidinobutryric acid, 2-amino-3- guanidinopropionic acid, canavanine, homoarginine, thio-citrulline, histidine, 2- fluoro-l-histidine, 2-fluoro-l-histidine, 2-fluoro-l-histidine, (2-furyl)-alanine,3-( 1 -pyrazolyl)-alanine, 3-(2-tetrazolyl)-alanine, 3-(3-thienyl)-alanine, 3-(2-thienyl)-alanine, 3-(1 ,2,4-triazol-1 -yl)-alanine, and (4-thiazolyl)-alanine;preferably the amino acid variant of R (arginine) is selected from the group consisting of: 5-methyl-arginine, lysine, histidine, c-gamma-hydroxy arginine, citrulline, 2-amino-4-guanidinobutryric acid, 2-amino-3-guanidinopropionic acid, canavanine, homoarginine, thio-citrulline, 2-fluoro-l-histidine, 2-fluoro-l-histidine, 2-fluoro-l-histidine, (2-furyl)-alanine, 3-(1-Pyrazolyl)-alanine, 3-(2-Tetrazolyl)-alanine, 3-(3-thienyl)-alanine, 3-(2-thienyl)-alanine, 3-(1 ,2,4-Triazol-1-yl)-alanine, and (4-thiazolyl)-alanine;more preferably the amino acid variant of R (arginine) is selected from the group consisting of 5-methyl-arginine, c-gamma-hydroxy arginine, citrulline, 2-amino-4-guanidinobutryric acid, 2-amino-3-guanidinopropionic acid, canavanine, homoarginine, thio-citrulline, lysine, and histidine;the amino acid variant of W (tryptophan) is selected from the group consisting of: 7-hydroxy-l-tryptophan, 3-(4H-thieno[3,2-b]pyrrol-6-yl)-L-alanine, 4-fluoro-tryptophan, 4-hydroxy-tryptophan , 4-amino-l-tryptophan, 6-chloro-l-tryptophan, 3-(3-benzothienyl)-alanine, 6-bromo-tryptophan, 7-chloro-tryptophan, 6-fluoro-l-tryptophan, 5-Fluoro-tryptophan, 5-Hydroxy-tryptophan, beta-hydroxy-tryptophane, 5-Methoxy-tryptophan, 5-Methyl-tryptophan, 6-Methyl-tryptophan, 2-hydroxy-tryptophan, 6-hydroxy-tryptophan, 6-amino-7-hydroxy-l-tryptophan, leucine, isoleucine, valine, proline, phenylalanine, methionine, histidine, and tyrosinepreferably the amino acid variant of W (tryptophan) is selected from the group consisting of: 7-hydroxy-l-tryptophan, 3-(4H-thieno[3,2-b]pyrrol-6-yl)-L-alanine, 4-fluoro-tryptophan, 4-hydroxy-tryptophan, 4-amino-l-tryptophan, 6-chloro-l-tryptophan, 3-(3-benzothienyl)-alanine, 6-bromo-tryptophan, 7-chloro-tryptophan, 6-fluoro-l-tryptophan, 5-Fluoro-tryptophan, 5-Hydroxy-tryptophan, beta-hydroxy-tryptophane, 5-Methoxy-tryptophan, 5-Methyl-tryptophan, 6-Methyl-tryptophan, 2-hydroxy-tryptophan, 6-hydroxy-tryptophan, 6-amino-7-hydroxy-l-tryptophan, phenylalanine, histidine, and tyrosine;the amino acid variant of L (leucine) is selected from the group consisting of: alpha-amino-2-indanacetic acid, isoleucine, tert-leucine, allo-isoleucine, norleucine, 5-bromo-L-isoleucine, homoleucine, 5,5,5-trifluoro-leucine, 3- methyl-L-alloisoleucine, 4-hydroxy-L-isoleucine, 4,5-dihydroxy-isoleucine, betahydroxyleucine, 6-hydroxy-L-norleucine, (4r)-5-oxo-l-leucine, 5-oxo-l- norleucine, (4s)-5-fluoro-l-leucine, 2-aminobutyric acid, 2-aminoheptanoic acid, diethylalanine, norvaline, (2s)-2-amino-4,4-difluorobutanoic acid, hydroxynorvaline, valine, 3-fluoro-valine, 3-hydroxy-l-valine, 4-oxo-l-valine, 3- hydroxy-l-valine, alanine, proline, phenylalanine, histidine, methionine, tryptophan, serine, threonine, and tyrosine;preferably the amino acid variant of L (leucine) is selected from the group consisting of: alpha-amino-2-indanacetic acid, isoleucine, tert-leucine, allo- isoleucine, norleucine, 5-bromo-L-isoleucine, homoleucine, 5,5,5-trifluoro- leucine, 3-methyl-L-alloisoleucine, 4-hydroxy-L-isoleucine, 4,5-dihydroxy- isoleucine, beta-hydroxyleucine, 6-hydroxy-L-norleucine, (4r)-5-oxo-l-leucine, 5-oxo-l-norleucine, (4s)-5-fluoro-l-leucine, 2-aminobutyric acid, 2- aminoheptanoic acid, diethylalanine, norvaline, (2s)-2-amino-4,4- difluorobutanoic acid, hydroxynorvaline, valine, methionine, 3-fluoro-valine, 3- hydroxy-l-valine, 4-oxo-l-valine, and 3-hydroxy-l-valine.

16. The peptide of any one of the preceding claims, wherein the amino acid variant is the conservative substitute amino acid, whereinthe conservative substitute amino acid of K (lysine) is D- or L-histidine or D- or L-arginine, preferably arginine;the conservative substitute amino acid of R (arginine) is D- or L-lysine or D- or L-histidine, preferably D- or L-lysine;the conservative substitute am ino acid of W (tryptophan) is a D- or L- amino acid selected from the group consisting of: alanine, leucine, isoleucine, valine, proline, phenylalanine, methionine, histidine, and tyrosine; preferably the conservative substitute amino acid W (tryptophan) is a D- or L- amino acidselected from the group consisting of phenylalanine, histidine, and tyrosine;the conservative substitute amino acid of L (leucine) is a D- or L- amino acid selected from the group consisting of: alanine, isoleucine, norleucine, valine, proline, phenylalanine, tryptophan, serine, threonine, and methionine; preferably the conservative substitute amino acid of L (leucine) is a D- or L- amino acid selected from the group consisting of isoleucine, valine, methionine, and norleucine.

17. The peptide of any one of the preceding claims, whereinthe conservative substitute amino acid of K (lysine) is D- or L-arginine;the conservative substitute amino acid of R (arginine) is D- or L-lysine;the conservative substitute am ino acid of W (tryptophan) is a D- or L- amino acid selected from the group consisting of phenylalanine, histidine, and tyrosine;the conservative substitute amino acid of L (leucine) is a D- or L- amino acid selected from the group consisting of isoleucine, valine, methionine, and norleucine.

18. The peptide of any one of the preceding claims, wherein the peptide comprises an amino acid sequence selected of the group consisting of SEQ ID NO: 1-151, or a derivative thereof, wherein up to 200 amino acids, preferably up to 100 amino acids, more preferably up to 50 amino acids are added to the N terminus and / or C terminus of a sequence of SEQ ID NO: 1-151.

19. The peptide of any one of the preceding claims, wherein the peptide comprises more than one sequence of SEQ ID NO: 1-151, preferably an n-mer, wherein n is an integer between 2 and 20, preferably 2 and 10.

20. The peptide of any one of the preceding claims, wherein the peptide is a diastereomer comprising at least one D-amino acid.

21. The peptide of any one of the preceding claims, wherein the amino acids of thepeptide comprise or consist of a racemic mixture of D- and L-amino acids.

22. The peptide of any one of the preceding claims, wherein the amino acids of X1 to X11 of the sequence of SEQ ID NO: 1 comprises or consist of a racemic mixture of D- and L-amino acids.

23. A polynucleotide encoding the peptide of any one of claims 1 -22.

24. A composition comprising the peptide of any one of claims 1-22 or the polynucleotide of claims 23.

25. The composition of any one of claims 24 further comprising a pharmaceutically acceptable carrier.

26. The composition of claim 24 or 25 further comprising a compound selected from the group consisting of an antibiotic, lysozyme, adjuvant, vaccines, immunostimulant and combinations thereof.

27. The peptide of any one of claims 1 -22, or the composition of any one of claims 24-26, for use in treating and / or preventing an infection and / or inflammation in a subject or on a subject.

28. A method for treating or preventing a microbial infection and / or inflammation in or on a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the peptide of any one of claims 1- 22, or the composition of any one of claims 23-26.

29. The peptide or the composition for use according to claim 27, or the method of claim 28, further comprising administering to the subject at least one antibiotic agent, wherein the antibiotic is administered simultaneously or sequentially with the disclosed peptide.

30. The peptide or the composition for use according to claim 27 or 29, or the method of claim 28 or 29, wherein the infection and / or inflammation is caused by one or more microorganisms.

31. A non-in vivo method of inhibiting growth of one or more microorganisms, the method comprises contacting the microorganisms with an effective amount of the peptide of any one of the preceding claims 1-22, or the composition of any one of claims 24-26.

32. A non-in vivo method of inhibiting biofilm formation and / or dissolution of a biofilm, the method comprises contacting the microorganisms with an effective amount of the peptide of any one of the preceding claims 1-22, or the composition of any one of claims 24-26.

33. The method of claims 31 or 32, wherein the step of contacting the microorganisms with an effective amount of the peptide or the composition comprises applying the peptide or the composition to an article or a surface of an article.

34. The peptide or the composition for use according to claim 30, or the method of claim 30, or the methods of any one of claims 31-33, wherein the one or more microorganisms are selected from the group consisting of bacteria, fungi, viruses, archaea, protozoa, and combinations of two or more thereof.

35. The peptide or the composition for use according to claim 30, or the method of claim 30, or the methods of any one of claims 31-33, wherein the one or more microorganisms are selected from the group consisting of bacteria, fungi, and a combination thereof.

36. The peptide or the composition for use according to claim 30, or the method of claim 30, or the methods of any one of claims 31-33, wherein the bacteria are selected from the group consisting of Gram-positive bacteria, Gram-negative bacteria, and a combination thereof.

37. The peptide or the composition for use according to claim 30, or the method of claim 30, or the methods of any one of claims 31-33, wherein the bacteria are from a genus selected from the group consisting of Pseudomonas, Escherichia, Staphylococcus, Bacillus, Gardnerella, Neisseria, and a combination thereof;preferably the bacteria are from a species selected from the group consisting of Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis, Bacillus subtilis, Gardnerella vaginalis and Neisseria gonorrhoeae and a combination thereof;more preferably the bacteria are from the species Staphylococcus aureus and / or Bacillus subtilis.

38. The peptide or the composition for use according to claim 30, or the method of claim 30, or the methods of any one of claims 31-33, wherein the fungus / fungi is / are from the genus Candida and / or Aspergillus, preferably the fungus / fungi is / are is of the species Candida albicans and / or Aspergillus niger.

39. A method for identifying a compound, preferably the compound is a peptide, having antimicrobial activity, the method comprises the steps of:mixing a bacterial suspension with the compound to be tested;incubating the mixture comprising the bacterial suspension and the compound to be tested, wherein incubation time and incubation temperature are adjusted to the bacteria included in the bacterial suspension;quantifying bacterial growth in the mixture by defining a minimum inhibitory concentration (MIC), wherein MIC is defined as the lowest concentration of the compound or peptide that inhibits bacterial growth to a non-detectable level, wherein preferably bacterial growth is quantified by measuring density or metabolic activity of the bacteria in the mixture.

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