Carbosilane dendrimer and use thereof to eliminate pathogens in suspension or in biofilm

Abstract

The invention relates to a cationic carbosilane dendrimer comprising a cyclotetrasiloxane core, wherein each silicon of the core is bonded by means of a carbosilane structure (CH2)2{Si(Me)2}(CH2)2S(CH2)2 to a peripheral structure comprising an ammonium group bonded by means of an alkyl chain (CH2)2 to a hydroxyl group. The invention also relates to the use of the dendrimer to remove at least one pathogenic species, either in suspension or adhered to a surface, forming a biofilm, and to a transport vehicle of at least one drug. The invention further relates to the use of the dendrimer as a drug or in the prevention and / or treatment of at least one disease caused by at least one pathogen.

Description

[0001]DESCRIPTION CARBOSILANE DENDRIMER AND ITS USE FOR THE ELIMINATION OF PATHOGENIC MICROORGANISMS IN SUSPENSION OR IN BIOFILMS ART SECTOR The present invention relates to the eradication of cells in suspension and adhered to a surface, including mixed biofilms (yeasts and bacteria in a common culture), using a cationic carbosilane dendrimer that has ammonium groups modified with a hydroxyl function on its periphery (compound BDNG001). PRIOR STATE OF THE ART The elimination of pathogenic microorganisms, especially those of greatest concern and high clinical relevance, is an increasingly complex process due to the emergence of microbial resistance. Therefore, the treatments employed require the use of highly toxic compounds or extremely high concentrations to achieve complete eradication, especially if these microorganisms are eukaryotic or present in the form of microbial biofilms.Therefore, it is essential to develop new alternatives capable of promoting the elimination of potentially problematic microorganisms, minimizing the environmental impact and, in particular, the side effects in people receiving these antimicrobial therapies. The use of dendritic systems with antibacterial and antifungal activity is widely recognized in the literature (de la Mata, FJ WIREs Nanomed Nanobiotechnoly 2022, e1871). Likewise, their antibiofilm activity has been observed with promising results (Heredero-Bermejo, I et al. Pharmaceutics. 2020, 12 (10): 918; Gómez-Casanova, N. et al. J Fungi (Basel). 2021, 7 (7): 574; Gómez-Casanova, N. et al. Pharmaceutics 2022, 14, 1604). Generally, these compounds are systems that present positive charges on the periphery together with a hydrophobic skeleton, such as carbosilane (CBS), which facilitates interaction with the bacterial membrane.An important advantage of dendritic systems is that they are polyfunctional molecules with a well-defined structure (de la Mata, FJ WIREs Nanomed Nanobiotechnol.2022, e1871). The formation of these molecules is usually carried out by a series of repetitive steps in which the degree of functionalization of the dendritic system is increasingly increased, giving rise to molecules of a globular nature with branches that arise from a central multivalent group. The process of treating and eliminating yeasts is quite complex and complicated, and this is especially due to the fact that they are eukaryotic cells. Therefore, most of the antifungals currently used are highly toxic to the cells of the treated person, which can sometimes also become targets of these treatments (Kagan, S. et al. Antimicrob Agents Chemother. 2012, 56(11):5603-11).In addition to the above, biofilms, especially mixed biofilms formed by yeasts and bacteria, are an even bigger problem, because their treatment requires very high concentrations of compound to be completely destroyed (Lara HH, et al. Pathogens. 2020, 9 (10): 784). The present invention therefore arises from the need to provide an innovative solution to eliminate microorganisms, both in their planktonic form and in their biofilm state. In this way, the invention provides a new compound capable of actively acting as an active ingredient in antiseptics and / or disinfectants, being effective both for the treatment of a single species and for the treatment of microbial communities made up of different species.DESCRIPTION OF THE INVENTION The compound referred to in the invention is a CBS dendritic system, similar to those described in WO2014016460, but in this invention it is a cationic dendritic system with hydroxyl groups on the periphery linked to the ammonium group by an alkyl chain. In particular, a first object of the invention is a cationic carbosilane dendrimer (hereinafter also referred to as BDNG001) comprising a cyclotetrasiloxane core, where each silicon in the core is linked by a carbosilane structure (CH2)2{Si(Me)2}(CH2)2S(CH2)2 to a peripheral structure comprising an ammonium group linked by an alkyl chain (CH2)2 to a hydroxyl group, according to the following formula:. The term "dendritic compound" in the present invention refers to a highly branched macromolecule in which the growth units, branches, or ramifications have a carbosilane skeleton. In particular, the dendrimer object of the present invention is cationic. Therefore, it may comprise counterions to neutralize its charge. Thus, in particular embodiments of the invention, the claimed dendrimer will comprise at least one anion selected from a group consisting of at least one halide (preferably chloride, bromide, or iodide), hydroxide, triflate, nitrate, and sulfate, as well as any combination thereof.Another object of the invention is the use of the claimed carbosilane dendrimer (or a pharmaceutical composition comprising it) as an antibacterial or antifungal agent, that is, for the elimination of pathogenic microorganisms (including yeasts and bacteria), either suspended in a liquid medium (such as planktonic cells) or adhered to a surface, in a biofilm state. The term "biofilm" refers to a set of highly organized cells that are adhered to a surface, which may be biotic or abiotic. Biofilms may be formed by a single species of microorganism, or several, surrounded by an extracellular matrix. Eradicating a biofilm is extremely complicated, especially if it has already established itself, and this difficulty is even greater in the case of mixed biofilms, that is, those formed by more than one species.Their treatment is severely limited because the compounds are either toxic or high concentrations of the biocide are required to eliminate them. Furthermore, their presence increases the development of resistance, another major problem encountered today. Thus, one of the main advantages of the invention is that it has proven effective in treating mixed biofilms, that is, those of different microorganisms such as yeast and bacteria. The compound's effectiveness against biofilms has been demonstrated using colorimetric and agar plating methods. The claimed dendrimer is completely soluble in water and does not exhibit any type of interaction with the media required for the study of microorganisms in vitro. Additionally, it is a compound with low cytotoxicity.In particular, the pharmaceutical composition comprising the dendrimer object of the invention may additionally comprise at least one second active ingredient selected from a group consisting of an antibiotic, anti-inflammatory and antiviral, as well as any combination thereof. Also object of the invention is a method for the elimination of at least one biofilm constituted by a single species of microorganisms or by at least two different species of microorganisms, either during its formation or once formed, characterized in that it comprises applying a solution (preferably aqueous) of the dendrimer object of the invention to the biofilm for a time preferably between 1 hour and 72 hours.Additionally, the object of the invention is the cationic carbosilane dendrimer object of the invention for its use as a medicament, as well as for its use in the prevention and / or treatment of at least one disease caused by at least one pathogenic microorganism. Likewise, the object of the invention is the use of the claimed dendrimer for the manufacture of a medicament for the treatment of at least one disease caused by at least one pathogenic microorganism. Finally, the object of the invention is the use of the claimed dendrimer as a transport vehicle for at least one drug, preferably with antibacterial and / or antifungal properties. NOMENCLATURE IN BIOFILMS The nomenclature defined below for biofilms has been previously standardized in various aforementioned publications, as well as in a book chapter (Gómez-Casanova, N., Candida and Candidiasis, 2022, chapter in IntechOpen).Dendritic compounds with these characteristics have been evaluated using the resazurin colorimetric method based on cellular metabolism to quantify biofilm cell viability (quantitative method) and the dropplate method (seeding on agar plates, qualitative method). Nomenclature for the treatment of forming biofilms: ^CMIB: Minimum Inhibitory Concentration of Biofilm, defined as the minimum concentration capable of inhibiting the formation of a biofilm, but which might not kill the cells. It is determined using the resazurin colorimetric method.^ CMBB or CMFB: Minimum Bactericidal or Fungicidal Concentration of Biofilm, defined as the minimum concentration capable of preventing the formation of a biofilm and killing all the cells. It is determined by the drop plate method.Nomenclature for the treatment of previously established biofilms: ^MBDC: Minimal Biofilm Damaging Concentration, defined as the minimum concentration capable of severely damaging an established biofilm but which may not kill all the cells. It is determined by the colorimetric method with resazurin. ^MBEC: Minimal Biofilm Eradication Concentration, defined as the minimum concentration capable of completely eradicating a previously established biofilm. In this case, all the cells that make up the biofilm would be dead. This is the most difficult objective to achieve in in vitro and in vivo studies of new compounds. It is determined by the drop plate method. DESCRIPTION OF THE FIGURESFigure 1. Percentage viability of BDNG001 in previously established C. albicans biofilms.Figure 2. Percentage viability of BDNG001 in previously established S. aureus biofilms. Figure 3.Percentage viability of BDNG001 in previously established mixed biofilms of C. albicans and S. aureus.Figure 4. Percentage cell viability in the HeLa cell line of BDNG001 at 24 hours.Figure 5. Percentage cell viability in the HeLa cell line of BDNG001 at 48 hours. EXAMPLES The synthesis procedures of the cationic dendrimer BDNG001 are described below, as well as the study of its antifungal, antibacterial and antibiofilm activity against some pathogenic microorganisms. Example 1.- Synthesis of the dendrimer BDNG001 (G0Si4O4Si(S-NMe2OHBr)4). This compound can be obtained from G0Si4O4Si(S-NMe2)4 (0.150 g, 0.13 mmol) (Heredero-Bermejo et al. Pharmaceutics 2020, 12, 918) by reaction with Br(CH2)2OH (0.077 g, 0.62 mmol), without ruling out other agents that introduce the ethanol group, preferably in THF (tetrahydrofuran) as a solvent, or others such as DMF (dimethylformamide), acetone, DMSO (dimethyl sulfoxide), heating for several days (e.g.3 days at 60ºC). The product BDNG001 is obtained as a pale yellow solid. Example 2.- Microbial activity of BDNG001 in planktonic cells The use of the compound of the invention, BDNG001, a dendritic system with –OH groups on its periphery, is proposed for the treatment of microorganisms, both prokaryotic (bacteria) and eukaryotic (yeast-like fungi), as an alternative to traditionally used drugs against said microorganisms. The compound is suitable for the evaluation of its antibacterial activity based on ISO 20776-1:2006. For this purpose, the bacteria were grown in Mueller Hinton liquid medium and, after 24 hours of incubation, the amount of microorganism was adjusted to an inoculum of 108 CFU / mL in bacteria. Next, 2 mL of the adjusted inoculum was added to 8 mL of sterile distilled water. Finally, 100 µL of compound, 100 µL of twice concentrated Mueller Hinton medium and 5 µL of the adjusted inoculum were added to a multiwell plate.The plates were incubated for 24 hours at 37 °C. After this time, the absorbance was quantified at 630 nm. On the other hand, the standardized methods for the study of antifungal susceptibility are based on the European Committee on Antimicrobial Susceptibility Testing (EUCAST). For this purpose, the yeasts were grown on a Sabouraud agar plate. Then, several colonies were collected and homogenized in sterile distilled water until reaching a density of 0.5 McFarland (106 CFU / mL) (inoculum). Afterwards, a 1:10 dilution was made in twice concentrated RPMI-1640 medium, supplemented with MOPS and glucose. Finally, 100 µL of the compound and 100 µL of the adjusted inoculum were added to a multiwell plate. The plates were incubated for 24 hours at 37°C. After this time, they were analyzed at 530 nm. Since this compound is water-soluble, it does not require any procedure to improve its solubility during the testing process.Both bacteria and yeasts were tested at concentration gradients up to 512 mg / L. Each concentration was performed in triplicate, as well as the untreated bacterial control. Control wells of water, culture medium and compound were also incorporated to control for possible contamination or interference from the compound in the absorbance reading. These tests included a Candida albicans strain from the Spanish Type Strain Collection (CECT1002), a Staphylococcus aureus strain (CECT240) and an Escherichia coli strain (CECT515). Example 3.- Activity of BDNG001 on forming and established bacterial and fungal biofilms The use of the compound of the invention, BDNG001, a dendritic system with –OH groups on its periphery, is proposed for the treatment of bacterial and fungal biofilms as an alternative to traditionally used drugs.The compound is suitable for the evaluation of its activity against developing biofilms, as well as established biofilms. The procedures and nomenclature commonly used to test the antibiofilm activity are collected in Heredero-Bermejo, I et al. Pharmaceutics.2020,12(10):918; Fernandez, J. et al. Polymers (Basel). 2021, 13(13):2127; Gómez-Casanova, N. et al. J Fungi (Basel). 2021, 7(7):574; Gómez-Casanova, N., Candida and Candidiasis, 2022 Chapter in IntechOpen). For these tests, the yeast C. albicans (CECT 1002) was used as a representative eukaryotic strain, and the bacterium S. aureus (CECT240) was used as a representative prokaryotic strain. Biofilms can be treated before they form, to prevent biofilm formation, or once they have established themselves. To treat forming biofilms in S. aureus (bacteria), cells were grown in Mueller Hinton medium. The following day, the inoculum was adjusted to 0.5 McFarland ratio.Next, a 1:100 dilution was made in Tryptic Soy Broth medium supplemented with twice concentrated glucose. Finally, 50 µL of BDNG001 compound and 50 µL of the bacterial suspension were added to a Nunc flat-bottom multiwell plate. The plates were incubated for 24 hours at 37 °C. After that time, the biofilms were carefully washed with phosphate-buffered saline (PBS). Then, 100 µL of PBS was added to each well along with 20 µL of 0.01% resazurin (previously filter-sterilized). The microplates were incubated for 20 hours in the dark and measured at 570 nm and 600 nm. These data provided the CMIB value. Finally, 5 µL of each well was plated on plate count agar (PCA) and incubated at 37°C for 24 hours. These data provided the CMBB value. Furthermore, standardized methods for studying developing biofilms in yeasts present slight differences.Candida was grown overnight in YPD (yeast extract, peptone, and dextrose) medium and adjusted to 0.5 McFarland in RPMI-1640 medium supplemented with MOPS and glucose. 50 µL of BDNG001 compound and 50 µL of the fungal suspension were then added to a Nunc flat-bottom multiwell plate. The plates were incubated for 48 hours at 37 °C. After that time, the biofilms were carefully washed with PBS. Then, 100 µL of PBS was added to each well along with 20 µL of 0.01% resazurin. The multiwell plates were incubated for 20 hours in the dark and measured at 570 nm and 600 nm. These data provided the CMIB value. Finally, 5 µL of each well was plated on Sabouraud agar plates and incubated at 37°C for 24 hours. These data provided the CMFB value. Concentration gradients were tested for both bacteria and yeast up to 512 mg / L (the highest concentration tested).Each concentration was performed in triplicate, as was the untreated bacterial control. Control wells containing water, culture medium, and compound were also added to control for possible contamination or interference from the compound in the absorbance reading. For the treatment of previously established S. aureus biofilms, cells were grown overnight in Mueller Hinton medium. The following day, the inoculum was adjusted to 0.5 McFarland. A 1:100 dilution was then made in Tryptic Soy Broth medium supplemented with glucose. Then, 100 µL of the bacterial suspension was added to a flat-bottom multiwell Nunc plate. The plates were incubated for 24 hours at 37 °C. After this time, the biofilms were carefully washed with PBS and treated with compound BDNG001. The multiwell plates were again incubated for 24 hours at 37 ºC.Finally, after this time, the biofilms were carefully washed again with PBS. Then, 100 µL of PBS was added to each well along with 20 µL of 0.01% resazurin. The multiwell plates were incubated for 20 hours in the dark and measured at 570 and 600 nm. These data provided the MCBD value. Finally, 5 µL from each well were plated on PCA agar plates and incubated at 37 °C for 24 hours. These data provided the MCEB value. On the other hand, previously established standardized methods for studying yeast biofilms again present slight differences. Candida was grown in YPD medium overnight and adjusted to 0.5 McFarland in RPMI-1640 medium supplemented with MOPS and glucose. Finally, 100 µL of the fungal suspension was added to a flat-bottom multiwell Nunc plate. The plates were incubated for 48 hours at 37°C.After this time, the biofilms were carefully washed with PBS and treated with the concentration gradient of compound BDNG001. The plates were again incubated for 48 hours at 37 °C. After this time, the wells were washed with PBS. Then, 100 µL of PBS was added to each well along with 20 µL of 0.01% resazurin. The multiwell plates were incubated for 20 hours in the dark, and the absorbance was quantified at 570 nm and 600 nm. These data provided the CMDB value. Finally, 5 µL from each well was seeded on Sabouraud agar plates and incubated at 37 °C for 24 hours. These data provided the EBMC value. Concentration gradients were tested for both bacteria and yeast up to 1024 mg / L (the maximum concentration studied). Each concentration was performed in triplicate, as was the untreated bacterial control.Control wells of water, culture medium and compound were also incorporated to control possible contamination or interference of the compound in the reading of the absorbance. Example 4.- Activity of BDNG001 on mixed biofilms in formation and established The use of the compound of the invention, BDNG001, a dendritic system with -OH groups in its periphery, is proposed for the treatment of mixed biofilms as an alternative to drugs in traditional use. The compound BDNG001 was tested on mixed eukaryotic + prokaryotic biofilms, with C. albicans being the eukaryotic representative and S. aureus being a prokaryotic microorganism. The results obtained were based, as previously mentioned, under the nomenclature described by the chapter Gómez-Casanova, N., Candida and Candidiasis, 2022 Chapter in IntechOpen and various articles collected in the literature (Heredero-Bermejo, I et al. Pharmaceutics.2020,12(10):918; Fernandez, J. et al.Polymers (Basel).2021, 13(13):2127; Gómez-Casanova, N. et al. J Fungi (Basel). 2021,7(7):574). For the treatment of mixed biofilms in formation, the inoculum of each species must be initially adjusted separately. First, the S. aureus species (bacteria), previously grown in Mueller Hinton liquid medium, was adjusted to 0.5 McFarland. Then, a 1:100 dilution was made. Secondly, C. albicans (yeast), previously grown in YPD medium, was adjusted to 0.5 McFarland in medium. Finally, both species were mixed in equal parts, leaving the inoculum adjusted (1:1). The experiment was carried out using RPMI-1640 medium. Once the mixture was homogenized, 50 µL of the inoculum suspension and 50 µL of compound were added to a flat-bottom multiwell Nunc plate. The plates were incubated for 24 hours at 37°C. After this time, the biofilms were carefully washed with PBS.Next, 100 µL of PBS was added to each well along with 20 µL of 0.01% resazurin (previously filter-sterilized). The multiwell plates were incubated for 20 hours in the dark, and absorbance was assessed at 570 nm and 600 nm. These data provided the CMIB value. Finally, 5 µL from each well was plated on PCA agar and Sabouraud agar plates and incubated at 37 °C for 24 hours. These data provided the CMBB / CMFB value. Concentration gradients were tested up to 512 mg / L (maximum concentration tested). For the treatment of previously established mixed biofilms, the inoculum of each species must be initially adjusted separately. First, the species S. aureus (bacteria), previously grown in Mueller Hinton liquid medium, at 0.5 McFarland. A 1:100 dilution was then made. Secondly, C. albicans (yeast), previously grown in YPD medium, was adjusted to 0.5 McFarland.Finally, both species were mixed in equal parts (1:1). The experiment was performed using RPMI medium. Once the mixture was homogenized, 100 µL of suspension was inoculated into a flat-bottom multiwell Nunc plate. The plates were incubated for 24 hours at 37 °C. After that time, the biofilms were carefully washed with PBS and treated with compound BDNG001. The multiwell plates were again incubated for 24 hours at 37 °C. After that time, the biofilms were carefully washed again with PBS. Then, 100 µL of PBS was added to each well along with 20 µL of 0.01% resazurin (previously sterilized by filtration). The multiwell plates were incubated for 20 hours in the dark, and absorbance was quantified at 570 nm and 600 nm. These data provided the CMDB value.Finally, 5 µL of each well were plated on PCA agar and Sabouraud agar plates and incubated at 37 °C for 24 hours. These data provided the MCEB value. Concentration gradients were tested up to 1024 mg / L (maximum concentration studied). In both procedures, treatment of mixed biofilms in formation and previously established mixed biofilms, each concentration was performed in triplicate, as well as the control of untreated bacteria. Control wells of water, culture medium and compound were also incorporated to control for possible contamination or interference of the compound in the absorbance reading. Example 5.- Biocompatibility of BDNG001 The biocompatibility of the main compound of the invention, BDNG001, was studied. For this purpose, HeLa cells (ATCC® CCL-2™) were used.The assays were performed in 24-well plates (NUNC™) in Dulbecco's Modified Eagle medium supplemented with 10% fetal bovine serum and 1% of an antibiotic mixture composed of: 10,000 U penicillin, 10 mg streptomycin and 25 μg AmB per mL. The cells were adjusted to a density of 1 × 10. 4 cells / well in 500 μL of fresh medium. The plates were incubated at 37 °C and 5% CO2 for 5 days until a confluent monolayer was obtained. After the incubation time, the medium was removed and the concentration gradient of the compound diluted in fresh medium was added. The control contained only fresh medium. The plates were incubated for 24 and 48 hours. Afterwards, the medium was removed and the wells were carefully washed with PBS. Finally, 500 μL of fresh medium and 50 μL of tetrazolium microculture (MTT, 5 mg / mL) were added to each well. The plates were again incubated for 4 hours at 37 °C and 5% CO2. ºC. After incubation, the medium was removed and 500 μL of DMSO was added to dissolve the crystals. Finally, the absorbance of the wells was measured in a reader at 570 nm. These experiments were also performed in triplicate and repeated three times. To assess these results, the percentage of viability of the HeLa cells was calculated. Percentage viability is understood as: OD570 compound / OD570 control × 100 (the control is considered as 100% viability). They were considered non-cytotoxic when only a reduction in viability <10% was obtained, values ​​between 10–25% were considered as low cytotoxic and values ​​between 25–40% were considered as moderately cytotoxic levels. RESULTSThe activity tests of the dendrimer object of the present invention have been carried out preferably using water as a solvent, although the possibility of using other solvents is not ruled out.The use of water, however, eliminates the use of solvents that are inherently harmful to microorganisms. This characteristic is extremely important, especially in the fungal field, where most antifungals used in clinical practice are poorly soluble in water. This property of the dendrimer object of the invention favors its easy handling and use. The concentration ranges studied preferably reach maximum concentrations of 512 mg / L for planktonic cells and developing microbial biofilms and up to 1024 mg / L for previously established microbial biofilms. In the trials carried out, the antimicrobial activity of the main compound of the invention, BDNG001, was tested against bacterial planktonic cells (S. aureus and E. coli) and yeast (C. albicans). All strains used belong to the stock of the Spanish Type Strains Collection (CECT).The minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and minimum fungicidal concentration (MFC) values ​​are shown in Table 1. The results shown in this table support the good antimicrobial activity, especially against C. albicans. Table 1. MIC, MBC and MFC values ​​(mg / L) of different microorganisms (one yeast and two bacteria) using the main compound of the invention, BDNG001. CECT: Spanish Type Strain Collection. Strains CECT MIC MBC / MFC. albicans CECT1002 2 2S. aureus CECT240 2-4 4E. coli CECT515 16 16-32 The main compound of the invention has been tested on forming biofilms and previously established biofilms of eukaryotic (C. albicans) and prokaryotic (S. aureus) microorganisms. The compound is easily soluble in water and no aggregates are observed in contact with the culture media used.The results obtained were very satisfactory as can be seen in Table 2, which shows the values ​​obtained for a forming biofilm: Minimum Biofilm Inhibitory Concentration (MIIC), Minimum Biofilm Fungicidal Concentration (MFFC), and Minimum Biofilm Bactericidal Concentration (MBCC); and values ​​for a previously established biofilm: Minimum Biofilm Damaging Concentration (MBDC) and Minimum Biofilm Eradication Concentration (MBEC). Figures 1 and 2 show the viability percentage for each concentration tested in previously established biofilms of C. albicans and S. aureus. Table 2. Values ​​of MIIC, MIIC, MFC, MFC, MFC, and MFC (mg / L) obtained for compound BDNG001. Forming biofilmsEstablished biofilms MFC MFC / MFC MFC MFC MBC. albicans CECT1002 4-8 8 16 16S.aureus CECT240 4 4 64 64The compound of the invention BDNG001 has been tested on mixed biofilms in formation and previously established mixed biofilms of C. albicans (yeast) together with S. aureus (bacteria). The results obtained were extremely satisfactory as can be seen in Table 3, which indicates the values ​​obtained from a mixed biofilm in formation of: CMIB and CMBB / CMFB; and values ​​for a formed biofilm: CMDB, CMEB. Figure 3 shows the percentage of viability in previously established mixed biofilms for each concentration tested with BDNG001.Table 3. Values ​​of CMIB, CMFB / CMBB, CMDB and CMEB (mg / L) obtained for the compound BDNG001 in mixed biofilm treatments. MIXED BIOFILM OF C. albicans CECT1002 + S. aureus CECT240. These results are highly relevant. Very few studies have been conducted on mixed biofilms. The dendrimer object of the present invention has managed to completely eradicate a previously established biofilm of two completely different species, at low concentrations. Finally, the cytotoxicity produced after 24 and 48 hours of incubation of the compound with the HeLa cell line was evaluated. This invention guarantees the absence of toxicity at the main concentrations with an effect on biofilms, including some of the most difficult to eliminate: mixed biofilms. These results are represented in Figures 4 and 5. These figures show the viability percentages, demonstrating the good biocompatibility of the dendrimer object of the present invention.

Claims

CLAIMS 1. Dendrímero carbosilano catiónico que comprende un núcleo ciclotetrasiloxano, where each silicon in the core is linked by a carbosilane structure (CH2)2{Si(Me)2}(CH2)2S(CH2)2 a una estructura periférica que comprende un grupo amonio unido mediante una cadena alquílica (CH2)2 a un grupo hidroxilo, de acuerdo a la siguiente formula:

2. Dendrímero carbosilano, de acuerdo a la reivindicación 1, donde la carga de dicho dendrimer is neutralized by at least one anion selected from a group consisting of at least one halide, hydroxide, triflate, nitrate and sulfate, as well as any combination thereof.

3. Dendrímero carbosilano, de acuerdo a la reivindicación 2, donde el haluro es seleccionado de un grupo que consiste en cloruro, bromuro y yoduro.

4. Uso de un dendrímero carbosilano, de acuerdo a una cualquiera de las Claims 1 to 3, for the elimination of at least one species of pathogenic microorganism, where said microorganism is either in suspension or adhered to a surface, forming a biofilm 5. Uso, de acuerdo a la reivindicación 4, donde dicho microorganismo es seleccionado from a group consisting of yeasts and bacteria.

6. Uso, de acuerdo a la reivindicación 4 o 5, donde la biopelícula está constituida por una única especie de microorganismos o por al menos dos especies distintas de microorganisms.

7. Método de eliminación de al menos una especie de microorganismo patógeno, caracterizado por que comprende aplicar al microorganismo patógeno una disolución del dendrímero definido según una cualquiera de las reivindicaciones 1 a 3.

8. Método de eliminación de al menos una biopelícula constituida por una única species of microorganisms or at least two different species of microorganisms, either during their formation or once constituted, characterized by comprising aplicar a la biopelícula una disolución del dendrímero definido según una cualquiera de las reivindicaciones 1 a 3.

9. Método de acuerdo a la reivindicación 7 u 8, donde la aplicación de la disolución del dendrimer takes place over a period of between 1 hour and 72 hours.

10. Método de acuerdo a una cualquiera de las reivindicaciones 7 a 9, donde la solution is an aqueous solution.

11. Dendrímero carbosilano catiónico según una cualquiera de las reivindicaciones 1 a 3 for use as a medicine.

12. Dendrímero carbosilano catiónico según una cualquiera de las reivindicaciones 1 a 3 para su uso en la prevención y / o tratamiento de al menos una enfermedad causada por at least one pathogenic microorganism.

13. Uso de una composición farmacéutica caracterizada porque comprende un carbosilane dendrimer according to any one of claims 1 to 3 for the elimination of at least one species of pathogenic microorganism, wherein said microorganism is either in suspension or adhered to a surface, forming a biofilm.

14. Uso de una composición de acuerdo a la reivindicación 13, donde dicha composition additionally comprises at least one second active ingredient selected from a group consisting of an antibiotic, anti-inflammatory and antiviral, as well as as any of its combinations.

15. Uso de un dendrímero carbosilano según una cualquiera de las reivindicaciones 1 a 3 as a vehicle for transporting at least one drug.

16. Uso, de acuerdo a la reivindicación 15, donde el fármaco es un fármaco con antibacterial and / or antifungal properties.

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