PHARMACEUTICAL COMPOSITION COMPRISING TRANS - CINNAMALDEHYDE AND ITS USE IN THE TREATMENT OF INFECTIONS
This invention pertains to an anti-microbial, in particular anti-bacterial, more particularly against Gram negative bacteria, and/or anti-fungal composition comprising as active blend trans-cinnamaldehyde and a potentiating agent. In particular this composition is intended for preventing and/or treating microbial infection in an animal.
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This application is a Continuation of U.S. application Ser. No. 15/188,131, filed on Jun. 21, 2016, which is a Divisional of U.S. application Ser. No. 13/992,137, filed on Aug. 21, 2013, which is a national phase of PCT International Application No. PCT/EP2011/072389 filed on Dec. 9, 2011. This application also claims benefit to U.S. Provisional Application No. 61/421,362 filed on Dec. 9, 2010 and Application No. 10306388.9 filed in Europe on Dec. 9, 2010, the entire contents of which are hereby incorporated by reference.
This invention pertains to an anti-microbial, in particular anti-bacterial, more particularly against Gram−bacteria, and/or anti-fungal and/or anti-viral composition comprising as active blend trans-cinnamaldehyde and a potentiating agent, and to the potentiating agent. In particular this composition is intended for preventing and/or treating microbial infection in an animal.
The arrival of penicillin followed by streptomycin in the 40s opened the era of anti-bacterials. The discovery of anti-bacterial compounds such as penicillin, aminosides, macrolides and quinolones have been one of the biggest achievement of the modern medicine for the treatment of bacterial infections.
However, there is a growing need for new compounds or compositions allowing fighting bacterial infections. This need is in particular due to the fact that there is more and more microbes which are exhibiting various drug resistance to known anti-microbial compounds or compositions.
For example in the case of bacteria it becomes a major problem to find novel efficient anti-bacterial compositions to prevent or treat bacterial infections. For example among Gram+bacteria exhibiting drug resistance can be cited Staphylococcus, in particular Staphylococcus aureus, Enterococcus, in particular Enterococcus faecalis and Enterococcus cloacae, and Propionibacter, in particular Propionibacter acnes, and among Gram−bacteria exhibiting drug resistance can be cited Escherichia, in particular Escherichia coli, Pseudomonas, in particular Pseudomonas aeruginosa, and Acinetobacter, in particular Acinetobacter baumanii, Serratia, in particular Serratia marscescens, Citrobacter, in particular Citrobacter freundii, Klebsiella, in particular Klebsiella pneumonia, and Enterobacter, in particular Enterobacter aerogenes.
A particularly significant case is Staphylococcus aureus for which more than 95% of the Staphylococcus aureus strains are penicillin resistant and more than 60% are also resistant to its methicilline derivative (MRSA). Moreover some strains are resistant to vancomycin (VRSA).
According to WHO, the ratio of methicillin-resistant Staphylococcus aureus strains which became mipiromicin (an anti-bacterial inhibiting protein synthesis) resistant increased from 2.7% to 65% in three years time. This shows that the action of classical anti-bacterial analogs may be quickly countered by multiple resistance mechanisms from bacteria.
This drug resistance is even more preoccupying as at present it is not confined to hospital but is also disseminated outside. For example, there is a very important prevalence of infections of person over 65 years old caused by Staphylococcus aureus. Thus, the proportion of microbial infections, in particular bacterial, pneumonia, endocarditis, osteoarticular or urinary infections, developed by persons over 65 years old and linked with Staphylococcus aureus is particularly preoccupying.
Gram negative bacteria, in particular enterobacteria and Pseudomonas aeruginosa, are naturally resistant to, often at low level of, most hydrophobic and/or high molecular weight anti-microbial agents, for example such as penicillin G, penicillin M, macrolides, rifampicin, fusidic acid, novobiocine, vancomycine, as this anti-microbials cannot cross the external membrane wall of the bacteria.
Apparition and propagation of microbial strains resistant to almost all or even to all the known anti-microbial agents becomes a major health issue.
There is clearly a growing need for novel anti-microbial, in particular anti-fungal and/or anti-bacterial compounds, in particular to fight with Gram+ and/or Gram−bacteria.
Thus, one aim of the invention is to provide active anti-microbial agents, in particular anti-fungal and/or anti-bacterial, more particularly having a large spectra and/or efficient with resistant microbes, even more particularly to non-natural resistant and/or to natural resistant microbes.
The “natural resistance” may be as defined pages 15, 16 and 17 of the Communiqué 2006 (Edition of January 2006) of the “Comite de l′Antibiogramme de la Societe Francaise de Microbiologie”. In infectious pathology, a bacteria is said «resistant» when it may bear a concentration much higher than the concentration which is possible to reach in vivo by a treatment. Thus, another aim of the invention is to obtain anti-microbial agents exhibiting a high activity, in particular at low level, on their targets.
Known anti-microbial agents may exhibit the following drawbacks they may have an insufficient activity, in particular at low level and/or when interfering agent are present, they may be insufficiently active or not active at all on some or on any drug resistant microbes, in particular on drug resistant bacteria such as drug resistant Staphylococcus aureus, for example such as MRSA, VRSA; drug resistant enterobacter, such as NDM-1, or New Dehli Metallo-beta-lactamase type 1; they may presenting undesirable side effects, they may not be well tolerated by the organism, their use may lead to bacteria having drug resistance, in particular crossed drug resistance, they may be expensive, and/or difficult to obtain.
More particularly cinnamon essential oils, especially those comprising trans-cinnamaldehyde, or trans-cinnamaldehyde alone are known to be a potent anti-microbial, however, they lead to undesirable effects, such as neurotoxicity, which may lead to death in case of high dose, hepatotoxicity, irritation of the skin, sensitization of the skin, phototoxicity and/or photoallergy.
Thus, the invention aims to composition solving all or part of these problems. In particular the invention aims to anti-microbial agents exhibiting high activity, in particular at low level and/or when interfering agents are present; activity on some or as many as possible microbes, in particular drug resistant microbes, in particular activity on:
-
- bacteria, for example drug resistant bacteria such as drug resistant Staphylococcus aureus, for example such as MRSA, VRSA, drug resistant enterobacter, such as NDM-1; and at least one bacteria such as those disclosed below;
- fungi, for example
- Epidermal, dermal and/or keratinous appendage fungi, in particular Candida, Trichophyton, Malassezia, and Microsporum,
- Systemic, in particular non-opportunistic disease, more particularly due to Blastomyces, Coccidioides, and opportunistic disease due to Aspergillus, Candida albicans, and Cryptococcus,
- viruses, such as HIV; herpes viruses, the hepatitis B and C viruses, and influenza A and B viruses, in particular enveloped viruses.
presenting as few as possible side effects well tolerated by the organism; leading to as few as possible or not leading at all to drug resistance, in particular to cross drug resistance; reproducible; obtained through a process easy to follow analytically, for example with HPLC and/or GC; cheap; and/or easy to obtain.
An “anti-microbial” is a substance that kills or inhibits the growth of microorganisms such as bacteria, fungi, or viruses. Antimicrobial drugs either kill microbes (microbiocidal) or prevent the growth of microbes (microbiostatic). Disinfectants and antiseptics are antimicrobial substances used on non-living objects or outside the body.
By «anti-bacterial active», is meant a compound or a composition exhibiting bacteriostatic or bactericidal properties, in particular in vitro, for example in a composition such as a pharmaceutical composition, a food composition or a cosmetic composition, or for disinfecting industrial plants or livestock farming, or in vivo, more particularly with animals or human beings.
Antibiotics are a class of medication used specifically for treating bacterial infections by helping the organism to fight the bacterial infection.
By «resistant bacteria», is meant a bacteria resistant or less sensitive than usually expected to at least one classic antibiotic and/or anti-bacterial drug, while this bacteria belongs to a species which should be sensitive or a priori sensitive. The classic antibiotic and/or anti-bacterial drug may be chosen from the compounds belonging to families listed below.
By «multi-resistant bacteria» is meant, a bacteria resistant to several antibiotic(s) and/or anti-bacterial(s), in particular for which the species should be sensitive or a priori sensitive, “non-natural drug resistance”, more particularly a bacteria presenting at least two non-natural drug resistance. More particularly, the expression “multi-resistant” may apply to a bacterium or a stock of bacteria resistant to all antibiotics tested in at least two antibiotic classes.
Antiviral drugs are a class of medication used specifically for treating viral infections. Like antibiotics, viruses may evolve to resist the antiviral drug.
By “anti-fungal” is meant a compound or a composition exhibiting fungistatic or fungicidal properties, in particular in vitro, for example in a composition such as a pharmaceutical composition, a food composition or a cosmetic composition, or for disinfecting industrial plants or livestock farming, or in vivo, more particularly with animals or human beings.
An anti-fungal may be active on moulds, yeast and/or dimorphic fungi, in particular on a subject, such as an animal, and in particular a human being.
A substance or a composition is bacteriostatic/virustatic/fungistatic when the bacteria/virus/fungi multiplication is suspended or stopped. Experimentally the Minimum Inhibitory Concentration (MIC) is the lowest concentration of the substance or composition where no bacterial/viral/fungical growth is observed after 18 to 24 h of contact in favourable conditions to bacterial/viral/fungical growth.
A substance or a composition is bactericidal/virucidal/fungicidal when it definitively destroys the vitality of bacteria/virus/fungi. Experimentally, the logarithmic fall of bacteria/virus/fungi population is measured. The batericidial/virucidal/fungicidal effect is defined as a fall of 3 Log of the bacteria/virus/fungi population. The “bactericidal effect” may also be defined as in example 6.
By «nosocomial infection» is meant all microbial infections resulting from treatment in an hospital or a healthcare service unit, in particular with the apparition of symptoms happening at least 25 h, more particularly at least 48 h, after the subject is admitted in the hospital or within 30 days after discharge.
A genotoxic product is a product that shows an effect on the genetic material of a cell. The genotoxicity may be measured by Ames test, in particular according to ICH guidelines. A carcinogen product is a product that shows an effect on the genetic material of a cell, this effect is transmissible through mitosis and may leads to apparition of tumours, in particular measured in compliance with the OECD guideline 474.
“Not carcinogen” and/or “non genotoxic” means that at the dosage at which this the composition is used, no carcinogenic and/or no genotoxic effect is shown.
The expression “to potentiate” means that the anti-microbial, in particular anti-bacterial effect or antiviral effect, of the composition of the invention is higher, faster or leads to much less, or no, induction of resistance than with trans-cinnamaldehyde alone, in particular there is much less, or no, resistance induced when potentiated.
By “Interfering agents” is meant organic or mineral compounds which in contact to the anti-microbial agent reduces or avoids the anti-microbial effects. By example, it is well-known that chlorine or iodine are inactivated by proteins (in particular albumin). As examples of interfering agents, one can notably cite bovine albumine and/or sheep erythrocytes.
By “the composition is free of X” is meant that the amount of X is lower than 10 ppm by weight compared to the total weight of the composition, in particular lower than 1 ppm by weight compared to the total weight of the composition, and more particularly lower than 0.1 ppm by weight compared to the total weight of the composition, and even lower than 0.01 ppm by weight compared to the total weight of the composition, more particularly its means that 0 ppm of X is present in the composition.
The «synergy» is be calculated as follows:
FICindex=(MICA/B/MICA)+(MICB/A/MICB)
where
-
- FICindex is the index of the Fractional Inhibitory Concentration,
- MICA=MIC of the A compound alone,
- MICB=MIC of the B compound alone,
- MICA/B=MIC of the A compound, in the mixture A+B,
- MICB/A=MIC of the B compound, in the mixture A+B.
This formula allows determining the effect, as when:
-
- FICindex is less or equal to 0.5 there is a synergic effect,
- FICindex is more than 0.5 and less or equal to 1 there is an additive effect,
- FICindex is more than 1 and less or equal to 4 an indifferent effect, and
- FICindex is more than 4 there is an antagonistic effect.
Trans-cinnamaldehyde may comprise an amount of cis-cinnamaldehyde of less than 20% by weight, in particular less than 10% by weight, more particularly less than 5% by weight, even less than 2% by weight, very particularly less than 1% by weight. Following an embodiment, trans-cinnamaldehyde is free of cis-cinnamaldehyde.
CNM is sometimes used as abbreviation of trans-cinnamaldehyde.
By “terpenoïd” is meant in the invention derivatives issues from isoprene which are biologically obtained from condensation of C5 units, in particular leading to hemiterpenes, monoterpenes and sesquiterpenes. In this description, cinnamaldehyde and cinnamaldehyde derivatives are excluded from the “terpenoïd” definition.
By “excipient” is meant all compounds which do not belong to active blend and which is not anti-microbial activity or which is used for other reasons than its potential anti-microbial activity.
For the growth curves, the following symbols are used:
Following a first aspect, the invention has for subject matter a composition, in particular a pharmaceutical composition, comprising:
-
- a synergistic active blend comprising, or consisting of:
- trans-cinnamaldehyde and
- a potentiating agent comprising or consisting of:
- at least one terpenoïd, chosen from mono and sesquiterpenoïds and/or
- at least one derivate from trans-cinnamaldehyde, belonging to the group of phenylpropanoids, with a molecular weight below 240 g/mol,
- optionally phenyl propane derivatives
- in particular the potentiating agent comprises six compounds or less,
- optionally a drug, in particular an antibiotic or an antiviral drug,
- optionally a carrier.
- a synergistic active blend comprising, or consisting of:
The composition is preferably a synthetic one which consequently is chemically well defined and shows reproducible properties, what is essential in the pharmaceutical field. In addition, the composition is not carcinogen and preferably further non genotoxic. To this end, the composition preferably comprises amounts of coumarin and/or safrole lower than 1% by weight compared to the total weight of the composition, even more particularly the composition is free of coumarin and/or safrole, still more particularly free of coumarin and safrole.
In a preferred embodiment, coumarin content is less than 0.5% by weight, particularly less than 0.1% by weight, more particularly less than 0.01% by weight compared to the total weight of the composition. In a preferred embodiment, safrole content is less than 0.5% by weight, particularly less than 0.1% by weight, more particularly less than 0.01% by weight compared to the total weight of the composition.
The composition may comprise eugenol in an amount of less than 0.5% by weight, more particularly of less than 0.1% by weight, more particularly less than 0.01% by weight compared to the total weight of the composition. Following an embodiment, the composition is free of eugenol.
The composition is preferably free of coumarin, safrole and eugenol.
This low amount of coumarin, safrole and/or eugenol, or even the absence of coumarin, safrole and/or eugenol, may lead to a composition exhibiting low or no carcigenocity and/or genotoxicity, more particularly no carcigenocity.
It has been surprisingly discovered that by using potentiating agent a synergy is obtained with the consequence that the antimicrobial effect (namely not only the antibacterial effect but also the antiviral or the antifungal effect) of trans-cinnamaldehyde is potentiated. As a consequence, the effective amount of trans-cinnamaldehyde in the composition, in particular in a drug, can be lowered resulting in a composition, in particular a drug, with reduced toxicity and preferably improved efficacy.
In addition, it has been surprisingly discovered that by using potentiating agent the composition, in particular the drug, has a broad spectrum of activity, also on Gram−bacteria.
In particular, the composition of the invention shows activity on most of possible microbes, including drug resistant microbes, in particular activity on:
-
- bacteria, for example drug resistant bacteria such as drug resistant Staphylococcus aureus, for example such as MRSA, VRSA, drug resistant enterobacter, such as NDM-1; and at least one bacteria such as those disclosed below;
- fungi, for example
- Epidermal, dermal and/or keratinous appendage fungi, in particular Candida, Trichophyton, Malassezia, and Microsporum,
- Systemic, in particular non-opportunistic disease, more particularly due to Blastomyces, Coccidioides, and opportunistic disease due to Aspergillus, Candida albicans, and Cryptococcus,
- viruses, such as HIV; herpes viruses, the hepatitis B and C viruses, and influenza A and B viruses, in particular enveloped viruses.
The composition of the invention is consequently a wide spectrum antibacterial and/or antifungal and/or antiviral drug.
The anti-microbial effect of the composition is effective even at low content in trans-cinnamaldehyde and/or when interfering agent are present.
The anti-microbial effect of the composition is also observed for anaerobic bacteria.
Moreover, the potentiating agent advantageously increases the MIC at least to one or more bacteria as defined below and/or increases the kinetics of the activity. In particular, it has been discovered that the active blend of the invention is able to decrease bacterial division capabilities at sub-MIC concentrations while trans-cinnamaldehyde is not.
In addition, it has been surprisingly discovered that the composition of the invention reduces the log reduction time in comparison to trans-cinnamaldehyde alone, meaning in particular that the composition of the invention acts more quickly and/or shows a bactericidal/virucidal/fungicidal effect.
Furthermore, it has been surprisingly discovered that by using potentiating agent the composition, in particular the drug, induces less resistance. Thus, the potentiating agent advantageously decreases the resistance induction, in particular to zero, of trans-cinnamaldehyde.
The composition of the invention further shows very interesting results on viruses, in particular on enveloped viruses. It is believed by the inventors that the composition of the invention may act on membranes of the bacteria/viruses/fungi.
The potentiating agent may comprise one to five terpenoïds, in particular one, two or three terpenoïds, chosen from monoterpenoïds, and sesquiterpenoïds.
More particularly, the terpenoïd(s) may come belong to the following families:
-
- monoterpenoïds (10 carbon atoms-skeleton) such as
- acyclic monoterpenoïds, in particular regular or irregular acyclic monoterpenoïds, more particularly myrcane, even more particularly neryl acetate, linalyl acetate, citral, citronellol, citronellal, geranial, neral, geraniol, linalool;
- bicyclic monoterpenoïds, in particular camphene, thuyane, pinane, more particularly borneol, alpha pinene, incensol, isopenocamphone, monoterpenes, pinene; and
- menthanes, in particular 1,8-cineole, alpha terpineol, ascaridiole, carvacrol, carvone, menthol, menthone, piperitone, pulegone, alpha pinene, cuminaldehyde, terpenyl acetate, limonene, terpin-1-en-4-ol, thujan-4-ol, and thymol
- Sesquiterpenoïds (15 carbon atoms-skeleton), such as
- aromadendranes, in particular viridiflorol and thujone;
- beta santalanes, in particular santalols;
- bisabolanes, in particular bisabololoxydes and zingiberene;
- daucanes, in particular carotol;
- eremophilanes, in particular furanoeudesma-1,3diene;
- caryophyllanes, in particular beta-caryophyllene; and
- and patchoulanes, in particular patchoulol.
- monoterpenoïds (10 carbon atoms-skeleton) such as
The terpenoïd(s) present(s) in the potentiating agent are in particular chosen from menthane, acyclic monoterpenoïds and caryophyllane, still more particularly they are chosen from cineol, linalool and beta-caryophyllene. The terpenoïd(s) present(s) in the potentiating agent is preferably linalool and optionally cineole and/or beta-caryophyllene
Phenylpropane derivatives are biologically obtained from phenylpropane and leads to C6-C3 derivatives or C6-C1 derivatives, and lactone-corresponding molecules. Phenylpropane derivatives are in particular selected from the group consisting of apiole, coniferyl benzoate, chavicol, cinnamein, vanillin and benzyl benzoate. Benzyl benzoate is preferred.
The terpenoïds and phenylpropane derivatives present in the potentiating agent may comprise, or consists of, benzyl benzoate and cineol; benzyl benzoate and linalool; benzyl benzoate and beta-caryophyllene; cineole and linalool; cineole and beta-caryophyllene; linalool and beta-caryophyllene; benzyl benzoate, cineol and linalool; benzyl benzoate, cineol, and beta-caryophyllene; benzyl benzoate, linalool and beta-caryophyllene; cineol, linalool and beta-caryophyllene; and benzyl benzoate, cineol, linalool and beta-caryophyllene. The terpenoïds and phenylpropane derivatives present in the potentiating agent preferably comprises, or consists of, benzyl benzoate, cineol, linalool and beta-caryophyllene.
The benzyl benzoate may range from 0.1 to 25% by weight, in particular from 0.15 to 5% by weight, and more particularly from 0.15 to 3% by weight compared to the total weight of the active blend. According to an embodiment, the amount of benzyl benzoate ranges from 0.2 to 1% by weight, and more particularly is around 0.35% by weight compared to the total weight of the active blend.
The cineol may range from 0.1 to 25% by weight, in particular from 0.25 to 7% by weight, and more particularly from 0.3 to 3.5% by weight compared to the total weight of the active blend. According to an embodiment, the amount of cineol ranges from 0.5 to 2.5% by weight, and more particularly is around 1.04% by weight compared to the total weight of the active blend.
The linalool may range from 0.1 to 25% by weight, in particular from 0.5 to 6% by weight, and more particularly from 0.75 to 5% by weight compared to the total weight of the active blend. According to an embodiment, the amount of linalool ranges from 1 to 5% by weight, and more particularly is around 2.45% by weight compared to the total weight of the active blend.
The beta-caryophyllene may range from 0.1 to 25% by weight, in particular from 0.5 to 10% by weight, and more particularly from 0.75 to 5% by weight compared to the total weight of the active blend. According to an embodiment, the amount of beta-caryophyllene ranges from 1 to 3.5% by weight compared to the total weight of the active blend, and more particularly is around 1.75% by weight compared to the total weight of the active blend.
The potentiating agent may comprise an amount of terpenoïd(s) and phenylpropane derivatives ranging from 1 to 50% by weight, in particular from 1 to 25% by weight, more particularly from 1 to 15% by weight, even more particularly from 3 to 10% by weight, still more particularly from 3.5 to 8% by weight compared to the total weight of the active blend.
The potentiating agent comprises at least one, in particular 1, 2, 3, 4 or 5, derivate from trans-cinnamaldehyde. These derivates may correspond to Formula I:
wherein
-
- X represents —CH2OH, —CH2OY, —CHO, —COOH, —COOZ, —CO-Hal,
- Y represents Ra or —CORa with Ra being an alkyl comprising 1 to 6 carbon atoms, such as methyl or ethyl,
- Z represents an alkyl comprising 1 to 6 carbon atoms, such as methyl or ethyl,
- R1 and R2 represent independently, H, —OH, an alkyl comprising 1 to 6 carbon atoms, such as methyl or ethyl, or an halogen atom, for example chosen from F, Cl and Br,
- R3 and R4 represent independently, H, an alkyl comprising 1 to 6 atoms, such as methyl or ethyl, an alkoxy comprising 1 to 6 carbon atoms, such as methoxy or ethoxy, or an halogen atom, for example chosen from F, Cl and Br,
- Hal represents an halogen atom, for example chosen from F, Cl and Br,
and wherein when X is —CHO then at least one of R1, R2, R3 and R4 does not represent H.
In particular when X is —CHO one of R1, R2, R3 and R4 does not represent H and three represent H, and when X is —CH2OH, —CH2OY, —COOH or —COOZ then all or three of R1, R2, R3 and R4 represent H.
The trans-cinnamaldehyde derivate may be chosen from:
-
- a substituted trans-cinnamaldehyde:
- on the aromatic cycle, such as ortho, meta or para, by alkyl groups, such as methyl, ethyl, propyl, alkoxy group, such as methoxy or ethoxy, an halogen atom, such as F, CI or Br, and/or
- on the lateral chain, such as alkyl group, such as methyl, ethyl, propyl, or an halogen atom, such as F, Cl or Br on the double bond, or
- a compound corresponding to trans-cinnamaldehyde wherein the aldehyde function is replaced by another function, in particular such as acid, the corresponding ester, in particular methyl, ethyl and propyl ester, alcohol, the corresponding ester, in particular acetate or propionate.
- a substituted trans-cinnamaldehyde:
The potentiating agent may comprise at least one, two, three or four derivates from trans-cinnamaldehyde, preferably two derivatives. The derivates present in the potentiating agent preferably comprises trans-cinnamaldehyde, substituted on the aromatic cycle by R3 and/or R4, in which at least one radical R3 or R4 is not H, and a compound corresponding to the trans-cinnamaldehyde wherein the aldehyde function is replaced by another function, advantageously an esterified alcohol (X is —CH2OY and Y is —CORa with Ra is as defined above, such as methyl or ethyl). In a preferred embodiment, the derivates present in the potentiating agent are trans-2-methoxycinnamaldehyde and cinnamyl acetate.
The trans-2-methoxycinnamaldehyde may range from 0.1 to 30% by weight, in particular from 1 to 20% by weight, and more particularly from 2 to 15% by weight compared to the total weight of the active blend. According to an embodiment, the amount of trans-2-methoxycinnamaldehyde ranges from 2.5 to 8% by weight, and more particularly is around 5.35% by weight compared to the total weight of the active blend.
The cinnamyl acetate may range from 0.1 to 30% by weight, in particular from 0.5 to 15% by weight, and more particularly from 0.75 to 10% by weight compared to the total weight of the active blend. According to an embodiment, the amount of cinnamyl acetate ranges from 1 to 5% by weight, and more particularly is around 2.5% by weight compared to the total weight of the active blend.
The amount of derivate(s) of trans-cinnamaldehyde in the potentiating agent may range from 1 to 50% by weight, in particular from 2 to 20% by weight, more particularly from 4.5 to 12% by weight compared to the total weight of the active blend.
The trans-cinnamadehyde derivate(s), in particular comprising, or consisting of, trans-2-methoxycinnamaldehyde and/or cinnamyl acetate, may range from 1 to 50% by weight, in particular from 1 to 40% by weight, more particularly from 1 to 17.5% by weight, even more particularly from 3 to 10% by weight, still more particularly from 3.5 to 8% by weight compared to the total weight of the active blend.
In a specific embodiment, the potentiating agent consists of:
-
- at least one terpenoïd and phenylpropane derivatives in an amount ranging from 1 to 50% by weight, in particular from 1 to 25% by weight, more particularly from 1 to 15% by weight, even more particularly from 3 to 10% by weight, still more particularly from 3.5 to 8% by weight compared to the total weight of the active blend,
- at least one derivate from trans-cinnamaldehyde in an mount ranging from 1 to 50% by weight, in particular from 1 to 40% by weight, more particularly from 1 to 17.5% by weight, even more particularly from 3 to 10% by weight, still more particularly from 3.5 to 8% by weight compared to the total weight of the active blend,
in particular wherein the total amount of the compounds of these two species ranges from 3 to 70% by weight, more particularly from 4 to 40% by weight, even more particularly from 5 to 25% by weight, still more particularly from 6 to 15% by weight compared to the total weight of the active blend.
In a specific embodiment, the potentiating agent comprises at least:
-
- one terpenoïd selected from menthane, in particular cineole, acyclic monoterpenoïds, in particular linalool, and caryophyllane, in particular beta-caryophyllene
- one phenylpropane derivative, in particular benzyl benzoate,
- trans-cinnamaldehyde, substituted on the aromatic cycle by R3 and/or R4, in which at least one radical R3 or R4 is not H, preferably trans methoxycinnamaldehyde and
- a compound of formula I wherein X is —CH2OY, Y is —CORa with Ra is as defined above, preferably cinnamyl acetate
in the above mentioned amounts.
The following active blends are preferred:
The contents are expressed in % by weight compared to the total weight of the active blend. The numerical value is expressed by meant plus or minus 10%, for example 5 means in the range of 4.5 to 5.5%.
The active blend may comprise an amount of trans-cinnamaldehyde ranging from 30 to 97% by weight, in particular from 60 to 96% by weight, even more particularly from 75 to 95% by weight, still more particularly from 85 to 94% by weight compared to the total weight of the active blend.
Following one embodiment, the active blend ranges from 0.1 to 10% by weight compared to the total weight of the composition, in particular when the composition is intended to be used as a topical product.
Following another embodiment, the active blend ranges from 0.1 to 70% by weight compared to the total weight of the composition, in particular when the composition is intended to be used as a systemic product.
Following an embodiment, this composition comprises as active compounds, more particularly as anti-microbial compounds, only the synergistic active blend.
Following an embodiment, the composition, in particular the pharmaceutical composition, further comprises at least one, and in particular one, antibiotic.
The antibiotics which may be used in the present invention, in particular in the pharmaceutical composition, more particularly for use for preventing and/or treating bacterial infection(s), are numerous and may be gathered by families according to their spectrum of action, of their chemical structure, like according to their mode of action on the bacteria.
According to a particular mode of realization, the antibiotic or at least one of the antibiotic is selected among:
- 1. antibiotics acting on the bacterial wall, in particular, interfering with the synthesis of peptidoglycane,
- 2. antibiotics operating the membranes of the cells, external membrane and/or cytoplasmic membrane,
- 3. antibiotics acting on the synthesis of proteins, in particular on bacterial ribosome,
- 4. antibiotics blocking the synthesis of messenger RNA,
- 5. antibiotics acting on DNA, for example cut of the bits of DNA and unfolding of the DNA or inhibition of the replication of DNA,
- 6. antibiotics acting by competitive inhibition, in particular an antibiotic interfering with the folate metabolism,
- 7. one of their pharmaceutically acceptable salts, and
- 8. one of their combinations.
As example, the antibiotics acting on the bacterial wall can be selected among (A) the fosfomycin (or phosphomycin), (B) an antibiotic of the family of the glycopeptides, such as the vancomycin, the teicoplanin (or teichoplanin), the ristocetin, or the avoparcin, or (C) an antibiotic of the family of the beta-lactam antibiotics.
The family of beta lactamins is in particular represented by (A) the penams, in particular the group G of penicillin G, the group M of antistaphylococcic penicillins, the group has amino-benzylpenicillin (ampicillin), the group of acyl-ureido-penicillins, the group of the amidino-penicillins and the group of the inhibitors of the betalactamases, (B) the penems and the carbapenems, such as the meropenem (or meropenem) and the imipenem, (C) the cephems, in particular the cephalosporins of 1st, 2nd and 3rd generation, and (D) the monobactas.
The beta-lactam antibiotics have as targets of proteins present on the external face of the membrane cytoplasmic and called proteins binding penicillin (PLP).
As example, the antibiotics operating the membranes of the cells can be selected among the family of the polymyxins, the family of the gramicidins, and the tyrocidin. Still as example, the antibiotics acting on bacterial ribosome, can be selected among the fusidic acid, the family of the aminosides, the family of phenicoles, the family of the tetracyclines, the family of the oxazolidinones and groups it macrolides, lincosamides and synergistins (or streptogramins).
As for antibiotics blocking the synthesis of messenger RNA, they include in particular the family of the rifamycins, represented by the rifamycin SV and the rifampicin. The rifamycins are fixed on under unit B of RNA polymerase and prevent the initiation of the synthesis of mRNA.
Always as example the antibiotics acting on DNA can be selected among the family of the quinolones, the family of the fluoroquinolones, and the products nitrated such as the nitro-imidazoles oxyquinoleines, nitrofurans or nitro-imidazoles.
Lastly, the antibiotics interfering with the metabolism of the folates include the family of sulphamides and the trimethoprim.
According to a particular mode of realization, the antibiotic or at least one of the aforesaid antibiotic is selected among the meropenem, the rifampicin and the tigecyclin.
According to a particular mode of realization, the antibiotic or one of the aforesaid antibiotic is the meropenem or the rifampicin.
More particularly, the antibiotic may be chosen from the following:
-
- aminosides,
- betalactamins, as betalactamins cephalosporins, betalactamins penicillins, and other betalactamins (carbapenems, monobactame),
- cyclins, such as doxycyclin, limecyclin, metacyclin, minocyclin, tetracyclin, oxtetracyclin, tigecyclin,
- glycopeptides, such as teicoplanin and vancomycin, and polypeptides,
- macrolides and macrolides like, such as lincosamides, ketolides and synergistins,
- quinolones, in particular fluoroquinolones,
- anti-bacterial peptides, such as gramicidin,
- phages, and
- others, such as fusidic acid, noxytiolin, daptomycin, fosfomycin, oxazolidinone, phenicoles, polymyxins, rifampicin,
It has been surprisingly discovered that the active blend of the invention is able to potentiate the antibiotics, advantageously meaning that the effect and/or the scope of the antiobitic is increased. Furthermore, the combination active blend of the invention plus antibiotic can be efficient against bacteria which are resistant to the antibiotic of the combination. Thus, the active blend of the invention can be used to re-sensibilize bacteria towards antiobiotic.
It has been surprisingly discovered that the antibiotics is able to potentiate the active blend of the invention, advantageously meaning that the effect and/or the scope of the active blend of the invention is increased.
The synergy effect is particularly observed with antibiotics with other target than the membrane (gentamicin, amikacin, erythromycin and clindamycin).
The composition, in particular the pharmaceutical composition, preferably comprises the antiobiotic(s) at its normal doses or at a reduced dosis (in comparison to the normal doses known by the skilled person).
Following an embodiment, the composition, in particular the pharmaceutical composition, further comprises at least one, and in particular one, antiviral drug.
The antiviral drug can in particular chosen from: entry inhibitors, fusion inhibitors, Integrase inhibitor, Interferon type III, Interferon type II, Interferon type I, Interferon, Nucleoside analogues, Protease inhibitor, Reverse transcriptase inhibitor, Synergistic enhancer (antiretroviral),
In particular, the antiviral drug can in particular chosen from: Abacavir, Aciclovir, Adefovir, Amantadine, Amprenavir, Rintatolimod, Atazanavir, Emtricitabine/tenofovir/efavirenz, Boceprevir, Darunavir, Delavirdine, Didanosine, Docosanol, Edoxudine, Efavirenz, Emtricitabine, Enfuvirtide, Entecavir, Famciclovir, Fomivirsen, Fosamprenavir, Foscarnet, Ganciclovir, lbacitabine, combination of inosine, acetamidobenzoic acid, and dimethylaminoisopropanol, Idoxuridine, Imiquimod, Indinavir, Inosine, Lamivudine, Lopinavir, Loviride, Maraviroc, Moroxydine, Methisazone, Nelfinavir, Nevirapine, Oseltamivir, Peginterferon alfa-2a, Penciclovir, Peramivir, Pleconaril, Podophyllotoxin, Raltegravir, Ribavirin, Rimantadine, Ritonavir, Pyramidine, Saquinavir, Stavudine, Tea tree oil, Tenofovir, Tenofovir disoproxil, Tipranavir, Trifluridine, Trizivir, Tromantadine, Truvada, Valaciclovir (Valtrex), Valganciclovir, Vicriviroc, Vidarabine, Viramidine, Zalcitabine, Zanamivir (Relenza), Zidovudine,
The composition, in particular the pharmaceutical composition, preferably comprises the antiviral drug(s) at its normal doses or at a reduced dosis (in comparison to the normal doses known by the skilled person).
The composition, in particular the pharmaceutical composition, may comprise a carrier. The carrier may range from 0.1 to 99.9% by weight, preferably from 0.5 to 99.9% by weight, more preferably from 1 to 99.5% by weight, more preferably from 1 to 80% by weight, compared to the total weight of the composition.
In particular, the carrier is such that it allows to analytically follow the process to obtain the composition, in particular the pharmaceutical composition, for example with HPLC and/or GC analysis, more particularly capillary analysis.
In an embodiment, the carrier comprises or consists of, less than ten excipients, more particularly less than eight, even more particularly less than five excipients, more particularly less than four excipients, still more particularly less then three excipients, and even more particularly one excipient.
When the excipient is a compound bearing at least a charge “one excipient” may mean one compound under different salt forms.
Following one embodiment, the carrier represents 95 to 99.9% by weight compared to the total weight of the composition, in particular the remaining is the active blend, more particularly the composition is intended for an external use, for example meaning external to the skin or to the mucosa of the subject.
Following another embodiment, the carrier represent 0.5 to 50% by weight compared to the total weight of the composition, in particular the remaining is the active blend, more particularly the composition is intended for an internal use.
The carrier may be solid, liquid, gel, or pasty.
The carrier may comprise, or consists of, one or more excipient, in particular pharmaceutically acceptable, and optionally one or more additive such as preservatives, vitamins, minerals.
The invention has for second object the composition of the invention for use as a medicament.
Following one embodiment, the medicament is free of classic antibiotics and/or antiviral drugs, in particular as disclosed in the description.
Following another embodiment, the medicament comprises at least one classic antibiotic and/or antiviral drug, in particular as disclosed in the description.
The invention has for third object the composition for use for preventing and/or treating microbial, in particular bacterial and/or fungal and/or viruses, infection of a subject, advantageously even in the presence of interfering agents. This may also be worded as preventing and/or treating disease induce by microbes, such as bacteria and/or fungus and/or viruses. The bacteria and/or fungus and/or viruses advantageously also include drug-resistant bacteria and/or fungus and/or viruses more particularly to antibiotics and/or antiviral drugs, even more particularly to one or several of the classic antibiotics and/or antiviral drugs disclosed in this description.
A subject may be an animal such as production animals, for example cattle, swine and poultry, pets, for example dog or cat, or a human being.
The pharmaceutical composition may have a microbicidal activity, in particular a bactericidal activity, more particularly when interfering substance(s) is(are) present, such as bovine albumine and/or sheep erythrocytes.
The bacteria may be aerobic or anaerobic.
Interestingly, the medicament of the invention shows a wide spectrum of activity on all kind of bacteria, including Gram−bacteria, which can further be resistant or multi-resistant.
As was said before, more and more bacteria become drug resistant to one or several of these anti-bacterial agents, in particular to antibiotics, more particularly to classic antibiotics, for example such as those listed in the description.
Principal actual anti-bacterial agents are more active on, or even are specific of, Gram+ or Gram−bacteria. More particularly, most of the known anti-bacterial are active on, or even are specific of, Gram+bacteria. There is thus a need for anti-bacterial compositions, in particular pharmaceutical compositions, able to stop the development or to destroy the vitality of Gram−bacteria, and advantageously also of Gram+bacteria.
In particular, the compositions, more particularly the pharmaceutical compositions, according to the invention, present an activity toward Gram− and Gram+bacteria. They can be efficient toward non drug resistant and/or drug resistant bacteria, in particular toward multi-resistant bacteria.
In particular, the compositions, more particularly pharmaceutical compositions, are active toward drug-resistant Gram−bacteria strains such as:
-
- Pseudomonas, and more particularly P. aeruginosa;
- Acinetobacter, and more particularly A. baumanii;
- Escherichia, and more particularly E. coli;
- Enterobacter and more particularly E. aerogenes and or E. cloacae;
- Serratia, in particular Serratia marscescens;
- Citrobacter, in particular Citrobacter freundii; and/or
- Klebsiella, in particular Klebsiella pneumonia.
It has also been shown that the compositions, more particularly pharmaceutical compositions, may be active toward drug-resistant Gram+bacteria strains such as:
-
- Staphylococcus, in particular S. aureus;
- Enterococcus, in particular E. faecalis; and or
- Propionibacter, in particular Propionibacter acnes.
Compositions, more particularly pharmaceutical compositions, may be active toward anaerobic bacteria, in particular:
-
- Bacteroides, such as B. fragilis and B. thetaiotaomicron;
- Eggerthella, such as E. lenta;
- Peptostreptococcus, such as P. micros, P. spp, and P. anaerobius;
- Clostridium, such as C. perfringens and C. difficile; and/or Micromonas.
In a first embodiment the bacteria is of the group Pseudomonas, in particular drug-resistant Pseudomonas, more particularly at least one drug resistance, even more particularly two, three, four, five or six drug resistance chosen among:
-
- fluoroquinolones resistance;
- cephalosporins resistance in particular 1st, 2nd or 3rd generation;
- production of a penicillinase, meaning betalactamins penicillins resistance, in particular in case of hyperproduction of chromosomic cephalosporinase;
- production of extended spectrum betalactamase (ESBL, ex: types PER-1 or GES-2);
- production of a metallo-betalactamase, in particular of V1M-2 type;
- lack of porin, in particular D2 porin, which may lead to betalactamins other than penicillins and cephalosporins resistance; and
- aminosides resistance.
In a second embodiment the bacteria is of the group Acinetobacter, in particular drug-resistant Acinetobacter, more particularly at least one drug resistance, even more particularly two, chosen among:
-
- multiresistance;
- Vietnamese expanded Spectrum betalactamase VEB-1; and
- production of a metallo-betalactamase, in particular of VIM-4 type.
In a third embodiment the bacteria is of the group Escherichia, in particular drug-resistant Escherichia, more particularly at least one drug resistance, even more particularly two, chosen among:
-
- fluoroquinolones and quinolones resistance;
- cephalosporins resistance in particular 1st, 2nd or 3rd generation;
- production of extended spectrum betalactamase (ESBL);
- production of a metallo-betalactamase, ex: NDM-1 type;
- production of a carbapenemase, ex KPC-2 type; and
- production of a penicillinase.
In a fourth embodiment the bacteria is of the group Staphyloccocus, in particular drug-resistant Staphyloccocus, more particularly at least one drug resistance, even more particularly two, chosen among:
-
- methicillin resistance;
- aminosides resistance; in particular tobramycin/kanamycin resistance: KT phenotype; and
- fluoroquinolones resistance.
In a fifth embodiment the bacteria is of the group Enteroccocus, in particular drug-resistant Enteroccocus, more particularly at least one drug resistance, even more particularly two, chosen among:
-
- aminosides resistance; and
- macrolides and apparented macrolides resistance.
In a sixth embodiment the bacteria is of the group Enterobacter, in particular drug-resistant Enterobacter, more particularly at least one drug resistance, more particularly production of extended spectrum betalactamase resistance (ESBL).
-
- production of extended spectrum betalactamase (ESBL, ex: types PER-1 or GES-2); and
- production of a carbapenemase, ex KPC-2 type.
In a seventh embodiment the bacteria is of the group Propionibacter, in particular drug-resistant Propionibacter, more particularly at least one drug resistance, even more particularly two.
In an eighth embodiment the bacteria is of the group Serratia, in particular drug-resistant Serratia, more particularly at least one drug resistance, even more particularly two, chosen among:
-
- production of a carbapenemase, ex KPC-2 type; and
- production of extended spectrum betalactamase (ESBL, ex: types SME-1 or SME-2);
In an ninth embodiment the bacteria is of the group Citrobacter, in particular drug-resistant Citrobacter, more particularly by production of a carbapenemase, ex KPC-2 type.
In an tenth embodiment the bacteria is of the group Klebsiella, in particular drug-resistant Klebsiella, more particularly at least one drug resistance, even more particularly two, chosen among:
-
- production of a carbapenemase, ex KPC-2 type or VIM; and
- production of extended spectrum betalactamase (ESBL, ex: type OXA 48).
According to another embodiment, the composition may be active toward bacteria from 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th embodiments and their mixture, for example mixture of 2, 3, 4, 5, 6, 7, 8, 9 and even the 10 embodiments.
The composition may be active, bacteriostatic and/or bactericidal, on Gram− and/or Gram+bacteria, in particular to drug resistant bacteria, and more particularly to at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or 11, in particular to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or 11, bacteria chosen from:
Gram−bacteria, for example such as:
-
- Pseudomonas, for example Pseudomonas aeruginosa, in particular drug resistant, more particularly VIM-2, GES-2 or PER-1;
- Acinetobacter, for example Acinetobacter baumanii, in particular drug resistant, more particularly VIM-4;
- Escherichia, for example Escherichia coli, in particular drug resistant, more particularly NDM-1 or KPC-2;
- Enterobacter, for example Enterobacter aerogenes, in particular drug resistant;
- Serratia, for example Serratia marscescens, in particular drug resistant, more particularly KPC-2, SME-2 or SME-1;
- Citrobacter, for example Citrobacter freundii, in particular drug resistant, more particularly KPC-2; and
- Klebsiella, for example Klebsiella pneumonia, in particular drug resistant, more particularly KPC-2;
and/or to Gram+bacteria, for example such as: - Staphylococcus, for example Staphylococcus aureus, in particular drug resistant;
- Enterococcus, for example Enterococcus faecalis and Enterococcus cloacae, in particular drug resistant, more particularly Enterococcus cloacae GES-5, KPC-2; and/or
- Propionibacter, for example Propionibacter acnes, in particular drug resistant.
The composition may be active, fungistatic or fungicidal, to fungi, in particular to drug resistant fungi, and more particularly to at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or 11 fungi chosen from:
-
- Epidermal, dermal and/or keratinous appendage fungi, in particular Candida, Trichophyton, Malassezia, and Microsporum,
- Systemic, in particular non-opportunistic disease, more particularly due to Blastomyces, Coccidioides, and opportunistic disease due to Aspergillus, Candida albicans, and Cryptococcus.
The composition may be active towards viruses, in particular HIV, herpes viruses, the hepatitis B and C viruses, and influenza A and B viruses. The composition is particularly active on enveloped viruses.
The pharmaceutical composition may be for use for anti-microbial prevention and/or treatment of a subject, for example for preventing and/or treating a disease and/or an infection caused by one or several, in particular two or three, bacteria such as the ones disclosed in the description, more particularly drug resistant and/or multiresistant bacteria; and/or fungus; and/or viruses. Following an embodiment the bacteria is an anaerobic bacteria.
The infections may be chosen from urinary system infection, respiratory system infection, digestive system infection, central nervous system infection, skin and soft tissues infection, bone infection, articulations infection, muscles infection, vasculary system infection, diabetic foot and eschar.
The pharmaceutical composition may thus be used for prevention and/or treatment of syndromes or diseases, in particular induced by a microbial infection, in particular such as bacterial and/or fungus and/or viruses infection, more particularly by bacteria and/or fungus and/or viruses disclosed in the instant description, wherein the syndrome and/or disease may concern:
-
- gastroenterology sphere, in particular syndromes and/or disease linked to digestive tube, more particularly Crohn disease (specifically in view of the good results obtained on anaerobic bacteria) and peptic ulcer,
- dermatology, in particular syndromes and/or disease linked to skin, more particularly diabetic foot and eschars,
- systemic infections which can be treated via systemic administration
- diseases linked to infections of urinary system,
- diseases linked to infections of respiratory system,
- diseases linked to infections of digestion system,
- diseases linked to infections of central nervous system,
- diseases linked to infections of skin and soft tissues,
- diseases linked to infections of bones, articulations an muscles,
- diseases linked to infections of vascular system,
- septic shock,
- AIDS
- Herpes
- Hepatitis B and C
More particularly, the pharmaceutical composition is for use for preventing or treating, in particular treating, infections, in particular bacterial infections, leading to bacteremia, Crohn disease, peptic ulcer, diabetic foot and/or eschars, more particularly when it is due to resistant or multi-resistant bacteria.
Drug-resistant bacteria and/or fungus and/or viruses appear more and more frequently, in particular in hospitals, where they may lead to nosocomial infections, which may cause nosocomial syndrom. Thus fighting against this bacteria and/or fungus and/or viruses allows preventing and/or treating at least some nosocomial infections.
The pharmaceutical composition may also be used for treatment and/or prevention of syndromes met in the following aetiology:
-
- infections passing through intact epithelial barrier, such as infections by inhalation or by ingestion;
- infections passing through discontinuous epithelial barrier, such as infections by biting, cuttings, wounds, injection, transplantation, transfusion;
- infections linked to the use of an invasive medical device, such as a prosthesis or a stent;
- surgery, peri-surgery and post-surgery infections;
- infections resulting from burns.
The pharmaceutical composition may also be used for treating and/or preventing the travel of bacteria, in particular such as Staphyllococcus aureus, in the different flora of the subject, in particular cutaneous flora, buccal flora and nose-throat flora.
The composition may be intended to be applied on skin and/or mucosa, on a device intended to contact skin and/or mucosa, and/or on a device intended to break the epidermal barrier.
Following one aspect, the invention has for subject matter a process for prophylaxis or for preventing microbial, such as bacterial and/or fungal and/or viral, infections, in particular nosocomial, comprising the step of applying the composition, in particular the pharmaceutical composition, on the skin or on the mucosa surface where the epithelium barrier is or risks to be broken or damaged, such as by the use of a catheter, or presenting a break allowing the passage of microbes, such as bacteria and/or fungus.
The composition, especially the pharmaceutical composition, may be formulated for topic or systemic administration, per os or parenteral administration. In particular, for:
-
- injection administration, such as pulmonary, intraveinous, subcutaneous intramuscular and/or intraperitoneal formulation; the composition may thus be hydrophobic or hydrophilic.
local administration (rectal, cutaneous),
oral administration.
The composition may be presented as a liquid, a paste, a powder, a pomade, an emulsion, a cream, a gel, a tablet, a gelule.
The composition may be under hydrophobic form, meaning being free of water and or polar solvent.
When under the gel form, the viscosity may go from 500 to 2000, in particular from 750 to 1500 centipoise (cP). It may be measured at 25° C. with a CPE52 rotor at 250 rpm.
The composition, in particular as a pomade, a gel or a cream, may comprise polyethylene glycol, in particular macrogol 400, medium chains triglycerides and/or soja oil. The composition, may also comprise colloidal silica.
When under an emulsion form, the composition may comprise a gelling agent, which may be a polymer, such as polyvinyl pyrrolidone, in particular povidone, for example POVIDONE K30®. The emulsion may comprise a surfactant, for example such as a polysorbate, polyglyceryl oleate or capric/caprylic acids glycerides.
A hydrophilic composition may comprise a gelling agent and/or a surfactant, in particular such as disclosed above.
The intravenous formulation may comprise polyvinylpyrrolidone.
In a further object of the invention, the composition is used as a cleaning agent, in particular to clean medical devices which are intended to be in contact with the skin or mucosa, and more particularly when potential breaches may exist. For example this composition may be used on skin surface where an infusion is to be done or on skin or mucosa surface where a catheter is used, in particular urinary catheter.
In a further object of the invention, the composition is used as a cleaning, preventing or protecting agent against microbially induced corrosion.
The composition may also be used as a conservative, in particular for food or cosmetic composition.
The composition may also be used for cleaning and/or protecting a surface from a secondary colonisation.
Following a specific embodiment, the active blend and/or the composition, in particular as pharmaceutical composition or as a conservative, is bactericidal.
Following an embodiment, the composition, in particular pharmaceutical, comprises a large spectrum antiseptic, such as chlorhexidine, chlorine or iodine, in particular polyvidone iodine, such as betadine.
Following another embodiment, the invention has for subject matter a patch comprising a composition, in particular a pharmaceutical composition, according to the invention.
According to another aspect, the invention has for subject matter the use of the active blend for the preparation of a composition, in particular a pharmaceutical composition, in particular such as disclosed above.
According to still another aspect, the invention has for subject matter a treatment of microbial infection, in particular such as disclosed above, in which an efficient amount of active blend is delivered to a subject.
According to another aspect, the invention has for subject matter the use of a potentiating agent as defined above for improving the anti-microbial, in particular anti-bacterial and/or anti-fungal, activity of cinnamaldehyde, in particular in terms of decreasing the resistance induction, in particular to zero, increasing the MIC at least to one or more bacteria as defined above and/or increasing the kinetics of the activity.
The medicament can be administered at a dosage regime of one, two or three times per day. The dose can be determined by the doctor on the basis of his general knowledge. Initial PK/PD models in healthy CD-1 and Balb/c mice of both sexes suggested a maximal tolerated dose of 300 mg/kg with no clinically relevant adverse effect and no major biochemical and haematological change after 15×100 mg doses. Preliminary kinetic data suggest a rapid elimination of the active blends of the invention in particular active blend 9, with a remaining MRSA inhibition by plasma during 2 hours, in line with bibliographic data on the main compound, cinnamaldehyde (1.7 h).
The following examples are intended to illustrate the invention and in no way to limit it.
EXAMPLES Example 1: Bacterial StrainsThe tested strains have been isolated from various samplings from human patients (blood, urines, pulmonary aspiration, etc) They have been isolated from patients non-infected at their admission to the hospital and who have developed an infection after at least 48 h of hospitalisation. The studied strains are as follows:
Following a method allowing the dissolution of the active blend in a Mueller Hinton gelose the following MIC has been measured with the active blend 9 disclosed in table 1. Said active blend 9 having been tested at concentrations shown in the following table:
Antibacterial activity appears to be quite constant on every strain, regardless to the type of cell wall structure and to the presence of antibiotic resistance. One will note, with big interest, that active blend 9 is also active against anaerobic bacteria. In addition, active blend 9 is significantly more active on anaerobic bacteria than on aerobic bacteria.
Example 3: Antibacterial Activity of Active Blends 1 to 8Following a method allowing the dissolution of the active blend in a Mueller Hinton gelose the following MIC has been measured with pure trans-cinnamaldehyde (CNM) and with the active blends 1 to 8 disclosed in table 1 The MICs (%) of trans-cinnamaldehyde and of active blends 1 to 8 are shown in the following table.
Antibacterial activity appears to be quite constant on every strain, regardless to the type of cell wall structure and to the presence of antibiotic resistance.
Example 4: Efficacy of Active Blend 9Active blend 9 was thus tested against 123 strains isolated from nosocomial representing the main resistances mechanisms).
These results confirm the wide-spectrum efficiency of active blend 9 as the blend is active against all strains tested. MIC levels are quite constant and differences within one genus are never higher than one dilution.
While most genera have an in vitro MIC of 300 mg/L, some are especially susceptible (Acinetobacter and Burkholderia) while some others are little less susceptible (Serratia, Pseudomonas and Enterococcus).
Example 5: Bactericidal Activity of Active Blend 9Bacteria Numbering
The numberings are done by successive dilutions of tenth of the samples. Each dilution (100 μl) is spread on a Mueller Hinton gelose. The numbering is done on a Petri dish which contains between 15 and 150 colonies. The numbering threshold is thus 150 UFC/ml.
Neutralising Power of D/E
To stop the activity of the active blend after a defined time, 100 μl of the mixture active blend+bacteria is taken and diluted in 900 μl of a neutraliser dilutant in order to block the anti-bacterial action of the active blend.
The used neutraliser is the «Neutralizing broth for neutralizing and testing disinfectants and antiseptics» from Dey and Engley marketed by Criterion, with the following formula:
Glucose (10 g), lecithine (7 g), caseine peptone (5 g), Tween 80 (5 g), sodium thiosulfate (6 g), bipotassic phosphate (3.3 g), sodium bisulfite (2.5 g), yeast extract (2.5 g sodium), thioglycollate (1 g), monopotassic phosphate (0.1 g) and bromocresol purple (20 mg). The obtained powder is dissolved in a litre of deionised water, and the, after heating and dissolution, the medium is sterilised by going in an oven at 121° C. for 15 minutes. The final pH is 7.6±0.2.
A neutraliser control is done as follows:
A mixture active blend+neutraliser (half and half) is contacted with the bacterial inoculum. After 48 h a 37° C. of incubation, bacterial numbering should not be less than 50% of the control numbering. This allows demonstrating the absence of activity of the neutraliser and in particular the fact that the neutraliser blocks completely the anti-bacterial action of the active blend, avoiding the carry over phenomenon.
Bactericidal Activity Measure of the Active Blends
After 15, 30, 45 and 60 minutes of contacting active blend+bacteria, 100 μl of the mixture product/bacteria is taken and added to 900 μl neutraliser, then two dilution (10th and 100th) are done with cysteinated Ringer. The, 100 μl of each of the dilution is spread on a Mueller Hinton gelose. The dishes are then incubated 48 h and survivors are numbered.
The dose of active blend is corresponding to MIC x 4 of each of the tested strain.
Results Expression
By definition bactericidal effect is obtained if a minimum fall of 3 logarithms is observed from the starting inoculum. A graph of the bactericidy linking the bacteria number with the contact time with the active blend may be drawn; this allows to check if the bactericidy is intense and fast or slow.
Results:
For example, the active blend 9 shows a bactericidy on the 8132, 8239, 8154, 9004 strains. The results are shown in the following tables.
Bactericidal effect of active blend 9 was then analyzed with killing—curves for 60 strains at 4× the MIC, and compared to CNM at 4× the MIC and active blend 9 at 1% (positive control). Results are expressed in minutes necessary to reduce bacterial counts by 1 log.
Active blend 9 showed a mean lag of 50.8 minutes at 4×MIC and 15.6 minutes at 1% (Table 19). For seven strains, reduction of count was not observed at 4× the MIC of active blend 9 but active blend 9 was bactericidal for all strains at 1%.
Example 6: Kinetics StudiesGrowing capacities of 4 bacteria (NDM-1 E. coli, Multiresistant Enterococcus sp, VIM-2 P. aeruginosa and OXA-48 K. pneumoniae) in contact with active blend 9 was tested at three different concentrations: MIC, MIC/2, MIC/4, MIC/8 and MIC/16.
The results are reported in
Active blend 9 shows a bactericidal effect at MIC on K. pneumonia only while the other strains present a stable count of bacteria at MIC. Surprisingly, for the four strains, count of bacteria are stable during 12 hours at MIC/2 and 4 hours at MIC/4, indicating that active blend 9 is able to decrease bacterial division capabilities at sub-MIC concentrations.
Example 7: Internal SynergyFollowing a method allowing the dissolution of the active blend in a Mueller Hinton gelose the MIC has been measured with pure components of the active blends 1 to 9.
Table 9 shows that only cinnamaldehyde (CNM) is highly effective on all the strains. 3 compounds present no activity at the tested concentrations. The other compounds show specific activities with either narrow, intermediate or wide spectrum for caryophyllen, cinnamaldehyde acetate (CNM-A) and linalool respectively.
Following the same method, the MIC has been measured with pure trans-cinnamaldehyde (CNM) and active blends 1 to 9. In table 10, the results for active blends 1 to 8 are compared to active blend 9 (the most active) and are expressed in terms of dilution (value=Log 2 (CMI active blend or component/CMI active blend 9)).
The FIC index with reference to active blend 9 has been calculated and reported in the table below:
These results show that active blend 9 is the more potent blend. This proves that although the non CNM compounds do not present an activity at the tested concentration as such, their presence in the blend potentiate the antibacterial properties of CNM. All the products from active blend 9 formula are involved in the activity of the blend. Indeed, even when compounds removed are inactive against some bacteria, corresponding blends are less effective or present a narrower spectrum than active blend 9. These results confirm the importance of using blends, either for increasing efficacy or for decreasing toxicity. Indeed MIC measured with active blend 9 are, in overall, lower than those of natural compounds.
Example 8: Activity on Viruses of Active Blend 9The activity of active blend 9 (M9) on VIH has been tested and compared to the activity of AZT.
One notes a good inhibition of P24 by active blend 9.
Example 9: Antibacterial GelThe active blend 9 is solubilised in 93 ml of Macrogol 400 (Lutreol 400). Then colloidal silica (6 g) is added.
The mixture is homogenised to obtain a non flowing gel.
This gel may be used for example for preventing bacterial and/or fungal infections, for example in case of use of a catheter.
Said catheter may be covered by said gel and/or the surface susceptible to have an epithelial breach may be covered with said gel.
The ability of destroying bacteria is measured by an inoculation of a strain of MRSA in the cream, and then viable bacteria are measured.
Results are shown in the following table and
The reduction log time is 4 h10.
Such a gel allows thus the decrease of the amount of bacteria.
Example 10: Interfering SubstancesThis example (in accordance with NF EN 13727 has been performed with the active blend 9.
Interfering substances tested are a mixture of bovine albumine (3.0 g/l) and sheep erythocytes (3.0 ml/1).
Example 11: Treatment of Infected MiceThe test is performed on Balb/c mice aged 10 to 12 weeks.
They are infected by intraperitoneal injection of 4.107 Staphylococcus aureus, methicilline resistant, in a growing phase or with E. coli.
The day after, a daily treatment is done as follows:
-
- Intact: no infection
- Placebo: no treatment, and
- Active blend 9 at daily treatment of 50 and 100 mg/kg/j (one or two administration(s) per day).
The data are summarised in the following table.
Survival rates were improved by active blend 9 at 100 mg/kg/j on both strains model. This parameter was enhanced by 7 days follow up of health scores and bodyweights. Survival time of mice that did not survive the challenge was quite unchanged (MRSA) or slightly increased (E. coli at 100 mg/kg/bid). Excess death at maximal dose on E. coli suggests toxic effects in moribund mice. Overall table variations shows that for each strain model, two groups receiving intraperitoneal treatment at 100 mg/kg recovered from challenge (health score, sum of bodyweight).
Example 12: Interaction with AntiobioticsInteractions between active blend 9 and amikacin or colistine have been explored. Synergistic effects were evaluated with fractional inhibitory concentration (FIC) indexes and antibiotic—active blend 9 combinations were classified as synergistic (FIC <0.5) additional (FIC index between 0.5 and 1), indifferent (between 1 and 4) or antagonistic (>4). For each combinations several dose of antibiotic and active blend 9 were tested, leading to extensive evaluation of the combination and the impact of relative ratios. 56 FIC indexes were then calculated for every combination tested. These FIC index are represented in the tables below.
This formulation is thereafter diluted in water.
Claims
1. A method for preventing or treating microbial infection, comprising administering to a subject an active blend comprising trans-cinnamaldehyde and a potentiating agent,
- wherein the antimicrobial effect of trans-cinnamaldehyde is potentiated by said potentiating agent,
- wherein the active blend is free of coumarin or safrole or is free of coumarin and safrole,
- wherein the potentiating agent comprises: at least one terpenoid, chosen from mono and sesquiterpenoids, and at least one derivate from trans-cinnamaldehyde, either being a substituted trans-cinnamaldehyde or a compound corresponding to trans-cinnamaldehyde wherein the aldehyde function is replaced by another function, and optionally phenylpropane derivative,
- and wherein the potentiating agent comprises six compounds or less.
2. The method according to claim 1 wherein the antimicrobial effect of trans-cinnamaldehyde is an anti-bacterial or anti-fungal or anti-viral effect.
3. The method of claim 1, in which the potentiating agent consists of 6 compounds or less.
4. The method of claim 1, wherein the terpenoid is chosen from menthanes, acyclic monoterpenoids and caryophyllanes.
5. The method of claim 4, wherein the terpenoid is chosen from cineol, linalool and beta-caryophyllene.
6. The method of claim 1, wherein the phenylpropane derivative is selected from the group consisting of apiole, coniferyl benzoate, chavicol, cinnamein, vanillin and benzyle benzoate.
7. The method of claim 1, wherein it comprises at least one trans-cinnamaldehyde derivate corresponding to Formula I:
- wherein X represents —CH2OH, —CH2OY, —CHO, —COOH, —COOZ, —CO-Hal, Y represents Ra or —CORa with Ra being an alkyl comprising 1 to 6 carbon atoms, Z represents an alkyl comprising 1 to 6 carbon atoms, R1 and R2 represent independently, H, —OH, an alkyl comprising 1 to 6 carbon atoms, or an halogen atom R3 and R4 represent independently, H, an alkyl comprising 1 to 6 atoms, an alkoxy comprising 1 to 6 carbon atoms or an halogen atom, Hal represents an halogen atom,
- and wherein when X is —CHO then at least one of R1, R2, R3 and R4 does not represent H.
8. The method of claim 7, wherein the trans-cinnamaldehyde derivate is chosen from trans-2-methoxycinnamaldehyde and cinnamyle acetate.
9. The method of claim 8, wherein the amount of trans-cinnamaldehyde derivate ranges from 1 to 50% by weight compared to the total weight of the active blend.
10. The method of claim 1, wherein the potentiating agent comprises:
- at least one terpenoid and phenylpropane derivative in an amount ranging from 1 to 50% by weight compared to the total weight of the active blend,
- at least one derivate from trans-cinnamaldehyde in an amount ranging from 1 to 50% by weight compared to the total weight of the active blend.
11. The method of claim 1, for preventing or treating infections resulting from drug-resistant Gram negative bacteria strains.
12. The method of claim 11, for preventing or treating infections resulting from 1, 2, 3, 4, 5, 6 or 7 drug-resistant Gram negative bacteria strains selected from:
- Pseudomonas;
- Acinetobacter;
- Escherichia;
- Enterobacter;
- Serratia;
- Citrobacter; and
- Klebsiella.
13. The method of claim 1, wherein the infection is selected from the group consisting of urinary system infection, respiratory system infection, digestive system infection, central nervous system infection, skin and soft tissues infection, bone infection, articulations infection, muscles infection, vasculary system infection and systemic infections.
14. The method of claim 13 for preventing or treating diseases or syndromes selected from the group consisting of bacteremia, Crohn disease, peptic ulcer, diabetic foot and eschars, septic shock, AIDS, Herpes, Hepatitis B and C and Influenza.
15. The method of claim 1, wherein the subject is an animal.
16. The method of claim 15, wherein the animal is chosen among cattle, swine, poultry, and pets.
17. The method of claim 1, wherein an antibiotic is further administrated.
18. The method of claim 17, wherein the antibiotic is selected among:
- antibiotics acting on the bacterial wall;
- antibiotics operating the membranes of the cells, external membrane or cytoplasmic membrane;
- antibiotics acting on the synthesis of proteins;
- antibiotics blocking the synthesis of messenger RNA;
- antibiotics acting on DNA;
- antibiotics acting by competitive inhibition;
- one of their pharmaceutically acceptable salts, and
- one of their combinations.
Type: Application
Filed: Oct 11, 2022
Publication Date: Feb 16, 2023
Applicant: SEPTEOS (Paris)
Inventor: Nichola TESSE (Vaucresson)
Application Number: 17/963,836
