PHARMACEUTICAL FORMULATION WITH CINEOL AND AMOXICILLIN

Description

[0001] The present invention falls within the field of medicine, particularly bacterial infections. It relates to new treatments and new pharmaceutical formulations particularly suited for medical or veterinary use in combating bacterial infections, especially antibiotic-resistant infections. Technological background of the invention

[0002] Antibiotics are natural or synthetic substances that have bactericidal or bacteriostatic activity. Their widespread introduction after World War II was one of the most significant therapeutic advances of the 20th century. Antibiotic treatments have increased life expectancy by more than ten years, more than any other medical treatment. However, the widespread, even excessive, use of certain antibiotics, including for preventive and curative purposes, as dietary supplements in animal feed, fish farming, veterinary and human medicine, and as pesticides for plant treatment, has introduced selective pressure that has led to the development of antibiotic-resistant microorganisms and a general decline in therapeutic efficacy.In a hospital setting, this leads to an increased risk of nosocomial infections, particularly due to a lack of appropriate treatment against certain multi-resistant germs.

[0003] In its latest report (April 2014) on antibiotic resistance, the WHO emphasizes that "unless the many stakeholders involved act urgently and in a coordinated manner, the world is heading towards a post-antibiotic era, where common infections and minor injuries that have been treated for decades could once again kill." Resistance to certain so-called last-resort treatments is already a reality. For example, resistance to treatments for life-threatening infections caused by a common intestinal bacterium, Klebsiella pneumoniae has spread to all regions of the world. The same is true for resistance to one of the most widely used antibacterial drugs in the treatment of urinary tract infections caused by E. coli- fluoroquinolones. Recently, the failure of last-resort treatment for gonorrhea - third-generation cephalosporins - has been confirmed in South Africa, Australia, Austria, Canada, France, Japan, Norway, the United Kingdom, Slovenia and Sweden.

[0004] To address this threat, the WHO is calling in particular for the development of new diagnostic products, new antibiotics and other essential tools so that healthcare professionals can maintain their lead over the progression of resistance.

[0005] Amoxicillin is a bactericidal β-lactam antibiotic belonging to the aminopenicillin family, indicated for the treatment of bacterial infections caused by susceptible organisms. Amoxicillin is the most commonly used antibiotic, particularly in children, because it is well absorbed orally, has a broad antimicrobial spectrum, and is inexpensive. Amoxicillin is used to treat various infectious diseases, including those of the lungs, bronchi, nose, throat, ears, blood, digestive or urinary tract, genital tract, gums, and teeth.

[0006] Amoxicillin is often used in combination with another molecule, clavulanic acid, a β-lactamase inhibitor. β-lactamase is an enzyme produced by bacteria resistant to β-lactam antibiotics. By inhibiting β-lactamases, clavulanic acid prevents the inactivation of amoxicillin by β-lactamases, thus allowing it to retain its activity against resistant bacteria that produce β-lactamases.

[0007] However, in recent years we have witnessed the emergence of particularly resistant bacterial germs, notably extended-spectrum β-lactamase (ESBL) bacteria, which are no longer, or only partially, susceptible to classic β-lactamase inhibitors such as clavulanic acid. WO2016 / 041958 discloses a composition comprising a cineole, amoxicillin / clavulanic acid mixture with synergistic properties against various resistant bacterial strains, but without peanut oil.

[0008] Therefore, it is essential to find solutions to restore the effectiveness of amoxicillin against resistant bacteria, in particular extended-spectrum β-lactamase (ESBL) producing bacteria. Summary of the invention

[0009] The inventors discovered that cineole increases the effectiveness of amoxicillin, particularly against resistant bacteria. They demonstrated that the combination of amoxicillin and cineole creates a synergistic effect that significantly enhances the antibacterial activity of amoxicillin. They thus developed a new combination of molecules including amoxicillin, cineole, and possibly clavulanic acid, enabling effective combating of resistant bacteria, especially those resistant to the combination of amoxicillin and clavulanic acid.

[0010] The inventors have also developed a pharmaceutical formulation that significantly increases the antibacterial activity of amoxicillin, alone or in combination with clavulanic acid, particularly against bacterial germs resistant to the combination of amoxicillin and clavulanic acid.

[0011] The subject matter of the present invention is defined by the claims.

[0012] Thus, the present invention relates, in a first aspect, to a pharmaceutical formulation in powder form comprising, or consisting essentially of, cineole, amoxicillin, a β-lactamase inhibitor and a pharmaceutically acceptable oil, said pharmaceutically acceptable oil being peanut oil.

[0013] Preferably, the β-lactamase inhibitor is clavulanic acid.

[0014] The formulation according to the invention may comprise between about 5 mg and about 100 mg of cineole per gram of powder, preferably between about 10 mg and about 50 mg of cineole per gram of powder, more preferably between about 20 mg and about 40 mg of cineole per gram of powder, and most preferably about 33 mg of cineole per gram of powder.

[0015] The formulation according to the invention may also comprise between about 20 mg and about 500 mg of amoxicillin per gram of powder, preferably between about 50 mg and about 300 mg of amoxicillin per gram of powder, more preferably between about 150 mg and about 200 mg of amoxicillin per gram of powder, and most preferably about 167 mg of amoxicillin per gram of powder.

[0016] The formulation according to the invention may further comprise between about 2 mg and about 50 mg of oil per gram of powder, preferably between about 10 mg and about 25 mg of oil per gram of powder, even more preferably between about 15 mg and about 20 mg of oil per gram of powder, and most particularly preferably about 17 mg of oil per gram of powder.

[0017] The formulation according to the invention may finally comprise between about 1 mg and about 100 mg of β-lactamase inhibitor, preferably clavulanic acid, per gram of powder, preferably between about 5 mg and about 50 mg of β-lactamase inhibitor, preferably clavulanic acid, per gram of powder, more preferably between about 15 mg and about 25 mg of β-lactamase inhibitor, preferably clavulanic acid, per gram of powder, and most particularly preferably about 21 mg of β-lactamase inhibitor, preferably clavulanic acid, per gram of powder.

[0018] The formulation according to the invention may include in particular between about 5 mg and about 100 mg, preferably between about 10 mg and about 50 mg, of cineole per gram of powder; and / or between about 20 mg and about 500 mg, preferably between about 50 mg and about 300 mg, of amoxicillin per gram of powder; and / or between about 2 mg and about 50 mg, preferably between about 10 mg and about 25 mg, of oil per gram of powder; and / or optionally between about 1 mg and about 100 mg, preferably between about 5 mg and about 50 mg, of β-lactamase inhibitor, preferably clavulanic acid, per gram of powder.

[0019] Preferably, the formulation according to the invention comprises between about 20 mg and about 40 mg, preferably about 33 mg, of cineole per gram of powder; between about 150 mg and about 200 mg, preferably about 167 mg, of amoxicillin per gram of powder; between about 15 mg and about 20 mg, preferably about 17 mg, of oil per gram of powder; and / or optionally between about 15 mg and about 25 mg, preferably about 21 mg of β-lactamase inhibitor, preferably clavulanic acid, per gram of powder.

[0020] The formulation according to the invention may have an amoxicillin / cineole mass ratio of between 2 and 8, preferably between 3 and 7, more particularly preferably between 4 and 6, and most particularly preferably an amoxicillin / cineole mass ratio of about 5.

[0021] The formulation according to the invention may have an amoxicillin / oil mass ratio of between 5 and 15, preferably between 7 and 13, more particularly preferably between 8 and 12, and most particularly preferably an amoxicillin / oil mass ratio of about 10.

[0022] The formulation according to the invention may have a cineole / oil mass ratio of between 0.1 and 5, preferably between 0.5 and 4, more particularly preferably between 1 and 3, and most particularly preferably a cineole / oil mass ratio of about 2.

[0023] The formulation according to the invention may have an amoxicillin / β-lactamase inhibitor mass ratio of between 5 and 11, preferably between 6 and 10, more particularly preferably between 7 and 9, and most particularly preferably, an amoxicillin / β-lactamase inhibitor mass ratio of about 8.

[0024] The formulation according to the invention can be intended for oral administration, preferably after suspension in an aqueous solvent.

[0025] The formulation according to the invention can be packaged in a single-dose container, preferably a single-dose container containing between approximately 1 g and approximately 150 g of powder, more preferably between approximately 1 g and approximately 50 g of powder, even more preferably between approximately 1 g and approximately 10 g of powder, and more particularly preferably approximately 3 g of powder.

[0026] The formulation according to the invention may further comprise at least one pharmaceutically acceptable excipient or carrier, preferably selected from the group consisting of a sweetener, a flavoring agent, an anti-caking agent, a lubricant, a disintegrant, and a mixture thereof.

[0027] In a second aspect, the present invention also relates to the formulation as described above for use in the treatment of an infectious pathology in a subject, preferably an animal or a human.

[0028] Preferably, the said infectious pathology is an infectious pathology of bacterial origin, more preferably an infectious pathology caused by a bacterium resistant to antibiotics of the β-lactam family.

[0029] Preferably, the formulation is administered or intended to be administered to the subject for a period of 1 day to 4 weeks and at a dose of 3 to 30 grams per day, in one or more doses.

[0030] The present invention also relates, in a third aspect, to a method for manufacturing the pharmaceutical formulation according to the invention comprising: obtaining a wetting solution by mixing cineole and a pharmaceutically acceptable oil; and wetting a powder comprising amoxicillin with the wetting solution to obtain a powder preparation comprising amoxicillin, cineole and oil.

[0031] Optionally, the process may further include mixing the powdered preparation comprising amoxicillin, cineole, and oil with a powder comprising a β-lactamase inhibitor, preferably clavulanic acid; and / or the addition of sweetener, flavouring and / or lubricant, and the mixing of these to obtain a homogeneous powder; and / or the sieving of the powder thus obtained; and / or the packaging of the sieved powder in single-dose containers.

[0032] Preferably, the powder comprising amoxicillin and / or the powder comprising a β-lactamase inhibitor, preferably clavulanic acid, further comprise a disintegrant and / or an anti-caking agent.

[0033] Preferably, the bacterial infection is caused by a bacterium resistant to antibiotics, preferably to antibiotics of the β-lactam family.

[0034] The subject matter is an animal, preferably a mammal, and most particularly preferred a human.

[0035] The present invention also relates to a formulation according to the invention for use in the treatment of a bacterial infection in a subject, preferably a bacterial infection caused by an antibiotic-resistant bacterium.

[0036] Preferably, the bacterial infection is selected from the group consisting of cystitis, in particular recurrent acute cystitis, bacterial sinusitis, in particular acute maxillary sinusitis, otitis, in particular acute otitis media, bronchitis, in particular chronic and / or acute bronchitis, bronchopneumopathies, in particular chronic and / or acute bronchopneumopathies, pyelonephritis, upper gynecological infections, periodontitis, severe stomatological infections, in particular abscesses, phlegmons and cellulitis, animal bites, bone and joint infections, in particular osteomyelitis, preferably said bacterial infection is cystitis, in particular cystitis caused by a bacterium resistant to β-lactam antibiotics. Brief description of the drawings

[0037] Figure 1 :Study of the antibacterial activity of rabbit serum treated with a combination of amoxicillin, clavulanic acid, and cineole. A (AMC without cineole): After oral administration of a single dose of a composition comprising amoxicillin (1.5 g) and clavulanic acid (186.5 mg) to 3 rabbits, the percentage of serum inhibition at times T0, T1h, T2h, T3h, and T6h was calculated against a strain of Escherichia coli multidrug-resistant ESBL. B (AMC with cineole): after oral administration of a single dose of a composition comprising amoxicillin (1.5 g), clavulanic acid (186.5 mg) and cineole (300 mg) to 3 rabbits, the percentage of serum inhibition at times T0, T1h, T2h, T3h and T6h was calculated against a strain of Escherichia coli multi-resistant ESBL. Figure 2Monitoring of mean serum amoxicillin concentrations over 24 hours. A (without cineol): After oral administration of a single dose (12 g) of a composition containing amoxicillin (2 g) and clavulanic acid (250 mg), serum amoxicillin concentrations were monitored for 24 hours in 12 healthy volunteers. B (with cineol): After oral administration of a single dose (12 g) of a composition containing amoxicillin (2 g), clavulanic acid (250 mg), and cineol (400 mg), serum amoxicillin concentrations were monitored for 24 hours in 12 healthy volunteers. Figure 3 :Monitoring of mean plasma amoxicillin concentrations over 7 days. A (cineole-free): Following oral administration of a single 12 g dose of a composition containing amoxicillin (2 g) and clavulanic acid (250 mg), the 12 volunteers received maintenance doses (3 g) of the same composition containing amoxicillin (500 mg) and clavulanic acid (62.5 mg) three times daily for 7 days. Plasma amoxicillin concentrations were monitored for 7 days in the 12 healthy volunteers. B (with cineole): after oral administration of a single dose (12 g) of a composition comprising amoxicillin (2 g), clavulanic acid (250 mg) and cineole (400 mg), the 12 volunteers receive, 3 times a day for 7 days, maintenance doses (3 g) of the same composition comprising amoxicillin (500 mg), clavulanic acid (62.5 mg) and cineole (100 mg).Plasma amoxicillin concentration was monitored for 7 days in the 12 healthy volunteers. Figure 4 : Spectroscopic study of amoxicillin complex formation. A (AMX alone): Mass spectrometry analysis of amoxicillin alone in aqueous solution. B (AMX with cineole): Mass spectrometry analysis of amoxicillin with cineole in aqueous solution. Detailed description of the invention

[0038] The inventors demonstrated that combining amoxicillin with cineole produces a synergistic effect that significantly increases the efficacy of amoxicillin, particularly against resistant bacteria. Specifically, they observed that cineole, at sub-therapeutic concentrations, protects amoxicillin from the inhibitory effect of β-lactamases, an enzyme produced by bacteria resistant to amoxicillin.

[0039] The inventors have therefore developed a new therapeutic combination including amoxicillin, cineole and possibly clavulanic acid, enabling effective combating of resistant bacterial germs, in particular germs resistant to the combination of amoxicillin and clavulanic acid.

[0040] The inventors also developed a powdered pharmaceutical formulation combining cineole, amoxicillin, and clavulanic acid. They observed that this combination significantly increased the antibacterial activity of amoxicillin, thus enabling the effective treatment of patients with infections caused by resistant bacteria.

[0041] This new formulation also boasts excellent preservation properties. In particular, the use of oil in the formulation allows for the highly effective binding of cineole, a highly volatile agent, to the powdered base. This enables a single formulation for the administration of the active compounds, which, after suspension in an aqueous solvent, can be easily administered orally. Having a single medication, especially one administered orally, greatly increases treatment adherence and, consequently, its effectiveness.

[0042] The inventors further demonstrated that amoxicillin, in the presence of cineole, forms a complex of at least three amoxicillin molecules. The formation of this complex makes the amoxicillin molecules less accessible to β-lactamases, thus increasing their therapeutic efficacy.

[0043] Thus, according to a first aspect, the present invention relates to a pharmaceutical formulation in powder form comprising, or consisting essentially of, cineole, amoxicillin, and a pharmaceutically acceptable oil.

[0044] According to a second aspect, the present application discloses a therapeutic combination (not claimed) comprising cineole and amoxicillin for use in the treatment of a bacterial infection in a subject. The disclosed combination may further include a β-lactamase inhibitor, preferably clavulanic acid. In particular, the disclosed combination may be a combination preparation for the simultaneous, separate, or sequential use of the active ingredients of the combination, preferably cineole, amoxicillin, and clavulanic acid. Definitions

[0045] In this document, the term "approximately" refers to a range of values ​​within ±10% of the specified value. For example, "approximately 50" includes values ​​within ±10% of 50, i.e., values ​​in the range of 45 to 55. Preferably, the term "approximately" refers to a range of values ​​within ±5% of the specified value. It is understood that values ​​preceded by the term "approximately" are also to be considered as specifically described in this application.

[0046] The term "consists essentially of", as used here, refers to a formulation according to the invention comprising no other active ingredient than those mentioned in said formulation, in particular no other antibiotics or β-lactamase inhibitors.

[0047] As used here, the term "pharmaceutically acceptable excipient or carrier" refers to any substance other than an active ingredient present in a pharmaceutical formulation. Its addition is intended, in particular, to impart a specific consistency, or other specific physical or taste characteristics, to the final product, while avoiding any interaction, especially chemical, with the active ingredient(s).

[0048] As used here, the term "active ingredient" refers to a molecule with a therapeutic effect. In particular, cineole, amoxicillin, and β-lactamase inhibitors are active ingredients.

[0049] As used herein, the term "therapeutic effect" refers to an effect induced by an active ingredient, a composition according to the invention, or a combination according to the invention, capable of preventing or delaying the onset of a bacterial infection, or of curing or reducing the effects of a bacterial infection.

[0050] As used here, the term "antibacterial effect" refers to an effect induced by an active ingredient, a composition according to the invention, or a combination according to the invention, capable of reducing in a subject the quantity and / or concentration of bacteria responsible for bacterial infection.

[0051] As used herein, the term "synergistic effect" refers to a composition or combination according to the invention having a therapeutic and / or antibacterial effect greater than the sum of the therapeutic and / or antibacterial effects of all the active ingredients present in said composition or combination when taken individually. The presence of such an effect can be assessed by calculating the fractional inhibitory concentration index (fic-index) as illustrated in Example 1.

[0052] As used here, the term "treatment" refers to any action aimed at improving the medical status of a person with a bacterial infection. Treatment may aim to improve the patient's condition, that is, to reduce the infection or some of its symptoms, or to eradicate the infection or some of its symptoms. Treatment may also prevent or slow the progression of a bacterial infection. Furthermore, treatment may have a prophylactic or preventive effect, that is, prevent or delay the onset of the bacterial infection.

[0053] As used here, the terms "quantity" and "dose" are equivalent and can be used interchangeably.

[0054] As used here, the term "therapeutically effective amount" refers to an amount of active ingredient, composition according to the invention, or combination according to the invention, sufficient to induce a therapeutic effect. Alternatively, the term "therapeutically effective amount" may refer to an amount of active ingredient, composition according to the invention, or combination according to the invention, sufficient to induce an antibacterial effect. It is evident that the amount to be administered can be adjusted by a person skilled in the art, depending on the individual being treated, the nature of the bacterial infection, etc. In particular, the doses and administration regimens depend on the nature, stage of development, and severity of the bacterial infection being treated, as well as the weight, age, and general health of the individual being treated, or the judgment of the prescribing physician.

[0055] As used here, the term "sub-therapeutic quantity" refers to an amount of active ingredient insufficient to induce a therapeutic effect on its own. Alternatively, the term "sub-therapeutic quantity" can refer to an amount of active ingredient insufficient to induce an antibacterial effect on its own.

[0056] As used here, the terms "therapeutic combination" and "combination preparation" refer to an association of active ingredients, preferably cineole, amoxicillin and optionally clavulanic acid, which may each be formulated separately or in one or more formulations for simultaneous, separate, sequential administration, or a mixture of these modes of administration when the combination includes more than two active ingredients.

[0057] As used here, the term "simultaneous" refers to a combination according to the invention in which the active ingredients of the combination are used or administered simultaneously, i.e., at the same time.

[0058] As used herein, the term "sequential" refers to a combination according to the invention in which the active ingredients of the combination are used or administered sequentially, that is, one after the other. Preferably, when administered sequentially, all the active ingredients are administered within an interval of at most about 1 hour, preferably at most about 10 minutes, and even more preferably at most about 1 minute.

[0059] As used herein, the term "separate" refers to a combination according to the invention in which the active ingredients of the combination are used or administered at separate times of the day. Preferably, when administration is separate, the active ingredients are administered at intervals of approximately 1 hour to approximately 15 hours, preferably from approximately 1 hour to approximately 8 hours, and even more preferably from approximately 1 hour to approximately 5 hours. The cinema

[0060] As used here, the term "cineole," "eucalyptol," or "1,8-cineole" refers to a cyclic ether (CAS No. 470-82-6) belonging to the monoterpene group, that is, terpenoids with ten carbon atoms. Cineole is a natural, organic, and colorless compound that can be extracted from the essential oils of certain eucalyptus species (for example... Eucalyptus polybractea ), rosemary (for example Rosmarinus officinalis ), mugwort (for example Artemisia vulgaris), but also essential oils of wormwood, bay leaf, sage, basil, and camphor leaves ( Cinnamomum camphora Cineol can be used in any pharmaceutically acceptable form. As used here, "pharmaceutically acceptable" refers to a molecule, compound, or composition suitable for pharmaceutical administration. It is preferably used in a purified form. In its purified form, cineole is a liquid. Amoxicillin

[0061] Amoxicillin (CAS No. 26787-78-0) is a bactericidal β-lactam antibiotic belonging to the aminopenicillin family and is currently indicated for the treatment of bacterial infections caused by susceptible organisms. β-lactams are a large class of antibiotics that includes penicillin derivatives, cephalosporins, monobactams, and carbapenems. β-lactams are characterized by the presence of a β-lactam ring in their molecular structure, which confers their bactericidal activity.

[0062] In the formulation according to the invention, amoxicillin may be in any pharmaceutically acceptable form. Amoxicillin may thus be in the form of a pharmaceutically acceptable salt, in particular a sodium or potassium salt, in anhydrous or hydrated form, preferably as a trihydrate, or a mixture of these forms. In a preferred embodiment, the formulation according to the invention comprises amoxicillin trihydrate. Pharmaceutically acceptable oil

[0063] The pharmaceutical formulation according to the invention also comprises a pharmaceutically acceptable oil. As used here, the term "oil" refers to a phase consisting of fatty substances that are liquid at room temperature and do not mix with water.

[0064] The oil used in the formulation disclosed in this document may be any pharmaceutically acceptable oil, that is, any oil whose toxicological data are compatible with oral administration to a subject. In an unclaimed aspect, the oil is preferably selected from the group consisting of animal, mineral, vegetable, synthetic oils and mixtures thereof.

[0065] The oil used in the combination is not an essential oil. As used here, the term "essential oil" refers to a concentrated, hydrophobic liquid containing volatile aromatic (fragrant) compounds from a plant. Essential oils can be obtained by mechanical extraction, cold pressing, volatile solvent extraction, supercritical CO2 extraction, steam distillation, or dry distillation.

[0066] The oil can be used in the formulation according to the invention for its adsorptive properties. As used here, the term "adsorbent" refers to an excipient capable of binding liquid molecules, for example cineole molecules, to a solid support, for example powder, in a pharmaceutical formulation.

[0067] In an unclaimed aspect, the disclosed formulation includes a mineral oil. Mineral oils are obtained by distilling coal, petroleum, or certain oil shales. Mineral oils include, among other things, hydrocarbons, alkanes, and paraffins. In particular, the mineral oil may be selected from the group consisting of refined paraffins, microcrystalline waxes, ozokerites, ceresins, petrolatum, and mixtures thereof.

[0068] In another unclaimed aspect, the disclosed formulation includes a synthetic oil. In particular, the synthetic oil may be selected from the group consisting of silicone oils, synthetic waxes, synthetic mono-, di- and triglycerides, for example caprylic / capric triglycerides, and a mixture thereof.

[0069] In yet another unclaimed aspect, the disclosed formulation includes an animal oil. This animal oil may, for example, be selected from the group consisting of mink oil, sperm whale oil, whale oil, seal oil, emu oil, ox foot oil, fish oils, in particular anchovy oil, sardine oil, capelin oil, herring oil, salmon oil, sprat oil, cod oil, blue whiting oil, pilchard oil, tuna oil, shark oil, and a mixture thereof.

[0070] In a preferred embodiment, the formulation according to the invention comprises a vegetable oil, said vegetable oil being peanut oil. Other unclaimed examples of vegetable oils include, but are not limited to, wheat germ, corn, sunflower, shea, castor, sweet almond, macadamia, apricot, soybean, cottonseed, alfalfa, poppy seed, pumpkin seed, sesame, pumpkin seed, avocado, hazelnut, grape seed, blackcurrant seed, evening primrose, millet, barley, quinoa, olive, peanut, rye, safflower, candlenut, passionflower, rosehip, coconut, argan, rapeseed, copra, flaxseed, walnut, cashew nut, neem, pistachio, rice, camelina, sacha inchi, borage, hemp, pea, jojoba, and other oils. neem, black cumin, perilla, and a mixture of these. β-lactamase inhibitor

[0071] The pharmaceutical formulation according to the invention further comprises one or more β-lactamase inhibitors. These inhibitors can block β-lactamase activity in various ways, for example by acting as a suicide substrate that binds irreversibly to these enzymes, as is notably the case for clavulanic acid and sulbactam.

[0072] The β-lactamase inhibitor can be any pharmaceutically acceptable β-lactamase inhibitor. This inhibitor is preferably selected from the group consisting of clavulanic acid, sulbactam, tazobactam, aztreonam, avibactam, pharmaceutically acceptable salts thereof, and mixtures thereof.

[0073] In a preferred embodiment, the formulation according to the invention comprises clavulanic acid. Clavulanic acid (CAS No. 58001-44-8) is commonly used in combination with β-lactam antibiotics, particularly amoxicillin.

[0074] In the formulation according to the invention, clavulanic acid may be in any pharmaceutically acceptable form, preferably in the form of a pharmaceutically acceptable salt, in particular in the form of a potassium salt of clavulanic acid. Pharmaceutical formulation

[0075] According to a first aspect, the present invention relates to a pharmaceutical formulation in powder form comprising, or consisting essentially of, cineole, amoxicillin, a β-lactamase inhibitor and a pharmaceutically acceptable oil, said pharmaceutically acceptable oil being peanut oil.

[0076] As used here, the term "powder" refers to a fractionated state of matter; it is a solid state present in the form of small particles.

[0077] Preferably, the powder particles have a diameter less than or equal to about 5 mm, more preferably less than or equal to about 2.5 mm, and most preferably less than or equal to about 1.25 mm.

[0078] The powder of the pharmaceutical formulation according to the invention is preferably a dry powder. As used here, the term "dry powder" refers to a powder having a moisture content of 20% or less, preferably 15% or less, and even more preferably 10% or less.

[0079] The pharmaceutical formulation according to the invention preferably meets the requirements of the European Pharmacopoeia (8th edition), particularly in terms of impurities and / or microbiological quality.

[0080] In particular, the bacterial count present in the powder of the pharmaceutical formulation according to the invention is preferably less than 10,000 CFU (Colony Forming Units) per gram of powder, preferably less than 1,000 CFU per gram of powder, and most preferably less than 100 CFU per gram of powder. Preferably, the powder of the pharmaceutical formulation according to the invention does not contain Escherichia coli. Regarding other germs, the level of fungi and yeasts is preferably less than 1000 CFU per gram of powder, preferably less than 100 CFU per gram of powder, and most preferably less than 50 CFU per gram of powder.

[0081] The pharmaceutical formulation according to the invention comprises, or essentially consists of, cineole, amoxicillin, and a pharmaceutically acceptable oil, as defined above.

[0082] Preferably, amoxicillin and cineole are present in the formulation according to the invention at concentrations allowing the administration of doses sufficient to achieve a therapeutic and / or antibacterial effect, preferably a synergistic effect, as illustrated in the examples below. To achieve such an effect, amoxicillin and cineole can be administered in therapeutically effective or sub-therapeutic amounts.

[0083] In a preferred embodiment, cineole and amoxicillin are both administered in therapeutically effective amounts.

[0084] In another preferred embodiment, amoxicillin is administered in a therapeutically effective amount and cineole is administered in a sub-therapeutic amount.

[0085] In yet another preferred embodiment, amoxicillin is administered in a sub-therapeutic amount and cineole is administered in a therapeutically effective amount.

[0086] In yet another preferred embodiment, amoxicillin and cineole are both administered in sub-therapeutic amounts.

[0087] Preferably, the oil is present in the formulation according to the invention at a concentration sufficient to allow the adsorption of cineole to the powder of the formulation.

[0088] According to one embodiment, the pharmaceutical formulation according to the invention comprises, or essentially consists of, between about 5 mg and about 100 mg, preferably between about 10 mg and about 50 mg, more preferably between about 20 mg and about 40 mg, of cineole per gram of powder; and / or between about 20 mg and about 500 mg, preferably between about 50 mg and about 300 mg, more preferably between about 150 mg and about 200 mg, of amoxicillin per gram of powder; and / or between about 2 mg and about 50 mg, preferably between about 10 mg and about 25 mg, more preferably between about 15 mg and about 20 mg, of oil per gram of powder.

[0089] Preferably, the pharmaceutical formulation according to the invention comprises, or consists essentially of, between about 5 mg and about 100 mg of cineole, between about 20 mg and about 500 mg of amoxicillin, and between about 2 mg and about 50 mg of oil per gram of powder.

[0090] Preferably, the pharmaceutical formulation according to the invention comprises, or essentially consists of, between about 10 mg and about 50 mg of cineole, between about 50 mg and about 300 mg of amoxicillin, and between about 10 mg and about 25 mg of oil per gram of powder.

[0091] Most preferably, the pharmaceutical formulation according to the invention comprises, or consists essentially of, between about 20 mg and about 40 mg, preferably about 33 mg, of cineole per gram of powder, between about 150 mg and about 200 mg, preferably about 167 mg, of amoxicillin per gram of powder, between about 15 mg and about 20 mg, preferably about 17 mg, of oil per gram of powder.

[0092] The pharmaceutical formulation according to the invention comprises, or essentially consists of, cineole, amoxicillin, a pharmaceutically acceptable oil and a β-lactamase inhibitor, as defined above.

[0093] Preferably, the β-lactamase inhibitor is selected from clavulanic acid, sulbactam, tazobactam, and aztreonam. Even more preferably, the β-lactamase inhibitor is clavulanic acid.

[0094] Thus, the pharmaceutical formulation according to the invention may comprise, or consist essentially of, cineole, amoxicillin and a pharmaceutically acceptable oil in proportions such as described above, and a β-lactamase inhibitor present at a concentration sufficient to permit its administration in a therapeutically effective amount or in a sub-therapeutic amount.

[0095] According to one embodiment, the pharmaceutical formulation according to the invention comprises, or essentially consists of, cineole, amoxicillin and a pharmaceutically acceptable oil in proportions as described above, and between about 1 mg and about 100 mg, preferably between about 5 mg and about 50 mg, more preferably between about 15 mg and about 25 mg, and most preferably about 20.8 mg, of β-lactamase inhibitor, preferably clavulanic acid, per gram of powder.

[0096] In another particular embodiment, the pharmaceutical formulation according to the invention comprises, or essentially consists of, cineole, amoxicillin and a pharmaceutically acceptable oil, wherein the proportion of cineole is between about 0.02 mg and about 0.5 mg, preferably between about 0.1 mg and about 0.3 mg, of cineole per gram of amoxicillin, and even more preferably the proportion of cineole is equal to about 0.2 mg of cineole per gram of amoxicillin; and / or the proportion of oil is between about 0.01 mg and about 0.5 mg, preferably between about 0.05 mg and about 0.2 mg, of oil per gram of amoxicillin, and even more preferably the proportion of oil is equal to about 0.1 mg of oil per gram of amoxicillin.Preferably, the pharmaceutical formulation according to the invention further comprises a β-lactamase inhibitor, preferably clavulanic acid, the proportion of β-lactamase inhibitor being between about 0.01 mg and about 0.5 mg, preferably between 0.1 mg and about 0.2 mg, of β-lactamase inhibitor per gram of amoxicillin, and even more preferably the proportion of β-lactamase inhibitor is equal to about 0.125 mg of β-lactamase inhibitor per gram of amoxicillin.

[0097] In particular embodiments, the amoxicillin / cineole mass ratio is between 2 and 8, preferably between 3 and 7, more particularly preferred between 4 and 6. In a preferred embodiment, the amoxicillin / cineole mass ratio is about 5.

[0098] In particular embodiments, the amoxicillin / β-lactamase inhibitor mass ratio, preferably clavulanic acid, is between 5 and 11, preferably between 6 and 10, more particularly preferably between 7 and 9. In a preferred embodiment, the amoxicillin / β-lactamase inhibitor mass ratio, preferably clavulanic acid, is about 8.

[0099] In particular embodiments, the amoxicillin / oil mass ratio is between 5 and 15, preferably between 7 and 13, more particularly preferred between 8 and 12. In a preferred embodiment, the amoxicillin / cineole mass ratio is about 10.

[0100] In particular embodiments, the cineole / oil mass ratio is between 0.1 and 5, preferably between 0.5 and 4, more particularly preferred between 1 and 3. In a preferred embodiment, the cineole / oil mass ratio is about 2.

[0101] The pharmaceutical formulation according to the invention may further comprise at least one other active ingredient.

[0102] Thus, in a particular embodiment, the pharmaceutical formulation according to the invention comprises, or essentially consists of, cineole, amoxicillin, a pharmaceutically acceptable oil, optionally a β-lactamase inhibitor, preferably clavulanic acid, as defined above, and another active ingredient. Preferably, the additional active ingredient of the formulation according to the invention is another antibiotic, in particular a β-lactam antibiotic, and / or another β-lactamase inhibitor, and / or an antifungal, and / or an antiparasitic, and / or an analgesic.

[0103] The pharmaceutical formulation according to the invention may comprise, or consist substantially of, cineole, amoxicillin, a pharmaceutically acceptable oil, and optionally a β-lactamase inhibitor, preferably clavulanic acid, in quantities such as described above and another active ingredient present at a concentration sufficient to permit its administration in a therapeutically effective or sub-therapeutic amount. Excipients

[0104] The pharmaceutical formulation according to the invention may further comprise at least one pharmaceutically acceptable excipient or carrier in addition to the pharmaceutically acceptable oil.

[0105] This pharmaceutically acceptable excipient or support for the formulation according to the invention is preferably selected from the group consisting of a sweetener, a flavoring agent, an anti-caking agent, a lubricant, a disintegrant, and a mixture thereof.

[0106] Thus, in a particular embodiment, the pharmaceutical formulation according to the invention further comprises at least one disintegrant.

[0107] As used here, the term "disintegrant" refers to an excipient that improves disintegration, that is, the separation of molecules present in the pharmaceutical formulation in a liquid medium, preferably an aqueous medium, and their homogeneous suspension.

[0108] Preferably, the disintegrant is selected from the group consisting of microcrystalline celluloses, crosslinked carboxymethyl starches, crosslinked polyvinylpyrrolidones and crosslinked carboxymethylcelluloses.

[0109] As used here, the terms "crosslinked carboxymethylcellulose" and "croscarmellose" are equivalent and can be used interchangeably.

[0110] In a preferred embodiment, the pharmaceutical formulation according to the invention comprises between about 10 mg and about 1000 mg, preferably between about 100 mg and about 600 mg, more preferably between about 450 mg and about 550 mg, for example about 513.7 mg, of disintegrant per gram of powder.

[0111] In particular, the formulation may include microcrystalline cellulose, preferably in the form of a powder consisting of particles having a diameter of approximately 20 to approximately 200 µm, more preferably a diameter of approximately 50 to approximately 100 µm. In particular, the microcrystalline cellulose is selected from microcrystalline celluloses of the Avicel®, Emcocel® and Vitacel® type, or mixtures thereof, preferably the microcrystalline cellulose is selected from Avicel PH 101, 102, 103, 104, 112, 113, 301 and 302 microcrystalline celluloses, and most preferably the microcrystalline cellulose is of the Avicel PH 112 type.

[0112] Thus, in a particular embodiment, the pharmaceutical formulation according to the invention comprises between about 100 mg and about 800 mg, preferably between about 300 mg and about 500 mg, even more preferably between about 400 mg and about 420 mg, for example about 413.7 mg, of microcrystalline cellulose per gram of powder.

[0113] Alternatively, the formulation may include croscarmellose. In particular, the croscarmellose may be in powder form, consisting of particles with an average diameter of less than 36 µm. Preferably, the croscarmellose is croscarmellose sodium.

[0114] In another particular embodiment, the pharmaceutical formulation according to the invention comprises between about 10 mg and about 250 mg, preferably between about 50 mg and about 150 mg, even more preferably between about 80 mg and 120 mg, for example about 100 mg of croscarmellose per gram of powder.

[0115] According to a particularly preferred embodiment, the pharmaceutical formulation according to the invention comprises microcrystalline cellulose and croscarmellose as defined above. Thus, the pharmaceutical formulation according to the invention may comprise between approximately 100 mg and approximately 800 mg, preferably between approximately 300 mg and approximately 500 mg, and even more preferably between approximately 400 mg and approximately 420 mg, for example approximately 413.7 mg, of microcrystalline cellulose per gram of powder; and / or between approximately 10 mg and approximately 250 mg, preferably between approximately 50 mg and approximately 150 mg, and even more preferably between approximately 80 mg and 120 mg, for example approximately 100 mg of croscarmellose per gram of powder.

[0116] The pharmaceutical formulation according to the invention may also include at least one anti-caking agent type excipient.

[0117] As used here, the term "anti-caking agent" refers to an excipient that limits the clumping of particles in a powdered product and thus ensures its fluidity.

[0118] Preferably, the anti-caking agent is selected from the group consisting of talc, silica and its derivatives, sodium carbonate, ammonium carbonate, ammonium bicarbonate, magnesium carbonate, sodium ferrocyanide, potassium ferrocyanide, calcium ferrocyanide, or mixtures thereof.

[0119] Preferably, the anti-caking agent is selected from the group consisting of silica and its derivatives, in particular silica, colloidal silica, silicon dioxide, calcium silicate, magnesium silicate, sodium aluminum silicate, potassium aluminum silicate, calcium aluminum silicate, zinc silicate, aluminum silicate, or mixtures thereof.

[0120] In a particularly preferred manner, the anti-caking agent is silica, preferably amorphous synthetic silica, in particular syloid ®< Al-1 FP.

[0121] In a preferred embodiment, the pharmaceutical formulation according to the invention comprises between about 20 mg and about 500 mg, preferably between about 50 mg and about 300 mg, more preferably between about 150 mg and about 200 mg, for example about 180 mg of anti-caking agent, preferably silica, per gram of powder.

[0122] The pharmaceutical formulation according to the invention may also include at least one lubricant-type excipient.

[0123] As used here, the term "lubricant" refers to an excipient intended to facilitate the manufacturing steps of the powder, particularly through its slippery, non-stick and anti-friction effects.

[0124] Preferably, the lubricant is selected from the group consisting of magnesium stearate, aluminum stearate, calcium stearate, sodium stearate, zinc stearate, sodium stearyl fumarate, propylene glycol, glyceryl monostearate, or mixtures thereof. Even more preferably, the lubricant is magnesium stearate.

[0125] According to a preferred embodiment, the pharmaceutical formulation according to the invention comprises between about 1 mg and about 40 mg, preferably between about 2 mg and about 15 mg, more preferably between about 4 mg and about 10 mg, for example about 6 mg of lubricant, preferably magnesium stearate, per gram of powder.

[0126] The pharmaceutical formulation according to the invention may also include at least one sweetener-type excipient.

[0127] As used here, the term "sweetener" refers to an excipient intended to change the taste of a pharmaceutical formulation by giving it a sweet flavor.

[0128] Preferably, the sweetener is selected from the group consisting of acesulfame potassium (E950), alitame (E956), aspartame (E951), cyclamate (E952), neotame, saccharin (E954), acesulfame aspartame salt (E962), sucralose, thaumatin, polyols, brazzein, curculin, glycyrrhizin, hydrogenated starch hydrolysate, mabinlin, miraculin, monelline, pentadine, stevia, tagatose, trehalose, isomaltulose, erythritol, and mixtures thereof. Aspartame is particularly preferred.

[0129] According to a preferred embodiment, the pharmaceutical formulation according to the invention comprises between about 1 mg and about 60 mg, preferably between about 3 mg and about 20 mg, even more preferably the pharmaceutical formulation of the invention comprises between about 8 mg and about 15 mg, for example about 12 mg of sweetener, preferably aspartame, per gram of powder.

[0130] The pharmaceutical formulation according to the invention may also include at least one flavoring-type excipient.

[0131] As used here, the term "flavoring" refers to an excipient intended to change the taste of a pharmaceutical formulation by giving it an aroma.

[0132] Preferably, the flavoring is selected from the group consisting of strawberry, raspberry, cherry, banana, lemon, orange, peach, apple, caramel, or mixtures thereof. Even more preferably, the flavoring is a mixture of lemon, strawberry, and peach flavors.

[0133] According to a preferred embodiment, the pharmaceutical formulation according to the invention comprises between about 1 mg and about 100 mg, preferably between about 10 mg and about 50 mg, more preferably between about 20 mg and about 40 mg, for example about 30 mg, of flavoring per gram of powder.

[0134] To prevent the risk of microbiological contamination, the pharmaceutical formulation according to the invention may also include a preservative agent.

[0135] Preservatives usable in the formulation according to the invention include, but are not limited to, benzoic acid and its sodium or potassium salts such as sodium benzoate; parabens such as methylparaben, propylparaben or butylparaben; sorbic acid and its sodium or potassium salts such as potassium sorbate; quaternary ammonium compounds such as benzalkonium chloride; mercurial derivatives such as phenylmercury salts (acetate, borate or nitrate) or thiomersal; and a combination thereof.

[0136] The pharmaceutical formulation according to the invention may also include a pH buffer type excipient, such as various acids and their salts, such as citric acid, sodium citrate and succinic acid.

[0137] The formulation may also include a colorant, particularly to increase its acceptability to children. Preferably, the colorant is used to enhance the credibility of the flavor (for example, a pink colorant for a strawberry flavor).

[0138] In a particularly preferred embodiment, the pharmaceutical formulation according to the invention does not comprise a detergent. The term "detergent" (or "surfactant"), as used here, refers to an amphiphilic molecule possessing surface-active properties. Detergents commonly used in pharmaceutical formulations include, for example, ionic detergents such as sodium dodecyl sulfate (SDS), deoxycholate, and cholate; non-ionic detergents such as Triton X-100, n-Dodecyl β-D-Maltopyranoside (DDM), digitonin, Tween 20, and Tween 80; and amphoteric detergents such as 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) or 3-[dimethylammonio]-1-propanesulfonate.

[0139] In another preferred embodiment, the pharmaceutical formulation according to the invention is in the form of a powder comprising, or consisting essentially of, or consisting of, cineole, amoxicillin, a pharmaceutically acceptable oil, and at least one sweetener, flavoring, anti-caking agent, lubricant and / or disintegrant as defined above and, optionally, a β-lactamase inhibitor, preferably of clavulanic acid, as defined above.

[0140] Preferably, the pharmaceutical formulation according to the invention is in the form of a powder comprising, or consisting essentially of, or consisting of, cineole, amoxicillin, and oil in the proportions described above, and between approximately 1 mg and approximately 60 mg of sweetener, preferably aspartame, and / or between approximately 1 mg and approximately 60 mg of flavoring, and / or between approximately 20 mg and approximately 500 mg of anti-caking agent, preferably silica, and / or between approximately 1 mg and approximately 40 mg of lubricant, preferably magnesium stearate, and / or between approximately 10 mg and approximately 1000 mg of disintegrant, preferably a mixture of microcrystalline cellulose and croscarmellose, per gram of powder, and optionally, a β-lactamase inhibitor, preferably clavulanic acid, in the proportions described above.

[0141] Preferably, the pharmaceutical formulation according to the invention is in the form of a powder comprising, or consisting essentially of, or consisting of, cineole, amoxicillin, and oil in the proportions described above, and between approximately 3 mg and approximately 20 mg of sweetener, preferably aspartame, and / or between approximately 10 mg and approximately 50 mg of flavoring, and / or between approximately 50 mg and approximately 300 mg of anti-caking agent, preferably silica, and / or between approximately 2 mg and approximately 15 mg of lubricant, preferably magnesium stearate, and / or between approximately 100 mg and approximately 600 mg of disintegrant, preferably a mixture of microcrystalline cellulose and croscarmellose, per gram of powder, and, optionally, a β-lactamase inhibitor, preferably clavulanic acid, in the proportions described above.

[0142] Most preferably, the pharmaceutical formulation according to the invention is in the form of a powder comprising, or consisting essentially of, or consisting of, cineole, amoxicillin, and oil in the proportions described above, and between approximately 8 mg and approximately 15 mg, for example approximately 12 mg, of sweetener, preferably aspartame, and / or between approximately 20 mg and approximately 40 mg, for example approximately 30 mg, of flavoring per gram of powder, and / or between approximately 150 mg and approximately 200 mg, for example approximately 180 mg, of anti-caking agent, preferably silica, and / or between approximately 4 mg and approximately 8 mg, for example approximately 6 mg, of lubricant, preferably magnesium stearate, and / or between approximately 450 mg and approximately 550 mg, for example approximately 513.7 mg, of disintegrant, for example approximately 100 mg of croscarmellose and approximately 413.7 mg of microcrystalline cellulose, and, optionally, a β-lactamase inhibitor,preferably clavulanic acid, in the proportions described above. Packaging, stability and route of administration of the formulation according to the invention

[0143] The inventors have demonstrated that the pharmaceutical formulation according to the invention is particularly stable. The stability of a product is characterized by the fact that the quality of said product does not vary over a given time interval under the influence of various environmental factors, in particular temperature and humidity (ICH standard). The absence of variation can be observed for a final percentage of amoxicillin, cineole, and optionally clavulanic acid that has not varied, preferably, by more than 5% from the initial values.

[0144] Thus, in a particular embodiment, the pharmaceutical formulation according to the invention is stable for at least 18 months, preferably for at least 24 months, even more preferably for at least 36 months at a temperature of about 25°C and under a relative humidity of at least about 60%.

[0145] In another particular embodiment, the pharmaceutical formulation according to the invention is stable for at least 6 months, preferably for at least 12 months, even more preferably for at least 24 months at a temperature of about 30°C and under a relative humidity of at least about 65%.

[0146] In yet another particular embodiment, the pharmaceutical formulation according to the invention is stable for at least 3 months, preferably for at least 6 months, even more preferably for at least 12 months at a temperature of about 40°C and under a relative humidity of at least about 75%.

[0147] Climate chambers, well known to those skilled in the art, allow the pharmaceutical formulation according to the invention to be subjected to the temperature and relative humidity conditions mentioned above.

[0148] As used here, the term "relative humidity" refers to the ratio of the partial pressure of water vapor contained in the air to the saturation vapor pressure (or vapor tension) at the same temperature.

[0149] The pharmaceutical formulation according to the invention is intended for oral administration.

[0150] Preferably, the pharmaceutical formulation is administered after suspension in an aqueous solvent and mixing, preferably water. Particularly preferred, the pharmaceutical formulation is dissolved extemporaneously. The pharmaceutical formulation according to the invention is stable after suspension in an aqueous medium for at least 48 hours, preferably for at least 72 hours, and even more preferably for at least 96 hours at a temperature of approximately 20°C or lower and at a relative humidity of approximately 15% or lower.

[0151] After suspension in an aqueous solvent, preferably water, the pharmaceutical formulation of the invention can form a solution with a slightly acidic pH. Preferably, the pH of the solution formed by suspending the pharmaceutical formulation of the invention in an aqueous solvent is between approximately 5 and approximately 7.

[0152] The pharmaceutical formulation according to the invention can be packaged in a single-dose or multi-dose container, preferably in a single-dose container.

[0153] In a preferred embodiment, the pharmaceutical formulation according to the invention is packaged in a single-dose container containing between approximately 1 g and approximately 150 g of powder, more preferably between approximately 1 g and approximately 50 g of powder, even more preferably between approximately 1 g and approximately 10 g of powder, and more particularly preferably approximately 3 g of powder.

[0154] When the pharmaceutical formulation is packaged in a unit-dose container, said unit-dose containers may be subsequently packaged in a box. A box may contain, for example, between 3 and 31 unit-dose containers, preferably between 5 and 21 unit-dose containers, and even more preferably between 7 and 14 unit-dose containers.

[0155] In another embodiment, the pharmaceutical formulation according to the invention is packaged in a multi-dose container. Said container can contain, for example, between approximately 10 grams and approximately 500 grams of powder, preferably between approximately 20 grams and approximately 200 grams of powder, even more preferably between approximately 30 grams of powder and approximately 100 grams of powder, and most particularly preferably approximately 50 grams of powder.

[0156] When the pharmaceutical formulation is packaged in a multi-dose container, this container may be further packaged in a box, optionally accompanied by a dosing device, for example a spoon, allowing a predetermined quantity of powder to be dispensed, preferably from approximately 1 mg to approximately 30 mg of powder, even more preferably from approximately 2 mg to approximately 20 mg of powder, and even more preferably from approximately 3 mg to approximately 12 mg of powder. In particular, the dosing device may allow for the dispensing of approximately 3 g, approximately 6 g, approximately 9 g, approximately 12 g, approximately 15 g, and / or approximately 18 g of powder. Combination of cineole and amoxicillin

[0157] In a second unclaimed aspect, the present application discloses a combination comprising, or consisting substantially of, cineole and amoxicillin for use in the treatment of a bacterial infection in a subject.

[0158] Preferably, in the disclosed combination, amoxicillin and cineole are administered at doses that achieve a therapeutic and / or antibacterial effect, preferably synergistic, as illustrated in the examples below. To achieve such an effect, amoxicillin and / or cineole may be administered in therapeutically effective or sub-therapeutic amounts.

[0159] In one aspect, cineole and amoxicillin are both administered in therapeutically effective amounts.

[0160] In another aspect, amoxicillin is administered in a therapeutically effective amount and cineole is administered in a sub-therapeutic amount.

[0161] In yet another aspect, amoxicillin is administered in a sub-therapeutic amount and cineole is administered in a therapeutically effective amount.

[0162] In yet another aspect, amoxicillin and cineole are both administered in sub-therapeutic amounts.

[0163] According to one aspect, cineole may be administered to the subject at a dose of between approximately 0.1 mg / kg / day and approximately 50 mg / kg / day, preferably between approximately 0.5 mg / kg / day and approximately 20 mg / kg / day, and most preferably between approximately 1 mg / kg / day and approximately 10 mg / kg / day of subject's weight; and / or amoxicillin may be administered to the subject at a dose of between approximately 5 mg / kg / day and approximately 200 mg / kg / day, preferably between approximately 10 mg / kg / day and approximately 100 mg / kg / day, and most preferably between approximately 15 mg / kg / day and approximately 50 mg / kg of subject's weight per day.

[0164] Preferably, the cineole of the combination disclosed in this application is administered to the subject at a dose of between approximately 0.1 mg / kg / day and approximately 50 mg / kg / day and the amoxicillin of the disclosed combination is administered to the subject at a dose of between approximately 5 mg / kg / day and approximately 200 mg / kg of subject's weight per day.

[0165] Preferably, the cineole of the combination disclosed in this application is administered to the subject at a dose of between approximately 0.5 mg / kg / day and approximately 20 mg / kg / day and the amoxicillin of the disclosed combination is administered to the subject at a dose of between approximately 10 mg / kg / day and approximately 100 mg / kg of subject's weight per day.

[0166] In a particularly preferred manner, the cineole of the combination disclosed in this application is administered to the subject at a dose of between approximately 1 mg / kg / day and approximately 10 mg / kg / day and the amoxicillin of the disclosed combination is administered to the subject at a dose of between approximately 15 mg / kg / day and approximately 50 mg / kg of subject's weight per day.

[0167] In one aspect, the combination disclosed in this application comprises, or essentially consists of, cineole, amoxicillin, and a β-lactamase inhibitor, preferably selected from clavulanic acid, sulbactam, tazobactam, and aztreonam, and mixtures thereof, for use in the treatment of a bacterial infection in a subject. Preferably, the β-lactamase inhibitor is clavulanic acid.

[0168] Thus, the cineole and amoxicillin in the disclosed combination may be administered to the subject in the amounts described above, and the β-lactamase inhibitor may be administered to the subject in a therapeutically effective amount or in a sub-therapeutic amount.

[0169] According to one particular aspect, the cineole and amoxicillin of the combination according to the invention are administered to the subject at doses as described above and the β-lactamase inhibitor, preferably clavulanic acid, is administered to the subject at a dose of between about 0.1 mg / kg / day and about 50 mg / kg / day, preferably between about 0.5 mg / kg / day and about 20 mg / kg / day, and most particularly preferred between about 1 mg / kg / day and about 10 mg / kg of subject's weight per day.

[0170] In another particular aspect, the combination disclosed in this application comprises, or essentially consists of, cineole and amoxicillin, the proportion of cineole being between about 0.02 mg and about 0.5 mg, preferably between about 0.1 mg and about 0.3 mg, of cineole per gram of amoxicillin, and even more preferably the proportion of cineole is about 0.2 mg of cineole per gram of amoxicillin. Preferably, the disclosed combination further comprises a β-lactamase inhibitor, preferably clavulanic acid, the proportion of β-lactamase inhibitor being between about 0.01 mg and about 0.5 mg, preferably between 0.1 mg and about 0.2 mg, of β-lactamase inhibitor per gram of amoxicillin, and even more preferably the proportion of β-lactamase inhibitor is about 0.125 mg of β-lactamase inhibitor per gram of amoxicillin.

[0171] The combination disclosed in this application may further include at least one other active ingredient. Preferably, the additional active ingredient of the disclosed combination is another antibiotic, in particular a β-lactam antibiotic, and / or another β-lactamase inhibitor, and / or an antifungal, and / or an antiparasitic, and / or an analgesic.

[0172] Thus, cineole, amoxicillin, and optionally a β-lactamase inhibitor, preferably clavulanic acid, of the disclosed combination may be administered to the subject in quantities as described above and the other active substance(s) may be administered to the subject in therapeutically effective or sub-therapeutic amounts. Packaging and route of administration of the combination according to the invention

[0173] The active ingredients of the combination disclosed in this application may be administered to the subject simultaneously, sequentially, separately, or in a mixture of these modes of administration.

[0174] When active ingredients of the disclosed combination are administered sequentially or separately, the time interval(s) are preferably chosen to enable the desired therapeutic effect to be achieved, preferably with a synergistic effect between cineole and amoxicillin.

[0175] Preferably, the active ingredients of the disclosed combination are administered simultaneously. When the active ingredients of the disclosed combination are administered simultaneously, it is preferably done by administering a single formulation comprising all the active ingredients of the combination.

[0176] When the disclosed combination contains more than two active ingredients, some active ingredients may be administered simultaneously, some active ingredients may be administered sequentially, and / or some active ingredients may be administered separately. For example, when the disclosed combination contains three active ingredients, two active ingredients may be administered simultaneously and the third administered sequentially or separately, preferably sequentially.

[0177] In one particular aspect, the disclosed combination includes cineol and amoxicillin administered to the subject simultaneously, preferably from a pharmaceutical composition comprising cineol and amoxicillin, alternatively from two pharmaceutical compositions, one comprising cineol and the other comprising amoxicillin.

[0178] In another particular aspect, the disclosed combination includes cineole and amoxicillin administered to the subject sequentially or separately, preferably sequentially, from two pharmaceutical compositions, one comprising cineole and the other comprising amoxicillin.

[0179] In yet another specific aspect, the disclosed combination includes cineole, amoxicillin, and a β-lactamase inhibitor, preferably clavulanic acid, administered to the subject simultaneously. The cineole, amoxicillin, and β-lactamase inhibitor may be administered to the subject simultaneously from: (a) a pharmaceutical composition comprising cineol, amoxicillin and a β-lactamase inhibitor; (b) a pharmaceutical composition comprising cineol and a pharmaceutical composition comprising amoxicillin and a β-lactamase inhibitor; (c) a pharmaceutical composition comprising amoxicillin and a pharmaceutical composition comprising cineol and a β-lactamase inhibitor; (d) a pharmaceutical composition comprising a β-lactamase inhibitor, and a pharmaceutical composition comprising cineol and amoxicillin; or (e) a pharmaceutical composition comprising cineol, a pharmaceutical composition comprising amoxicillin, and a pharmaceutical composition comprising a β-lactamase inhibitor.

[0180] In yet another particular aspect, the disclosed combination comprises cineol, amoxicillin and a β-lactamase inhibitor, preferably clavulanic acid, administered to the subject sequentially or separately, preferably sequentially, from a pharmaceutical composition comprising cineol, a pharmaceutical composition comprising amoxicillin, and a pharmaceutical composition comprising a β-lactamase inhibitor.

[0181] In yet another specific aspect, the disclosed combination comprises cineole, amoxicillin, and a β-lactamase inhibitor, preferably clavulanic acid, in which the amoxicillin and the β-lactamase inhibitor are administered to the subject simultaneously and the cineole is administered to the subject sequentially or separately, preferably sequentially, from: (a) a pharmaceutical composition comprising cineole, a pharmaceutical composition comprising amoxicillin, and a pharmaceutical composition comprising a β-lactamase inhibitor; or (b) a pharmaceutical composition comprising cineole and a pharmaceutical composition comprising amoxicillin and a β-lactamase inhibitor.

[0182] In yet another specific aspect, the disclosed combination comprises cineol, amoxicillin, and a β-lactamase inhibitor, preferably clavulanic acid, in which the cineol and the β-lactamase inhibitor are administered to the subject simultaneously and the amoxicillin is administered to the subject sequentially or separately, preferably sequentially, from: (a) a pharmaceutical composition comprising cineole, a pharmaceutical composition comprising amoxicillin, and a pharmaceutical composition comprising a β-lactamase inhibitor; or (b) a pharmaceutical composition comprising amoxicillin and a pharmaceutical composition comprising cineole and a β-lactamase inhibitor.

[0183] In yet another specific aspect, the disclosed combination comprises cineole, amoxicillin, and a β-lactamase inhibitor, preferably clavulanic acid, in which amoxicillin and cineole are administered to the subject simultaneously and the β-lactamase inhibitor is administered to the subject sequentially or separately, preferably sequentially, from: (a) of a pharmaceutical composition comprising cineole, a pharmaceutical composition comprising amoxicillin, and a pharmaceutical composition comprising a β-lactamase inhibitor; or (b) of a pharmaceutical composition comprising a β-lactamase inhibitor, and a pharmaceutical composition comprising cineole and amoxicillin.

[0184] In one aspect, amoxicillin and a β-lactamase inhibitor, preferably clavulanic acid, are administered to the subject simultaneously, preferably from a pharmaceutical composition comprising amoxicillin and a β-lactamase inhibitor.

[0185] In another aspect, cineol, amoxicillin and optionally a β-lactamase inhibitor, preferably clavulanic acid, are administered to the subject simultaneously, preferably from a pharmaceutical composition comprising cineol, amoxicillin and optionally a β-lactamase inhibitor, or from a pharmaceutical composition comprising cineol and a pharmaceutical composition comprising amoxicillin and optionally a β-lactamase inhibitor.

[0186] In yet another aspect, cineole is administered to the subject sequentially or separately, preferably sequentially with respect to amoxicillin and / or, optionally, with respect to the β-lactamase inhibitor, preferably clavulanic acid.

[0187] The active ingredients of the combination can be administered to the subject by the same or different routes. The routes of administration generally depend on the pharmaceutical formulations used. The active ingredients of the combination according to the invention are preferably administered to the subject parenterally or enterally, preferably enterally, and even more preferably orally or rectally. Most preferably, the active ingredients of the combination are administered to the subject orally.

[0188] In one particular aspect, amoxicillin and, optionally, a β-lactamase inhibitor, preferably clavulanic acid, are administered to the subject orally and cineole is administered to the subject rectally.

[0189] In a preferred aspect, amoxicillin, cineole and, optionally, a β-lactamase inhibitor, preferably clavulanic acid, are administered to the subject orally.

[0190] The active ingredients of the combination, in particular amoxicillin, cineole, and possibly a β-lactamase inhibitor, preferably clavulanic acid, may be administered to the subject in the form of any pharmaceutical formulation, preferably compatible with parenteral or enteral administration, more preferably compatible with oral or rectal administration, particularly preferably compatible with oral administration.

[0191] The active ingredients of the combination, in particular amoxicillin, cineole, and possibly a β-lactamase inhibitor, preferably clavulanic acid, can thus be formulated in the form of tablets, capsules, softgels, granules, powder, suspensions, emulsions, solutions, polymers, nanoparticles, microspheres, suppositories, rectal capsules, enemas, gels, pastes, ointments, creams, plasters, potions, injectables, implants, sprays or aerosols.

[0192] In one aspect, the active ingredients of the combination according to the invention are formulated in powder form, preferably powders for oral aqueous suspensions.

[0193] In one particular aspect, the active ingredients are formulated in the form of suppositories or rectal capsules.

[0194] In another particular aspect, cineole is formulated as an oil, or as a suppository or rectal capsule, preferably as an encapsulated oil, and the other active ingredients are formulated as powders, preferably powders for oral aqueous suspensions. Treatment of an infectious disease in a subject

[0195] In another aspect, the present invention also relates to a pharmaceutical formulation according to the invention for use in the treatment of an infectious disease in a subject. It also relates to a pharmaceutical formulation according to the invention for the preparation of a drug intended for the treatment of an infectious disease. Furthermore, it relates to a treatment method comprising the administration of a therapeutically effective amount of the formulation according to the invention to a subject in need, in particular a subject suffering from an infectious disease.

[0196] The present application relates to a combination as disclosed herein for use in the treatment of infectious pathology, preferably a bacterial infection, in a subject...

[0197] Preferably, the infectious pathology is of bacterial origin; even more preferably, the infectious pathology is caused by one or more bacteria resistant to antibiotics, particularly to antibiotics of the β-lactam family. Preferably, the term "antibiotic-resistant bacterium" refers to a bacterium that is, depending on the case, insensitive or slightly sensitive to antibiotics. Similarly, a bacterium resistant to β-lactam antibiotics is insensitive or slightly sensitive to antibiotics of this family. A bacterial infection caused by a bacterium resistant to β-lactam antibiotics will therefore not be effectively treated with antibiotics of this family.

[0198] In a preferred embodiment, the infectious pathology is caused by a bacterium resistant to amoxicillin. Preferably, the bacterium responsible for the infectious pathology is at least partially resistant to a combination of amoxicillin and clavulanic acid.

[0199] In another preferred embodiment, the bacterium responsible for the infectious pathology is a β-lactamase-producing bacterium. β-lactamases are serine-active inactivating enzymes (classes A, C, and D) or metalloenzymes (class B) whose substrates are β-lactam antibiotics. A distinction is made between narrow-spectrum β-lactamases and extended-spectrum β-lactamases (ESBLs), which are capable of inhibiting a large number of different β-lactam antibiotics, particularly penicillins, including amoxicillin, first-, second-, third- (e.g., cefotaxime, ceftazidime), and fourth-generation (e.g., cefepime) cephalosporins, and monobactams (e.g., aztreonam).

[0200] In a particularly preferred embodiment, the bacterium responsible for the infectious pathology is an extended-spectrum β-lactamase (ESBL)-producing bacterium. Specifically, an ESBL-producing bacterium that is insensitive to clavulanic acid. Alternatively, the bacterium produces ESBLs that are only partially inhibited by clavulanic acid. ESBL-producing bacteria can be detected by tests well known to those skilled in the art, such as the double-disc test, the combined-disc method, and the ESBL E-test.

[0201] The bacterial infection treated by the pharmaceutical formulation of the invention can be selected from the group consisting of cystitis, in particular recurrent acute cystitis, bacterial sinusitis, in particular acute maxillary sinusitis, otitis, in particular acute otitis media, bronchitis, in particular chronic and / or acute bronchitis, bronchopneumopathies, in particular chronic and / or acute bronchopneumopathies, pyelonephritis, upper gynecological infections, periodontitis, severe stomatological infections, in particular abscesses, phlegmons and cellulitis, animal bites, bone and joint infections, in particular osteomyelitis, endocarditis, pericarditis, septicemia, and skin infections.Preferably, said bacterial infection is cystitis, more preferably cystitis resistant to β-lactam antibiotics, more preferably cystitis resistant to amoxicillin, and most preferably cystitis resistant to a combination of amoxicillin and clavulanic acid.

[0202] The terms "subject" and "patient" are equivalent and may be used interchangeably within the context of the present invention. As used herein, the term "subject" refers to an animal, preferably a mammal, and even more preferably, to a human.

[0203] Thus, in a particular embodiment, the subject to whom the formulation or combination according to the invention is administered is a human being. This may be a newborn, a child, an adolescent, an adult, or an elderly person. As used herein, the term "newborn" refers to a human being less than 12 months old, preferably less than 6 months old, and even more preferably less than 3 months old. As used in the present invention, the term "child" refers to a human being aged 1 to 12 years, preferably a human being aged 1 to 8 years, and even more preferably a human being aged 1 to 5 years. As used herein, the term "adult" refers to a human being aged 12 to 60 years, preferably a human being aged 15 to 60 years, and even more preferably a human being aged 18 to 60 years.As used in the present invention, the term "elderly person" refers to a human being aged 60 years or older, preferably to a human being aged 65 years or older, and even more preferably to a human being aged 70 years or older.

[0204] In a preferred embodiment, the subject to whom the formulation according to the invention is administered is an adult human.

[0205] In the field of veterinary applications, the subject of the invention may be a non-human animal, preferably a pet or farm animal, more preferably an animal selected from the group consisting of dogs, cats, cattle, sheep, rabbits, pigs, goats, equines, rodents, in particular hamsters and guinea pigs, non-human primates and poultry, preferably broiler chickens, laying hens, breeding roosters and hens, guinea fowl, turkeys, quail, ducks, geese and pigeons.

[0206] In a particularly preferred embodiment, the present invention relates to a formulation according to the invention in the treatment of cystitis resistant to β-lactam antibiotics in a human. Dosage

[0207] The pharmaceutical formulation according to the invention, used in the treatment of an infectious disease, preferably a bacterial infection, in a patient, can be administered as a single dose (one administration) or as multiple doses (multiple administrations), depending on the patient, their age, their state of health, and the infection being treated. When multiple doses (multiple administrations) of the pharmaceutical formulation according to the invention are administered, these can be spread over one or more days. The pharmaceutical formulation according to the invention can thus be administered as a single dose (one administration) per day of treatment. Preferably, the patient receives multiple doses (multiple administrations) of the pharmaceutical formulation according to the invention per day of treatment.Preferably, the subject receives three doses (three administrations) of the pharmaceutical formulation according to the invention per day of treatment administration, preferably morning, noon, and evening.

[0208] The number of doses (or administrations) received by the patient per day of treatment may also vary over time. Thus, periods may alternate between periods where the patient receives a single dose (one administration) per day of treatment and periods where the patient receives several doses (several administrations) per day of treatment, preferably three doses (three administrations) spaced morning, noon, and evening. In particular, the patient may receive a single dose (one administration), generally called a "loading dose," on the first day of treatment before receiving several doses (several administrations), called "maintenance doses," per day of treatment on subsequent days, preferably three maintenance doses (three administrations) per day of treatment, preferably morning, noon, and evening.

[0209] The loading dose, generally administered to the subject on the first day of treatment, preferably as a single dose, is a dose of the formulation according to the invention that is preferably greater than or equal to the maintenance doses; more preferably, the loading dose is greater than or equal to two maintenance doses when the subject receives at least two maintenance doses per day of treatment. Preferably, the loading dose is between approximately 5 g and approximately 20 g, more preferably between approximately 10 g and approximately 15 g, and most preferably approximately 12 g. Preferably, the maintenance dose is between approximately 1 g and approximately 15 g, more preferably between approximately 3 g and approximately 6 g, and most preferably approximately 3 g or approximately 6 g.

[0210] The pharmaceutical formulation according to the invention can be administered daily, every other day, or once a week, preferably daily. The frequency of administration of the formulation or combination depends on various parameters and can be readily determined by a person skilled in the art, based, for example, on the overall expected duration of administration of the pharmaceutical formulation or combination, the age of the patient, and / or the severity of the infection, preferably bacterial, to be prevented or treated. The frequency of administration of the pharmaceutical formulation or combination can also change over time in a given patient, particularly depending on the progression of their infection and / or their overall health.

[0211] In particular, the pharmaceutical formulation according to the invention can be administered to the subject for a period ranging from 1 day to approximately 3 months, 2 months, 1 month, 3 weeks, 2 weeks, 1 week, 5 days, 3 days, 2 days, preferably for a period of approximately 2 to approximately 21 days, and even more preferably for a period of approximately 7 to approximately 14 days. In a particularly preferred embodiment, the pharmaceutical formulation of the invention is administered to the subject for a period of approximately 7 days. Alternatively, the pharmaceutical formulation according to the invention can be administered for the entire duration of the infection. In the case of a urinary tract infection, the formulation according to the invention can be administered until sterile urine is obtained in the subject.

[0212] In a particular embodiment, the pharmaceutical formulation according to the invention can be administered for several months, possibly for several years, for example in the case of chronic bacterial infections.

[0213] The pharmaceutical formulation according to the invention can be administered to the subject at a dose of approximately 1 to approximately 150 grams, preferably approximately 1 to approximately 50 grams, even more preferably approximately 2 to approximately 30 grams, and most preferably approximately 3 to approximately 20 grams per day of treatment. For example, approximately 9 grams of the pharmaceutical formulation according to the invention can be administered to the subject per day of treatment. In another example, approximately 18 grams of the formulation according to the invention can be administered to the subject per day of treatment. Pharmaceutical composition of cineole and amoxicillin

[0214] The application also relates, in another unclaimed aspect, to a pharmaceutical composition comprising, or consisting essentially of, cineole, amoxicillin and a pharmaceutically acceptable excipient or carrier, and optionally a β-lactamase inhibitor.

[0215] In a preferred embodiment, the β-lactamase inhibitor is selected from clavulanic acid, sulbactam, tazobactam and aztreonam, preferably the β-lactamase inhibitor is clavulanic acid.

[0216] In one particular aspect, the disclosed composition comprises, or consists substantially of, cineole, amoxicillin, optionally a β-lactamase inhibitor, preferably clavulanic acid, and another active ingredient. Preferably, the additional active ingredient in the disclosed composition is another antibiotic, in particular a β-lactam antibiotic, and / or another β-lactamase inhibitor, and / or an antifungal, and / or an antiparasitic, and / or an analgesic.

[0217] Preferably, the disclosed composition comprises, or consists essentially of, active ingredients, preferably amoxicillin and cineole, present at concentrations sufficient to allow the administration of doses to achieve a therapeutic and / or antibacterial effect, preferably a synergistic effect, as illustrated in the examples below. To achieve such an effect, amoxicillin and cineole may be administered in therapeutically effective or sub-therapeutic amounts.

[0218] In one aspect, cineole and amoxicillin are both administered in therapeutically effective amounts.

[0219] In another aspect, amoxicillin is administered in a therapeutically effective amount and cineole is administered in a sub-therapeutic amount.

[0220] In yet another aspect, amoxicillin is administered in a sub-therapeutic amount and cineole is administered in a therapeutically effective amount.

[0221] In yet another aspect, amoxicillin and cineole are both administered in sub-therapeutic amounts.

[0222] When present in the composition according to the invention, the β-lactamase inhibitor and / or the additional active ingredient can be administered in therapeutically effective or sub-therapeutic amounts.

[0223] The disclosed composition may comprise, or consist substantially of, between about 5 mg and about 100 mg, preferably between about 10 mg and about 50 mg, even more preferably between about 20 mg and about 40 mg, most preferably about 33 mg, of cineole per gram of composition; and / or between about 20 mg and about 500 mg, preferably between about 50 mg and about 300 mg, even more preferably between about 150 mg and about 200 mg, most preferably about 167 mg, of amoxicillin per gram of composition; and / or optionally between about 1 mg and about 100 mg, preferably between about 5 mg and about 50 mg, even more preferably between about 15 mg and about 25 mg, and most preferably about 21 mg, of β-lactamase inhibitor, preferably clavulanic acid per gram of composition.

[0224] Preferably, the disclosed composition comprises, or consists substantially of, between about 5 mg and about 100 mg of cineole, between about 20 mg and about 500 mg of amoxicillin, and optionally between about 1 mg and about 100 mg of β-lactamase inhibitor, preferably clavulanic acid, per gram of composition.

[0225] Preferably, the disclosed composition comprises, or consists substantially of, between about 10 mg and about 50 mg of cineole, between about 50 mg and about 300 mg of amoxicillin, and optionally between about 5 mg and about 50 mg of β-lactamase inhibitor, preferably clavulanic acid, per gram of composition.

[0226] Most preferably, the disclosed composition comprises, or consists substantially of, between about 20 mg and about 40 mg, preferably about 33 mg, of cineole, and between about 150 mg and about 200 mg, preferably about 167 mg, of amoxicillin, and optionally between about 15 mg and about 25 mg, and most preferably about 21 mg, of β-lactamase inhibitor, preferably clavulanic acid, per gram of composition.

[0227] The disclosed composition may be in the form of tablets, capsules, softgels, granules, powder, suspensions, emulsions, solutions, polymers, nanoparticles, microspheres, suppositories, rectal capsules, enemas, gels, pastes, ointments, creams, plasters, potions, injectables, implants, sprays, or aerosols. Preferably, the disclosed pharmaceutical composition is in powder form, more particularly a powder for oral suspension.

[0228] Preferably, the technical characteristics of the powder are as defined previously for the pharmaceutical formulation according to the invention.

[0229] The disclosed composition also includes at least one pharmaceutically acceptable excipient or carrier. A person skilled in the art can readily determine the excipients required for a composition based on the chosen pharmaceutical form.

[0230] In particular, when the disclosed composition is in powder form, the pharmaceutically acceptable excipient or carrier of the disclosed composition is preferably selected from the group consisting of a sweetener, a flavoring agent, an anti-caking agent, a lubricant, a disintegrant, an adsorbent, and a mixture thereof.

[0231] Preferably, when the composition according to the invention is in powder form, the composition according to the invention comprises at least one sweetener, preferably aspartame, a flavoring, an anti-caking agent, preferably silica, a lubricant, preferably magnesium stearate, a disintegrant, preferably a mixture of croscarmellose and microcrystalline cellulose, and an adsorbent, preferably oil. Such a composition may further comprise a preservative, a colorant, and / or a pH buffer.

[0232] Thus, in a preferred embodiment, the composition according to the invention is in the form of a powder comprising, or consisting essentially of, or consisting of, cineole, amoxicillin, and at least one sweetener, flavoring agent, anti-caking agent, lubricant, disintegrant and / or adsorbent as defined above and a β-lactamase inhibitor, preferably clavulanic acid, as defined above.

[0233] Preferably, the composition is a powder comprising, or consisting essentially of, or consisting of, cineole and amoxicillin in the proportions described above, and between approximately 1 mg and approximately 60 mg of sweetener, preferably aspartame, and / or between approximately 1 mg and approximately 60 mg of flavoring, and / or between approximately 20 mg and approximately 500 mg of anti-caking agent, preferably silica, and / or between approximately 1 mg and approximately 40 mg of lubricant, preferably magnesium stearate, and / or between approximately 10 mg and approximately 1000 mg of disintegrant, preferably a mixture of microcrystalline cellulose and croscarmellose, and / or between approximately 2 mg and approximately 50 mg of adsorbent, preferably oil, per gram of powder, and optionally a β-lactamase inhibitor, preferably acid clavulanic, in the proportions described above.

[0234] The composition according to the invention may, in particular, include at least one adsorbent-type excipient. As used here, the term "adsorbent" refers to an excipient capable of binding liquid molecules, for example cineole molecules, to a solid support, for example a powder, in a pharmaceutical composition. Preferably, the adsorbent is a pharmaceutically acceptable oil. The pharmaceutically acceptable oil is as described above.

[0235] In another particularly preferred embodiment, the composition according to the invention does not include a detergent. The detergent is as described previously in the chapter on excipients.

[0236] The application also relates to a disclosed pharmaceutical composition for use in the treatment of an infectious disease, preferably a bacterial infection, in a subject.

[0237] It also relates to a treatment method comprising the administration of a therapeutically effective amount of the disclosed pharmaceutical composition to a subject in need of it, in particular a subject suffering from an infectious pathology, preferably a bacterial infection.

[0238] The embodiments and definitions described for the use of a disclosed combination or pharmaceutical formulation according to the invention in the treatment of an infectious pathology, preferably a bacterial infection, in a subject, as well as concerning the dosage of this combination or formulation, are also to be taken into consideration in this aspect.

[0239] Preferably, the infectious pathology is caused by one or more bacteria resistant to antibiotics, preferably to antibiotics of the β-lactam family, even more preferably to amoxicillin and particularly preferably to one or more bacteria at least partially resistant to a combination of amoxicillin and clavulanic acid.

[0240] In a particularly preferred embodiment, the present application relates to a pharmaceutical composition as disclosed in this application for use in the treatment of cystitis, in particular cystitis resistant to antibiotics, preferably to antibiotics of the β-lactam family, further preferred to amoxicillin and particularly preferred to a combination of amoxicillin and clavulanic acid. Kit

[0241] The application also relates, in another unclaimed aspect, to a kit for the treatment of an infectious disease, preferably a bacterial infection, in a subject comprising: (a) a pharmaceutical composition comprising, or consisting substantially of, cineole and a pharmaceutical composition comprising, or consisting substantially of, amoxicillin; (b) a pharmaceutical composition comprising, or consisting substantially of, cineole and a pharmaceutical composition comprising, or consisting substantially of, amoxicillin and a β-lactamase inhibitor, preferably clavulanic acid; (c) a pharmaceutical composition comprising, or consisting substantially of, amoxicillin and a pharmaceutical composition comprising, or consisting substantially of, cineole and a β-lactamase inhibitor, preferably clavulanic acid; (d) a pharmaceutical composition comprising, or consisting substantially of, a β-lactamase inhibitor, preferably clavulanic acid, and a pharmaceutical composition comprising, or consisting substantially of, cineole and amoxicillin;or (e) a pharmaceutical composition comprising, or consisting substantially of, cineole, a pharmaceutical composition comprising, or consisting substantially of, amoxicillin, and a pharmaceutical composition comprising, or consisting substantially of, a β-lactamase inhibitor, preferably clavulanic acid; and (f) optionally, a guide containing instructions for the use of such a kit.

[0242] Preferably, each item included in the kit is in a separate container, receptacle, and / or package.

[0243] In a particular aspect, at least one pharmaceutical composition of the disclosed kit further includes at least one other active ingredient. Preferably, the additional disclosed active ingredient is another antibiotic, in particular a β-lactam antibiotic, and / or another β-lactamase inhibitor, and / or an antifungal, and / or an antiparasitic, and / or an analgesic.

[0244] Preferably, the active ingredients of the kit compositions according to the invention, in particular amoxicillin and cineole, are present at concentrations allowing the administration of doses sufficient to achieve a therapeutic and / or antibacterial effect, preferably a synergistic effect when said active ingredients are present in the same composition or when, present in different compositions, they are used simultaneously, sequentially, or separately. To achieve such an effect, cineole and amoxicillin can be administered in therapeutically effective or sub-therapeutic amounts.

[0245] In a preferred aspect, cineole and amoxicillin are both administered in therapeutically effective amounts.

[0246] In another preferred aspect, amoxicillin is administered in a therapeutically effective amount and cineole is administered in a sub-therapeutic amount.

[0247] In yet another preferred aspect, amoxicillin is administered in a sub-therapeutic amount and cineole is administered in a therapeutically effective amount.

[0248] In yet another preferred aspect, amoxicillin and cineole are both administered in sub-therapeutic amounts.

[0249] When present in the compositions of the kit according to the invention, the β-lactamase inhibitor and / or the additional active ingredient can be administered in therapeutically effective or sub-therapeutic amounts.

[0250] The pharmaceutical composition of the disclosed kit containing cineole may include between approximately 5 mg and approximately 100 mg, preferably between approximately 10 mg and approximately 50 mg, more preferably between approximately 20 mg and approximately 40 mg, most preferably approximately 33 mg, of cineole per gram of composition.

[0251] The pharmaceutical composition of the disclosed kit comprising amoxicillin may include between approximately 20 mg and approximately 500 mg, preferably between approximately 50 mg and approximately 300 mg, more preferably between approximately 150 mg and approximately 200 mg, most preferably approximately 167 mg, of amoxicillin per gram of composition.

[0252] When a pharmaceutical composition of the disclosed kit includes a β-lactamase inhibitor, preferably of clavulanic acid, said composition may include between about 1 mg and about 100 mg, preferably between about 5 mg and about 50 mg, more preferably about 15 mg and about 25 mg, and most preferably about 21 mg, of β-lactamase inhibitor per gram of composition.

[0253] The compositions of the disclosed kit may be in the form of tablets, capsules, softgels, granules, powder, suspensions, emulsions, solutions, polymers, nanoparticles, microspheres, suppositories, rectal capsules, enemas, gels, pastes, ointments, creams, plasters, potions, injectables, implants, sprays or aerosols.

[0254] Preferably, the disclosed pharmaceutical compositions are in the form of powders, suppositories or rectal capsules, preferably in powder form, even more preferably in dry powder form, more particularly powders for oral suspensions.

[0255] When cineole is the only active ingredient in a composition, it may be in oil form, particularly encapsulated oil.

[0256] The pharmaceutical composition(s) of the disclosed kit preferably include at least one pharmaceutically acceptable excipient or carrier. A person skilled in the art can readily determine the excipients required for a composition based on the chosen pharmaceutical form. Pharmaceutically acceptable excipients or carriers are as defined above for the pharmaceutical composition.

[0257] The application relates, in yet another unclaimed aspect, to a kit disclosed for use in the treatment of an infectious pathology, preferably a bacterial infection, in a subject.

[0258] The embodiments and definitions described for the use of a disclosed formulation or combination in the treatment of an infectious pathology in a subject, as well as concerning the administration and dosage of this formulation or combination, are also to be taken into consideration in this aspect.

[0259] Preferably, the infectious pathology is caused by one or more bacteria resistant to antibiotics, preferably to antibiotics of the β-lactam family, even more preferably to amoxicillin and particularly preferably by one or more bacteria at least partially resistant to a combination of amoxicillin and clavulanic acid.

[0260] In one aspect, the present application relates to a pharmaceutical composition as disclosed in this application for use in the treatment of cystitis, in particular cystitis resistant to antibiotics, preferably to β-lactam antibiotics, further preferred to amoxicillin and particularly preferred to a combination of amoxicillin and clavulanic acid. Pharmaceutical formulation manufacturing process

[0261] The present invention also relates, in a third aspect, to a method for manufacturing the pharmaceutical composition or formulation according to the invention, comprising: obtaining a wetting solution by mixing cineole and a pharmaceutically acceptable oil; wetting a powder comprising amoxicillin with the wetting solution to obtain a powder preparation comprising amoxicillin, cineole and oil.

[0262] The present invention also relates to the pharmaceutical composition or formulation obtained by the process according to the invention.

[0263] As used here, the term "wetting solution" refers to a solution intended to wet or moisten a dry powder. The wetting process can be carried out, in particular, by spraying the wetting solution onto the powder to be wetted.

[0264] The step of obtaining a wetting solution for the process according to the invention can be carried out by mixing cineole and a pharmaceutically acceptable oil as described above in oil-to-cineole mass proportions of between about 0.1 and about 1, preferably between about 0.2 and about 0.8, and even more preferably between about 0.4 and about 0.6. Most preferably, the oil-to-cineole mass proportion is about 0.5. For example, about 50 mg of oil is mixed with about 100 mg of cineole, or about 16.7 mg of oil is mixed with about 33.3 mg of cineole.

[0265] Preferably, the mixture of cineole and oil to obtain a wetting solution is carried out in a closed chamber, preferably at a temperature not exceeding about 20°C to avoid evaporation of the cineole, but sufficient to maintain the mixture in a liquid state.

[0266] In a particular embodiment, the powder comprising amoxicillin and intended to be moistened by the wetting solution also comprises a β-lactamase inhibitor, preferably selected from the group consisting of clavulanic acid, sulbactam, tazobactam, aztreonam and pharmaceutically acceptable salts thereof, even more preferably the β-lactamase inhibitor is clavulanic acid.

[0267] Preferably, the powder containing amoxicillin, intended to be moistened by the wetting solution, also comprises at least one pharmaceutically acceptable excipient or carrier, preferably selected from the group consisting of a sweetener, a flavoring agent, an anti-caking agent, a lubricant, a disintegrant, and a mixture thereof. Even more preferably, the powder containing amoxicillin comprises at least one sweetener, a flavoring agent, an anti-caking agent, a lubricant, and a disintegrant as defined above.

[0268] In another particular embodiment, the powder comprising amoxicillin and intended to be moistened by the wetting solution, also comprises clavulanic acid, silica, colloidal silica, aspartame, croscarmellose, microcrystalline cellulose, magnesium stearate and a flavoring as defined above.

[0269] Preferably, the process according to the invention further comprises a step of compacting the powder containing amoxicillin before it is moistened with the wetting solution. The granules obtained by compaction are preferably calibrated and then fractionated before being moistened with the wetting solution.

[0270] In another embodiment, the manufacturing process for the pharmaceutical composition or formulation according to the invention comprises: obtaining a wetting solution by mixing cineole and a pharmaceutically acceptable oil; wetting a powder comprising amoxicillin with the wetting solution to obtain a powder preparation comprising amoxicillin, cineole and oil; mixing the preparation obtained in the previous step with a powder comprising a β-lactamase inhibitor, preferably selected from the group consisting of clavulanic acid, sulbactam, tazobactam, aztreonam and pharmaceutically acceptable salts thereof, even more preferably the β-lactamase inhibitor being clavulanic acid; sieving the powder thus obtained.

[0271] In this process, the powder comprising amoxicillin and / or the powder comprising the β-lactamase inhibitor, preferably clavulanic acid, may further comprise a disintegrant as described above. Preferably, two disintegrants, in particular microcrystalline cellulose and croscarmellose. In a preferred embodiment, the powder comprising amoxicillin and the powder comprising a β-lactamase inhibitor, preferably clavulanic acid, comprise two disintegrants, microcrystalline cellulose and croscarmellose.

[0272] The powder containing amoxicillin and / or the powder containing a β-lactamase inhibitor, preferably clavulanic acid, may further include an anti-caking agent as described above. In a preferred embodiment, both the powder containing amoxicillin and the powder containing a β-lactamase inhibitor, preferably clavulanic acid, include an anti-caking agent, preferably silica.

[0273] In a particularly preferred embodiment, the powder comprising amoxicillin and the powder comprising a β-lactamase inhibitor, preferably clavulanic acid, both comprise microcrystalline cellulose, croscarmellose, and silica.

[0274] Thus, the process according to the invention may include a sieving step followed by mixing the disintegrant(s) and the anti-caking agent. Alternatively, the disintegrant(s) and the anti-caking agent may be mixed before the mixture is sieved. In particular, microcrystalline cellulose, croscarmellose, and silica may be mixed in proportions of approximately 1000 mg to approximately 1500 mg, preferably approximately 1241 mg, of microcrystalline cellulose to approximately 200 mg to approximately 400 mg, preferably approximately 300 mg, of croscarmellose, and approximately 400 mg to approximately 600 mg, preferably approximately 540 mg, of silica.

[0275] The process according to the invention may also include a step of mixing these excipients with the powder comprising amoxicillin and / or with the powder comprising clavulanic acid, preferably one part of these excipients is mixed with the powder comprising amoxicillin and the other part of these excipients with the powder comprising clavulanic acid.

[0276] In another preferred embodiment, the powder comprising a β-lactamase inhibitor, preferably clavulanic acid, further comprises colloidal silica, preferably in a mass proportion relative to clavulanic acid of between about 0.7 and about 1.3, preferably between about 0.9 and about 1.1, more preferably about 1. Preferably, the mixture of clavulanic acid and colloidal silica is made before the addition of any other excipient or mixture with the powder comprising amoxicillin.

[0277] Preferably, the process according to the invention further comprises a step of compacting the powder containing amoxicillin before it is moistened with the wetting solution. The granules obtained by compaction are preferably calibrated and then fractionated before being moistened with the wetting solution.

[0278] Similarly, the process according to the invention preferably includes a step of compacting the powder comprising a β-lactamase inhibitor, preferably clavulanic acid, before it is mixed with the amoxicillin powder moistened by the wetting solution. The granules obtained by compaction are preferably calibrated and then fractionated before being moistened with the wetting solution.

[0279] During the step of wetting the powder containing amoxicillin with the wetting solution, the wetting solution may be used at a rate of approximately 100 to approximately 200 mg, preferably approximately 150 mg, of wetting solution for a powder containing approximately 400 to approximately 600 mg, preferably approximately 500 mg, of amoxicillin.

[0280] During the mixing step of the amoxicillin, cineole and oil preparation with the powder comprising a β-lactamase inhibitor, preferably clavulanic acid, the amoxicillin, cineole and oil preparation is mixed with a powder comprising approximately 550 mg to approximately 750 mg, preferably approximately 650 mg, of β-lactamase inhibitor.

[0281] Optionally, the process of the invention may include, after the step of mixing the amoxicillin cineole and oil preparation with the powder comprising clavulanic acid, a further step of adding a sweetener, a flavoring, and / or a lubricant as described above. Preferably, a sweetener, a flavoring, and a lubricant are added. Preferably, the sweetener is aspartame and the lubricant is magnesium stearate.

[0282] During this step, these additional excipients may be added in proportions of about 30 mg to about 50 mg, preferably about 36 mg, of sweetener, preferably aspartame, about 15 mg to about 25 mg, preferably about 18 mg, of lubricant, preferably magnesium stearate, and / or about 70 mg to about 110 mg, preferably 90 mg, of flavoring per 3 grams of pharmaceutical formulation.

[0283] After the step of adding a sweetener, a flavoring and / or a lubricant, the resulting preparation is mixed until a homogeneous powder is obtained.

[0284] In a preferred embodiment, the final sieving step is carried out with a sieve grid having openings of a diameter of at most 5 mm, preferably at most 2.5 mm, even more preferably at most 1.25 mm.

[0285] Optionally, the process of the invention may include an additional step of conditioning the sieved powder obtained by the process of the invention in a single-dose or multi-dose container, preferably in a single-dose container.

[0286] In a preferred embodiment, the sieved powder obtained by the process of the invention is packaged in a single-dose container containing between approximately 1 g and approximately 150 g of powder, more preferably between approximately 1 g and approximately 50 g of powder, even more preferably between approximately 1 g and approximately 10 g of powder, and more particularly preferably approximately 3 g of powder.

[0287] In another embodiment, the sieved powder obtained by the process of the invention is packaged in a multi-dose container comprising, for example, between approximately 10 grams and approximately 500 grams of powder, preferably between approximately 20 grams and approximately 200 grams of powder, even more preferably between approximately 30 grams and approximately 100 grams of powder, and most particularly preferably approximately 50 grams of powder.

[0288] Optionally, the process of the invention may include an additional step of secondary conditioning of the primary conditioning of the sieved powder obtained by the process of the invention.

[0289] In a preferred embodiment, the unit-dose containers containing the sifted powder are subsequently packaged in a box. In particular, a box may contain, for example, between 3 and 31 unit-dose containers, preferably between 5 and 21 unit-dose containers, and even more preferably between 7 and 14 unit-dose containers.

[0290] In another embodiment, the multi-dose container holding the sifted powder is further packaged in a box, optionally accompanied by a dosing device, for example a spoon, for dispensing a predetermined quantity of powder, preferably from approximately 1 mg to approximately 30 mg, even more preferably from approximately 2 mg to approximately 20 mg, and most preferably from approximately 3 mg to approximately 12 mg. In particular, the dosing device may allow for dispensing approximately 3 g, approximately 6 g, approximately 9 g, approximately 12 g, approximately 15 g, and / or approximately 18 g of powder. Molecular complex

[0291] Amoxicillin is an antibiotic susceptible to β-lactamases. β-lactamases, produced by bacteria resistant to amoxicillin, recognize and inactivate the β-lactam ring of amoxicillin. When dissolved, preferably in an aqueous solvent, amoxicillin can transiently form complexes of two amoxicillin molecules. The formation of these complexes is too transient to have any protective effect against β-lactamases. In contrast, when the pharmaceutical formulation of the invention is dissolved, stable amoxicillin complexes comprising at least three amoxicillin molecules are formed and protect the antibiotic from the action of β-lactamases. The formation of these complexes can also be achieved by dissolving amoxicillin in the presence of cineole.

[0292] Thus, in a final unclaimed aspect, the application also relates to a molecular complex comprising more than two amoxicillin molecules arranged linearly or in a ring and interacting with each other by non-covalent bonds.

[0293] Preferably, the molecular complex disclosed here is formed solely from amoxicillin molecules.

[0294] The molecular complex disclosed herein consists of at least three molecules of amoxicillin, preferably three to six molecules of amoxicillin, more preferably three or four molecules of amoxicillin, and most preferably four molecules of amoxicillin.

[0295] The amoxicillin molecules of the molecular complex disclosed here can be arranged linearly or in a ring. Preferably, they are arranged in a ring so that each amoxicillin molecule interacts with two other amoxicillin molecules.

[0296] In one particular aspect, amoxicillin molecules can freely switch from a linear complex organization to a ring complex organization by breaking or forming non-covalent bonds.

[0297] The molecular complex disclosed herein can be obtained by dissolving amoxicillin in the presence of cineole in an aqueous solvent. Preferably, the molecular complex disclosed herein is obtained by dissolving amoxicillin in the presence of cineole in an aqueous solvent without the use of detergent.

[0298] In one particular aspect, the molecular complex disclosed here is obtained by dissolving the pharmaceutical formulation of the invention in an aqueous solvent.

[0299] Preferably, the amoxicillin molecules of the molecular complex disclosed here are not recognized by β-lactamases. Thus, the molecular complex disclosed here can be used in the treatment of bacteria considered resistant to amoxicillin.

[0300] In one particular aspect, the molecular complex disclosed herein can be obtained in aqueous medium when the mass ratio of amoxicillin to cineole is between about 0.01 and about 1000, preferably between about 0.1 and about 100, even more preferably between about 1 and about 10, and most particularly preferred when the mass ratio of amoxicillin to cineole is about 5.

[0301] This application also relates to the use of the molecular complex disclosed herein as a medicinal product. It also relates to the molecular complex disclosed herein for use in the treatment of an infectious disease in a subject. Furthermore, it relates to the molecular complex disclosed herein for the preparation of a medicinal product for the treatment of an infectious disease. It also relates to a treatment method comprising the administration of a therapeutically effective amount of the molecular complex disclosed herein to a subject in need, in particular a subject suffering from an infectious disease.

[0302] The embodiments described for the formulation or combination disclosed herein are also to be taken into account in this aspect.

[0303] Although they have different meanings, the terms "comprising", "having", "containing" and "consisting of" may be substituted for one another throughout the description of the invention.

[0304] Other features and advantages of the invention will become clearer from the following examples, which are given by way of illustration and not limitation. Examples Example 1 - Study in vitro of the antibacterial activity of the combination of amoxicillin and cineole Materials and methods Biological material, culture medium and antimicrobial agents

[0305] The six strains tested in this study are purified clinical isolates identified at the bacteriology laboratory of the Hassan II University Hospital (CHU, Fez, Morocco). Three of the six bacterial strains tested are strains of Escherichia coli BLSE (P956, P933 and P7847) and the three other bacterial strains tested are strains of Klebsiella pneumoniae BLSE (H1878, H2001 and H1893).

[0306] For each test performed, precultures of the bacterial strains (previously frozen at -20°C) were prepared for 24 hours at 37°C. From these precultures, bacterial inocula at 2 x 10⁷ < CFU (Colony Forming Unit) / ml were prepared by adjusting the optical density to 540 nm.

[0307] Mueller Hinton (MH) agar and liquid culture media were supplied by BIOKAR (France). The agar was used for strain growth, and the liquid, supplemented with 20% (v / v) glycerol, was used for strain preservation. Both media were prepared according to the supplier's instructions.

[0308] Amoxicillin (AMX) and cineole were supplied by Sigma Aldrich (France). A stock solution of AMX (400 µg / ml) was prepared by dissolving 40 mg of the antibiotic in 100 ml of sterile distilled water. A series of dilutions were performed from this stock solution. The cineole concentrations used were prepared by emulsifying pure cineole in 0.2% (v / v) agar according to the method described by Remmal A et al (J. Essent. Oil. Res [Book], 1993, 5: pp. 1179-1184). Evaluation of the percentages of inhibition of the association of AMX with cineole

[0309] Partial inhibitory concentrations (PICs) are the concentrations of antibacterial agents that inhibit the growth of a given percentage (90%, 75%, 50%, 40%...) of the bacterial population studied. The PICs of AMX and cineole against the six bacterial strains used were determined by a microdilution technique in microplates based on monitoring bacterial growth by measuring optical density (Casey JT et al, J Microbiol Meth [Book], 2004, 58: pp. 327-334; Patton T et al, J Microbiol Meth [Book], 2006, 64: pp. 84-95).

[0310] Sterile U-shaped microplates with 96 wells and a capacity of 200 µl were used. For each microplate, two control rows were prepared: ➢ One row containing 200 µl of MH liquid medium (Sterility control and negative control). ➢ One row containing 150 µl of MH liquid medium and 50 µl of bacterial inoculum at 2x10⁷ < CFU / ml (Positive control).

[0311] For AMX and cineole, two rows were prepared, each containing 100 µl of liquid MH medium, 50 µl of bacterial inoculum at 2 x 10⁷ < CFU / ml, and 50 µl of decreasing concentrations of antimicrobial agents. The AMX concentrations used were: 50-25-12.5-6.25-3.1-1.6-0.8-0.4-0.20-0.10-0.05-0.025 µg / ml. The cineole concentrations used were: 100-50-25-12.5-6.25-3.1-1.6-0.8-0.4-0.20-0.10-0.05 µl / ml. Optical densities (OD) were determined at 540 nm using a microplate spectrophotometer (Versamax, Molecular Devices, USA). OD was measured at time t=0 and after 22 hours of incubation at 30°C. The percentage of inhibition of the antimicrobial agents against each of the six strains was calculated from Casey JT et al (J Microbiol Meth [Book], 2004, 58: pp. 327-334) using the following formula: 1 − % d ' inhibition = DO T 22 − DO T 0 DO C 22 − DO C 0 × 100

[0312] With DO T0 = DO of the test cup at time t=0, DO T22 = DO of the test cup after 22h of incubation, DO C0 = DO of the positive control cup at time t=0, DO C22 = DO of the positive control cup after 22h of incubation.

[0313] After determining the CIPs of AMX and cineole alone, the antibacterial activity of the AMX-cineole combination was evaluated. The same microdilution technique in microplates was used. The AMX-cineole combinations were prepared in sterile tubes as follows: ➢ AMX CIP 50% + CIP cineol 40% ➢ AMX CIP 50% + CIP cineol 30% ➢ AMX CIP 50% + CIP cineol 20% ➢ AMX CIP 25% + CIP cineol 40% ➢ AMX CIP 25% + CIP cineol 30% ➢ AMX CIP 25% + cineole CIP 20%

[0314] The fractional inhibitory concentration index (FIC-index) expressing the degree of synergy of the AMX-cineol combination was calculated according to the following formula (Odds FC et al, J Antimicrob Chemoth [Book], 2003, 52: p. 1). FIC − index = CIP AMX + Cinéol CIP AMX seule + CIP AMX + Cinéol CIP Cinéol seul The AMX-cineol association is considered: ➢ Synergistic when the FIC-index is ≤ 0.5 ➢ Additive when 0.5 < FIC-index < 1 ➢ Indifferent when 1 < FIC-index < 2 ➢ Antagonistic when FIC-index > 2. Results Determination of the CIPs of amoxicillin and cineole

[0315] The antibacterial activity of AMX against the three strains of E . coli and the three strains of K. pneumoniae is comparable (see Table 1). The minimum inhibitory concentrations (MIC, minimum concentration required to achieve 100% inhibition) are on the order of 50 µg / ml (p>0.05). Cineol also has comparable antibacterial activity against the three strains of E . coli and the three strains of K. pneumoniae (see table 2). The MICs are in the order of 100 µl / ml (p>0.05). Table 1: Partial inhibitory concentrations of Amoxicillin Strains CIP 50% (µg / ml) CIP 25% (µg / ml) Escherichia coli P956 1,1±0,25 0,7±0,14 P933 1,3±0,24 0,5±0,10 P7847 1,5±0,13 0,8±0,17 Klebsiella pneumoniae H1878 1,6±0,17 0,9±0,09 H2001 1,3±0,12 0,6±0,14 H1893 1,7±0,09 0,8±0,12 Table 2: Partial inhibitory concentrations of cineole Strains CIP 40% (µl / ml) CIP 30% (µl / ml) CIP 20% (µl / ml) Escherichia coli P956 6,2±0,20 2,6±0,12 1,2±0,16 P933 6,8±0,14 2,3±0,09 1,3±0,24 P7847 6,5±0,10 2,7±0,12 1,2±0,11 Klebsiella pneumoniae H1878 7,1±0,12 2,9±0,12 0,4±0,12 H2001 6,9±0,12 3,1±0,10 0,7±0,09 H1893 6,8±0,09 2,7±0,09 0,6±0,12 Evaluation of the inhibition percentages of the amoxicillin and cineole combination

[0316] The combination of AMX and cineole yields higher inhibition percentages than the sum of the two alone (see Tables 3 and 4). Indeed, the 50% concentration of AMX (1.1-1.7 µg / ml) combined with the 40% concentration of cineole (6.2-7.1 µg / ml) resulted in 100% growth inhibition for all tested strains. Table 3: Percentages of inhibition of the associations of AMX (CIP 50%) with cineole (CIP 40%, CIP 30% and CIP 20%) Strains AMX + Cinéol CIP 50% + CIP 40% AMX + Cinéol CIP 50% + CIP 30% AMX + Cinéol CIP 50% + CIP 20% Escherichia coli P956 100,0±0,0 92,5±0,56 80,4±0,69 P933 100,0±0,0 95,2±0,75 81,5±0,59 P7847 100,0±0,0 90,8±1,29 79,4±0,47 Klebsiella pneumoniae H1878 100,0±0,0 90,2±0,68 82,8±0,94 H2001 100,0±00 93,7±0,80 81,2±0,46 H1893 100,0±0,0 90,5±0,25 85,2±0,41 Table 4: Percentages of inhibition of AMX associations (CI 25%) with cineole (CI 40%, CI 30% and CI 20%) Strains AMX + Cinéol CIP 25% + CIP 40% AMX + Cinéol CIP 25% + CIP 30% AMX + Cinéol CIP 25% + CIP 20% Escherichia coli P956 72,0±0,38 62,1±0,22 55,3±0,75 P933 73,1±0,45 59,4±0,32 51,2±0,48 P7847 70,1±0,46 63,3±0,36 53,4±0,60 Klebsiella pneumoniae H1878 74,4±0,28 61,2±0,26 56,3±0,38 H2001 72,4±0,25 60,2±0,77 53,9±0,67 H1893 71,1±0,29 64,4±0,34 52,6±0,35

[0317] This powerful synergy was confirmed by calculating the fractional index (FIC-index, see Table 5) for the MICs of amoxicillin and cineole. For the six bacterial strains tested, these indices ranged from 0.08 to 0.10. These results, below 0.5, demonstrate a strong synergistic effect between amoxicillin and cineole. Table 5: FIC-index of the amoxicillin and cineole combination Strains CMI 100% FIC-Index Result Alone Combinations E. coli P956 50-100 1,1-6,2 0,08 Synergy E. coli P933 50-100 1,3-6,8 0,09 Synergy E. coli P7847 50-100 1,5-6,5 0,10 Synergy K. pneumoniae H1878 50-100 1,6-7,1 0,10 Synergy K. pneumoniae H2001 50-100 1,3-6,9 0,10 Synergy K. pneumoniae H1893 50-100 1,7-6,8 0,10 Synergy Discussion

[0318] The results clearly show that the tested strains, highly resistant to AMX, become susceptible to minimal concentrations of AMX when AMX is combined with cineole. Thus, 100% growth inhibition of all tested bacterial strains was achieved with the AMX-cineole combination (IC50% + IC40%). This combination provides antibacterial activity similar to that of AMX alone or cineole alone, at concentrations significantly lower than the MIC of either AMX or cineole. Indeed, comparing the concentrations used for 100% inhibition, the concentration of AMX used in combination with cineole is 25 times lower than that of AMX alone, and the concentration of cineole used in combination with AMX is 15 times lower than that of cineole alone.

[0319] The bactericidal activity of this combination of AMX and cineole results from a strong synergy of action, as demonstrated by the calculation of the fraction index (index less than or equal to 0.1).

[0320] All of these trials demonstrate the effectiveness of the combination of AMX and cineole against ESBL-resistant bacteria and its value in the fight to contain bacterial resistance. Example 2 - Study in vitro of the effect of cineole on the inhibition of amoxicillin by β-lactamases

[0321] This study is based on an enzymatic assay that first involves contacting AMX combined with cineole with β-lactamases. The antibacterial activity of this antibiotic was then verified on a strain of Escherichia coli sensitive to AMX. Materials and Methods Biological material, culture medium and antimicrobial agents

[0322] A bacterial strain of Escherichia coli (ATCC 8739) sensitive to amoxicillin was provided by the National Institute of Hygiene (INH-Rabat).

[0323] For each test performed, precultures of this strain of E . coli Precultures of 24h at 37°C were prepared from this bacterial strain (previously frozen at -20°C). From these precultures, bacterial inocula of 3.3x10⁶ CFU / ml were prepared by adjusting the optical density to 540 nm.

[0324] Mueller Hinton agar and liquid culture media were supplied by BIOKAR (France). The composition of this medium has been described previously. Both media were prepared according to the supplier's instructions.

[0325] Antibiotic-loaded discs (diameter Ø = 6 mm) were used for the enzyme test in agar medium (AMX: Amoxicillin and AMC: Amoxicillin + clavulanic acid). They were provided by the National Institute of Hygiene (INH-Rabat). The AMX used for the enzyme test in liquid medium was provided by Sigma Aldrich (France).

[0326] Cineol was supplied by Sigma Aldrich (France). Its preparation was carried out by emulsification in 0.2% (v / v) agar agar according to the method described by Remmal et al (J. Essent. Oil. Res [Book], 1993, 5: pp. 1179-1184).

[0327] The β-lactamase powder was supplied by Sigma Aldrich (France). It was solubilized, according to the supplier's instructions, to 10 mg / ml in 0.1 M Tris HCl at pH 7, containing 0.1% gelatin. The β-lactamase (0.03 U / ml) thus prepared was stored between 2 and 8°C. Experimental protocol in agar medium

[0328] In agar medium, the sub-inhibitory concentration of cineole (the highest concentration at which cineole does not induce inhibition) was determined by the microdilution method described previously. It is on the order of 0.002 µl / ml.

[0329] Starting from a pre-culture of E . coliFor a 24-hour sensitivity test, eighteen Petri dishes of MH agar were inoculated by surface spreading. Six control dishes and two test dishes were prepared as follows: Dishes #1 and #2 correspond to the controls for strain sensitivity to antibiotics: Control 1: Three discs loaded with AMX alone at 20 µg / ml were aseptically placed on the surface of plate #1. Control 2: Three discs loaded with AMC alone at 30 µg / ml were aseptically placed on the surface of plate #2.

[0330] Boxes #3 and #4 correspond to controls of β-lactamase activity on antibiotics: Control 3: Three discs loaded with AMX at 20 µg / ml were aseptically placed on the surface of plate #3. 10 µl of β-lactamase at 0.03 U / ml were added to the surface of each of these three discs. Control 4: Three discs loaded with AMC at 30 µg / ml were aseptically placed on the surface of plate #4. 10 µl of β-lactamase at 0.03 U / ml were added to the surface of each of these three discs.

[0331] Boxes #5 and #6 correspond to controls of the antibacterial activity of antibiotics combined with cineole (sub-inhibitory concentration): Control 5: Three discs loaded with AMX at 20 µg / ml were aseptically placed on the surface of plate #5. 10 µl of cineole at 0.002 µl / ml were added to the surface of each of these three discs. Control 6: Three discs loaded with AMC at 30 µg / ml were aseptically placed on the surface of plate #6. 10 µl of cineole at 0.002 µl / ml were added to the surface of each of these three discs.

[0332] Boxes #7 and #8 correspond to tests of the effect of cineole on the inhibition of amoxicillin by β-lactamases: Test 1: Three discs loaded with AMX at 20 µg / ml were aseptically placed on the surface of plate #7. To the surface of each of these three discs, 10 µl of β-lactamase at 0.03 U / ml and 10 µl of cineole at 0.002 µl / ml were added. Test 2: Three discs loaded with AMC at 30 µg / ml were aseptically placed on the surface of plate #8. To the surface of each of these three discs, 10 µl of β-lactamase at 0.03 U / ml and 10 µl of cineole at 0.002 µl / ml were added.

[0333] After incubating the 8 Petri dishes for 18 hours at 37°C, the diameters of the inhibition halos were measured. Experimental protocol in liquid medium

[0334] The MIC of AMX is 100% against the strain of Escherichia coli sensitive, determined by the microdilution method described above, is on the order of 6 µg / ml.

[0335] From a 24-hour preculture of the strain of E. colisensitive, bacterial inocula at 3.3x10 6< CFU / ml were prepared by adjusting the optical density to 540 nm.

[0336] Five control tubes and one test tube were prepared as described in Table 6 (see below).

[0337] Tube 1 is the sterility control for the medium. Tube 2 is the positive control for bacterial strain viability. Tube 3 is the control for strain sensitivity to AMX. Tube 4 is the control for non-inhibition of bacterial growth by a sub-inhibitory concentration of cineole. Tube 5 is the control for AMX degradation in the presence of β-lactamase. Tube 6 is the test for the effect of cineole on the inhibition of amoxicillin by β-lactamases. Table 6: Contents of control and test tubes for the liquid medium study Tubes Liquid Medium MH Bacterial inoculum (3.3 10 6< UFC / ml) AMX (6 µg / ml) Cineol (0,002 µl / ml) β-lactamase (0.03 U / ml) Tube #1: 1000 µl - - - - Tube #2: 970 µl 30 µl - - - Tube #3: 940 µl 30 µl 30 µl - - Tube #4: 940 µl 30 µl - 30 µl - Tube #5: 930 µl 30 µl 30 µl - 10 µl Tube #6: 900 µl 30 µl 30 µl 30 µl 10 µl

[0338] The OD was measured at time t=0, and after 22 hours of incubation at 30°C. The percentage of inhibition was then calculated according to the formula described previously: 1 − % d ' inhibition = DO T 22 − DO T 0 DO C 22 − DO C 0 × 100

[0339] With DO T0 = DO of the test tube at time t=0, DO T22 = DO of the test tube after 22h of incubation, DO C0 = DO of the positive control tube at time t=0, DO C22 = DO of the positive control tube after 22h of incubation. Results

[0340] In an agar medium (see table 7 below) Table 7: Diameters of Escherichia inhibition halos. Susceptible coli (in mm). Treatment Halo diameters (mm) * Treatment Halo diameters (mm) * Witness 1: AMX alone 15 ± 1.0 Witness 2: AMC alone 16 ± 0.4 Control 3: AMX + β-lactamase 6 ± 0.0 Control 4: AMC + β-lactamase 13 ± 0.5 Witness 5: AMX + Cineol 17 ± 0.3 Witness 6: AMC + Cineol 18 ± 0.9 Test 1: AMX+ β-lactamase+ Cinéol 12 ± 1.0 Test 2: AMC + β-lactamase + Cineol 15 ± 0.9 * Average of three halo inhibition values

[0341] Measurements of the diameters of the inhibition halos show that: The strain of E . coliThe assay used was highly sensitive to AMX (20 µg) and to the combination of AMX and clavulanic acid (AMC 30 µg), with halo diameters of 15 and 16 mm, respectively. The addition of cineole at a subinhibitory concentration (0.002 µl / ml) slightly increased the size of the inhibition halos, to 17 mm for AMX (20 µg) and to 18 mm for AMC (30 µg). The addition of β-lactamases (0.03 U / ml) suppressed the inhibition halo of AMX (20 µg) (6 mm being the disc diameter) and reduced that of AMC (30 µg) to approximately 13 mm. The addition of β-lactamases (0.03 U / ml) and cineole (0.002 µl / ml) considerably reduces β-lactamase-induced inhibition with halo diameters of approximately 12 mm for AMX (20 µg) and 15 mm for AMC (30 µg).

[0342] In a liquid medium (see table 8 below) Table 8: Percentages of inhibition of growth of the susceptible Escherichia coli strain. Treatment % inhibition Witness 4: Cineol alone 0 % ± 0,0 Witness 3: AMX alone 100 % ± 0,0 Control 5: AMX + β-lactamase 0 % ± 0,0 Test 1: AMX + β-lactamase + Cinéol 83,4 % ± 1,1

[0343] Table 8 shows that: The cineole concentration used (0.002 µl / ml) was well below the inhibitory threshold; there was no growth inhibition (Control 4). In the presence of β-lactamases (0.03 U / ml), the percentage of growth inhibition of the susceptible strain by AMX (6 µg / ml) was zero (Control 5). In the presence of both β-lactamase (0.03 U / ml) and cineole (0.002 µl / ml), the percentage of growth inhibition of the susceptible strain by AMX (6 µg / ml) was approximately 83.4% (Test 1). Discussion

[0344] The study conducted in an agar medium showed that: In the presence of cineole, at a sub-inhibitory concentration, the antibacterial activity of amoxicillin (AMX) and clavulanic acid (AMC) was enhanced (inhibition zone diameters larger than those obtained with the antibiotics alone). Thus, the activity of AMX and AMC is increased in the presence of cineole (at sub-inhibitory concentrations). In the presence of β-lactamases, the inhibition induced by AMX (20 µg) was negligible, and that induced by AMC (30 µg) was significantly reduced. This is due to the hydrolysis of the amoxicillin β-lactam ring. β-lactamase is only partially inhibited by clavulanic acid. In the presence of β-lactamases and cineole, the antibacterial activity of AMX and AMC was increased (diameters of the inhibition halos obtained were comparable to those obtained with the antibiotics alone).Since cineole was used at a sub-inhibitory concentration, the increase in antibacterial activity could be explained by a reduced effectiveness of β-lactamases on amoxicillin in the presence of cineole.

[0345] The study conducted in a liquid environment showed that: In the presence of β-lactamase, AMX completely loses its antibacterial activity, demonstrating the complete degradation of this antibiotic by hydrolysis of the β-lactam ring. In the presence of β-lactamases and cineole, the percentage of AMX inhibition is approximately 83.4%. These results show that in the presence of cineole at a sub-inhibitory concentration, AMX acquires protection against β-lactamase, which is no longer able to maintain its activity. The antibacterial activity of AMX is thus preserved.

[0346] These results show that amoxicillin combined with cineole is only weakly susceptible to β-lactamases. While not directly related to this theory, this could be due to complexation of amoxicillin molecules in the presence of cineole, preventing β-lactamases from attacking the β-lactam ring of AMX. Example 3 - Study of the antibacterial activity of rabbit serum treated with a combination of amoxicillin, clavulanic acid and cineole Materials and Methods Biological material, culture medium and antimicrobial agents

[0347] A bacterial strain of Escherichia coli Multidrug-resistant ESBL was provided by the National Institute of Hygiene (INH-Rabat). Precultures were prepared from the bacterial strain (previously frozen at -20°C) at 24 hours at 37°C. From these precultures, bacterial inocula of 2 x 10⁷ CFU / ml were prepared by adjusting the optical density to 540 nm.

[0348] Mueller-Hinton (MH) agar and liquid media were supplied by BIOKAR (France). The composition of these media has been described previously. Both media were prepared according to the supplier's instructions.

[0349] The six rabbits used in this study were female New Zealand White rabbits, supplied by a specialist breeder. They were 70 to 75 days old and weighed approximately 2 kg. They were randomly divided into two groups of three rabbits and fed ad libitum with an industrial fattening-type feed. Experimental Protocol

[0350] Rabbits in the first group received a single dose of AMC containing 1.5 g of amoxicillin and 186.5 mg of clavulanic acid. Rabbits in the second group received a single dose of AMC combined with cineole containing 1.5 g of amoxicillin, 186.5 mg of clavulanic acid, and 300 mg of cineole.

[0351] The solutions of the two treatments, reconstituted with purified water, were administered to the rabbits by gastric gavage.

[0352] After administration of the treatment, the rabbits were immobilized using a suitable restraint device. Their ears were exposed to an infrared lamp to dilate the marginal and central auricular veins. Blood samples of 0.5 ml were then taken from the central auricular vein at time T=0 (T0) (before administration), at time T1 (after one hour), at time T2 (after two hours), at time T3 (after three hours) and at time T6 (after six hours).

[0353] The microdilution method, described previously, was used to determine the percentage of inhibition of the serum samples.

[0354] Sterile, U-shaped microplates with 96 wells and a capacity of 200 µl were used. The negative control consisted of 200 µl of MH liquid medium, and the positive control of 150 µl of MH liquid medium and 50 µl of bacterial inoculum at 2 x 10⁷ CFU / ml. For each serum sample collected at a given time, two wells were prepared, each containing 100 µl of MH liquid medium, 50 µl of bacterial inoculum at 2 x 10⁷ CFU / ml, and 50 µl of serum.

[0355] For each sample, an OD reading was taken at t=0 and after incubating the microplates for 22 hours at 30°C. The percentage of inhibition for the different serum samples was calculated using the formula described previously: 1 − % d ' inhibition = DO T 22 − DO T 0 DO C 22 − DO C 0 × 100

[0356] With DO T0 = DO of the test cup at time T0, DO T22 = DO of the test cup after 22h of incubation, DO C0 = DO of the positive control cup at time t=0, DO C22 = DO of the positive control cup after 22h of incubation. Results

[0357] There figure 1 presents the monitoring of the percentages of inhibition of bacterial growth by the serum samples of rabbits from the group treated with AMC alone (amoxicillin / clavulanic acid combination) and from the group treated with AMC boosted by cineole over time.

[0358] Before treatment administration (time T0), the antibacterial activity of rabbit serum samples from both treatments is almost zero. One hour after treatment administration (Ti), the inhibition induced by the serum of rabbits treated with AMC alone is approximately 40% ± 1.2, while that induced by the serum of rabbits treated with AMC and cineole is approximately 47% ± 2.1. After two hours of treatment, the inhibition percentages are 50% ± 2.5 for the sera of rabbits treated with AMC alone and 54% ± 2.9 for those treated with AMC boosted by cineole. After three hours of treatment, the percentages of inhibition decrease to 41% ± 3.2 for rabbits treated with AMC alone and to 48% ± 1.6 for rabbits treated with AMC boosted by cineole.Finally, after six hours of treatment, the percentages of inhibition fall to 16% ± 1.3 for the sera of rabbits treated with AMC alone, while they are still around 44% ± 1.5 for rabbits treated with AMC boosted with cineole.

[0359] Thus, the percentages of inhibition obtained with the serum samples from rabbits in the batch treated with the combination of amoxicillin, clavulanic acid and cineole are significantly higher than those obtained with the serum samples from rabbits in the reference batch treated with the combination of amoxicillin and clavulanic acid (p<0.05). Example 4 - Pharmaceutical development of a formulation comprising amoxicillin, clavulanic acid and cineole Materials and methods

[0360] All raw materials, active ingredients and excipients used are of pharmaceutical grade.

[0361] This formulation is characterized by the presence of cineole, a volatile active ingredient. To stabilize this volatile compound, several excipients or combinations of excipients with adsorptive properties were tested (see Table 9 below).

[0362] The process tested for Form No. 8 (Peanut Oil) involves introducing a wetting step of the granule obtained with the mixture of the other active ingredients, amoxicillin and clavulanic acid, and excipients with an oily phase including cineole. The wetting step may be followed by a lubrication, mixing, and sieving step. Table 9: Adsorbent composition of the tested formulations Adsorbents Tested formulations Form 1 Form 2 Form 3 Form 4 Form 5 Form 6 Form 7 Form 8 Levillite √ √ √ Tixosil √ √ Talc √ Aerosil √ √ Dimethicone syloid √ silyloid √ Starcap √ Peanut oil √

[0363] All raw materials were sieved before mixing on different grids of the FREWITT sifter.

[0364] The raw materials were mixed in a HOBART type mixer.

[0365] The bag filling was carried out using the MARCHESINI bagging machine.

[0366] The temperature and relative humidity of the manufacturing room were respectively in the range of < 20 °C and < 15%.

[0367] The wetting solution, prepared for the cineole stabilization process, was prepared in a closed chamber to prevent any evaporation.

[0368] For each form tested, a complete quality control including the dosage of the three active ingredients (amoxicillin, clavulanic acid and cineole) was carried out: on the final mixture, at the end of manufacturing (ten sampling points) at time T0 and after 24h, 48h and 72h of exposure to a temperature < 20 °C and for an ambient humidity < 15%. on the sachet at the end of filling (ten sampling points)

[0369] Active ingredient concentrations (%) between 95% and 105% are considered compliant. The homogeneity of the mixture is verified by calculating the coefficients of variation (CV%) of the individual concentrations obtained for the ten sampling points. Coefficients of variation less than or equal to 2% are considered compliant. Results

[0370] The average amoxicillin and clavulanic acid levels are within acceptable limits for all tested formulations (forms 1 to 8). Regarding cineole (see Table 10 below), formulations 1 to 7 exhibit a cineole content of less than 95% in the final mixture, a content that decreases significantly after 24, 48, and 72 hours of exposure to a temperature < 20°C and ambient humidity < 15%.

[0371] Form 8 (peanut oil), on the other hand, stabilizes cineole. Indeed, the cineole content in this form is between 95% and 105% in the final mixture and remains within this range after 24, 48, and 72 hours of exposure to a temperature < 20°C and ambient humidity < 15%.

[0372] The homogeneity of the final mixture and the filling in the sachet is also compliant for formula 8, the values ​​of the coefficients of variation (CV%) of the contents not exceeding 2%. Table 10: Cineole content (%) according to the different adsorbents tested Feasibility studies Cineole content (%) / Coefficient of variation (%) Final mix Bag / end of filling Stability of the final mixture After 24 hours After 48 hours After 72 hours Form 1 81,9 % / 0,9 % 69,8 % / 1,5 % 75,2 % / 1,7 % 62,2 % / 0,9 % 57,8 % / 0,7 % Form 2 90,9 % / 1,7 % 75,6 % / 1,1 % 81,0 % / 1,5 % 70,4 % / 1,3 % 62,0 % / 1,5 % Form 3 89,0 % / 0,9 % 72,3 % / 1,6 % 74,2 % / 1,3 % 70,1 % / 1,1 % 59,3 % / 1,6 % Form 4 94,1 % / 0,6 % 92,1 % / 0,8 % 86,3 % / 1,3 % 80,7 % / 1,8 % 72,5 % / 1,3 % Form 5 75,9 % / 1,6 % 72,6 % / 1,2 % 60,8 % / 1,0 % 55,7 % / 1,6 % 50,8 % / 1,1 % Form 6 90,9 % / 1,3 % 86,2 % / 1,3 % 84,2 % / 1,5 % 77,2 % / 1,2 % 70,3 % / 1,7 % Form 7 93,9 % / 0,9 % 91,6 % / 0,7 % 88,1 % / 1,6 % 80,3 % / 1,4 % 74,2 % / 1,5 % Form 8 102,3 % / 0,8 % 102,1% / 1,2 % 102,1 % / 1,0 % 101,8 % / 1,0 % 101,7 % / 1,3 %

[0373] Furthermore, the tests quantifying impurities of the three active ingredients (amoxicillin, clavulanic acid, and cineole) in the final mixture, exposed to a temperature < 20°C and ambient humidity < 15% for 24, 48, and 72 hours, all met the acceptance criteria (results not shown). The results of the other quality tests of the final mixture and the sachet dispensing also all met the acceptance criteria (results not shown). Example 5 - Example of a pharmaceutical formulation according to the invention

[0374] Component name Quantity per 3g single-dose sachet Amoxicillin trihydrate corresponding to a quantity of 500 mg of amoxicillin 1:1 mixture of potassium clavulanate and hydrated colloidal silica corresponding to a quantity of clavulanic acid of 62.5 mg Cineol 100 mg Syloid ®< A1 FP 540 mg Aspartame 36 mg Peanut oil 50 mg Croscarmellose sodium 300 mg Avicel ®< PH112 microcrystalline cellulose 1241 mg* Magnesium stearate 18 mg Flavoring 90 mg *: The amount of microcrystalline cellulose is adjusted so that the total weight is 3 g.

[0375] This formulation was obtained by the following process: STEP 1: Sieving and pre-mixing

[0376] Avicel PH 112, Croscarmellose sodium and Syloid Al are mixed after sieving. STEP 2: Compaction 2-1 Amoxicillin

[0377] Amoxicillin trihydrate is mixed with a portion of the premixed powder obtained in step 1 and then compacted. The resulting granules are then calibrated and fractionated. 2-2 Clavulanic Acid

[0378] Clavulanic acid is mixed with a portion of the powder premix obtained in step 1 and then compacted. The resulting granules are then calibrated and fractionated. STEP 3: Granulation 3-1 Preparation of the Wetting Solution (S1)

[0379] The wetting solution is obtained by mixing cineole and peanut oil in a closed container and then fractionated. 3-2 Mooring

[0380] The fractions of the compacted mixture containing amoxicillin obtained in step 2 are moistened with the fractions of solution S1 and then mixed. The fractions of the compacted mixture containing clavulanic acid obtained in step 2 are added. 3.3.- Mixture

[0381] The different fractions are combined and mixed. STEP 4: Lubrication

[0382] Aspartame and the flavoring composition are mixed after sieving. Magnesium stearate is then added. STEP 5: Sieving and final mixing

[0383] The final powder is sifted and then mixed for a few minutes. STEP 6: Distribution

[0384] The final mixture is distributed into sachets. STEP 7: Secondary packaging in boxes Example 6 - Stability study of a powdered pharmaceutical formulation comprising amoxicillin, clavulanic acid, cineole, and peanut oil Materials and Methods

[0385] The stability of the pharmaceutical formulation of example 5 was tested under three different conditions (see table 11). Table 11: Experimental conditions for the stability study of the formulation Temperature Relative humidity (RH) Study duration Stability study under condition 1 25 °C ± 2 °C 60% ± 5% 24 months Stability study under condition 2 30 °C ± 2 °C 65% ± 5% 12 months Stability study under condition 3 40 °C ± 2 °C 75% ± 5% 6 months

[0386] Climate chambers with controlled temperature and relative humidity were used to maintain the formulations under the chosen conditions.

[0387] For condition 1, quality control was carried out every 3 months during the first year and every 6 months during the second year (0, 3, 6, 9, 12, 18 and 24 months).

[0388] For conditions 2 and 3, a quality control was carried out every 3 months (0, 3, 6, 9 and 12 months, for condition 2 and 0, 3 and 6 months for condition 3).

[0389] This quality control check covered: The organoleptic qualities, water content, and pH of the suspension were monitored. The three active ingredients (amoxicillin, clavulanic acid, and cineole) were determined. Impurities in the three active ingredients were quantified. A microbiological test was performed. Results

[0390] In all three conditions, throughout the study, the levels of the three active ingredients were measured between 95% and 105% (see tables 12 to 14 below), demonstrating the stability of the composition. Table 12: Results of the stability study under condition 1 T 0 T3 T 6 T9 T12 T18 T24 AMX content (%) 100,6 % 100,4 % 100,3 % 100,3 % 100,1 % 99,7 % 99,4 % Clavulanic acid content (%) 100,2 % 100,1 % 100,1 % 99,8 % 99,7 % 99,5 % 99,0 % Cineole content (%) 100,8 % 100,5 % 100,4 % 100,2 % 99,9 % 99,5 % 99,2 % Table 13: Results of the stability study under condition 2 T 0 T3 T 6 T9 T12 AMX content (%) 100,6 % 100,2 % 99,9 % 99,6 % 99,2 % Clavulanic acid content (%) 100,2 % 99,8 % 99,5 % 99,4 % 99,1 % Cineole content (%) 100,8 % 100,3 % 100,0 % 99,6 % 99,3 % Table 14: Results of the stability study under condition 3 T 0 T3 T 6 AMX content (%) 100,6 % 100,0 % 99,3 % Clavulanic acid content (%) 100,2 % 99,6 % 98,9 % Cineole content (%) 100,8 % 99,7 % 99,0 %

[0391] The quantification of impurities in the three active ingredients also complies with the acceptance criteria for all three conditions (results not shown).

[0392] The other parameters controlled (water content, appearance of the powder and reconstituted suspension, pH of the reconstituted suspension and microbiological control) all comply with the acceptance criteria (results not shown).

[0393] Thus, the formulation of the invention retains its chemical, physical, organoleptic and microbiological properties under the three tested conditions. Example 7 - Pharmacokinetic study in humans Experimental Protocol :

[0394] 48 healthy volunteer subjects were recruited for these experiments.

[0395] In the first experiment, volunteers, divided into two groups of 12, received orally either 12 g of the composition from Example 5 (comprising a total of 2 g amoxicillin, 250 mg clavulanic acid, and 400 mg cineole) or 12 g of an identical composition but without cineole. Blood samples were taken every half hour for the first 3 hours, then hourly until 6 hours, then at 8 hours, 10 hours, and 24 hours. The plasma concentration of amoxicillin in the samples was measured by chromatography, and the serum kinetics of amoxicillin were analyzed.

[0396] In a second experiment, the volunteers, divided into two groups of 12, initially received the same doses as in the previous experiment before receiving maintenance doses of 3 g of the same compositions three times a day for one week. This meant either three 3 g doses per day of the composition from Example 5 (each dose containing 500 g amoxicillin, 62.5 mg clavulanic acid, and 100 mg cineole), or three 3 g doses per day of an identical composition but without cineole. Blood samples were taken every 24 hours (at minimum concentration) and two hours later, at the time of peak concentration (at t = 26 h, 50 h, 72 h, etc.) for seven days. The plasma concentration of amoxicillin in the samples taken was measured by chromatography, and the serum kinetics of amoxicillin were analyzed. Amoxicillin kinetics during 24 hours after administration of a dose unique :

[0397] The curves for monitoring mean serum concentrations of amoxicillin obtained with volunteers treated with a combination of amoxicillin and clavulanic acid or with a combination of amoxicillin, clavulanic acid and cineole overlap almost perfectly, despite strong inter-individual variability (cf. figure 2 They exhibit the same pattern during the absorption phase, and maximum concentrations (Cmax) are reached at approximately two and a half hours for both formulations. Concentrations begin to decline from two and a half hours onward for both formulations, and the two elimination curves remain almost parallel between the time of administration and 24 hours later.

[0398] For all the pharmacokinetic parameters studied, namely the area under the curve (AUC 0-t), the maximum concentration (Cmax), the time to peak (Tmax) and the half-life (t 1 / 2), the values ​​obtained are not significantly different between the two conditions tested (see table 15 below).

[0399] The relative bioavailability of amoxicillin in the cineole-containing form compared to the non-cineole form is: F (AUC) = 0.888.

[0400] All of these results lead to the conclusion that the two compositions studied have the same bioavailability. Table 15: Pharmacokinetic parameters of serum amoxicillin Settings AUC 0-t (ng.h / ml) Cmax (ng / ml) Tmax (hour) t 1 / 2 (hour) Composition of AMX and clavulanic acid 56161 ± 26760 11991 ± 1371 2,73 ± 0,68 1,66 ± 0,45 Composition of AMX, clavulanic acid and cineole 50417 ± 12186 11322 ± 3171 2,38 ± 0,40 1,67 ± 0,44 Amoxicillin kinetics during repeated dose administration (7 days) :

[0401] The curves for monitoring mean serum concentrations of amoxicillin obtained with volunteers treated with a combination of amoxicillin and clavulanic acid or with a combination of amoxicillin, clavulanic acid and cineole overlap almost perfectly (cf. figure 3 ).

[0402] These results show that the two compositions studied have the same pharmacokinetic behavior when administered repeatedly. Example 8 - Clinical trial on patients with susceptible bacterial infection

[0403] The objective of this clinical trial was to evaluate, in patients with urinary tract infections caused by bacteria susceptible to amoxicillin, the efficacy of a pharmaceutical formulation containing amoxicillin, clavulanic acid, and cineole compared to a formulation containing only amoxicillin and clavulanic acid. This randomized clinical trial was conducted on a population of 28 patients divided into two groups (or arms) of 14 patients each. Patients in each group were treated in parallel for 7 days. The patients were men and women over 20 years of age.

[0404] An antibiogram performed on the patients before the start of treatment confirmed that they all suffered from a urinary tract infection with susceptible bacterial germs. Criteria for evaluating effectiveness:

[0405] The effectiveness of the treatments is assessed by a cytobacteriological examination of the urine (ECBU) carried out at the end of treatment. Products and dosage:

[0406] Example 5 test formulation (amoxicillin clavulanic acid and cineole) comprising 500 mg of amoxicillin, 62.5 mg of clavulanic acid and 100 mg of cineole per 3 grams of powder (corresponding to the contents of one sachet).

[0407] Each patient eligible for treatment in the study was randomly assigned according to the randomization table: A loading dose of 12 grams (4 sachets in a single dose) of the test formulation on the first day, then 3 sachets per day of the same formulation (1 in the morning, 1 at midday, and 1 in the evening) for 6 days (arm A). A loading dose of 12 grams (4 sachets in a single dose) of the test formulation on the first day, then 6 sachets per day of the same formulation (2 in the morning, 2 at midday, and 2 in the evening) for 6 days (arm B).

[0408] Table 16 below summarizes the distribution of patients in the different arms as well as the nature of the bacterial infection detected during the preliminary urine culture before treatment. Results :

[0409] The treatment was effective in all cases (elimination of the initially detected bacteria) except for one patient on whom the treatment was ineffective in arm A (Patient No. 2).

[0410] The treatment has even proven effective in some more complex patients who are known to be difficult to sterilize in case of infection, namely patients with benign prostatic hyperplasia, diabetic patients, patients with urethral strictures, urinary diversions, bladder stones or bladder tumors.

[0411] In conclusion, the tested treatment (amoxicillin, clavulanic acid and cineole), just like the reference treatment (amoxicillin and clavulanic acid), makes it possible to effectively treat urinary tract infections with susceptible bacterial germs, including in difficult cases. Table 16: Patient characteristics and their distribution Patient No. Age (years) Sex Bacteria responsible for the patient's urinary tract infection Arm 1 65 Man E. coli B3 2 44 Man E. coli A5 3 80 Man E. coli A1 4 75 Women Staphylococcus hemolyticus B12 5 20 Man Staphylococcus epidermidis A14 6 66 Women E. coli A8 7 43 Women E. coli A7 8 46 Man Proteus mirabilis A4 9 66 Man Klebsiella pneumoniae B2 10 69 Man Staphylococcus agalactiae A6 11 46 Man E. coli A9 12 58 Man Staphylococcus agalactiae A2 13 52 Man Staphylococcus agalactiae B9 14 78 Women E. coli B6 15 77 Women E. coli A3 16 69 Man Streptococcus agalpactie B4 17 76 Man E. coli B1 18 54 Women E. coli B5 19 55 Women Klebsiella oxytoca B7 20 52 Women E. coli A10 21 74 Man E. coli B8 22 70 Man Enterococcus B10 23 40 Women Streptococcus agalactiae A11 24 85 Man Klebsiella pneumoniae A12 25 26 Man E. coli B11 26 33 Women E. coli A13 27 60 Man E. coli B14 28 90 Man Group D Streptococcus B13 Example 9 - Clinical trial on patients with resistant bacterial infection

[0412] The objective of this clinical trial was to evaluate, in patients with urinary tract infections caused by bacteria resistant to a combination of amoxicillin and clavulanic acid, the efficacy of a pharmaceutical formulation containing amoxicillin, clavulanic acid, and cineole. This clinical trial was conducted on a population of 23 patients treated for 7 days.

[0413] The formulation tested (see example 2) is a powder comprising 500 mg of amoxicillin, 62.5 mg of clavulanic acid and 100 mg of cineole per 3 grams of powder (corresponding to the contents of one sachet).

[0414] Each patient initially received a loading dose of 12 grams (4 sachets in one dose) of the test formulation on the first day, then 6 sachets per day of the same formulation (2 in the morning, 2 at midday and 2 in the evening) for 6 days.

[0415] Table 17 below summarizes the characteristics of the patients in this trial, including the nature of the bacterial infection detected during the preliminary urine culture before treatment. Results :

[0416] The treatment was effective in all cases (elimination of the initially detected bacteria) except for one patient (Patient No. 6).

[0417] The case of patient No. 22 is particularly interesting. Indeed, this patient presented with a urinary tract infection considered refractory to all available antibiotics tested for almost 20 years, including amoxicillin and clavulanic acid, and was cured by the formulation according to the invention.

[0418] In conclusion, the formulation according to the invention comprising amoxicillin, clavulanic acid and cineole has proven to be very effective against urinary tract infections caused by bacterial germs resistant to the reference treatment, namely a formulation comprising amoxicillin and clavulanic acid. Table 17: Patient Characteristics Patient Number Age Sex Bacteria responsible for the patient's urinary tract infection 1 37 Women Staphylococcus aureus 2 49 Man Pseudomonas aeruginosa 3 46 Man E. coli 4 22 Women E. coli + Candida albicans 5 82 Man E. coli 6 37 Man E. coli 7 46 Man Acinetobacter iwoffii 8 83 Women E. coli 9 71 Man E. coli 10 76 Man Klebsiella pneumoniae 11 76 Man E. coli 12 53 Man Klebsiella pneumoniae 13 66 Women E. coli 14 39 Women E. coli 15 84 Man E. coli 16 69 Man Klebsiella pneumoniae 17 77 Man Enterobacter cloacae 18 59 Man Klebsiella pneumoniae 19 21 Man E. coli 20 25 Women E. coli 21 71 Man E. coli 22 65 Women Klebsiella oxytoca 23 49 Women Proteus mirabilis Example 10 - Spectroscopic study of amoxicillin complex formation

[0419] Spectroscopic analyses were performed with solutions comprising 500 mg of amoxicillin and, where appropriate, 62.5 mg of clavulanic acid or 100 mg of cineole in solution in 100 ml of water.

[0420] Mass spectroscopy analysis of amoxicillin in aqueous solution (cf. figure 4-A The graph shows the presence of a main peak corresponding to molecular amoxicillin (peak at 349.06 amu) and another peak corresponding to amoxicillin in dimer form (peak at 731.17 amu). The peak amplitude of the amoxicillin dimer is significantly lower than that of the molecular amoxicillin peak. Thus, when alone in solution, amoxicillin is predominantly in its molecular form.

[0421] When amoxicillin is dissolved in the presence of cineole, in addition to the peaks already observed for amoxicillin alone, new peaks appear in mass spectrometry (cf. figure 4-B ). These peaks correspond to amoxicillin trimers (peak at 1134.24 amu) and amoxicillin tetramers (peak at 1499.34 amu).

[0422] However, when amoxicillin is dissolved in the presence of clavulanic acid, and in the absence of cineole, the spectroscopic profile of amoxicillin is unchanged (results not shown).

[0423] Thus, the addition of cineole allows the rearrangement of amoxicillin molecules in solution into oligomers of 3 to 4 amoxicillin molecules. This rearrangement is not observed when amoxicillin is only in the presence of clavulanic acid.

Claims

1. A pharmaceutical formulation in powder form comprising, or consisting substantially of, cineole, amoxicillin, a β-lactamase inhibitor and a pharmaceutically acceptable oil, said pharmaceutically acceptable oil being peanut oil.

2. The formulation according to claim 1, wherein said β-lactamase inhibitor is clavulanic acid.

3. The formulation according to any one of the preceding claims, wherein said formulation comprises between about 5 mg and about 100 mg of cineole per gram of powder.

4. The formulation according to any one of the preceding claims, wherein said formulation comprises between about 20 mg and about 500 mg of amoxicillin per gram of powder.

5. The formulation according to any one of the preceding claims, wherein said formulation comprises between about 2 mg and about 50 mg of oil per gram of powder.

6. The formulation according to any one of claims 2 to 5, wherein said formulation comprises between about 1 mg and about 100 mg of clavulanic acid per gram of powder.

7. The formulation according to any one of the preceding claims, wherein the amoxicillin / cineole mass ratio is comprised between 2 and 8, preferably between 3 and 7, more particularly preferably between 4 and 6.

8. The formulation according to any one of the preceding claims, wherein the amoxicillin / oil mass ratio is comprised between 5 and 15, preferably between 7 and 13, and more particularly preferably between 8 and 12.

9. The formulation according to any one of the preceding claims, wherein the cineole / oil mass ratio is comprised between 0.1 and 5, preferably between 0.5 and 4, more particularly preferably between 1 and 3.

10. The formulation according to any one of claims 1 to 9, wherein the amoxicillin / β-lactamase inhibitor mass ratio is comprised between 5 and 11, preferably between 6 and 10, more particularly preferably between 7 and 9.

11. The formulation according to any one of the preceding claims, wherein said formulation is intended for oral administration, preferably after suspension in an aqueous solvent.

12. The formulation according to any one of the preceding claims, wherein said formulation is packaged in a single-dose container, preferably a single-dose container containing between about 1 g and about 150 g of powder, preferably between about 1 g and about 10 g of powder.

13. The formulation according to any one of the preceding claims, wherein said formulation further comprises at least one pharmaceutically acceptable excipient or carrier, preferably selected from the group consisting of a sweetener, a flavoring agent, an anti-caking agent, a lubricant, a disintegrant, and a mixture thereof.

14. The formulation according to any one of the preceding claims for use in the treatment of an infectious disease in a subject, preferably an infectious disease of bacterial origin, more preferably an infectious disease caused by a bacterium resistant to β-lactam antibiotics.

15. The formulation for use according to claim 14, wherein the infectious pathology is selected from the group consisting of cystitis, in particular recurrent acute cystitis, bacterial sinusitis, in particular acute maxillary sinusitis, otitis, in particular acute otitis media, bronchitis, in particular chronic and / or acute bronchitis, bronchopneumonia, in particular chronic and / or acute bronchopneumonia, pyelonephritis, upper gynecological infections, periodontitis, severe stomatological infections, in particular abscesses, phlegmons and cellulitis, animal bites, bone and joint infections, in particular osteomyelitis, preferably said bacterial infection is cystitis, in particular caused by a bacterium resistant to β-lactam antibiotics.

16. The formulation for use according to claim 14 or 15, wherein the formulation is administered to the subject for a period of 1 day to 4 weeks, preferably for a period of about 7 days, at a dose of 3 to 30 grams per day, in one or more daily doses.

17. A method for manufacturing the pharmaceutical formulation according to any one of claims 1 to 13 comprising: - obtaining a wetting solution by mixing cineole and a pharmaceutically acceptable oil, said pharmaceutically acceptable oil being peanut oil; - wetting a powder comprising amoxicillin with the wetting solution in order to obtain a powder preparation comprising amoxicillin, cineole and oil; - mixing the powdered preparation comprising amoxicillin, cineole and oil with a powder comprising a β-lactamase inhibitor, preferably clavulanic acid; and / or - optionally, the powder comprising amoxicillin and / or the powder comprising a β-lactamase inhibitor, preferably clavulanic acid, further comprise a disintegrant and / or an anti-caking agent; and / or - optionally, adding a sweetener, flavoring and / or lubricant agent, and mixing the latter to obtain a homogeneous powder; and / or - optionally, sifting the resulting powder; and / or - optionally, packaging the sifted powder in single-dose containers.