MXPA98007066A - Broad spectrum antimicrobial peptides containing a trippethane triplete and methods of using my - Google Patents

Abstract

The present invention relates to a peptide comprising a tryptophan triplet, having between 10 and 34 amino acid residues in length and exhibiting antimicrobial activity, it is provided together with pharmaceutical and non-pharmaceutical compositions containing said peptides and methods that utilize

Description

LARGE SPECTRUM ANTIMICRQBIAN PEPTIDES CONTAINING TRIPTOPHAN TRIPLETE UM AND METHODS OF USE THEREOF PREFERENCE TO RELATED REQUESTS 5 This request is based on provisional application series 2 SO / 012 »392, filed on February 28, 1996.

BACKGROUND OF THE INVENTION 10 This invention relates generally to antimicrobial compounds and agents. Very specific »refers to novel broad-spectrum antimicrobial peptides containing a triplet of tryptophan and its derivatives that present antimicrobial activity with low immunogenicity. ¿. Methods of microbicidal inhibition or Microbial growth microbial using pharmaceutical and non-pharmaceutical compositions wherein said antimicrobial peptides are active component. 20 Infectious diseases have plagued human populations throughout history and continue to cause millions of deaths each year. Although the widely distributed use of vaccines and drug therapy has drastically reduced the mortality caused by disease Infectious diseases in developed countries, infectious diseases continue to be the leading cause of death in # third world countries. It is estimated that approximately 600 million people are infected with tropical diseases, reaching a figure of approximately 20 million deaths each year. Even in developed countries »infectious disease organisms spread to countless millions each year. Individuals with an immunodeficiency disease are particularly at risk for infectious diseases. The basic clinical manifestations # 10 are frequent, prolonged »severe» infections that are often caused by normally low pathogenicity organisms. The one disease signs of disease are recurrent bacterial or fungal infections. Clinical manifestations vary from light infections of the skin to systemic infections that threaten life. Common infectious organisms include Staphylococcus aureas » # Streptococcus pneu oniae »Escherichia coli, and several species of Pseuco onas, Candida, and Aspergí '11us. Due to the severity of many infectious diseases, scientists continue to investigating effective means of killing or controlling such organisms that "at the same time" show low in ungenicity. It has been speculated that the cate! na »a protein isolated from the leukocytes of the pig's blood, is the N-term fragment of a homologous pig of a novel family of antibacterial peptides. The cDNA sequence of a putative homologue of this porcine antibacterial protein rich in pro! ina / argi a was described by Joze Pungercar et al., "Molecular cloning of a putative hoology of pro / ine / arginine-rich antibacterial peptides from porc ne bone marraw," FEBS »336: 284-82 (1993). However, the potentially antimicrobial portion of this proposed 228 amino acid peptide has a length of 101 amino acids which is too long and too large for effective production and use as an antimicrobial agent. # ÍO BRIEF DESCRIPTION OF THE INVENTION Among the various objects of the invention, therefore, the provision of two antimicrobial peptides can be noted. of amplitude or spectrum having low and ungenicity that can be easily synthesized »the provision of the pharmaceutical and non-pharmaceutical antimicrobial compositions containing these peptides. as well as methods of inhibiting microbial growth and treating infectious diseases that use those compositions. Therefore »in a few words» the present invention is directed to a broad spectrum antimicrobial peptide containing a tryptophan triplet of at least 10 amino acid residues in length. Preferably » antimicrobial peptides of the invention have substantially the same sequence as defined by SEQ ID NO: 1. The present invention is further directed to compositions having an antimicrobial peptide of the invention as its active component. In yet another aspect of the invention, a method of microbicidal or microbysteatic inhibition of microbial growth in a microbial growth support environment is provided. Other objects and features in part will be evident and in part will be indicated later.

OR BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 shows the bactericidal activities of four synthesized peptides containing tryptophan triplets. E. coli cells that were grown to grow at a logarithmic stage were mixed with several peptide concentrations »l (x> 2 ()» 3 (*) and 4 (+) »in a pH buffer solution After incubation at 37 ° C for 10 minutes, 'samples were taken in aliquots', they were diluted and plated on L-agar which was incubated for 18 hours, then the number of colonies was counted.

DETAILED DESCRIPTION OF THE PREFERRED MODALITY In accordance with the present invention, the applicants have discovered a family of broad-spectrum but low-immunogenicity antimicrobial peptides which are distinguished because they contain a tryptophan triplet as a part of their active site. These peptides (herein referred to as "Tri Ripticin") are similar to the previously unappreciated active portion of an antibacterial peptide fragment rich in pro! na / argi i a de IOI amino acids isolated from pig blood leukocytes, known as cate! na Tryptidine is a relatively small peptide that can be easily synthesized »although it has potent antimicrobial activity. As used herein, the term "tr? Trythin" refers to a tryptophan-rich peptide containing a tryptophan triplet illustrated by those peptides having substantially the same sequence as SEQ ID NO: 1 and having antimicrobial activity. As used herein, the term "tryptophan rich" refers to the overrepresentation of tryptophan amino acids in an antimicrobial compound. The frequency of individual amino acids within a protein varies between each amino acid, with tryptophan being typically less frequent. The average frequency of tryptophan within a randomly selected protein is approximately VA. The abundance of tryptophan within an antimicrobial peptide rich in tryptophan is generally greater than 20%. A specific example of a tryptophan-rich peptide is the tri-tryptylin peptide shown as SEQ ID NO: 1 having a tryptophan content of about 23 JÍ.

# As used herein, the term "tryptophan triplet" refers to an amino acid sequence of three consecutive tryptophan residues. The tryptophan triplet is thought to be the rarest triplet sequence among the naturally occurring peptides. As used herein, the term "substantially the same sequence" refers to a sequence of peptides either identical to the peptide sequence defined by SEQ ID NO: 1 or having functional homology to it. It is understood that limited modifications can be made to the peptide which results in an increased function. It is also understood that limited modifications can be made without destroying the biological function of the peptide. Said minor modification of these sequences that do not destroy the antimicrobial activity of the peptides also falls within this definition. The modi cations can be included but not limited to additions, • deletions or substitutions of amino acid residues, substitutions with compounds that mimic the structure or function of the amino acids as well as the addition of portions chemicals such as amino and acetyl groups. Modifications may be deliberate or may be accidental such as through mutation is hosts that produce tritriptycin. All these modifications are included as long as the peptide retains its antimicrobial activity. As used herein, the term "antimicrobial activity" refers to the ability of a compound to # inhibit or irreversibly prevent the growth of a microorganism. Said inhibition or prevention can be through a microbicidal action or microbistatic inhibition. Therefore, the term "icrobicidal inhibition", as used herein refers to the ability of a compound to kill or irreversibly damage the target organism. The term "microbistatic inhibition" as used herein refers to the ability of the antimicrobial compound to inhibit the growth of the target microorganism without death. 0 Microbicidal or microbistatic inhibition of microorganisms in an environment that currently presents microbial growth (therapeutic treatment) or an environment at risk of sustaining microbial growth (prevention) is included within this definition. As used herein, the term "environment sustaining microbial growth" refers to any fluid »substance or organism in which microbial growth exists or may occur. Such environments can be, for example, human beings, animals, body fluids »or animal tissue, water, other liquids, food, food products» crops and various inanimate objects. The environment only needs to allow the subsistence of microbes. It does not need to promote microbial growth. The invention is directed to broad spectrum antimicrobial peptides containing a tryptophan triplet "compositions wherein said peptides are an active component" and methods of their use to inhibit or prevent microbial growth. The peptides are sequences of at least 10 amino acid residues »most preferably of between 10 and 34 amino acids in length» most preferably 13 amino acid residues in length. The antimicrobial peptides of these inventions are distinguished from many antimicrobial peptides by their amino acid sequence in tryptophan. In addition »unlike other tryptophan-rich peptides known in the art» such as indo! icidine »the Tritriptycin has a novel amino acid sequence that contains a tryptophan triplet. In a preferred embodiment of the invention, the antimicrobial peptide is a novel peptide substantially having the same sequence as defined by SEQ ID NO: 1. Trittriptycin exhibits broad spectrum antimicrobial activity. The test data show their effectiveness against various classes of organisms such as gram-negative bacteria, gram-positive bacteria, viruses and fungi. In addition »triiphyttin is effective against such diverse microorganisms such as Escherichia coli, Pseudomonas aeru inosa, Klebsiella pneumonieae, Staphylococcus epidermidis, Proteus mirabilis, and group D of Streptococcus. Based on these initial experimental results, it is believed that tritriptycin will be similarly effective against other organisms against which has not yet been tested. An additional feature of tritriptycin is its low price and high quality. Since tr ipytin is a relatively small peptide that occurs in nature, there is a reduced risk of producing an immune response in the host over other known antimicrobial peptides. Tritriptyline can be synthesized using conventional synthesis methods commonly used by one skilled in the art. For example, the antimicrobial peptide of the present invention can be chemically synthesized using an automated peptide synthesizer.

IO (such as one manufactured by Pharmacia LKB Biotechnology Co. »LKB Biolynk 4170 or Milligen, Model 9050 (Milligen, Mfordford, MA) following the method of Sheppard, and others» Journal of Chemical Society Perk I »page 538 (1981) In this procedure, N.N'-dicyclohexylcarbonate is added to amino acids whose amino functional groups are protected by 9-luoroenulmethoxycarbonyl groups (Fmoc) and or produce anhydrides of the desired amino acids. These inoperated F oc-a anhydrides can be used for peptide synthesis. An anhydride of Fmoc-am octa do corresponds to the residue of C-terminal amino acid binds to Ultrosin A resins through the carbo group. using di et laminopyridine as a catalyst. Next, the resin is washed with pyridine containing dimethyl Iformamide and the protecting group of the amino functional group of C-terminal acid is removed. The next The amino acid corresponding to the desired peptide is coupled to the C-terminal amino acid. The deprotection procedure is * repeated later. The successive desired amino acids are fixed in the same manner until the pep- tic chain of the desired sequence is formed. Protective groups other than acetoamide eti are then removed and the peptide is released with solvent. Alternatively, the peptides can be synthesized using nucleic acid molecules that modify the peptides of that invention into an appropriate expression vector that includes the coding nucleotide sequences. Said 0 DNA molecules can be easily prepared using an automated DNA sequencer and the well-known codon-amino acid ratio of the genetic code. Said DNA molecule can also be obtained as a genomic DNA or as a cDNA using ologonucl probes eotides and conventional hybridization methodologies. Said DNA molecules can be incorporated into expression vectors "including plasmids" which are adapted for the expression of DNA and the production of the polypeptide in a suitable host such as a bacterium, for example »Escherichia coli» yeast cells or mammalian cells . It is known that certain modifications of the primary sequence shown as SEQ ID NO: 1 can be made without completely suppressing the anti-chlamy activity of the peptide. The modifications include the removal and addition of amino acids. Specific examples of said modifications of the present invention are shown as SEQ IN NO: 2 and SEQ ID NO: 3. The peptides that contain other modifications can be synthesized by a person skilled in the art and tested for retention or increase of antimicrobial activity using the teachings described herein. Therefore »the potency of tritriptycin peptides can be modulated through several changes in the sequence or structure of amino acids. Another embodiment of the invention provides antimicrobial compositions having tr trythtycin as an active component. In one aspect of this invention tritriptycin is used as an active component in pharmaceutical compositions with a pharmaceutically acceptable carrier. A variety of vehicles, such as pH regulators »can be included in the composition where the antimicrobial activity is retained. Said pH regulators include pH regulated saline with phosphate »normal saline solution and Krebs ringer solution. Compounds such as EDTA and / or other chelating compounds »that are known to alter microbial membranes» can also be included in the composition. The many pharmaceutically acceptable carriers suitable for use in this invention are known to one skilled in the art. In another aspect of this invention tritriptycin can be used as an active component in various non-pharmaceutical compositions. In this aspect of the invention tritriptycin can be used, for example, as a food preservative or to inhibit potential microbial degradation. For example, shellfish and poultry routinely carry microbes that produce severe human disease. Those microbes can be inhibited with tritriptycin. Foods such as fruits and vegetables can be treated with tritriptycin to protect against microbial degradation. Tritripticin can be administered topically or by transgenic expression of the recombinant peptide. The tasgenic expression is known to a person skilled in the art and can easily be performed given the nucleic acid encoding the IO peptide shown as SEQ ID NO: 1. The invention also provides a method of microbicidal or microbial inhibition of microbial growth in a microbial growth support environment. The method includes administering a microbic quantity or microbistática effective tr tripticina to inhibit the -.j, microbial growth. Tritriptycin can be used as a disinfecting agent to sterilize or maintain microbial-free environments. Essentially any environment where microbial growth is undesirable can be treated with tritriptycin. Such environments include »for example» hospital surfaces »bathrooms and where food is prepared as well as water supplies. Tritriptycin can also be used as a disinfecting agent to sterilize or maintain microbial-free products. Examples of such products include baby towels »diapers» bandages »towels» makeup products »hair spray products» mouthwashes »eye drops and contact lens solutions. The effective amounts of tritriptycin to be administered will depend on the target microbe environment and the severity of the infection or growth. In yet another embodiment of the invention, a method of treating infectious diseases in a human being or an animal is provided. In this embodiment, tritriptythine is administered to a human being or an animal in an amount Icrobically or microbistically effective to inhibit microbial growth. Tryptidine can be administered to the host subject, for example, by intravenous injection, intrapeneal injection, orally, topically or by means of a nebulizer or aerosol spray composition. The lipid vehicles or emulsion preparations containing "Tritriptyline can also be used to administer tritriptycin to humans or animals. Specific modes of administration will depend on the target microorganism. The best administration mode for a particular application will be readily apparent to one skilled in the art. One skilled in the art will know the effective amount of triiptycline to be administered for a desired method given the teachings described herein. The following examples illustrate the invention. 25 EXAMPLE 1 A radial diffusion test was performed using double layer agarose as described by Lehrer and other »5" Ul transensitive assays for endogenous antimicrobial polypeptides. " J. I unol. Methods 137, 167-73 (1991), with minor modifications. In brief »2 x 10B bacterial growth cells from mid-log phase in a TSB medium were Wtf mixed with l.OJi agarose in phosphate pH regulator IO sodium at 10 M (pH 7.4) containing Tween 20 at 0.02 JI and TSB at 0.03 J4. The mixture was then emptied into a petri dish. When the agarose solidified, cavities with a diameter of 5 mm were made in the agarose and samples of the peptides in various concentrations were placed in each of the cavities. The box of petri was incubated at 37 ° C for 2 hours. The upper agar containing 1.0% agarose in pH buffer of F sodium phosphate at lOnM (pH 7.4) and TSB at SJ4 was emptied thereon and the petri dish was re-incubated at 37 ° C for 18 hours. The diameters of the inhibitor zones were measured for the quantification of inhibitory activities. As shown in Table 1, peptide 1 with the sequence of SEQ ID NO: 1 showed strong antibacterial effects on Escherichia coli when tested by the radial diffusion test. Several derivatives of this peptide » named peptides 2 »3 and 4» also shown in Table I were then synthesized and tested by radial diffusion »as described above» against E. col. Peptide 2 contained aginin at both ends and showed slightly higher activity than peptide 1. Peptide 3 'which shared 9 amino acids with peptide 1' showed detectable but significantly less activity than peptide 1. Peptide 4 which shared 7 residues of amino acids with peptide 1 had no detectable activity.

TABLE 1 PROOF OF ZONE INHIBITION WITH E. COLI AX PLO T.T, The antimicrobial effects of the above peptides on other gram-positive and gram-negative bacteria including Pseudomonas »Klebsiella. Staphylococcus Proteus and Streptococcus were then tested using the method described in Example 1. As shown in Table II, peptide 1 showed a wide range of antimicrobial activity on all tested organisms except Proteus mirabilis. Again »peptide 2 showed slightly higher activity in those same bacteria» while peptide 3 showed low but detectable activity and peptide 4 had no activity in any organism. i 10 TABLE 2 PROOF OF ZONE INHIBITION WITH TRAM-POSITIVE AND GRAM-NEGATIVE BACTERIA fifteen " EXAMPLE III To examine whether the antimicrobial activities of the peptides are bactep'oßtát cas or bactericides »was performed a bactericidal test for all peptides using Escherichia coli The cells were incubated with several concentrations of peptides in sodium phosphate pH regulator. After incubation for 10 minutes, samples were taken in aliquots (2 x 10ß) and mixed with several samples. concentrations of the peptides in 20 μl of sodium phosphate pH regulator. The mixtures were incubated at 37 ° C and the samples were extracted at 10 minute intervals. They were diluted with sodium phosphate pH regulator and plated with L-agar. After incubation during the night the plates 37 ° C »the number of colonies was counted. The results are ^ show in Figure 1. The number of cells that survived ~ was told later. As shown in figure 1, peptide 1 showed strong bactericidal activity and 97% of the cells were killed within 10 minutes at a concentration of 50 μg / ml. Peptide 2 showed a significantly lower bactericidal activity and peptides 3 and 4 showed minimal activity. These results suggest that the antimicrobial activity of the peptides shown by the radial diffusion test is bactericidal. However, it should be noted that the antimicrobial activity of peptide 2 by radial diffusion test was stronger than that of peptide 1 »while the bactericidal activity of peptide 2 is significantly less than that of peptide 1. Thus» the mechanism of antimicrobial activity of the peptide and peptide 2 seem to differ due to the modification of the arginine residues present at both ends of peptide 2.

EXAMPLE IV Since peptide 1 showed broad spectrum antimicrobial activity, the effect of the peptides on two fungi was tested, namely Aspergillus 11 us fumigates and Candida albicans. As shown in Table III »Peptide 1 inhibited the growth of Aspergillin and Candida at a concentration of 250 μg / ml and 10000 μg / ml, respectively. Peptide 2 also inhibits the growth of both fungal strains at a similar concentration. Peptide 3 and peptide 4 also showed growth inhibitory effects on Asperus' 11us fumigatus at a concentration of 500 μg / ml but had no inhibitory effect on Candida albicans. These results suggest that peptide 1 and peptide 2 could be effective antifungal drugs, although the concentration required would be higher than for bacteria.

TABLE III TEST OF FUNGICIDE MINIMUM INHIBITOR CONCENTRATION EXAMPLE V The minimum inhibitory concentration (MIC) was determined by cultivating fungal strains in a YG medium until the fungi reached the logarithmic phase. The cells were diluted in YG medium and then placed in 96-well plates along with a series of double dilutions of the peptides. The plate was then incubated at 37 ° C for 24 hours. The minimum inhibitory growth was defined as the lowest concentration at which there is no fungal growth. In view of the above, it will be seen that the various objects of the invention are achieved. Since various changes can be made in the above compositions and prior methods without departing from the scope of the invention, it is intended that all matter contained in the above description be interpreted as illustrative and not in a limiting sense.

» # LIST OF SEQUENCES (1. GENERAL INFORMATION: (ii) TITLE OF THE INVENTION: Broad Spectrum Antimicrobial Peptides Containing a Tryptophan Triplet and Methods of Using Them (iii) SEQUENCE NUMBER: 4 (iv) ADDRESS TO SEND CORRESPONDENCE (TO) RECIPIENT: Senniger »Powers» Leavitt £ Roedel (B) STREET: one Metropolitan Square »16th Floor (C) CITY: St. Louis (D) STATE: Missouri (E) COUNTRY: USA (F) POSTAL CODE: 63102 (iv) COMPUTER READABLE FORM: (A): TYPE OF MEDIUM: Flexible disk (B) COMPUTER: IBM COMPATIBLE PC (OR OPERATING SYSTEM: PC-DOS / MS-DOS (D) SOFTWARE: PatentIn Relay # 1.0 »Version # 1.25 (i) DATA OF THE CURRENT APPLICATION: (A) APPLICATION NUMBER: E.U.A. (B) DATE OF SUBMISSION: < C > CLASSIFICATION (Viii) INFORMATION ABOUT THE APPORTER / AGENT * (A) NAME: Blosser, G.H. (B) REGISTRATION NUMBER: 33,650 (C) REFERENCE NUMBER / CASE: SIU 7343 (ix) TELECOMMUNICATIONS INFORMATION (A) TELEPHONE: 314-231-5400 (B) TELEFAX: 314-231-4323 (2) INFORMATION FOR SEQ ID NO. i: • »(i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 13 amino acids (B) TYPE: amino acid (ii) TYPE OF MOLECULE: peptide (i) SEQUENCE DESCRIPTION: SEQ ID NO: i: «Val Arg Arg Phe Pro Trp Trp Trp Pro Phe Leu Arg Arg 1 5 10 20 (2) INFORMATION FOR SEQ ID NO. 2: (i) CHARACTERISTICS OF THE SEQUENCE: 25 (A) LENGTH: 14 amino acids (B) TYPE: amino acid (ii) TYPE OF MOLECULE: peptide (ix) SEQUENCE DESCRIPTION: SEQ ID NO: 2: Arg Arg Arg Phe Pro Trp Trp Trp Pro Phe Leu Arg Arg Arg 30 1 5 10 (2) INFORMATION FOR SEQ ID NO. 3: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 10 amino acids (B) TYPE: amino acid (ii) TYPE OF MOLECULE: peptide (ix) SEQUENCE DESCRIPTION: SEQ ID NO: 3: Arg Phe Pro Trp Trp Trp Pro Phe Leu Arg 1 5 10 (2) INFORMATION FOR SEQ ID NO (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 39 nucleotides (B) TYPE: nucleic acid (ix) SEQUENCE DESCRIPTION: SEQ ID NO: 4: TCA GGA GAT TTC CCT GGT GGT GGC CGT TCC TAC GAA GAC 39

Claims (4)

* NOVELTY OF THE INVENTION CLAIMS

1. A peptide comprising a tryptophan triplet »has between 10 and 34 amino acid residues in length and exhibits antimicrobial activity.

2. The peptide according to the re-indication jB 1 »further characterized in that said peptide comprises Substantially the sequence as identified by SEQ ID NO: 1 'has between 10 and 34 amino acid residues in length and exhibits antimicrobial activity.

3. The peptide according to the claim 1"further characterized in that said peptide comprises 15 substantially the sequence as identified by SEQ ID NO: 2 » ^ Do you have between 10 and 34 amino acid residues in length and ^ * presents antimicrobial activity. 4.- The peptide in accordance with the rei indication 1"further characterized in that said peptide comprises Substantially the same amino acid sequence as identified by SEQ ID NO: 3, has between 10 and 34 amino acid residues in length and exhibits antimicrobial activity. 5. The peptide according to claim 1, further characterized in that said peptide is low in oneness. 6. The peptide according to claim 1 further characterized in that said peptide exhibits antimicrobial activity effective against a class of organisms selected from the group consisting of gram-positive bacteria, gram-negative bacteria, and fungi. 5 7.- The peptide according to the claim 1"further characterized in that said peptide exhibits antimicrobial activity effective against an organism selected from the group consisting of E. coli. Pseudomonas aeruginosa. Klebsei lia pneumonieae, Staphylococcus epedermidis, Proteus maribil s, O Streptococcus group D, Aspergillus fumigatus, and Candida albicans. 8. The peptide according to claim 1, further characterized in that said antimicrobial activity is presented at a final concentration between 0.5 and 1000 μg / ml. 9. The peptide according to claim 1, further characterized in that said antimicrobial activity is presented at a final concentration between 0.5 and 500 μg / ml. 10. A synthetic peptide comprising a tryptophan triplet 0, exhibits antimicrobial activity and is produced by chemical synthesis. 11. The synthetic peptide according to claim 10, further characterized by having between 10 and 34 amino acid residues of longi. 12. A recombinant nucleic acid sequence encoding an antimicrobial peptide, said peptide containing a tryptophan triphenyl, has between 10 and 34 amino acid residues in length and exhibits antimicrobial activity. 13. An antimicrobial composition comprising a 5 peptide containing a tryptophan triplet as an active component "wherein said peptide has between 10 and 34 amino acid residues in length and exhibits antimicrobial activity. fl | 14.- The antimicrobial composition in accordance with 10 claim 13, further characterized in that said peptide exhibits low immunsgenicity. 15. The antimicrobial composition according to the re-indication 13"further characterized in that said peptide exhibits effective antimicrobial activity against a class of 15 organisms selected from the group consisting of gram positive bacteria »gram negative bacteria» viruses and fungi. • 16.- The antimicrobial composition according to the rei indication 15"further characterized in that said organisms are selected from the group consisting of E. coli» 20 Pseudomonas aeruginosa, Klebsei lia pneumonieae, Staphylococcus epedermidis, Proteus marib lis, Streptococcus group D, Aspergillus fumigatus »and Candida albicans. 17.- The use of a microbiocidally or microbistically effective amount of an antimicrobial peptide, 25 said peptide containing a tryptophan triplet and has between 10 and 34 amino acid residues in length to prepare a composition for the microbicidal or microbial inhibition of microbial growth. 18. The use according to claim 17 »further characterized in that the quantity crobicida or microbistá ca effective effective is at a final concentration of between 0.5 and 1000 μg / ml. 19. The use according to claim 17 »further characterized in that said peptide has antichrobial activity effective against a class of organisms selected from the group consisting of gram positive bacteria» gram negative bacteria »viruses and fungi. 20.- Use in accordance with re-indication 19, further characterized in that said organisms are selected from the group consisting of E. coli »Pseudomonas aeruginosa. 15 Klebsei lia pneumonieae. Staphylococcus epedermidis. Proteus maribi 1 is. Streptococcus group D »Aspergillus fumigatus» and • Candida albicans. «21. Use according to claim 17. further characterized in that the composition obtained comprising the peptide is administered to a human or animal. 22. The use according to claim 17. further characterized in that the obtained composition comprising the peptide is administered to foods or products to the imenticioß. 23. The use according to claim 17, further characterized in that the composition obtained that • comprises the peptide is administered to liquid. 2

4. The use according to claim 17. further characterized in that the obtained composition comprising the peptide is administered to an inanimate object. 5 25.- The use of an microbially or microbistically effective amount of an antimicrobial peptide »said peptide containing a tryptophan triplet and has between IO and 34 amino acid residues in length to prepare "B a composition for treatment of an infectious disease 10 a human being or an animal 26 .- The use of claim 25" characterized further because said peptide has low i ungem'cidad. 27. The use according to claim 25 »further characterized in that the amount microbiocidally or microbistáticamente effective is at a final concentration of between 0.5 and 1000 μg / ml. *