NL1008745C2 - New peptide conjugates useful for treating yeast, fungal and bacterial infections and tumors - Google Patents

Abstract

Conjugates of an antifungal peptide and a mucoadhesive polymer are new. The peptide has a domain of 10 - 25 amino acids in which most of the amino acids in one half of the domain are positively charged amino acids and most of the amino acids in the other half of the domain are uncharged amino acids.

Description

J

THERAPEUTICALLY ACTIVE COMPOUND FOR TREATMENT OF YEAST AND / ON MOLD INFECTIONS

The invention relates to a therapeutically active compound for the treatment of yeast and / or fungal and / or bacterial infections in the oral cavity, in the gastrointestinal tract, on the skin and in the vagina.

Candida species are oral commensals, but can cause infections under certain circumstances. This is called candidiasis. People particularly susceptible to oral candidiasis are, for example, denture wearers, immunocompromised patients and patients with decreased saliva production (xerostomia), for example due to Sjögren's syndrome or radiation therapy. Candida can further lead to vaginal candidiasis and lesions on the skin.

Three main categories of antifungal agents are currently available for the treatment of oral candidiasis: polygenes, such as amphotericin B, azoles, such as fluconazole, and disinfectants, such as chlorhexidine. A disadvantage of these classic antibiotics is the emergence of resistance. Resistances to amphotericin B and fluconazole have already been reported for Candida. Very resistant strains are also known of the bacteria Staphylococcus aureusf that can cause skin infections.

An additional problem, which arises in particular in the treatment of the oral cavity, is that the available amount of the antifungal or antibacterial agent falls below the effective concentration by swallowing and dilution by saliva. Similar availability problems can also arise in the other forms of Candida or bacterial infection, for example due to proteolytic degradation by micro-organisms from the oral flora.

The object of the invention is to solve both problems at the same time.

This is achieved by the invention by a therapeutically active compound for treatment of yeast and / or fungal infections of the oral cavity, gastrointestinal tract, skin and vagina, comprising a peptide having anti-fungal activity, consisting of a amino acid chain, which contains a domain of 10 to 25 amino acids, the majority of the amino acids of one half of the 5 domain being positively charged amino acids and the majority of the amino acids of the other half of the domain being uncharged amino acids linked to a mucoadhesive (co) polymer. The presence of the mucoadhesive (co) polymer ensures a longer residence time in the mouth. The mucoadhesive (co) polymer is preferably selected from xanthan gum, polyacrylic acid, cellulose derivatives, carrageenans and derivatives thereof, chitin and derivatives thereof, such as chitosan, scleroglucan and gums, such as guar gum, locust bean gum, etc. Also synthetic polymers with carboxyl groups or amine groups are suitable for this. Such mucoadhesive (co) polymers have the advantages that they increase and improve the bioavailability of the drug, that the agent has to be applied less often due to the longer residence time and that there is an optimal contact with the (co) polymer adsorbing surface.

In the compound of the invention, the peptides, which are cationic, have a Candida albicans-killing effect and also inhibit pathogenic microorganisms such as Streptococcus mutans and a methylline-resistant Staphylococcus aureus (MRSA). It has been found that the cationic peptides do not cause resistant mutants after 20 passages at a concentration near the MIC value. Furthermore, they work synergistically with classic antibiotics.

The action of the peptides is probably based on the fact that the cationic peptides in the negatively charged cell wall form amphipathic α-helices, which act as pores through which, inter alia, K + ions leak out of the cell.

The structure of the peptides which are part of the therapeutic compounds according to the invention has a number of variations. First, the domain; 100 874 δ * 3 form an α-helix, of which at least a majority of positions 1, 2, 5, 6, 9 (12, 13, 16, 19, 20, 23 and 24) contain a positively charged amino acid, position 8 is a positive or an uncharged amino acid and at least a majority of positions 3, 4, 7, 10, (11, 14, 15, 17, 18, 21, 22, 25) contain an uncharged amino acid. These peptides have a lateral amphipathicity, i.e. a maximum hydrophobic moment at 100 °. Simply put, these peptides are hydrophobic on the left and hydrophilic on the right or vice versa. These peptides are referred to herein as "type I".

Furthermore, the domain can form an α-helix, of which at least a majority of positions 1, 2, 5, 6, 9 (12, 13, 16, 19, 20, 23 and 24) contain an uncharged amino acid 15, position 8 a is positive or an uncharged amino acid and at least a majority of positions 3, 4, 7, 10, (11, 14, 15, 17, 18, 21, 22, 25) contain a positively charged amino acid. These peptides have a lateral amphipathicity, ie a maximum hydrophobic moment at 100 °. Simply put, these peptides are hydrophobic on the right and hydrophilic on the left or vice versa. These peptides are referred to herein as "type II" and are basically the mirror images of type I peptides.

In addition, the domain may form an α-helix, wherein at least a majority of positions 1 to 6 (or 7 or 8 or 9 or 10 or 11 or 12) contain an uncharged amino acid, and at position 7 (or 8 or 9 or 10 or 11 or 12 or 13) to 25 a positively charged amino acid 30. These peptides have a longitudinal amphipathicity, i.e. a minimal hydrophobic moment at 100®. These peptides are hydrophobic on their "top" and hydrophilic on their "bottom". Such peptides are designated "type III".

Conversely, the domain may form an e-helix, wherein at least a majority of positions 1 to 6 (or 7 or 8 or 9 or 10 or 11 or 12) contain a positively charged amino acid, and at position 7 (or 8 or 9 or 10 10 0874 5 · «4 or 11 or 12 or 13) through 25 an uncharged amino acid. These peptides also have a longitudinal amphipathicity, and thus a minimal hydrophobic moment at 100 °. These peptides are hydrophobic on their "bottom" and hydrophilic on their "top". Such peptides are referred to as "type IV".

Finally, the domain can form a so-called β-beach and at least a majority of positions 1, 3, 5, 7, 9 (11, 13, 15, 17, 19, 21, 23 and 25) are loaded positively. amino acid, and in at least a majority of positions 2, 4, 6, 8, 10, (12, 14, 16, 18, 20, 22, 24) an uncharged amino acid. Such a β-strand is laterally amphipathic and has a maximum hydrophobic moment at 180 °. The β-strand structure is flatter than the α-helix and is simply hydrophobic on the left and hydrophilic on the right or vice versa. These are "type V" peptides.

The positively charged amino acids are preferably selected from the group consisting of ornithine 20 (O), lysine (K), arginine (R) and histidine (H), while the uncharged amino acids are preferably selected from the group consisting from the aliphatic amino acids glycine (G), alanine (A), valine (V), leucine (L), isoleucine (I), the amino acids with a dipolar side chain 25 methionine (M), asparagine (N), glutamine (Q) , serine (S), threonine (T), the amino acids with an aromatic side chain phenylalanine (F), tyrosine (Y), tryptophan (W). Amino acids on the boundary between hydrophilic and hydrophobic can be selected from either group or from the other amino acids.

In principle, hardly any difference in activity can be observed when one of the positive amino acids and / or one of the uncharged amino acids has been replaced by a random amino acid. The majority of the 35 positively charged amino acids is therefore preferably the total number of positively charged amino acids minus 1, and the majority of the uncharged amino acids is preferably the total number of uncharged amino acids minus 1.

1003745 ^ 5

The domain can be part of a larger peptide, but it can also make up the entire peptide itself. When the domain is part of a larger peptide, the C-terminal and / or N-terminal 5 amino acids, which are then additionally present, may be random amino acids.

Particular preference is given to the following type I peptides: KRLFKELKFSLRKY (peptide 3) KRLFKKLLFSLRKY (peptide 10)

A preferred type III peptide has the following amino acid sequence: LLLFLLKKRKKRKY (peptide 11). The peptides which are part of the compounds of the invention may further contain modifications. These modifications are, for example, an N-terminal amidation, for example with acetic anhydride, or an alternative cleavage of the synthetic resin, whereby the C-terminus is modified. For the latter, a replacement of the C-terminal carboxylic acid group with an amide, ester, ketone, aldehyde or alcohol group can be envisaged. The possibility of modification depends on the end to which the (co) polymer is attached. This will usually be the N-terminal end. This is then no longer free for modification. For example, peptides with such a modification are: KRLFKELKFSLRKY amide (peptide 12) KRLFKELFSLRKY amide (peptide 13)

In addition to single peptides, oligomers can also be used. These are preferably linear oligomers of the peptides of the invention. The 100874 5¾ 6 link can be either head-to-head, tail-tail or tail-head, either by direct synthesis in vitro or in vivo and by translation from a transgenic cDNA library, or by post-synthetic enzymatic coupling. The formation of a transmembrane pore requires a minimum peptide length. Oligomers of the peptides according to the invention are doubly long and therefore, in principle, better able to span the entire phospholipid bilayer of the bacterial or eukaryotic cell membrane at once. This could further improve the action of the peptide. In addition, elongation of the peptides stabilizes the helical conformation. Usually a spacer has to be inserted. In direct synthesis of back-to-back coupled oligomers, a custom spacer can be incorporated using a chain of unnatural amino acids of appropriate length, for example, B-alanine, γ-aminobutyric acid, e-aminocaproic acid, etc. heterodifunctional coupling reagents, such as those commercially available to couple peptid-de-antigens to carrier proteins (e.g. 1-ethyl-3- [3-dimethylaminopropyl] carbodiimide (EDC), ifl-maleimidobenzoyl) -Ji-hydroxysuccinimide ester (MBS ), N-succinimidyl 3- [pyridyldithio] propionate (SPDD) etc. are used to make linear oligomers with an interposed spacer. Trivalent amino acids such as aspartic acid (D), glutamic acid (E), ornithine (O), lysine (K), serine (S), cysteine can be used for head-to-head and tail-to-tail couplings. Such oligomers include, for example: 30 KRKFHEKHHSHRGYC-CY GRH SHHKEHFKRK (peptide 14) YGRHSHHKEHFKRKC-CKRKFHEKHHSHRGY (peptide 15) ®N, * N- (KRKFHEKHHSHRGY) 2K-amide (peptide 16) ®LF * RLF * (peptide 17) eN, * N- (KRLFKKLKFSLRKY) 2K-amide (peptide 18) 35 Peptides 14 and 15 are obtained by synthesis of peptide 2 with an additional C-terminal and N-terminal cysteine, respectively, after which the oligomer is obtained by air oxidation. Peptides 16, 17 and 18 were obtained 1Q0GV4C, 7 using the Multi-Antigen Peptide (MAP) strategy, where the first amino acid is a lysine with Fmoc protection on both the α and e-amino groups as the first amino acid on the synthetic resin was used, thereby simultaneously synthesizing two identical amino acid chains (peptides 2, 3 and 9) on one lysine molecule.

The peptides described herein have little or no hemolytic activity in physiological buffers, such as PBS (phosphate buffered saline). Low activity against erythrocytes of human origin is indicative of low toxicity. This selectivity is essential for the use of these peptides as antibiotics.

All of the peptides described above can be covalently coupled to a mucoadhesive (co) polymer used for the treatment of yeast and / or mildew infections of the oral cavity, gastrointestinal tract, skin and vagina. Particularly preferred, however, is a peptide having the amino acid sequence KRLFKELKFSLRKY with xanthan gum covalently linked to it. An example of a method of covalently coupling xanthan gum to the peptides is given in Example 2.

The compounds of the invention can be used in or as an antifungal agent and in or as an anti-yeast infection agent. Their activity will be further illustrated in the accompanying examples.

The invention therefore further relates to the compounds for use in the treatment of yeast and / or fungal infections.

Also part of the invention is the use of the compounds for the manufacture of a medicament for the treatment of fungal infections and / or yeast infection.

The compounds of the invention can be used in various pharmaceutical administration forms. Particular preference is given to spray, ointment, gel and lozenges.

The invention is further illustrated in the accompanying examples, which are given for illustrative purposes only and not to limit the invention in any way.

EXAMPLES EXAMPLE 1 Peptide Synthesis

Peptides that are part of the compounds of the invention were chemically synthesized as described by Van 't Hof et al. (1991) and Helmerhorst et al. (1997). Peptides were synthesized using the T-bag method, which was adapted for 9-fluorenylmethoxycarbonyl ((Fmoc) chemistry). p-Ben-zyloxybenzyl alcohol resins to which the first N-Fmoc-protected amino acids have already been coupled were included in the T-bags. The coupling reactions were performed in N. N-dimethylformamide. After completion of the amino acid chain, it was cleaved from the resin and the side chain protecting groups were simultaneously removed with a mixture of 5% thioanisole, 5% phenol, 5% water and 85% trifluoroacetic acid. Purity analyzes were performed by reverse phase HPLC and showed a single peak with only few impurities (less than 5%).

All peptides were dissolved in 10 mM potassium phosphate buffer (PPB), pH 7.0, to a concentration of 2 mg / ml and stored at -80 ° C. The final pH of the stock solution was 6.0. The exact peptide concentrations used in the antibacterial assays were determined by amino acid analysis.

Table 1 lists the peptides 23 to 13 made in this manner. Peptides 1 and 2 from this table represent the histatin 5 and the C-terminal part thereof, respectively. Bold amino acids are changes from peptide 2.

1008745a 9

Table 1

PePtide sequence

1 DSHAKRHHGYKRKFHEKHHSHRGY

2 KRKFHEKHHSHRGY

5 3 KRLFKELKFSLRKY

4 KRLFKELLFSLRKY

5 KRLFKELKKSLRKY

6 KRLFKELLFSLRKY

7 OOLFOELLOSY

10 8 OOLFOELLOSLOOY

9 KRLFKKLKFSLRKY

10 KRLFKKLLFSLRKY

11 LLLFLLKKRKKRKY

12 KRLFKELFSLRKY amide 15 13 KRLFKELLFSLRKY amide

14 KRKFHEKHHSHRGYC-CYGRHSHHKEHFKRK

15 YGRHSHHKEHFKRKC-CKRKFHEKHHSHRGY

16 eN, fN- (KRKFHEKHHSHRGY) 2K-amide 17 ®N, "N- (KRLFKELKFSLRKY) 2K-amide 20 18" "," Ν- (KRLFKKLKFSLRKY) 2K-amide EXAMPLE 2

Manufacture of Xanthan Gum Peotide Conjugates Peptide 3 from Table 1 was covalently coupled to xanthan gum (Keltrol ™, Keko Inc., London) carbon black using 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDC) (Sigma, St. Louis, MO, USA), as described by Hattori et al., J. Food Sci. 61: 1171-1176 (1996).

1000745ë 10

Xanthan gum and peptide 3 were mixed and dissolved in water. The pH was adjusted to 4.75. EDC was added over a period of 30 minutes, after which the whole was incubated under room stirring for 5 hours at room temperature. The pH was kept at 4.75. The reaction was stopped by adding glycine while the pH was kept at 4.75. 10 mM HCl was used to adjust the pH. The final concentration of xanthan gum was 21.4 mM, of peptide 3 0.539 mM or 0.0539 mM and of EDC 52.1 mM, 21.4, 3.77 mM, 0.539 mM or 5.21 mM. The final glycine concentration was always 53.3 mM.

EXAMPLE 3 Analysis of the formed conjugates

The viscosity and elasticity of xanthan gum and the conjugate formed was determined using an oscillating capillary rheometer, Vilastic 3 Viscoelasticity Analyzer (Vilastic Scientific Inc. Austin TX, USA), as described by Van der Reijden et al., Arthritis & Rheumatism 39, 57-63 (1996).

The lowest concentrations of EDC (0.539 mM and 3.77 mM) resulted in an elasticity and viscosity comparable to that of xanthan gum alone. Higher concentrations reduced elasticity and viscosity.

Coupling efficiency was determined on SDS-PAGE. The higher the EDC concentration, the higher it was.

Based on the above it is quite easy for the expert to determine the optimal EDC concentration.

"100874 5 * * 11 EXAMPLE 4

Fungicidal activity

The antifungal activity of the conjugate was determined as described by Helmerhorst et al., Biochem. J. 326, 39-45 (1997). 25 µl of the solutions to be tested were diluted twice 8 times and then 25 µl Candida albicans ATCC 10231 was added. C. albicans had grown on Sabouraud dextrose agar (SDA, Oxoid, Hants, UK) at 30eC and diluted in 20 mM PPB, pH 7.0 to 107 colony forming units (cfu) per ml.

After 1 hour incubation at 37 ° C, the number of surviving cells was determined by diluting 100 times in PBS and 25 µl was plated on Sabourad dextrose agar. The percentage of viability was calculated by: number of viable cells _ * 100% number of viable cells at the lowest peptide concentration

The IC50 values were determined from the percent viability versus concentration graph.

Table 2 lists the tested solutions and their antifungal activity.

Table 2 Solution IC50 value (^ g / ml) 0.539 mM peptide 3 5 21.4 mM xanthan gum + 37.5 0.539 mM peptide 3 + 53.3 mM glycine 30 conjugate according to the invention from: 21.4 mM xanthan gum + 0.539 mM peptide 3 + 52.1 mM EDO + 35 53.3 mM glycine 1008745 * 12

From the above, it appears that the conjugate is less active than the free peptide, but still exhibits ample activity for practical application. In addition, the action of the xanthan gum 5 may compensate for the reduced effect by providing longer bioavailability.

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100874 5J

Claims (39)

1. J A therapeutically active compound for the treatment of yeast and / or fungal and / or bacterial infections of the oral cavity, skin and vagina, comprising a peptide with antifungal activity, consisting of a 5 amino acid chain, which contains a domain of 10 to 25 amino acids, wherein the majority of the amino acids of one half of the domain are positively charged amino acids and the majority of the amino acids of the other half of the domain are uncharged amino acids linked to a mucoadhesive (co) polymer. 2. Therapeutically active compound according to claim 1, characterized in that the mucoadhesive (co) polymer is selected from xanthan gum, polyacrylic acid, cellulose derivatives, carrageenans and derivatives thereof, chitin and derivatives thereof, such as chitosan, scleroglucan and gums, such as guar gum, locust bean gum, etc. Synthetic polymers with carboxyl groups or amine groups are also eligible. Therapeutically active compound according to claim 1 or 2, characterized in that the domain of the peptide forms an α-helix and at least a majority of positions 1, 2, 5, 6, 9 (12, 13, 16 , 19, 20, 23 and 24) a positively charged amino acid, in position 8 a positive or an uncharged amino acid and in at least 25 a majority of positions 3, 4, 7, 10, (11, 14, 15, 17, 18 , 21, 22, 25) contains an uncharged amino acid. Therapeutically active compound according to claim 3, characterized in that the positively charged amino acids of the peptide domain are selected from the group consisting of ornithine (O), lysine (K), arginine (R) and histidine (H). Therapeutically active compound according to claims 2-4, characterized in that the uncharged amino acids of the domain of the peptide are selected from the group consisting of the aliphatic amino acids glycine (G), alanine (A), valine ( V), leucine (L), isoleucine (I), the 1G 0 87 4 6 * 1 amino acids with a dipolar side chain methionine (M), asparagine (N), glutamine (Q), serine (S), threonine (T) , the amino acids with an aromatic side chain phenylalanine (F), tyrosine (Y), tryptophan (W). Therapeutically active compound according to claims 2-5, characterized in that the majority of the positively charged amino acids of the peptide domain is the total number of positively charged amino acids minus 1. Therapeutically active compound according to claims 2-6, characterized in that the majority of the uncharged amino acids of the domain of the peptide is the total number of uncharged amino acids minus 1. 8. Therapeutically active compound according to claims 2-7, characterized in that the domain constitutes the entire peptide. Therapeutically active compound according to claims 2-8, characterized in that the domain of the peptide has the following amino acid sequence: Therapeutically active compound according to claims 2-8, characterized in that the domain of the peptide has the following amino acid sequence: KRLFKELLFSLRKY (peptide 4). Therapeutically active compound according to claims 2-8, characterized in that the domain of the peptide has the following amino acid sequence: KRLFKELKKSLRKY (peptide 5). 12. Therapeutically active compound according to claims 2-8, characterized in that the domain of the peptide has the following amino acid sequence: KRLFKELLKSLRKY (peptide 6). Therapeutically active compound according to claims 2-8, characterized in that the domain of the peptide has the following amino acid sequence: OLOLFOELOSLOOY (peptide 7). Therapeutically active compound according to claims 2-8, characterized in that the domain of the peptide has the following amino acid sequence: OOLFOELLOSLOOY (peptide 8). Therapeutically active compound according to claims 2-8, characterized in that the domain of the peptide has the following amino acid sequence: KRLFKKLKFSLRKY (peptide 9). Therapeutically active compound according to claims 2-8, characterized in that the domain of the peptide has the following amino acid sequence: KRLFKKLLFSLRKY (peptide 10). Therapeutically active compound according to claim 1 or 2, characterized in that the domain of the peptide forms an α-helix and at least a majority of positions 1 to 6 (or 7 or 8 or 9 or 10 or 11 or 12) contains an uncharged amino acid, and at position 7 (or 8 or 9 or 10 or 11 or 12 or 13) through 25 a positively charged amino acid. Therapeutically active compound according to claim 1 or 2, characterized in that the domain of the peptide forms an α-helix and at least a majority of positions 1 to 6 (or 7 or 8 or 9 or 10 or 11 or 12) contains a positively charged amino acid, and at position 7 (or 8 or 9 or 10 or 11 or 12 or 13) through 25 an uncharged amino acid. Therapeutically active compound according to claim 17 or 18, characterized in that the positively charged amino acids of the peptide domain are selected from the group consisting of ornithine (0), lysine (K), arginine (R) and histidine (Η). A therapeutically active compound according to claim 17, 18 or 19, characterized in that the uncharged amino acids of the peptide domain are selected from the group consisting of the aliphatic amino acids glycine (G), alanine (A), valine (V), leucine (L), isoleucine (I), the amino acids with a dipolar side chain methionine (M), asparagine (N), glutamine (Q), serine 100874 5 * (S), the threonine (T), the amino acids with an aromatic side chain phenylalanine (F), tyrosine (Y), tryptophan (W). KRLFKELKFSLRKY (peptide 3). Therapeutically active compound according to claims 17-20, characterized in that the majority of 5 positively charged amino acids of the peptide domain is the total number of positively charged amino acids minus 1. Therapeutically active compound according to claims 17-21, characterized in that the majority of 10 uncharged amino acids of the peptide domain is the total number of uncharged amino acids minus 1. Therapeutically active compound according to claims 17-22, characterized in that the domain constitutes the entire peptide. Therapeutically active compound according to claims 17 and 19-23, characterized in that the domain of the peptide has the following amino acid sequence: LLLFLLKKRKKRKY (peptide 11). 25. Therapeutically active compound according to claim 1, characterized in that the domain of the peptide forms a so-called β-strand and in at least a majority of positions 1, 3, 5, 7, 9 (11, 13, 15, 17, 19, 21, 23 and 25) contains a positively charged amino acid, and in at least a majority of positions 2, 4, 6, 8, 10, 25 (12, 14, 16, 18, 20, 22, 24) an uncharged amino acid. A therapeutically active compound according to claim 25, characterized in that the positively charged amino acids of the peptide domain are selected from the group consisting of ornithine (O), lysine (K), arginine (R) and histidine (H). A therapeutically active compound according to claim 25, characterized in that the uncharged amino acids of the peptide domain are selected from the group consisting of the aliphatic amino acids glycine (G), alanine (A), valine (V) , leucine (L), isoleucine (I), the amino acids with a dipolar side chain methionine (M), asparagine (N), glutamine (Q), serine (S), threonine (T), 100874 5 «the amino acids with an aromatic side chain phenylalanine (F), tyrosine (Y), tryptophan (W). Therapeutically active compound according to claims 25-27, characterized in that the majority of the positively charged amino acids of the peptide domain is the total number of positively charged amino acids minus 1. 29. A therapeutically active compound according to claims 25-28, characterized in that the majority of the uncharged amino acids of the peptide domain is the total number of uncharged amino acids minus 1. A therapeutically active compound according to claims 25-29, characterized in that the domain constitutes the entire peptide. A therapeutically active compound according to claims 1-30, wherein the C-terminal carboxylic acid group is replaced by an amide, ester, ketone, aldehyde or alcohol group. 32. A therapeutically active compound according to claim 1 or 2, which compound consists of a peptide having the amino acid sequence KRLFKELKFSLRKY with xanthan gum covalently coupled thereto. 33. A therapeutically active compound according to any one of claims 1-32 for use in the treatment of yeast and / or fungal and / or bacterial infections of the oral cavity, skin and vagina. 34. Use of the therapeutically active compound according to claims 1-32 for the manufacture of a medicament for the treatment of yeast and / or fungal and / or bacterial infections of the oral cavity, skin and vagina. 35. Use of the therapeutically active compounds according to claims 1-32 for the manufacture of a medicament for the treatment of tumors. A pharmaceutical composition comprising one or more therapeutically active compounds according to claims 1-32 and one or more suitable excipients. 1008745¾ 4 A pharmaceutical composition according to claim 36 in the form of a spray, ointment, gel or lozenge. A therapeutically active compound according to any one of claims 1 to 33, use according to claim 34 or 35 or pharmaceutical composition according to claims 36 and 37, characterized in that the yeast infections are caused by Candida species, in particular Candida albicans. 39. Therapeutically active compound according to any one of claims 1-33, use according to claim 34 or 35 or pharmaceutical composition according to claims 36 and 37, characterized in that the bacterial infections are caused by Staphylococcus aureus, in particular methycillin resistant Staphylococcus aureus (MRSA). 100874 5 * 4 COOPERATION TREATY (PCT) REPORT ON NEWNESS CIRCUMSTANCES OF INTERNATIONAL TYPE IONENTIRKATION OF OE NATIONAL APPLICATION Reference of 0 · applicant or of the authorized LPS / eki / 7 Dutch application no. Name) NOTICE FOR THE TECHNICAL SCIENCES Oasjm of the request for a subject of intemaionaai type By oe insanse for Intematiortaai Onderzoen (ISA) to reconcile to net of an intema & onaJ type type no. SN 31154 NL I. CLASSIFICATION OF THE SUBJECT (if applicable) of different oe classifications. aJe dassificaoesymboien declarations) According to the International classification (IPC) Int.Cl.6: A 61 K 47/48, C 07 K 14/47 II. FIELDS OF TECHNIQUE EXAMINED _Requested minimum documentation __ Classification system _Classstficatiesvmoolen _1 Int.Cl.6: A 61 K, C 07 K Onaofzocfite other documentation o ca minimum documentation to the extent that such documents are in the prayers examined | included ! i i | Hl-i'N NO RESEARCH MCGEL1JK FOR CERTAIN CONCLUSIONS (comments oo supplementolad) i _ j IV.:_; LACK OF UNIT OF INVENTION (comments on supplement payload) "crn PCT.IS <V23'la i CS '95c