MX2008001651A - Antifungal peptides and methods of use thereof - Google Patents

Abstract

A method of treating fungal infections by treatment with CAP37 peptides and derivatives thereof, including peptide analogs having serine or threonine substitutions at least one of the two cysteine residues therein. Other substitutions of the amino acid residues of the peptide are also contemplated.

Description

ANTIMICTIC PEPTIDES AND METHODS FOR USING THE BACKGROUND OF THE INVENTION CAP37 (Mr 37 kDa cationic antimicrobial protein) was originally identified as a component of the oxygen independent annihilation mechanism of a human neutrophil (PMN) and was shown to have a strong bactericidal activity for Gram-negative organisms including Salmonella Typhimurium, Escherichia coli, and Presudomonas aeruginosae (Shafer et al., 1984, et al., 1986; Spitznagel 1990). Unlike its effect on protein Natural CAP37 has potent regulatory effects on host cells. It is an effective regulator of cells of the mononuclear phagocytic system such as monocytes (Pereira et al., 1990), icroglia (Pereira et al., 2-002) and macrophages (Larricks et al., 1991). It also regulates the functions of the corneo epithelium (Rúan and others, 2002), endothelial (Lee and others, 2002, Lee and others, 2003) and smooth muscle (Gonzalez and others, 2004). BRIEF DESCRIPTION OF THE INVENTION The analysis of the structure function of CAP37 allows delineating its antibacterial domain to a region corresponding to residues 20 to 44 of the natural molecule (Pereira and others, 1993). A peptide comprising this 25 amino acid sequence (CAP37 (20-44) nat) mimics the antimicrobial activity of the natural molecule (Pereira and Ref .: 190091 others, 1993) and extends its range of activity to include Staphylococcus aureus and Enterococcus faecalis, two Gram positive organisms. The bactericidal activity of the peptide was pH dependent, with a maximum activity obtained between pH 5.0 and 5.5. The replacement of the cysteine residues at positions 26 and 42 with cysteine residues (CAP37 (20-44) ser26 / 42) resulted in an inactive compound (Pereira et al., 1993). The in vivo experiments demonstrated the efficacy of CAP37 (20-44) na in attenuating the lethal effects of lipopolysaccharide E. coli (LPS) in a model of endotoxic shock-conscious rats (Bracket et al., 1997). Infections due to the various Candida species can result in disease manifestations on the scale of self-limiting superficial infections to systemic infections of lifelong treatment (Nola et al., 2003). In the recent past, there has been a dramatic increase in invasive fungal infections and Candida species that begin to contribute substantially to serious hospital acquired infections (Clark and Hajjeh, 2002, Hobson 2003, Ñola and others, 2003, Rapp 2004). The reasons for this are complex but can be attributed mostly to the latest advances in medicine that lead to increased survival of immunosuppressed people and the use of internal medical devices and catheters for the treatment of hospitalized patients (Clark and Hajjeh 2002; Ñola and others, 2003). The development of the incidence of resistance to available anti-fungal drugs also aggravates the clinical and public health problems associated with fungal infections. New drugs that have anti-fungal activity are necessary. It is to this object that the present invention is directed. BRIEF DESCRIPTION OF THE FIGURES Figures 1A-1F are a graph showing the Candida killing activity of four CAP37 peptides. The activity of CAP37 (20-44) nat, CAP37 (20-44) ser26, CAP37 (20-44) be 2, and CAP37 (20-44) ser26 / 42, 400 mg / ml (solid bar) and 200 mg / ml (diagonal bars) was tested against clinical isolates of Candida albicans (Fig. IA) A-155, (Fig. IB) A-22 and (Fig. IC) A-III (known as being sensitive to fluconazole) and isolated (Fig. ID) A-5, (Fig. 1E) A-20 y (Fig. 1F) A-46 (known as fluconazole resistant). The annihilated percentage is determined as detailed in the materials and methods section. The values were expressed as + standard error of the mean of three independent experiments carried out in triplicate. Figures 2A-2F are a graph showing the activity of CAP37 peptides in the form of a hyphae of C. albicans. The activity of CAP37 (20-44) nat, CAP37 (20-44) ser26, CAP37 (20-44) ser42, and CAP37 (20-44) ser26 / 42, at 400 mg / ml (solid bar) and 200 mg / ml (diagonal bars) was tested against forms of hyphae from clinical isolates of Candida albicans (Fig. 2A) A-155, (Fig. 2B) A-22 and (Fig. 2C) A-III, (Fig. 2D) A-5, (Fig. 2E) A -20 y (Fig. 2F) A-46. The annihilated percentage was determined as detailed in the materials and methods section. The values were expressed as + standard error of the mean from three different experiments carried out in triplicate. Figure 3 is a graph showing the activity of the CAP37 peptides against several Candida species. The fungicidal activity of CAP37 (20-44) nat / CAP37 (20-44) ser26, CAP37 (20-44) ser42, and CAP37 (20-44) ser26 / 42, was tested in C. guilliermondii, C. parapsilosis, C. pseudotropicalis, C. tropical, a blood isolate of C. albicans, two isolates of C. dubliniensis, three isolates of C. glabrata, one isolate of C. krueei and two isolates of S. cerevisiae. Peptide concentrations for C. guilliermondii and C. parapeilosis were 100 μg / ml (37.5 μM) while all other strains were tested with peptide concentrations at 400 μg / ml (150 μM). The values were expressed as + the standard error of the mean of 3 to 4 independent experiments carried out in triplicate. Figures 4A-4L show the activity of CAP37 determined with a vital dye FUN-1 and confocal microscopy. A fluconazole-sensitive C. albicans isolate (Figs 4A-4C), pseudotropicalis (Figs 4D-4F), was selected. isolated C. glabrata (Figs 4G-4I) and an isolate S. cerevisiae (Figs 4J-4L) to carry out these studies. The fungal isolates were incubated with the peptide CAP37 (20-44) nat, the peptide CAP37 (20-44) Ser2642 and in the absence of peptide. The cells that stained red are alive and those that stained uniformly green or greenish yellow are dead. Representative digital images of these independent experiments are shown. DETAILED DESCRIPTION OF THE INVENTION The anti-mycotic activity of peptides based on the natural sequence of the CAP37 antibiotic protein is demonstrated herein. The peptides can be used as anti-mycotic agents, particularly against Candida sp. CAP37 has traditionally been considered as a protein derived from PMN since it is constitutively expressed in the granules of this cell. However, more recently, the presence of an inducible form of CAP37 has been demonstrated (Lee et al., 2002, Pereira et al., 1997, Rúan et al., 2003, González et al., 2004, and Pereira et al., 2004). CAP37 can be expressed in endothelial cells that coat the vasculature in inflammatory mediated diseases such as Alzheimer's disease (Pereira et al., 1997) and atherosclerosis (Lee et al., 2002). In vitro studies indicate that induction is due to mediators inflammatories such as tumor necrosis factor -a (TNF-a), interieucin-1 (IL-1) and immodyodulatory substances such as LPS. In vivo studies use a rabbit model of S. aureus keratitis, demonstrating the very early induction of CAP37 in the corneal epithelium, and in the blood vessels that line the endothelium in the limbic circulation in rnse to infection (Rúa et al. 2002). Additionally, the expression of CAP37 has been demonstrated in squamous epithelial cells of the skin, cells that line the hair follicles, individual cells of the sebaceous glands and endothelial cells that line the blood vessels in rnse to lesions using a rat model. in vivo wound repair (Pereira et al., 2004). Epithelial antibiotics have been demonstrated on the skin (Nizet et al., 2001; S0rensen et al., 2002; Shirafuji et al., 1999; Oren et al., 2003), on the lining of the mucosal surfaces of the gastrointestinal tract (Froh, -Nilsson. et al, 1999; Fellermann and Stange, 2001), oral surfaces (Dale 2000; Dale 2001), rratory tract (Hiemstra 2001, Diamond et al., 2000; Huttner and Bevins, 1999) and genitourinary tract (Frohm-Nilsson et al. 1999; Mal and others, 2000). These antibiotic proteins are ideally located to serve as the first line of defense against invading pathogens. Whether the expression of CAP37 occurs in the mucous linings of the host in rnse to Candida infections or that induction of CAP37 is compromised on the mucosal and epithelial surfaces in patients with recurrent candidiasis suggests a physiological role for CAP37 in protecting the host against fungal infections is currently unknown. The study reported herein for the first time discloses that CAP37 peptides and analogs and derivatives based on the AT37 atural sequence possess potent antifungal activity and suggest broad spectrum anti-fungal activity for the peptide that was originally proposed. In a series of experiments, peptides based on the natural CAP37 sequence were used including CAP37 (20-44) nat and three peptide analogs (CAP37 (20-44) ser26 / CAP37 (20-44) ser2 »CAP37 (20-44) ) Ser2642 where the cysteine residues at positions 26 and / or 42 were replaced by serine residues as indicated in Table 1. The effect of the substitutions - in these two positions on the killing efficiency in vi tro - was investigated. on a scale of Candida species The findings demonstrate significant activity against C. albicane through the peptide based on the natural sequence (CAP37 (20-44) nat) and the two analogs where only one individual cysteine residue was replaced (CAP37 (20-44) ser26 and CAP37 (20-44) ser42) • The replacement of both cysteines significantly abrogated the activity against C. albicane and many of the Candida species tested. Since you do not want Being bound by theory, these data suggest that the intramolecular disulfide bond was important but the formation of a cyclic compound was not essential for anti-fungal activity since the intermolecular interactions between the two cysteine residues, which is possible with two substitutions mode -cysteine, resulted in a peptide that retained the annihilation activity. TABLE 1 Peptides and Derivatives CAP37 The activity of the CAP37 peptides (20-44) nat / CAP37 (20-44) Ser26 and CAP37 (20-44) ser42 against fluconazole-resistant mucosal isolates ((Fig. 2D) A-5, (Fig. 2E) A-20, (Fig. 2F) A-26 ) was attacked, with strong annihilation (less than 5% of survival) obtained against two of these Ufig isolates. 2D) a-5 and (Fig. 2E) A-20) and > 75% annihilation obtained against the isolate (Fig. 2F) A-46 (Figures 2A-2F). These findings are particularly relevant to the recent emergence of fluconazole-resistant isolates of Candida albicans and non-albicans species that will require treatment with new therapeutics with alternative novel mechanisms of action (Jabra-Rizk et al., 2004, Sullivan et al., 2004). . In addition to the important finding that the CAP37 peptides had activity against fluconazole-resistant mucosal isolates of C. albicans, Figure 3 indicates that the peptides showed potent activity against C. guilliermondii, and C. parapeilosis at a concentration of 37.5 μM. C. parapsilosis is the most common isolate of critically ill patients in intensive care units who typically have catheters and internal devices while undergoing treatment (Kuhn et al., 2004). Almost 100% of the starting inocula of C. peeudotropicalis, and C. tropicalie, were killed with peptide concentrations of 150 μM. C. tropicalee is being isolated in increasing blood cultures of patients with leukemias, other neoplasms of those in intensive care units (Warn et al., 2002). Of the peptides, CAP37 (20-44) eer42 was more effective against C. dublinieneie than CAP37 (20-44) na • C. dublinieneie is a newly identified species that rarely It is found in healthy people but tends to be found as the causative agent of oropharyngeal infections mainly in HIV-infected individuals (Sullivan et al., 2004). The peptides were not active against C. glabrata, C. krusel and S. cerevieiae and had more modest activity against the hyphae forms than C. albicane. The concentrations of peptide required to achieve the fungicidal activity are approximately twice those shown to kill the Gram negative isolates S. Typhimurium, E. coli and P. aeruginoea (Pereira et al., 1993). This range in activity is not uncommon among cationic antimicrobial peptides that tend to have specificity and potency towards certain bacterial species, fungi, parasites, and / or viruses. Apart from CAP37, PMN contains several cationic antimicrobial peptides that include defensins, hCAPl8 / LL37, protein that increases bactericidal permeability (BPI), and lactoferrin (Spitznagel, 1990: Ganz and Wells, 1997). Defensins are particularly active against C. albicane (Seisted et al., 1985; Hoover et al., 2003). Synthetic peptides based on the first cationic domain of the amino terminus of human lactoferrin demonstrated that they have Candida killing activities at micromolar levels (Lupetti et al., 2000). Antimicrobial peptides of human platelets have also been shown to have antimicrobial activities (Tang et al., 2002). Of the Seven peptides isolated from platelets, only the "Regulated on Normal Expressed and Secreted T Cell Activation" (RANTES) protein and platelet factor-4 (PF-4) exhibited Candida killing activity. Its activity as that of the CAP37 peptides was pH dependent, being most active at a pH of 5.5 (Tang et al., 2002). The active peptide concentrations used range from 150 μg to 1.0 mg / ml depending on the molecular weight of each peptide (Tang et al., 2002). Another well-known Candida-destroying molecule is salivary histatin-5 (Tsai and Bobek 1997, Edgerton et al., 2002). Salivary histatin-5 concentrations of 15 μM have been shown to kill between 80 and 100% of the blastopods C. albicane (Tsai and Bobek, 1996). Synthetic histamine analogues based on fungicidal domain of histamine-5 C term have been shown to be effective against C. albicane (Heimerhorst et al., 1997), C. krueei, fluconazole-resistant strains of C. glabrata and Aepergillue fumigatue (Helmerhorst et al., 1999). The use of natural cationic peptides as novel therapeutics in the treatment of infections is gaining enthusiasm in the scientific and biotechnology communities. The main drawback of conventional antibiotics is the rapidity with which microorganisms can gain multiple resistance patterns. The mechanism Exactly how these cationic antimicrobial peptides including those described herein, which kill microorganisms is not entirely known. However, without wishing to be restricted by theory, it is believed that the main bactericidal mechanism is the permeabilization of the membrane of the microorganism through purine channels and self-promoted absorption trajectories (Handcock, 1997). Cationic antimicrobial peptides seem not to be involved in the metabolic trajectories in organisms and thus can not be involved in common resistance mechanisms. Handcock (1997) has shown that cationic peptides do not include resistant mutants even after 20 passages in antibiotic concentrations close to the minimum inhibitory concentration. Clearly, although the mode of action of CAP37 peptides has not yet been determined, without wishing to be bound by theory, it seems to be different from the mechanism of action of azole-based drugs, since it annihilates fluconazole-resistant and sensitive strains equally. . MATERIALS AND METHODS Peptide Synthesis The peptides were synthesized through solid phase synthesis on a peptide synthesizer as previously described (Pereira et al., PNAS 1993). The purity of the peptides was verified. The mass of the peptide was confirmed through of mass spectrometry. The peptides were synthesized by predicting several findings of a synthetic peptide (CAP37 (20-44) nat) based on a natural amino acid sequence of CAP37 consisting of residues 20 to 44 that had a potent bactericidal activity for a number of organisms Gram negative (Pereira et al., PNAS 1993). An inactive analog of this peptide in which the cysteine residues at positions 26 and 42 were replaced by serine residues (CAP37 (20-44) ser26 / 42) (Pereira et al., PNSA 1993) was used as a peptide of inactive control. In addition, two other peptide analogs were synthesized. One peptide had the cysteine residue in position 26 replaced by a serine (CAP37 (20-44) ser26) and the other had the cysteine residue in position 42 replaced by a serine (CAP37 (20-44) Ser42) • Mycotic Care and Cultivation Appearance The isolates used in this study included Candida albicane (ATCC28367), three fluconazole-sensitive mucosal isolates of C. albicane, designated A-22, A-III and A-155, three isolates of mucosa resistant to fluconazole of C. albicane designated A-46, A-5 and A-20, and a blood isolate of C. albicane designated WDO. Isolates were stored frozen at -70 ° C and run on Sabouraud dextrose agar plates (Sigma, St. Louis, MO) and kept on plates at 4 ° C, with subculture on plates new ones of approximately every 10 days during the duration of the studies. Fluconazole resistance was evaluated. Three clinical isolates of C. glabrata, two clinical isolates of C. dublinieneie, and a single isolate of C. krusel, C. guilliermondii, C. parapeiloeie, C. pseudotropicalis, and C. tropical and two isolates of Saccharomycee cerevieiae They were also used in this study and were kept on Sabouraud dextrose agar plates at 49C. A single yeast colony was grown overnight at 33 SC in 1% phyton peptone broth (Becton Dickinson, Sparks MD) with 0.1% D-glucose (Sigma). Induction of the hypha To induce hyphal formation, a single colony of the C. albicane isolates A-5, A-20, A-22, A-46, A-ll and A-155 from an agar plate was transferred. of dextrose Sabouraud to 1% of peptone broth of phyton with 0.1% glucose and was cultivated overnight at 33 aC. An aliquot (500 μl) of the culture overnight was subcultured in 5 ml of RPMI-1540 (Cellgro Mediatech Inc., Rendon, VA) with 10% fetal calf serum (Invitrogen, Grand Island, NY) for 90 minutes to 37 SC. The formation of the hypha was determined under phase microscopy. Candida in vi trodeintructor The concentrated solutions of all the peptides were made at concentrations of 1 mg / ml in water free of Sterile endotoxin for irrigation (Baxter, Deerfield, IL). Subsequent dilutions were all made in tryptamine saline pH 5.5 (Shafer et al., Infect Immun, 1984). An aliquot (100 μl) of overnight culture from a single culture colony at 33 eC in 1% phytonone pentone broth with 0.1% glucose was subcultured in 5 ml of 1% phytonone pentone broth with 0.1 % glucose was incubated in a shaking water bath (80 oscillations per minute, Precision, Winchester, VA) for 90 minutes at 33 SC to produce a logarithmic culture. Cell cultures under these conditions were found to consist predominantly of blasto-spores as determined through phase contrast microscopy. The optical density was read and the culture was adjusted to 500 blasto-spores / 100 μl in tryptone saline pH 5.5 (Shafer et al., Infect Immun, 1984). To 100 μl of the organism suspension in a 96-well sterile polystyrene microtiter plate (Becton Dickinson, Franklin Lakes, NJ) were added 100 μl of the peptide (final concentrations of 400 μg / ml and 200 μg / ml) or 100 μl of tryptone saline. The above served as a control. The microtiter plate was incubated at 37 SC for 4 hours and 100 μl of the content of each well was placed on Sabouraud dextrose agar plates and incubated at 37 SC overnight. The colony forming units (cfu) were counted and the fungicidal activity expressed as the percentage annihilated and calculated according to the following equation: [(control cfu - cfu test) / cfu control] x 100 =% annihilated. The cfu control was indicated by the number of colonies present after 60 minutes of incubation in tryptone saline alone in the absence of peptide. The cfu test was determined by counting the number of colonies present after incubation in tryptone saline containing the peptide. Each experimental point was carried out in triplicate. Method for the Fungicide Eneay The viability of the yeast was also evaluated using the live / dead yeast viability kit FUN-1 (Molecular Probes, Eugene, OR). The methodology used was essentially in accordance with the protocol provided by the vendor for use with fluorescent microscopy. Briefly, the yeast cultures were prepared exactly as stated above. C. glabrata, C. pseudotropicalis, C. albicane (fluconazole-sensitive isolate) and S. cerevieiae (5 x 105 cfu in 100 μl tryptone saline) were incubated (37 aC for 2 hours) in the absence or presence of peptide (CAP37). (20-44) nat and CAP37 (20-44) Ser2642) at a final concentration of 400 μg / ml. At the end of the incubation period, the samples were transferred to microcentrifuge tubes and centrifuged (10,000 x g for 3 minutes at room temperature). The supernatant was removed and the granules were resuspended in 25 μl of GH solution (2% D-glucose) containing 10 mM Na-HEPES, pH 7.2) as described in the technical data sheet provided by the vendor. An operating solution of reagent FUN-1 (100 μl of 10 μM) was prepared from a concentrated solution of 10 mM and 25 μl (final concentration of 5 μM) were added to the yeast and incubated at room temperature for 30 minutes. A sample was placed (10 μl) on a microscope slide and staining was observed under a Leica TCS NT confocal microscope using Ar-488 and Kr-568 lasers and the water immersion objective 63x Plan APO 1.2 NA. The images were scanned and analyzed using the Leica TCS software. Eetaditic Analysis The data are presented as the mean _ + standard error of 3 or 4 independent experiments carried out in triplicate. RESULTS Peptide Synthesis The peptides synthesized from these studies are shown in Table 1. CAP37 (20-44) nat contains two cysteine residues corresponding to positions 26 and 42 of the natural CAP37 protein that forms a disulfide bridge in the molecule Natural CAP37 (Pohl et al., FEBS Lett 11990). Accordingly, the importance of these two cysteine residues was evaluated for peptide activity by replacing both cysteines with serine residues (CAP37 (20-44) ser26 / 42) or a single cysteine with a serine at position 26 (CAP37 (20-44) Ser26) or 42 (CAP37 (20-44) ser42) • The replacement of either or both cysteine residues confers on the peptide the inability to form a binding disulfide and therefore interferes with the possible formation of a cyclic structure. Without wishing to be bound by theory, the replacement of a cysteine in any position interferes with the formation of a cyclic structure but still allows for dimerization. Eetandarization of a destructive eneja of candida in vi tro C. albicans (ATCC 28367) was used to standardize the in vi tro annihilation test. Optimum growth conditions (temperature and time, 25SC for 5 h and 33aC for 90 minutes) were explored, blastopore numbers per cavity (200, 400, 500, 600, 800, and 1000 cfu), range of peptide concentrations (750, 500, 400, 200 and 1000 μg / ml) and the contact time between the peptide and Candida (1, 2 and 4 hr) for annihilation to occur. The data indicate that there was no significant difference between the use of Candida that had been grown for 5 h at 5 SC or for 90 minutes at 33 SC. The 90 minute incubation was more technically convenient and therefore was routinely used. The optimal number of cfu per cavity was determined as being 500 and an incubation time of 4 h at 37 eC between Candida of the peptide was required to obtain the optimal annihilation. The dose-dependent annihilation was obtained with varying concentrations of CAP37 (20-44) nat. However, the best distinction between active and inactive peptide was obtained at 400 μg / ml or 150 μm. Fungicidal activity of CAP37 peptides on strains of C. albicans senescent to fluconazole and resistant Peptides CAP37 (20-44) nat, CAP37 (20-44) ser26, and CAP37 (20-44) Ser42 / are strongly active against clinical isolates sensitive to fluconazole (Fig. IA) A-155, (Fig. IB) A-22 and (Fig. IC) A-III at 400 μg / ml, with a range of activity between 60% and 94% annihilated depending on the isolate (Figures 1A-1F). The isolate (Fig. 1A) A-155 appears to be more sensitive with 80-94% of annihilated organisms. There was no statistical difference between the activity of these three peptides for any given isolate. A marked contrast in the activity of these peptides was the lack of activity of CAP37 (20-44) ser26 / 42 where both cysteine residues were substantially through serine residues. The peptides CAP37 (20-44) nat, CAP37 (20-44) ser26, and CAP37 (20-44) Ser42 were highly active against fluconazole-resistant isolates (Fig. ID) A-5 and (Fig. 1E) A- 20 with a significant activity obtained even at lower concentrations (200 μg / ml or 75 μM) of the peptide. The highest concentrations (400 μg / ml) of the peptides were required for activity against the fluconazole-resistant isolate (Fig. 1F) A-46. While the peptide CAP37 (20-44) ser26 / 42 considered as inactive was moderately active against the isolates (Fig. ID) A-5 and (Fig. 1E) A-20. The effect of the peptides CAP37 (20-44) nat, CAP37 (20-44) STr26 > and CAP37 (20-44) ser42 on the fluconazole-resistant species (Fig. ID) A-5, (Fig. 1F) A-20, and (Fig. 1A) A-46 and the fluconazole-sensitive strain A-155 it was fungicidal rather than fungistatic since the viable colony counts in the presence of the peptides was less than the starting inoculum. Fungicidal activity of the CAP37 peptides in C. albicane hyphae The CAP37 peptides were less active in the hyphal forms of the clinical isolates of C. albicane when compared to their activities in the blasto- spores (Figures 2A-2F). When the activity was compared between peptides CAP37 (20-44) nat, CAP37 (20-44) ser26, and CAP37 (20-44) ser42, it might appear that greater activity was obtained with CAP37 (20-44) being 2 against hyphal forms than with the other CAP37 peptides. Fungicidal activity of peptide CAP37 on various types of Candida The effect of CAP37 peptides varied depending on the different Candida species tested (Figure 3). The CAP37 peptides were more effective in C. guilliermondii and C. parapeiloeie. Peptide concentrations as low as 100 μg / ml (37.5 μM) were fungicidal for these two species.

C. peeudotropicalie was also extremely sensitive to all CAP37 peptides. As with C. guilliermondii and C. parapeiloeie, some activity was also obtained with the peptide CAP37 (20-44) Ser26 / 42 although significantly lower than with the other three CAP37 peptides. C. tropicalee and the blood isolate of Candida were also annihilated through CAP37 (20-44) nat, CAP37 (20-44) ser26, and CAP37 (20-44) ser42 • The activity of all previous species was fungicide instead of fungiestática. The CAP37 peptides (CAP37 (20-44) nat, CAP37 (20-44) ser26, and CAP37 (20-44) ser2) were effective against both isolates of C. dubliniensis and an isolate of C. glabrata, although to a degree less. The CAP37 peptides were ineffective against two of the isolates C. glabrata, C. krueel and two isolates of S. cerevieiae. The antifungal activity of peptides CAP37 is fungicidal. Confocal microscopy and the fluorescent dye FUN-1 were used to evaluate the viability of the cells. A C. albicane isolate sensitive to fluconazole and C. peeudotropicalie that was sensitive to CAP3720-, and one of the isolates C. glabrata and S. cerevieiae were selected as examples of fungi that were not affected by the peptides as determined by the assay CFU. When using this technique, a good correlation was observed between the previous data using colony counts and the microscopic evaluation. The representative views of the confocal data (Figures 4A-4L) show that approximately 70-80% of all blastoconidia C. albicans stained green or yellowish green, indicate that most of the yeast cells were dead when incubated with the peptide CAP37 (20-44) nat. On the other hand the relatively inactive peptide (CAP37 (20-44) Ser26 / 42) showed only 30% of the blastoconidia to remain alive. The treatment of C. pseudotropicalie with the peptide CAP37 (20-44) nat showed an annihilation of more than 95% of the cell. Similar results were obtained with the inactive peptide (AP37 (20-44) Ser2642) confirming the results obtained with the colony forming unit assay. The studies carried out with C. glabrata and S. Cerevieiae imitated the data obtained with the test of the colony forming unit; approximately 10-15% of the cells were killed by the active peptide and virtually all the cells were viable in the presence of the inactive peptide. A control in which the yeast cells were incubated in the absence of the peptide had no effect on viability. UTILITY Peptides that can be used as anti-mycotic therapeutics in accordance with the present invention include peptides described herein as well as peptides described in the patent numbers of U.A.A. Nos. 6,107,460; 6,514,701; and 6,730,659; the specification of eachone of which is incorporated herein by reference in its entirety. As seen anywhere in the present, the invention contemplates in its preferred embodiments the use of CAP37 (20-44) na (linear, or cyclized between cysteine), CAP37 (20-44) ser26, CAP37 (20-44) ) ser42 and CAP37 (20-44) ser26 / 42 as antifungal treatments. The invention further comprises the use of peptides similar to peptides CAP37 (20-44) nat, CAP37 < 20-44) Ser26 CAP37 (20-44) ser42, and CAP37 (20-44) ser26 / 42 except where the three amino acids of the N term and the two amino acids of the C term are truncated (CAP37 (20-44) nat ( linear or cyclized between the cysteines), CAP37 (23-42) eer26, CAP37 (23-42) ser42, CAP37 and (23-42) Ser2642) • Alternaturalmente each of these three peptides substituted with serine may be substituted with threonine alternaturalmente , (ie, SEQ ID No. 9-14). In addition, the CAP37 peptide or peptide derivative used herein may comprise at least one of the following substitutions: phenylalanine replaced by tyrosine; glycine replaced by alanine; valine replaced by alanine, leucine or isoleucine; Alanine replaced by leucine, isoleucine, or valine; leucine replaced by alanine, isoleucine or valine; isoleucine replaced by valine, leucine or alanine, serine replaced by histidine, arginine or lysine; and threonine replaced by serine. As noted anywhere in the present, the peptide derivative may be a derivative of the peptide CAP37 (20-44) or CAP37 (23-42) modified as described above. In an alternative modality the peptide used may comprise CAP37 (120-146), ie, SEQ ID NO: 16. Alternatively, the complete CAP37 protein can be used in the antifungal treatment of the present invention. The present invention comprises a method of treating a fungal infection in a patient, subject or mammal, or prophylactically preventing a fungal infection in a patient, subject or mammal comprising administering to the patient, subject or mammal a therapeutically effective amount of a peptide described in the present. In addition, the peptide contemplated herein for use as an antifungal treatment may comprise a peptide having 20, 21, 22, 23, 24, or 25 residues and comprising the sequence (SEQ ID NO: 15). RH-X3-X4-X5-X6-X7-X8-X9-HX ?? -???? RX 3-X-X 4- -Xi6 -Xi8- i9-X20 where X3 and X3 is phenylalanine, tyrosine , arginine, lysine or histidine; X4 is selected from cysteine, serine, threonine, arginine, lysine or histidine; X5 and X6 are selected from glycine, alanine, arginine, lysine or histidine; X7 / Xii and Xi4 are selected from alanine, leucine, isoleucine, valine, arginine, leucine or histidine. Xg, X? and Xiß select from alanine, leucine, isoleucine and valine; Xiß is serine or threonine; X 9 is selected from serine, threonine, histidine, arginine and lysine; X20 is selected from cysteine, serine and threonine; R is arginine; H is histidine; and M is methionine, and wherein the peptide may comprise one, two, or three additional residues at the N-terminus and one or two additional residues at the C-terminus of the peptide and wherein in a modality for example X3-X7 could be arginine and Xn-X? They could be lysine. Any of the peptides described herein can be used alone or in combination with antifungal treatments. In any example of SEQ ID NO: 1-16 can be used alone or in combination as a "cocktail" of peptides, furthermore, when the peptides are conjugated to a polymer such as PEG, several of the peptides of SEQ ID NO: 1-16 can bind to the same PEG molecule. In addition, any of the peptides of the present invention can be dimerized, for example, to form homodimers or heterodimers, such as dimer CAP37 (20-44) nat, CAP37 dimer (20-44) Ser26 a dimer CAP37 (20 -44) ser42, or a dimer CAP37 (20-44) ser26-CAP37 (20-44) Ser42 for example. For intramolecular cyclization, a disulfide bridge is formed between the two cysteines. The dimerization of the peptides and the intramolecular cyclization between the thiol groups is well known in the art and a detailed explanation thereof is not made - needed in the present. However, an example of these procedures is shown in Technical Report TI-PEP05-0405 of Termo Electron 'Corp. 2005, included herein by reference in its entirety. The dimers could be linked through "intermolecular oxidation", for example, the thiol group attached to cys at position 26 could bind to the SH group of another peptide having a cys 26 giving a homodimer. Similarly one could cyclize the SH group in cys 42 with an SH group in a cys 42 of another peptide, giving a cys 42 homodimer. In an alternate third, the thiol groups between cys 42 and cys 26 could be linked to give a heterodimer. Other references showing the state of the art of cyclization and dimerization include J. Davies, J. Peptide Sci. 9: 471-501 (2003); P. Li and O. Soller. Dog. Top. In Med. Chem. 2: 325-341 (2002); and M. Hornef, et al., Nat. Immunol. 5 (8) 836-843 (2004) all of which are expressly incorporated by reference in their entirety. The present invention further comprises a DNA molecule having a nucleotide sequence that encodes a peptide having an amino acid sequence as defined in any of the amino acid sequences listed or described herein, in particular, those having residues of cysteine substituted at positions 26 or 42. The present invention comprises the use of peptides described herein and / or effective subunits of the both to treat fungal infections on the fly and to prophylactically treat an individual who may have a risk of such infection. The peptide used in the present invention, synthetically or recombinantly produced, can be used as a pharmaceutical composition when combined with a pharmaceutically acceptable carrier. Said composition may contain, in addition to the peptide and the carrier, diluents, fillers, salts, pH regulators, stabilizers, solubilizers, and other materials well known in the art. Suitable carriers, carriers and other components of the formulation are described, for example, in Remingtone 'Pharmaceutical Scienc-ee, (Mack Publishing Co., 1980 or latest edition). The term "pharmaceutically acceptable" means a non-toxic material that does not interfere with the effectiveness of the biological activity of the active ingredient (s). The characteristics of the carrier will depend on the administration route. The pharmaceutical composition of the invention may be in the form of a liposome in which the isolated peptide is combined, in addition to other pharmaceutically acceptable carriers, with amphiphatic agents such as lipids that exist in aggregated form such as micelle, insoluble monolayers, lipid crystals or Laminar layers in aqueous solution. Suitable lipids for the liposomal formulation include, without limitation, monoglycerides, diglycerides, sulfates, isolecithin, phospholipids, saponin, bile acids, and the like. The preparation of such liposomal formulations is within the skill of the person skilled in the art as described, for example, in the patent of US Pat. No. 4,235,871; U.A. Patent No. 4,501,728; U.A. Patent No. 4,837,028; and U.A. Patent No. 4,737,323, all of which are expressly incorporated herein by reference in their entirety. A therapeutically effective amount of a compound of the present invention refers to an amount effective in controlling, reducing or inhibiting a fungal infection. The term "control" is intended to refer to all procedures where there may be a deceleration, interruption, arrest, or arrest of the infection advance and does not necessarily indicate a total elimination of the symptoms of the infection. The term "therapeutically effective amount" further intends to define an amount that results from the improvement of any of the clinical parameters or symptoms characteristic of a fungal infection. The current dose will vary with the overall condition of the patient, the seriousness of the symptoms, and the contraindications. As used herein, the term "subject" or "patient" refers to a warm-blooded animal, in particular a mammal, that is afflicted with an infection mycotic It is understood that guinea pigs, dogs, cats, rats, mice, horses, goats, cattle, sheep, zoo animals, birds, primates and humans are examples of animals within the scope of the meaning of the term. A therapeutically effective amount of the compound used in the treatment described herein can be readily determined by the treating physician, as one skilled in the art, through the use of conventional techniques and observation of results obtained under analogous circumstances. In determining the therapeutically effective dose, a number of factors are set by the attending physician, including, but not limited to: mammalian spices; its size, age, and general health; the specific fungal disease or condition involved; the degree or implication the severity of the fungal disease or condition; the response of the individual subject; the particular compound administered; the mode of administration; the bioavailability characteristic of the preparation administered; the selected dose regimen; the use of the concomitant medication, and other relevant circumstances. A therapeutically effective amount of a compound of the present invention also refers to an amount of the compound that is effective in controlling or reducing fungal infection. A therapeutically effective amount of the The compositions of the present invention will generally contain sufficient active ingredient (ie, the peptide) to be distributed from about 0.1 μg / kg to about 100 mg / kg (active ingredient weight / patient's body weight). Preferably, the composition will distribute at least 0.5 μg / kg to 50 mg / kg and more preferably at least 1 μg / kg to 10 mg / kg. The practice of the method of the present invention comprises administering to a subject a therapeutically effective amount of the peptide in any suitable systemic or local formulation, in an amount effective to deliver the dosages listed above. A particularly preferred, effective dosage of the peptide to substantially inhibit fungal infection is 1 μg / kg to 1 mg / kg of the peptide. The dosage can be administered on a one-time basis or (for example) from 1 to 5 times per day or 1 or 2 times per week, or continuously via venous drip, depending on the desired therapeutic effect. As used herein, the term "therapeutically effective amount" means the total amount of each active component of the pharmaceutical composition or method that is sufficient to demonstrate a significant benefit to the patient, i.e., reduction of fungal infection. When an individual active ingredient is applied, administered alone, the term refers to that ingredient alone. When applied to a combination, the term refers to combined amounts of active ingredients that result in the therapeutic effect, whether administered in combination, serially or simultaneously. In practice of the method of treatment or use of the present invention, a therapeutically effective amount of the peptide composition is administered to a mammal having a fungal disease state. The peptide can be administered according to the method of the invention either alone or in combination with other therapies. The administration of the peptide used in the pharmaceutical composition or the practice of the method of the present invention can be carried out in a variety of conventional ways, including orally, through inhalation (eg, for fungal fungal infections), rectally, or through cutaneous, subcutaneous, intraperitoneal, vaginal, or intravenous injection. The oral formulations can be formulated in such a way that the peptide passes through a portion of the digestive system before being released, for example, it may not be released until it reaches the small intestine, or the colon. When a therapeutically effective amount of the peptide is administered orally, the peptide may be in the form of a tablet, capsule, powder, solution or elixir. When administered in the tablet form, the composition The pharmaceutical composition of the invention may additionally contain a solid carrier such as a gelatin or an adjuvant. The tablet, capsule and powder preferably contains between about 5 to 95% peptide. When administered in liquid form, a liquid carrier such as water, petroleum, animal or vegetable oils such as peanut oil, mineral oil, soy bean oil, or sesame oil, synthetic oils may be added. The liquid form of the pharmaceutical composition may further contain physiological saline, dextrose, or other saccharide solution, or glycols such as ethylene glycol, propylene glycol, or polyethylene glycol. When administered in liquid form, the pharmaceutical composition preferably contains from 0.005 to 95% by weight of the peptide. For example, 100-1000 mg of the active ingredient once or twice a day could be administered orally. For oral administration, the compounds can be formulated into solid or liquid preparations such as capsules, pills, tablets, dragees, fusions, powders, suspensions or emulsions. The solid unit dosage forms may be ordinary gelatin-type capsules containing, for example, surfactants, lubricants, and inert fillers such as lactose, sucrose, and corn starch, or they may be sustained release preparations.

In another embodiment, the compounds of this invention can be formed into tablets with conventional tablet bases such as lactose, sucrose, and corn starch in combination with binders, such as acacia, corn starch, or gelatin, disintegrating agents such as starch. of potato or alginic acid, or a lubricant such as stearic acid or magnesium stearate. Liquid preparations are prepared by dissolving the active ingredient in a pharmaceutically acceptable aqueous or non-aqueous solvent which may also contain suspending agents, sweetening agents, flavoring agents, and preservatives as is known in the art. For parenteral administration, the compounds can be dissolved in a physiologically acceptable pharmaceutical carrier and administered either as a solution or a suspension. Examples of suitable pharmaceutical carriers are water, saline, dextrose solutions, fructose solutions, ethanol, or oils of animal, vegetable or synthetic origin. The pharmaceutical carrier also has preservatives, and pH regulators as is known in the art. When a therapeutically etive amount of the peptide is administered by intravenous, cutaneous or subcutaneous injection, the peptide is preferably in the form of a parenterally acceptable solution or suspension. aqueous free of pyrogen. The preparation of such parenterally-acceptable peptide solutions has, due to pH, isotonicity, stability, and the like, and is within the scope of skill in the art. A preferred pharmaceutical composition for intravenous, cutaneous or subcutaneous injection should contain, in addition to the peptide, an isotonic vehicle such as sodium chloride injection, Ringer's injection, dextrose injection, dextrose and sodium chloride injection, citrate pH regulator, Lactated Ringer's injection pH 5.5, or other vehicle known in the art. The pharmaceutical composition of the present invention may also contain stabilizers, preservatives, pH regulators, antioxidants, or other additives known to those skilled in the art. As noted above, the compositions may also include an appropriate carrier. For topical use, any of the conventional excipients may be added in the formula of the active ingredients in a lotion, ointment, powder, cream, spray, or aerosol. For surgical implantation, the active ingredients can be combined with any of the well-known biodegradable or bioerodible carriers, such as polylactic acid and collagen formulations. Such materials may be in the form of solid implements, sutures, sponges, bandages, and the like. In any case, for local use of the materials, the active ingredients are usually present in the carrier or excipient in a weight ratio of about 1: 1000 to 1: 20,000, but is not limited to the ratios within this range. The preparation of compositions for local use is detailed in Remington's Pharmaceutical Sciences, latest edition, (Marck Publishing). In a preferred therapeutic method, the peptide composition is provided in an IV infusion on the scale of approximately 1 mg of the active ingredient / kg-4 mg / kg body weight once a day. As noted, preferred amounts and modes of administration are capable of being determined by one skilled in the art. One skilled in the art of preparation formulations can easily select the appropriate form and mode of administration depending on the particular characteristics of the selected compound, the infection to be treated, the state of the infection, and other relevant circumstances using the technology of formulation known in the art, described, for example, in Remington's Pharmaceutical Sciences, latest edition, (Marck Publishing). The pharmaceutical compositions can be manufactured using techniques known in the art. Typically the therapeutically effective amount of the peptide will be mixed with a pharmaceutically acceptable carrier.

The invention further includes a method for treating a topical fungal infection by topically applying an amount of peptide sufficient to treat the infection, for example 0.5-10%. Topical medication can take any number of external forms such as pastes, gels, creams, and ointments. Topical application can be achieved simply by preparing a solution of the compound to be administered, preferably using a solvent known to promote transdermal absorption such as ethanol or dimethyl sulfoxide (DMSO) with or without other excipients. Preferably topical administration will be achieved by using a patch either of the reservoir and of the porous membrane type or of a solid matrix variety. The amount of peptide in the pharmaceutical composition of the present invention will depend on the nature and severity of the condition to be treated, and on the nature of previous treatments the patient has experienced. Finally, the attending physician will decide the amount of peptide with which to treat each individual patient. Initially, the attending physician will preferably administer low doses of the peptide and observe the patient's response. The larger doses of the peptide can be administered until the optimal therapeutic effect is obtained for the patient, and at the point where the dosage can no longer be increased. Without wishing to be attached to a specific dosage, it is contemplated that the various pharmaceutical compositions used to practice the method of the present invention should contain about 0.1 mg to about 1000 mg of the peptide per kilogram of body weight per dose. The duration of intravenous therapy using the pharmaceutical composition of the present invention will vary, depending on the severity of the disease being treated and the potential idiosyncratic condition and response of each individual patient. It is contemplated that the duration of each application of the peptides will be on the scale of 1 to 2 hours and given once every 12 or 24 hours through continuous intravenous administration. Finally, the attending physician will decide the appropriate duration of intravenous therapy using the pharmaceutical composition of the present invention. Other antibiotics, intravenous fluids, cardiovascular and respiratory supports could also be provided if requested by the attending physician in a manner known to one skilled in the art. Mycotic diseases that can be treated by means of the peptide compositions described herein include but are not limited to Candida epp. , Saccharomycee cerevieiae, Hietoplaema capeulatum and other histoplasma species that cause histoplasmosis, Aepergillue fumigatue and other species (occurring mostly in the lung) cause Aspergillosis, and Cruptococcue neoformane (sometimes found in the lung but mostly in the central nervous system), which cause a disease known as cryptococcosis. Additional pharmaceutical methods can be used to control the duration of action of the peptide. Increased half-life and controlled-release preparations can be achieved through the use of polymers to conjugate, complex or absorb the peptide described herein. The controlled distribution and / or increased half-life can be achieved by selecting the appropriate macromolecules < for example, polysaccharides, polyesters, polyamino acids, homopolymers of polyvinylpyrrolidone, ethylvinylacetate, methylcellulose, or carboxymethylcellulose, and acrylamides such as N- (2-hydroxypropyl) methacrylamide and the appropriate concentration of macromolecules as well as the incorporation methods, with the order to control the release. Other possible methods useful in controlling the duration of action through controlled release and half-life preparations is the incorporation of the peptide molecule or its functional derivatives into particles of a polymeric material such as polyesters, polyamides, polyamino acids, hydrogels, poly (lactic acid), ethylene vinyl acetate copolymers, copolymer micelles of, for example, PEG and poly (1-aspartamide). The half-life of the peptides described herein can be extended by being conjugated to other molecules such as polymers using methods known in the art to form drug-polymer conjugates. For example, the peptide can be attached (eg, covalently) to inert polymer molecules known in the art, such as a polyethylene glycol (PEG) molecule in a method known as "pegylation". PEGylation can therefore extend the lifespan in vivo and thus the therapeutic effectiveness of the peptide molecule. PEGylation also reduces the potential antigenicity of the peptide molecule. PEGylation can also improve the subunit of the peptides thereby improving their therapeutic effect. The PEG used can be straight or branched chain. PEG molecules can be modified through functional groups, for example as shown in Harris et al., "Pegylation, A Novel Process for Modifying Phararmacokinetics", Clin Pharmacokinet, 2001: 40 (7); 539-551, and the amino group of the amino terminal residue of the peptide or an internal cysteine residue, or another amino acid having a linker group (eg, arginine, lysine, histidine, serine, threonine or methionine) which is therein can bind to it, where the PEG molecule can carry one or a plurality of one or more types of peptides, the peptide can carry more than one PEG molecule. By "pegylated peptide" is meant a peptide of the present invention having a polyethylene glycol moiety (PEG) covalently linked to an amino acid residue or linker group of the peptide. The PEG molecule can also be linked to the peptide through a linker comprising 1 to amino acids for example. By "polyethylene glycol" or "PEG" is meant a polyalkylene glycol compound or a derivative thereof, with or without coupling or derivatizing agents with coupling or activation portions (for example, with thiol, triflate, tresylate, azirdine, oxirane, or preferably with a portion of maleimide). Compounds such as PEG maleimido monomethoxy are illustrative or the activated PEG compounds of the invention. Other polyalkylene glycol compounds, such as polypropylene glycol, can be used in the present invention. Other suitable polymer conjugates include, but are not limited to, non-polypeptide polymers, charged or neutral polymers of the following types: dextrin, columnic acid or other carbohydrates based on polymers, biotin derivatives and dendrimers, for example. The term PEG also means that it includes other polymers of the class of polyalkylene oxides. PEG can be linked to any amino acid of the N-terminus of the peptide, and / or can be linked to a residue of amino acid in descending stream of the amino acid of the N-terminus such as lysine, histidine, tryptophan, aspartic acid, glutamic acid, serine, trionine, methionine and cysteine, for example, or other amino acids known to those skilled in the art. Pegylated cysteine peptides, for example, are created through the binding of polyethylene glycol to an SH group on a peptide cysteine residue. The chemically modified peptides contain at least one PEG portion, preferably at least two PEG portions, up to a maximum number of PEG portions linked to the peptide without abolishing the activity, for example, the PEG portion (s) bind to a residue of amino acid preferably at or near the N-terminus portion of the peptide. The PEG portion bound to the protein preferably is on the molecular weight scale of about 200 to 300,000 PM. Preferably the PEG portion will be in the form of about 1,000 to 8,000 PM, more preferably from 3,250 to 5,000 PM, more preferably at about 5,000 PM. The current number of PEG molecules covalently linked by chemically modified peptide of the invention can vary widely depending on the stability of the desired peptide (i.e., serum half-life). The peptide molecules contemplated for use in the present can be linked to PEG molecules using techniques shown, for example (but not limited to), in US Patent Nos. 4,179,337; 5,382,657; 5,972,885; 6,177,087; 6,165,509; 5,766,897; and 6,217,869; the specifications and figures of each of which is expressly incorporated herein by reference in its entirety. Alternatively, it is possible to trap the peptides in microcapsules prepared, for example, through coacervation techniques or through interfacial polymerization (for example, hydroxymethylcellulose, or gelatin-microcapsules and poly (methyltacrylate) microcapsules, respectively), in a system of colloidal drug distribution (eg, liposomes, albumin microspheres, microemulsions, nanoparticles, nanocapsules) or in macroemulsions. These techniques are described in the latest edition of Remington's Pharmaceutical Sciences. The U.A. Patent 4,789,734 describes methods for encapsulating biochemicals in liposomes and is therefore expressly incorporated by reference herein. Essentially, the material is dissolved in an aqueous solution, the appropriate phospholipids and lipids are added, along with surfactants if necessary, and dialyzed or sonicated material, as necessary. A review of the known methods is through G. Gregoriadis, Chapter 14. "Liposomes", Drug Carriere in Biology and Medicine, pp. 287- 341 (Academic Press, 1979). Microspheres formed of polymers or proteins are well known to those skilled in the art, and can be configured for passage through the gastrointestinal tract directly into the bloodstream. Alternatively, the agents can be incorporated and the microspheres, or microsphere composition, implanted for slow release over a period of time, in the scale from days to months. See, for example, the patents of E. U. A. Nos. 4,906,474; 4,925,673; and 3,625,214 which are incorporated by reference herein. When the composition to be used with an injectable material, it can be formulated in a conventional injectable carrier. Such carriers include pharmaceutically acceptable phosphate buffered pH regulated solutions which are preferably isotonic. For the reconstitution of a lyophilized product according to this invention, a sterile diluent may be used, which may contain materials generally recognized for physiological approach conditions and / or as required by governmental regulation. In this regard, the sterile diluent may contain a pH regulating agent to obtain a physiologically acceptable pH, such as sodium chloride, saline, phosphate buffered pH, and / or other substances that are physiologically acceptable and / or safe for use. In general, The material for intravenous injection in humans must conform to the regulations established by the Food and Drug Administration, which is available to experts in the field. The pharmaceutical composition may also be in the form of an aqueous solution containing several of the same substances as described above for the reconstitution of a lyophilized product. The compounds may also be administered as pharmaceutically acceptable acid addition salts or bases, formed through the reaction with inorganic acids such as hydrochloric acid, hydrobromic acid, perchloric acid, nitric acid, thiocyanic acid, sulfuric acid, and phosphoric acid, and organic acids such as formic acid, acetic acid, propionic acid, glycolic acid, lactic acid, pyruvic acid, oxalic acid, malonic acid, succinic acid, maleic acid and fumaric acid, or through reaction with an inorganic base such as hydroxide of sodium, ammonium hydroxide, potassium hydroxide, and organic bases such as mono-, di-, trialkyl and arylamines and substituted ethanolamines. As mentioned above, the compounds of the invention can be incorporated into pharmaceutical preparations that can be used for therapeutic purposes. However, the term "pharmaceutical preparation" is provided in the broader sense herein to include preparations containing a peptide composition according to this invention, used not only for therapeutic purposes but also for reagent or diagnostic purposes as is known in the art, or for tissue culture. The pharmaceutical preparation intended for therapeutic use should contain a "pharmaceutically acceptable amount" or "therapeutically acceptable amount" of a peptide, i.e., that amount necessary for preventive or curative health measures. If the pharmaceutical preparation is to be used as a reagent or diagnostic, then it must contain reagent amounts or diagnosis of a peptide. All of the test methods enumerated herein are convenient within the skill of one skilled in the art, given the teachings provided herein. While the invention is described herein in connection with certain embodiments in such a way that aspects thereof can be more readily understood and appreciated, it is not intended that the invention be limited to these particular embodiments. On the contrary, it is intended that all alternatives, modifications and equivalents are included within the scope of the invention as defined herein. In this way the examples described above, which include the preferred modalities, will serve to illustrate the practice of this invention, it being understood that the particulars are shown by way of example and for the purpose of illustrating the explanation of the preferred embodiments of the present invention only and are presented in order to also provide the principles and conceptual aspects of the invention. invention. Changes may be made in the formulation of the various compositions described herein or in the steps or sequence of steps of the methods described herein without departing from the spirit and scope of the invention as described herein. All references, articles and patent applications cited herein are hereby expressly incorporated by reference in their entirety. References cited Brackett, DJ. , M.R. Lerner, M.A. Lacquement, R. He, and H.A. Pereira 1997. A eynthetic lipopolysaccharide-binding peptide based on the neutrophil-derived protein CAP37 prevents endotoxin-induced responses in conscious rats. Infect. Immun. 65: 2803-2811. Clark, T.A. and R.A. Hajjeh 2002. Recent trends in the epidemiology of invasive mycoses. Curr. Opin. Infect. Dis. 15: 569-574. Dale, B.A. 2000. Periodontal epithelium: a newly recognized role in health and disease. Periodontal 30: 70-78. Dale, B.A., and S. Krisanaprakomkit. 2001. Defensin Antimicrobial peptides in the oral cavity. J. Oral Pathol. 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Claims (24)

1. CLAIMS Having described the invention as above, the content of the following claims is claimed as property: 1. A method for treating or inhibiting a fungal infection in a subject, characterized in that it comprises: administering a therapeutically effective amount of a peptide having 20- 25 amino acids and comprises SEQ ID NO: 15 or SEQ ID NO: 5 or a peptide derivative of SEQ ID NO: 5 wherein in the peptide derivative at least one of the cysteine residues is substituted with a serine or threonine residue .
2. 2. The method according to claim 1 characterized in that the peptide derivative comprises SEQ ID NO: 5 with a cysteine residue for the most part of the N term substituted with serine or threonine.
3. 3. The method according to claim 1 characterized in that the peptide derivative comprises SEQ ID NO: 5 with a cysteine residue for the most part of the C term substituted with serine or threonine.
4. 4. The method according to any of claims 1-3, characterized in that the peptide derivative of SEQ ID NO: 5 further comprises at least one of the substitutions comprising: a phenylalanine replaced with tyrosine; a glycine replaced with alanine; a valine replaced with alanine, leucine, or isoleucine; an alanine replaced with leucine, isoleucine or valine; a leucine replaced with alanine, isoleucine or valine; an isoleucine replaced with valine, leucine or alanine; a serine replaced with histidine, arginine, or lysine; and a threonine replaced with serine.
5. 5. The method according to any of claims 1-4, characterized in that the fungal infection treated or inhibited is caused by a Candida spp. , Saccharomyces cerevisiae, Histoplasma epp. , Aspergillus epp. , and Cryptococcue.
6. 6. The method according to any of claims 1-5, characterized in that the subject is a mammal.
7. The method according to any of claims 1-6, characterized in that the peptide has one to three additional amino acids at one end of the N terminus of the peptide and one to two additional amino acids at the terminus of the C terminus of the peptide.
8. 8. The method according to any of claims 1-7, characterized in that the peptide comprises the sequence (SEQ ID NO: 15): R-H-X3-X4-X5-X6-X7-Xβ-X9-H-X? ? _R-X? 3-X? -M-Xig-X? 7-Xifl-X19-X2O wherein: X3 is phe, tyr, arg, lys or his; X is cys, ser, thr, arg, lys or his; X5 is gly, ala, arg, lys or his; Xß is' gly, ala, arg, lys or his; X8 -X9, X? 7, and Xiß are ala, leu, ile or val; X7 / X11 and X14 are wing, leu, ile, val, arg, lys, or his; X? 3 is phe or tyr; Xi6 is to be or thr; X19 is to be, thr, his, arg or lys; X20 is to be, cys or thr; R is arg; H is his; and M is met.
9. 9. The method according to claim 8, characterized in that X is cys and X20 is ser or thr.
10. 10. The method according to claim 8 characterized in that X20 is c and X is ser or thr.
11. 11. The method of compliance with any of the claims 1-10, characterized in that the peptide is pegylated.
12. The method according to claim 11, characterized in that the peptide is covalently pegylated to a polyethylene glycol molecule through a linker molecule comprising from 1 to 15 amino acids.
13. 13. The use of a peptide for the preparation of a medicament for treating or inhibiting a fungal infection in a subject, wherein the peptide has 20-25 amino acids and comprises SEQ ID NO: 15 or SEQ ID NO: 5 or a derivative of peptide of SEQ ID NO: 5, wherein at least one of the cysteine residues in the peptide derivative is substituted with a serine or threonine residue.
14. The peptide of the medicament according to claim 13, characterized in that the cysteine residue for the most part at the N-terminus of the peptide derivative SEQ ID NO: 5 is substituted with serine or threonine.
15. 15. The peptide of the medicament according to claim 13, characterized in that the cysteine residue for the most part at the C terminus of the peptide derivative of SEQ ID NO: 5 is substituted with serine or threonine.
16. 16. The peptide of the medicament according to any of claims 13-15, characterized in that the peptide derivative of SEQ ID NO: 5 further comprises at least one of the substitutions comprising: a phenylalanine replaced with tyrosine; a glycine replaced with alanine; a valine replaced with alanine, leucine, or isoleucine; an alanine replaced with leucine, isoleucine or valine; a leucine replaced with alanine, isoleucine or valine; an isoleucine replaced with valine, leucine alanine; a serine replaced with histidine, arginine, or lysine; and a threonine replaced with serine.
17. 17. The peptide of the medicament according to any of claims 13-16, characterized in that the fungal infection treated or inhibited is caused by a Candida epp. , Saccharomyces cerevieiae, Hietoplaema epp. , Aepergillue epp. , and Cryptococcue.
18. 18. The peptide of the medicament according to any of claims 13-17, characterized in that the subject is a mammal.
19. 19. The peptide of the medicament according to any of claims 13-18, characterized in that the peptide has one to three additional amino acids at one end of the N terminus of the peptide and one to two amino acids 5 additional at the end of the C-terminus of the peptide.
20. 20. The peptide of the medicament according to any of claims 13-19, characterized in that the peptide comprises the sequence (SEQ ID NO: 15): RH-X3-X-X5-X6-X7-X8-X9-HX ?? -RX? 3-X? - -Xi6-X? 7-Xi8-Xi9- X20 Where: X3 is phe, tyr, arg, lys or his; X is cys, ser, thr, arg, lys or his; X5 is gly, ala, arg, lys or his; X6 is gly, ala, arg, lys or his; X8 -X9, X? , and Xiß are ala, leu, ile or val; 7 / X11 and X14 are wing, leu, ile, val, arg, lys, or his; X? 3 is phe or tyr; Xi6 is to be or thr; X19 is to be, thr, his, arg or lys; X2o is to be, cys or thr; R is arg; H is his; and M is met.
21. 21. The peptide of the medicament of claim 20, characterized in that X is cys and X20 is ser or thr.
22. 22. The peptide of the medicament of claim 20, characterized in that X20 is cys and X is ser or thr.
23. 23. The peptide of the medicament according to any of claims 13-22, characterized in that the peptide is pegylated. The peptide of the medicament of claim 23 characterized in that the peptide is covalently pegylated to a polyethylene glycol molecule through a linker molecule comprising from 1 to 15 amino acids.