FR3028180A1 - NOVEL COMPOSITIONS WITH BIOCIDAL PROPERTIES COMPRISING COMPOUNDS EXTRACTED FROM TROPICAL PLANTS - Google Patents

Abstract

The invention relates to a use of a composition comprising a combination of active agents selected from 2-dodecenal, nonanal, dodecanal or decanal, as a biocidal agent.

Description

FIELD OF THE INVENTION The present invention relates to the use of a composition comprising a combination of active agents chosen from 2-dodecenal, nonanal, dodecanal or decanal. , as a biocidal agent. Background of the invention Infections caused by pathogens are a major public health problem despite several decades of development and use of biocidal agent to destroy, repel or render harmless these pathogens. Among these pathogens, Staphylococcus aureus is a bacterium that causes many infections or food poisoning. It is particularly feared in hospitals, where it is often involved in so-called nosocomial infections. These infections are all the more dangerous because of the increasing resistance of certain strains of this bacterium to antibiotics. The pathogenicity of Staphylococcus aureus is due to the development by the bacterium of various substances with necrotic or toxic effects. This bacterium is responsible for some serious infections of foot wounds in diabetic patients. It is accepted that 5 to 10% of diabetics are at risk of amputation. In France, according to data from the PMSI database (Information Systems Medicalization Program), out of 15,353 people who were amputees in 2003, 7,955 (52%) were diabetic. In addition, the 10-year amputee survival rate is less than 10% and the median survival is less than 2.5 years (Diabetes Care 2006; 29 (10): 2252-6). Hospital management of foot lesions in diabetics is very expensive. However, the prevalence of diabetes, linked to the aging of the population and the development of obesity, should continue to increase in the years to come. It is now crucial to identify new treatments to combat the multi-resistance of certain pathogenic bacteria, especially Staphylococcus aureus. Alternatives to oral antibiotics may be particularly useful for pathologies such as bacterial infections of foot wounds in diabetic patients. Molecules, often from medicinal plants, have recently been studied for their antibacterial properties. Thus, in US 20120009255, it is shown that (E) -2-dodecenal extracted and isolated with petroleum ether Eryngium foetidium has properties for inhibiting gram-positive bacteria Staphylococcus aureus, Staphylococcus pyogenes, Enterococcus faecalis. The Minimum Inhibitory Concentration (MIC) is between 78 μg / ml and 313 μg / ml. In Ho et al. (Rev. Bras., Farmacogn., 22 (2), 2012, pp 277-283), the antibacterial properties of an essential oil extracted by hydrodistillation of Machilius zuihoenzis leaves were studied. Among the essential oil compounds, n-dodecanal and n-decanal showed antibacterial activity against gram-positive bacteria such as Bacillus cereus, Staphylococcus aureus, Staphylococcus epidermis. However, the activity of n-decanal is described as low, with a MIC greater than or equal to 1000 [tg / mi, n-dodecanal is more active with a MIC between 125 [tg / ml and 250 ug / ml. Finally, the possible antibacterial property of n-nonanal, another compound of the essential oil, has not been demonstrated. In Mahnoudi et al. (Jeobp, 12 (5), 2009, pp 574-579) n-decanal is also described as having low activity against Staphylococcus aureus.

Surprisingly, the Applicant has found that a composition comprising a combination of active agents, usually contained in an essential oil of Eryngium foetidium, exhibits biocidal agent properties and can be used to prevent or treat an infection caused by a pathogen. Thus, the present invention relates, in a first aspect, to the use of a composition comprising a combination of active agents selected from 2-dodecenal, nonanal, dodecanal or decanal, as a biocidal agent. According to a second aspect, the invention relates to a composition for its use for preventing or treating an infection caused by a pathogen, characterized in that it comprises a combination of active agents chosen from 2-dodecenal, nonanal, dodecanal or decanal. In another aspect, the invention relates to particular compositions. DETAILED DESCRIPTION OF THE INVENTION According to a first aspect, the present invention relates to the use of a composition comprising a combination of active agents chosen from 2-dodecenal, nonanal, dodecanal or decanal, as an agent. biocide. These various active agents are usually present in an essential oil extracted by hydrodistillation of Eryngium foetidium.

According to an advantageous embodiment, the combination of active agents used according to the invention does not consist in the decanal with the nonanal or in the decanal with the dodecanal. For the purpose of the present invention, the term "biocidal agent" designates a composition having properties enabling it to destroy or moderate the growth of a living organism, the concentration of active agents of which is between 0.2 g / ml and 25 μg / ml. The use of the composition according to the invention is particularly effective against a living organism chosen from a fungus, a virus or a bacterium.

Examples of fungi are Aspergillus niger and Candida albicans. The composition according to the invention can be used as antibacterial agent. For the purposes of the present invention, the term "antibacterial agent" refers to a compound having antibacterial properties enabling it to inhibit the proliferation of a bacterium. Without being bound by any theory, the inventors are of the opinion that the antibacterial properties of the composition according to the invention results from bactericidal rather than bacteriostatic properties. In particular, the composition according to the invention may be used as antibacterial agent for a bacterium, in particular a Gram-negative bacterium chosen from Escherichia coli and Salmonella enteritidis or a Gram-positive bacterium chosen from Bacilius cereus, Bacilius subtilis , Lactobacillus plantarum, Listeria monocytogenes and Staphylococcus aureus. According to a particularly preferred embodiment, the composition according to the invention can be used as an antibacterial agent against Staphylococcus aureus. Due to its biocidal properties, the composition according to the invention can advantageously be used for disinfection or preservation. It can thus be used as a disinfectant for medical equipment and / or implantable equipment. The composition, comprising a combination of active agents selected from 2-dodecenal, nonanal, dodecanal or decanal, may also be used for the prevention or treatment of an infection caused by a pathogen selected from a fungus, a virus, or bacteria. According to a preferred embodiment, the combination of active agents does not consist of the decanal with the nonanal or of the decanal with the dodecanal.

For the purposes of the present invention, the composition comprises the combination of active agents or the combination of active agents with formulation excipients, water and / or a solvent or diluent. The formulation excipients, the solvent and the diluent are chosen to be of suitable quality for the use of the composition. If the use of the composition is food (preservative) their quality will be food, if the use is for the disinfection of medical or implantable equipment, their quality is pharmaceutical, and even if their use is therapeutic. According to one particular embodiment, the combination of active agents is chosen from: 2-dodecenal and nonanal, 2-dodecenal and dodecanal, 2-dodecenal and decanal, Nonanal and dodecanal, 2-dodecenal and nonanal and dodecanal, 2-dodecenal and nonanal and decanal, 2-dodecenal and dodecanal and decanal, nonanal and dodecanal and decanal, 2-dodecenal and nonanal and dodecanal and decanal. The different active agents of the combination may be in one or the same formulation, that is to say they may be present simultaneously in the same composition. They may also be in separate forms so that they can be administered or applied not only simultaneously but also separately or sequentially. In particular, the composition according to the invention is particularly effective against an infection caused by a gram-positive bacterium. It is thus particularly effective against an infection caused by a Gram-positive bacterium mentioned above, and more particularly an infection caused by Staphylococcus aureus. Infections caused by Staphylococcus aureus include diarrhea, suppurative skin infections, acute myositis, pneumonia, endocarditis, urinary tract infections, phlebitis, certain types of enteritis, meningitis, osteomyelitis. According to a specific embodiment, the composition is used for the prevention or treatment of an infection caused by a pathogen in a mammal, in particular a human being, more particularly in a diabetic patient. When the composition is used for the prevention or treatment of an infection caused by a pathogen, said composition may further comprise at least one pharmaceutically acceptable excipient. For the purposes of the present invention, the term "pharmaceutically acceptable excipient" means a compound which is chemically and / or toxicologically compatible with the combination of active agents of the composition and with the mammal treated with the composition.

Whatever the use mentioned above, the composition is particularly effective for a total active agent concentration of from 0.2 to 1.1 g / ml to 1.1 g / ml, preferably from 1.1 g / ml to 1.1 g / ml. ml, more preferably still 1.6 1.1g / ml to 1.1g / ml. This low total concentration of active agents in the composition results from a synergy between the different active agents present in the composition. According to one particular embodiment, the composition used according to the invention comprises from 0.1 g / ml to 10 g / ml, preferably from 2 g / ml to 7 g / ml, more preferably from 5 g / ml to 10 g / ml. 1.1 g / ml at 5.5 g / ml of 2-dodecenal, in combination with at least one other active agent selected from dodecanal, nonanal, or decanal.

According to a particular embodiment, the composition according to the invention comprises from 0.1 to 1.1 g / ml to 1.1 g / ml, preferably from 0.5 to 1.1 g / ml to 2 to 1.1 g / ml, more preferably from 0.9 1.1 g / ml to 1.1 1.1 g / ml of nonanal in combination with at least one other active agent chosen from 2-dodecenal or dodecanal, or in combination with at least 2 other active agents chosen from decanal , 2-dodecenal or dodecanal. According to a particular embodiment, the composition according to the invention comprises from 0.1 to 1.1 g / ml to 1.1 g / ml, preferably from 0.5 to 1.1 g / ml to 2 to 1.1 g / ml, more preferably from 1.1 g / ml to 1.1 g / ml of dodecanal in combination with at least one other active agent chosen from 2-dodecenal or nonanal, or in combination with at least 2 other active agents chosen from 2 -dodecenal, nonanal or decanal. According to a particular embodiment, the composition according to the invention comprises from 0.1 to 1.1 g / ml to 1.1 g / ml, preferably from 0.5 to 1.1 g / ml to 2 to 1.1 g / ml, more preferably from 0.71 g / ml to 0.9 g / ml of decanal, in combination with 2-dodecenal, or in combination with at least 2 other active agents chosen from nonanal, 2-dodecenal or dodecanal. According to a particular embodiment, the composition used according to the invention comprises a combination of 2-dodecenal and an active agent chosen from nonanal, dodecanal or decanal. According to a particular embodiment, the composition according to the invention comprises a combination of nonanal and dodecanal. According to a specific embodiment, the composition used according to the invention comprises a combination of 2-dodecenal, nonanal, dodecanal and decanal. More specifically, the composition comprises from 0.1 to 1.1 g / ml to 1.1 g / ml of 2-dodecenal, from 0.1 to 1.1 g / ml to 5 g / ml of nonanal, from 0.1 to 1.1 g / ml. at 5 g / ml of dodecanal and from 0.1 g / ml to 10 g / ml of decanal. Preferably, it contains from 1.1 g / ml to 7 g / ml of 2-dodecenal, from 0.5 g / ml to 2 g / ml of nonanal, from 0.5 g / ml to 2.1 g / ml. g / m1 of dodecanal and from 0.5 to 1.1 g / ml of 1.1 to 1.1 g / ml of decanal. More preferably still, it contains from 1.1 g / ml to 5.5 g / ml of 2-dodecenal, from 0.9 g / ml to 1.1 g / ml of nonanal, 0.9 g / ml. 1.11 g / ml of dodecanal and 0.751 g / ml at 0.85 g / ml of decanal. It is understood that in these compositions, the total concentration of active agents is less than or equal to 1.1 g / ml. This low concentration of active agent allows the use of the composition as a preservative in a cosmetic product (soap, cream) or in a food product. In the first case, the composition may further comprise at least one excipient of physiologically acceptable formulation. In the second case, the composition may further comprise at least one food grade formulation excipient. For the purposes of the present invention, the term "excipient of physiologically acceptable formulation" denotes a compound whose physicochemical properties do not alter the physiology of the user of the composition. For the purpose of the present invention, the term "food grade formulation excipient" refers to a compound that is nontoxic to the user of the composition. In addition, the synergy resulting from the combination of active agents in the composition makes it possible to broaden the spectrum of living organisms against which said composition is effective. In particular, it makes it possible to widen the spectrum of bacteria, and their strain, against which the composition is effective. The composition can be applied identically regardless of the use mentioned above. It can thus be used by surface application, by spraying or by diffusion. More specifically, in the case where the composition is used for the prevention or treatment of an infection caused by a pathogen, said composition can be administered orally, intravenously, intramuscularly, subcutaneously, nasally, percutaneously or pulmonary, especially orally.

The use by surface application or the percutaneous administration of said composition may be carried out by formulating the composition in the form of cream or soap, or by impregnating a surface of a dressing, a wipe or a patch. or a stamp. Advantageously, the composition formulated as a cream or soap may be used to prevent or treat a bacterial infection of a foot wound of a diabetic patient. The volatility of the active agents of the composition makes it possible to formulate it in the form of a liquid solution in order to use it advantageously by spraying or by diffusion, using a spray or a diffuser. Such formulations also make it possible to administer the composition by nasal or pulmonary route. The composition can thus clean the air in a hospital environment or treat respiratory tract infections. To maximize the spray or diffusion zone, the composition may further comprise a suitable solvent. The formulation of the composition in liquid solution form also allows the administration of said composition intravenously, intramuscularly or subcutaneously. The composition may also be formulated as a capsule, tablet, dragees, granules, syrup, oral suspensions, potions, which allows the administration of said composition orally.

The effective amount of composition in the above-mentioned formulations can be evaluated according to the activity of living organisms, particularly bacteria, and toxicity results. The active agents of the composition according to the invention may be synthetic or may be obtained by distillation of a plant of the Apiaceae family selected from Aegopodium podagraria L., Bupleurum prealtum L., Bupleurum sibthorpianum S. S. var. Diversifolium, Cnidium silafolium, Coriandrum sativum, Ducrosia anethifolia, Ducrosia flabel4folia, Eryngium foetidum, Peucedanum alsaticum, Peucedanum cervaria, Lomatopodium staurophyllum and their mixture, preferably Eryngium foetidum. Preferably, the distillation is a hydrodistillation.

They may also be contained in an essential oil obtained by hydrodistillation of a plant of the Apiaceae family selected from Aegopodium podagraria L., Bupleurum prealuminum L., Bupleurum sibthorpianum S. S. var. Diversifolium, Cnidium silafolium, Coriandrum sativum, Ducrosia anethifolia, Ducrosia flabellifolia, Eryngium foetidum, Peucedanum alsaticum, Peucedanum cervaria, Lomatopodium staurophyllum and their mixture, preferably Eryngium foetidum. In the latter case, the essential oil is itself the composition. The invention also relates to a composition consisting of a combination of at least two active agents chosen from 2-dodecenal, nonanal, dodecanal or decanal. More particularly, the invention relates to the following compositions which contain as sole active agents the following combinations: 2-dodecenal and nonanal, 2-dodecenal and dodecanal, 2-dodecenal and decanal, Nonanal and dodecanal, 2-dodecenal and nonanal and dodecanal , 2-dodecenal and nonanal and decanal, 2-dodecenal and dodecanal and decanal, Nonanal and dodecanal and decanal, 2-dodecenal and nonanal and dodecanal and decanal. These compositions may be used as described above. The different active agents are present in this composition in the proportions defined previously.

The invention will be further described with the aid of the following illustrative examples and accompanying drawings in which: Figure 1 is a graph showing the antimicrobial activity of Composition 1 of Example 1 and each of the active agents isolated at their respective concentrations in Composition 1 of Example 1 against 5 different strains of Staphylococcus aureus. Figure 2 is a graph showing the antimicrobial activity of Composition 1 of Example 1 and each isolated active agent at a concentration of 12.5 ag / ml against 5 different strains of Staphylococcus aureus.

FIG. 3 is a graph showing the antimicrobial activity of the 2x compositions of Example 2 against two bacterial strains of Staphylococcus aureus RN 4220 and S 25. Examples Example 1: Obtaining a Composition Comprising (E) -2-dodecenal, dodecanal, nonanal, and decanal. A composition, denoted Composition 1, is obtained by hydrodistillation of Eryngium foetidum (leaf, root). Hydrodistillation process: 200 g of dry raw materials (aerial parts) are introduced into a 3 L reactor with 1.5 L of water and a few grains of pumice. The reactor is closed and equipped with a Clevenger. The whole is refluxed for 6 hours. The system is insulated with cotton strips wrapped in aluminum foil. The essential oil is recovered and then dried over MgSO4 through a pasteur pipette to give Composition 1. The hydrosol is also preserved, the whole is stored protected from light at 4 ° C. The concentration of active agents is analyzed by gas chromatography coupled with mass spectroscopy (GC-MS) and by gas chromatography with a flame ionization detector (GC-FID) Composition 1 is composed the following active agents: (E) -2-dodecenal, dodecanal, nonanal and decanal The total concentration of active agents in Composition 1 is 12.5 ag / ml, distributed as follows 5.125 μg / ml of (E) -2-dodecenal, 1 ag / ml of dodecanal, 0.9875 μg / ml of nonanal and 0.8 μg / ml of decanal. Example 2: Study of the antibacterial properties of composition 1 and comparison with each isolated active agent. Strains RN 4220 (laboratory strain), S 25 (strain isolated from a patient with osteoarticular infection), NSA 1385 and NSA 739 (strains isolated from foot wounds of diabetic patients), S 1799 (strain 5T80 MRSA IV Community Methylcillin resistant, currently diffusing in Europe) and S S385 (isolated strains of foot wounds of diabetic patients) were selected to test the antibacterial properties of the composition obtained in Example 1 and each isolated agent. Composition 1 and each isolated active agent (at two different concentrations) are tested according to the protocol described below. Control is performed by DMSO.

The first concentration of each isolated active agent corresponds to the concentration of said active agent in Composition 1. For example, the (E) -2-dodecenal is tested diluted in DMSO at a concentration of 5.125 μg / ml. The second concentration is identical for all the isolated active agents, it corresponds to the total concentration of active agents of Composition 1, that is to say to 12.5 μg / ml. Study protocol: For each strain, the protocol is identical and is the following. Pre-culture is carried out by adding a colony of Staphylococcus aureus in 13 ml of LB culture medium. The LB culture medium is then stirred for 2 h at 37 ° C. 0.5 11.1 of Composition 1, or active agent diluted in DMSO at the tested concentration, or DMSO for control is spot in the bottom of a 96-well flat-bottom plate. 100111 of the preculture is added to the multichannel pipette per well (there are 3 wells per sample, 3 control wells, and 3 LB wells. An Optical Density reading at 600 nm is made at T = Oh, 3h and 18h The results of this study are presented in Figures 1 and 2. The ordinate values are optical density measurements read at 600 nm, the value of the background noise of the plate being between 0.04 and 0.05 The lower the value, the lower the amount of bacteria remaining, these results show that the activity of Composition 1 is particularly strong on all these strains, in particular the growth is totally inhibited for all the strains tested. In contrast, the activity of each active agent tested in isolation depends both on the strains and on the concentration of the active agent, eg nonanal is more active against strains S RN4220 and S S25 at the concentration of 0.99 1.1g / mL only in con centration of 12.5 lg / mL Similarly, 2-dodecenal is less active than Composition 1 against strains S S385 and S S1799.

It therefore appears from the results that for some strains, such as S S385 and S S1799, the activity of Composition 1 (essential oil) is much stronger than the activity of each active agent tested in isolation. Thus, at this concentration (5.125 μg / ml), neither the (E) -2-dodecenal nor any of the other active agents tested is responsible for the activity of Composition 1 on these strains. The activity of Composition 1 thus comes from a synergistic effect between the different active agents of this Composition 1. Following these experiments, the media containing the strains that had been in contact with Composition 1 were cultured. for 24h. After 24 hours of incubation of these samples in an LB culture medium without Composition 1, no bacterial growth was observed. Composition 1 therefore has a bactericidal rather than a bacteriostatic action. Example 3: Obtaining 2x compositions, comprising a combination of 2 active agents.

Various compositions, denoted Compositions 2x, comprising active agents of Composition 1 are obtained from synthetic compounds (obtained from Sigma-Aldrich). These compositions are presented in the following Table 1. Table 1: Compositions 2x Compos Concent Concent ration in anal dodec Concent Concent itions 2x ration in (E) -2- ration in nonanal ration in decanal dodecenal 2a 5,125 tg / ml 1 tg / ml 0 0 2b 5,125 tg / ml 0 0, 9875 tg / ml 0 2c 5.125 tg / ml 0 0 0.8 tg / ml 2d 0 1 tg / ml 0.9875 tg / ml 0 2e 0 1 tg / ml 0 0.8 tg / ml 2f 0 0 0.9875 tg / ml 0.8 tg / ml EXAMPLE 4 Study of the Antibacterial Properties of the 2x Compositions The various 2x compositions are tested identically to the Composition 1 but only to the S RN4220 and S S25 bacterial strains. The results are shown in Figure 3. The study of the results presented in Figure 3 shows the synergistic effects between the active agents. Thus, it appears that the compositions 2a, 2b, 2c and 2d are particularly active on the bacterial strains S RN4220 and S S25. On the other hand, Compositions 2e and 2f do not exhibit the synergistic effects and are not active on bacterial strains S RN4220 and S S25.

Claims (13)

REVENDICATIONS1. Use of a composition comprising a combination of active agents selected from 2-dodecenal, nonanal, dodecanal or decanal, as a biocidal agent, 2. Use according to claim 1, wherein the active agent composition is not the combination of decanal and nonanal nor the combination decanal and 2-dodecenal. Use according to claim 1 or 2 as an antibacterial agent for Gram-positive bacterial strains, in particular Staphylococcus aureus. 4. Use according to any one of claims 1 to 3 for disinfection or preservation. 5. Use according to any one of claims 1 to 4 by surface application, by spraying or by diffusion. 6. Use according to any one of claims 1 to 5 as a disinfectant of medical equipment, or as a preservative in a cosmetic product or food product. Composition for use in the prevention or treatment of an infection caused by a pathogen selected from a fungus, a virus, or a bacterium, characterized in that it comprises a combination of active agents selected from the 2 -dodecenal, nonanal, dodecanal or decanal. The composition for use according to claim 7 wherein the combination of active agents is not the combination of decanal and nonanal nor the combination of decanal and 2-dodecenal. 9. Composition for its use according to claim 7 or 8, characterized in that the bacterium is Gram-positive, in particular Staphylococcus aureus. 10. Composition for use according to any one of claims 7 to 9, characterized in that it is intended for oral, intravenous, intramuscular, subcutaneous, nasal, percutaneous or pulmonary administration, in particular by the route of administration. oral. 11. A composition for use as claimed in any one of claims 7 to 10 in a diabetic patient. 12.Use according to any one of claims 1 to 6 or composition for use according to any one of claims 7 to 11 characterized in that the active agents are synthetic or contained in an essential oil obtained by distillation of a plant of the Apiaceae family selected from Aegopodium podagraria L., Bupleurum prealuminum L., Bupleurum sibthorpianum SS var. Diversifolium, Cnidium silaifolium, Coriandrum sativum, Ducrosia anethifolia, Ducrosia flabellifolia, Eryngium foetidum, Peucedanum alsaticum, Peucedanum cervaria, Lomatopodium staurophyllum, preferably Eryngium foetidum. 13.Composition comprising as sole active agents a combination of at least two active agents selected from 2-dodecenal, nonanal, dodecanal or decanal and at least one excipient.