TRIAZINE FORMULATIONS WITH A SECOND ACTIVE INGREDIENT AND SURFACTANT(S)

The present invention is relative to triazine formulations and the uses thereof. The invention more particularly relates to new aqueous suspensions of a triazine comprising one or more surfactant(s) having a particular HLB (Hydrophilic-Lipophilic Balance). The invention also relates to compositions comprising at least one triazine, preferably toltrazuril, and a further active ingredient (e.g., an antibiotic, anti-inflammatory agent, anthelmintic, endectocide, mineral or vitamin). The compositions preferably comprise one or more surfactant(s) having a HLB (Hydrophilic-Lipophilic Balance) comprised between 1 and 8. The compositions are adapted for parenteral and/or oral administration. The invention also relates to kits, methods of preparing such compositions, as well as the uses thereof, in non- human animals such a porcine, an ovine, a bovine, a canine, a feline or an avian for protecting against infectious diseases, such as protozoan disorders.

Background of the invention Triazines are commonly used in the veterinary industry to treat non-human animals against a variety of diseases. Triazines are broad- spectrum antimicrobials that inhibit both gram positive and gram-negative bacteria, as well as some protozoa, such as coccidia.

Triazines, such as toltrazuril and ponazuril, are authorized agents for the treatment and control of coccidiosis which are frequent parasitic infectious diseases caused by protozoans such as e.g., Eimeria or Isospora. Triazines are also effective against other protozoal parasites e.g., toxoplasma, Cryptosporidia, or sarcocystis. They are essentially administered to the animals by oral route. In this regard, EP116175 refers to a water miscible solution of a triazine for oral administration. DE19603954 proposes triazine granules for oral administration and DEI 9824483 relates to semi-solid preparations which are applied orally. EP2164496 relates to triazine-iron combination products. The triazine compound is administered orally, as a suspension.

WO01/26660 and US6,465,460 relate to a sodium salt of triazine compounds and to compositions for oral or parenteral administration. According to this patent the sodium salt allows the use of lower doses of the compound. Although different administration routes are mentioned, all of the experimental section is limited to intravenous and oral administration. Furthermore, the patent indicates that several administrations and/or a sustained release dose are required for maintaining appropriate blood levels. In particular, the patent proposes a starting dose and several maintenance doses, which need to be administered over several days.

Despite preliminary investigations on different routes (e.g., transdermal), oral administration is still preferred today because it is believed to provide the most appropriate pharmacokinetic profile of the drug, especially in young non- human animals. In particular, while the oral formulation requires substantive manipulation and cannot provide a strict dosage control, it is believed to ensure appropriate bioavailability and therapeutic efficacy of the triazine compounds.

Oral administration may, however, present drawbacks. Indeed, oral administration cannot be conveniently combined with other treatments which are given by injection (e.g., antibiotics, anti-inflammatory agents, anthelmintics, endectocides, minerals or vitamins) at early stages of growth of the non-human animal and therefore involves additional manipulation of the non-human animals. Oral administration may also prevents strict control of the dosage administered to each non-human animal.

It has been proposed in the art to combine a triazine with iron, in order to treat both coccidiosis and anemia in piglets. However, such a combination raises formulation and administration problems. Indeed, the triazine is usually administered orally, while the iron compound is typically injected. EP2164196 proposed oral compositions of both agents but, at the same time, recognizes that iron compounds administered orally exhibit low bioavailability and should be administered within 8 to 10 hours after birth, leading to a separate schedule of administration for both compounds. Moreover, it has been disclosed that depending on the dosage or formulation, iron compound administered by injection may cause side effects.

Accordingly, there is a need in the art for optimized triazine formulations/administration protocols, especially for optimized triazine/iron formulations that are efficient and convenient. Such optimized formulations should preferably contain a triazine (e.g., toltrazuril) and an iron complex in the same formulation, suitable for combined administration, with dosages, amounts, and pharmacokinetics adapted to provide effective protection of non-human animals, even shortly after birth.

The present invention solves the above technical problems by providing novel triazine formulations in the form of stable aqueous suspensions with antifoaming properties, which fulfill the conditions for a pharmaceutical use and which are adapted for oral and parenteral administration.

Summary of the invention

An object of the invention resides in a composition comprising a triazine compound, wherein said composition is an aqueous suspension and comprises one or more surfactant(s) having an HLB (Hydrophilic-Lipophilic Balance) comprised between 1 and 8.

Another object of the invention resides in a composition comprising at least one triazine compound and another active ingredient (preferably selected from e.g., antibiotics, anti- inflammatory agents, anthelmintics, endectocides, minerals or vitamins), wherein said composition is an aqueous suspension containing at least one surfactant.

Another object of the invention resides in a composition comprising at least one triazine compound and another active ingredient (preferably selected from e.g., antibiotics, antiinflammatory agents, anthelmintics, endectocides, minerals or vitamins), wherein said composition is an aqueous suspension containing one or more surfactant(s) having an HLB comprised between 1 and 8.

The compositions of the invention may optionally further comprise at least one hydrophilic solvent and/or one or several additional surfactant(s), and/or one or several acceptable excipients or carriers. The compositions are preferably adapted for oral or parenteral administration, preferably for injection such as subcutaneous, intramuscular, intradermal or intravenous injection.

Preferred triazine compound is a triazinetrione and, more preferably, toltrazuril. The further active ingredient may be selected, preferably, from antibiotics, anthelmintics, endectocides, anti-inflammatory agents, vitamins and/or minerals. Preferably, the further active ingredient is an iron complex such as, without limitation, iron dextran, iron dextran glucoheptonic acid, iron isomaltoside and iron carboxymaltose complexes. Even more preferably, the iron complex includes an aqueous colloidal solution of beta-ferric oxyhydroxide and dextran glucoheptonic acid (Gleptoferron commercialized under the trademark Gleptosil® or Ursoferran®).

A preferred object of the invention resides in a composition in the form of an aqueous suspension comprising (i) a triazine compound, (ii) an iron complex, and (iii) one or more surfactant(s) having an HLB comprised between 1 and 8.

Preferred surfactants having an HLB comprised between 1 and 8 include, without limitation, soybean lecithins, sorbitan esters, polyol esters, silicon emulsions, simethicone emulsions, propylene glycol monolaurate, propylene glycol monocaprylate, glyceryl monooleate, phospholipids, lauroyl polyoxylglycerides, linoleoyl polyoxylglycerides, oleoyl polyoxylglycerides, or polyoxyethylene alkyl ether. Most preferred surfactants are sorbitan monooleate, propylene glycol monolaurate and simethicone emulsion.

Preferred hydrophilic solvents are semi-polar to polar hydrophilic solvents such as, without limitation, diethylene glycol monoethyl ether (= transcutol®), glycerol, polyethlene glycol and propylene glycol.

The compositions of the invention may be prepared or stored in various containers, preferably glass or plastic containers. The plastic containers are preferably made such as, without limitation, of polypropylene, polyethylene, high density polyethylene (HDPE), polyethylene terephtalate, polyvinyl chloride, polyethylene-vinyl acetate, polyolefms, polyesters, polyamide, or polymers. Typically, the compositions of the invention are stored in a bottle or flask. In a specific embodiment, the compositions of the invention are stored in a multi-layered plastic polymeric container comprising at least one polyolefm.

Another object of the invention is a kit comprising a container, preferably a bottle or flask, comprising a composition as defined above, and a notice. The container typically comprises all of the ingredients in admixture. Alternatively, in a particular embodiment, the container comprises two (or more) separated compartments, each comprising distinct ingredient(s) of the composition, which are typically mixed prior to administration.

Another object of the invention is a kit comprising two containers, a first container comprising the triazine compound, for instance as a powder, and a second container comprising the other ingredient in an aqueous suspension with one or more surfactant(s) having an HLB comprised between 1 and 8. The content of the two containers are typically mixed prior to administration.

Another object of the invention relates to a method for preparing a composition as defined above, comprising providing a triazine, a further active ingredient, and a surfactant having an HLB comprised between 1 and 8, and mixing said ingredients, generally in an appropriate vehicle or carrier. Typically, the method comprises (i) providing a triazine compound, preferably in the form of a powder, (ii) providing the further active ingredient in an aqueous suspension with one or more surfactant(s) having an HLB comprised between 1 and 8, and (iii) mixing said ingredients in an appropriate container. The compositions of the invention may be prepared in advanced and stored in any appropriate container (flask, bottle, etc.). Alternatively, the compositions may be prepared extemporaneously, e.g., by mixing the ingredients shortly before administration.

A further object of the invention is a composition or kit as defined above for use to treat (e.g., prevent, retard, protect against, reduce or cure) an infectious disease in a non- human animal, preferably a parasitic infectious disease caused by a protozoan.

A further object of the invention relates to a method for treating (e.g., preventing, retarding, protecting against, reducing, or curing) an infectious disease in a non-human animal, preferably a parasitic infectious disease caused by a protozoan comprising the administration of an effective amount of a composition as defined above to the non- human animal. Administration may be a single administration. Alternatively, repeated administration may be performed. As indicated above, the composition may be contained in a single container, or in two (or more) containers which are mixed extemporaneously, for instance shortly prior to administration.

The invention is particularly suited for protecting porcine, ovine, bovine, canine, feline, or avian against parasitic infectious diseases caused by protozoans. It is particularly well adapted for the treatment of young non-human mammals such as a bovine or a swine and more preferably a piglet newborn to 10 days old, preferably newborn to 3 days, more preferably newborn to 48 hours.

Legend to the Figures

Figure 1 : Structure of preferred triazine compounds.

Detailed description of the invention

The present invention provides novel formulations comprising at least one molecule of the triazine family and, preferably, another active ingredient which is in the form of an aqueous suspension, does not foam and is suitable for treating parasitic infectious diseases caused by protozoans. More specifically, the invention provides a composition of triazine(s) and another active ingredient such as iron complex and a surfactant agent characterized by an HLB (Hydrophilic-Lipophilic Balance) comprised between 1 and 8.

The invention indeed proposes to formulate triazines and another active ingredient (e.g., antibiotics, anti-inflammatory agents, anthelmintics, endectocides, minerals or vitamins) as aqueous suspensions, which provide efficient pharmacokinetic profile. Suspensions are dispersions of the active principle(s) solid particles (insoluble or practically insoluble) in a liquid medium. Usually, the size of solid particles varies from one or few nanometers to 50 micrometers. It is thus a heterogeneous mixture made up of an external continuous liquid phase and an internal solid phase. The use of pharmaceutical or veterinary compositions in the form of aqueous suspensions has numerous advantages, for example: the absorption of active ingredients comprised in a suspension form is generally faster compared to a solid form (tablet for example) and the chemical stability of certain active ingredients is improved with a suspension (compared to a solution). However, suspensions are difficult to prepare because factors which control the physical stability of the suspensions are numerous. Generally, the crystals of active ingredient do not have the same density as the continuous phase of the suspension from where the observation of creaming or sedimentation when the suspension is left at rest. Indeed, the active particles precipitate at the end of a certain time. It is thus necessary, before use, to stir the suspensions in order to disperse the particles of the active ingredient throughout the fluid phase and thus to obtain a homogeneous sample. One of the major problems of aqueous suspensions is the foam formation when reconstituting liquid dosage forms after agitation, or during manufacturing processes. Such a formation of foams (gas bubbles dispersed in the liquid) is highly undesirable. In this regard, aqueous suspensions containing triazines tend to form a lot of foam which, in addition to aspect problems, does not allow to guarantee an exact dosage and can possibly present risks of gas embolism if bubbles of air are injected.

The present invention shows that efficient suspensions of a triazine and another active ingredient can be prepared by adding to the composition particular surfactants, preferably surfactants with a low HLB (preferably between 1 and 8). By using appropriate surfactants according to the invention, it is possible to prepare aqueous suspensions of triazine in which foam (if formed) disappears very quickly.

The inventors have surprisingly found that the addition of a surfactant having a low HLB to an aqueous suspension of a triazine with another active ingredient such as iron complex leads to a significant foaming reduction or a total foaming suppression.

The present invention thus provides aqueous suspensions of one or several triazine compound(s) and one or several further active ingredient(s) in a liquid medium containing one or more surfactant(s) with a low HLB. The compositions may in addition further comprise one or more hydrophilic solvent(s) and/or one or several additional surfactants, and/or one or several excipients or carriers.

Triazine compounds

The term "triazine(s)" designates a well-known class of active substances, especially against infections with coccidian. Typical triazines for use in a method or composition of the invention are compounds of formula A or B below, of any purity, preferably having a purity of at least 90%, as well as any salt, ester, racemate, isomer, or prodrug, thereof:

(formula B) wherein

R ¹ is R ³ -S0 ₂ - or -S- ,

R ² is alkyl, alkoxy, halogen or S0 ₂ N(CH ₃ ) ₂ ,

R ³ is an haloalkyl

R ⁴ and R ⁵ are independently from each other a hydrogen or CI atom; and

R ⁶ is fluorine or chlorine,

Triazine compounds include more preferably triazinediones (formula A) and

triazinetriones (formula B). Examples of triazinediones include, without limitation, clazuril (R ⁴ is CI, R ⁵ is H, R ⁶ is CI in formula A), diclazuril (R ⁴ is CI, R ⁵ is CI, R ⁶ is CI in formula A) or letrazuril (R ⁴ is CI, R ⁵ is CI, R ⁶ is F in formula A). Preferred 1,2,4- triazinediones are diclazuril and sulazuril.

Triazines for use in the present invention are, more preferably, triazinetriones of formula B, even more preferably wherein R ² is a CI to C4 alkyl or alkoxy group (e.g., methyl, ethyl, or n-propyl), and/or R ³ is a CI to C3 perfluoroalkyl group (e.g., trifluoromethy 1) . Specific and most preferred triazines of the invention are toltrazuril (R ¹ = R ³ -S- , R ² = CH ₃ , R ³ = CF ₃ in formula B) and ponazuril (R ¹ = R ³ -S0 ₂ - , R ² = CH ₃ , R ³ = CF ₃ in formula B).

Toltrazuril ( 1 -methyl-3 - [3 -methyl-4- [4-(trifluoromethyl)tio)phenoxil]phenyl] -1,3,5- triazine(lH,3H,5H)-2,4,6-trione) is widely used in porcine, ovine, bovine and avian for the prevention and treatment of coccidiosis, by oral administration. It is currently available on the market as Cevazuril ^R or Baycox ^R . Methods for the preparation of toltrazuril are disclosed in various patents such as US 4,219,552, US 5,219,853, EP 0 201 030, and EP 0 879 057. The chemical structure of toltrazuril is represented in formula (C) below:

(Formula C)

The chemical structures of further examples of triazine compounds for use in the present invention are depicted in Figure 1.

The triazine(s) are present in the composition of the invention preferably in the amount from 1% to 10% by weight of the total weight composition. Preferentially, the compositions contain from 1% to 6% of one or more triazine compound(s), and even more preferentially from 2% to 5% of one or several triazine compound(s). The compositions preferably contain only one triazine. The most preferred triazine compound according to the invention is toltrazuril, the percentage of which in the composition of the invention is comprised between 1% and 6% by weight, and even more preferably between 2 and 5%. The triazine(s) as defined is the present invention comprise(s) their salts, such as for example sodium salts. Other active ingredient(s)

As indicated above, the compositions of the invention preferably combine a triazine and another active agent, which may be selected preferably from antibiotics, anti- inflammatory agents, anthelmintics, endectocides, minerals or vitamins.

Antibacterial antibiotics are commonly classified based on their mechanism of action, chemical structure, or spectrum of activity. Most target bacterial functions or growth processes. Those that target bacterial cell wall (penicillins and cephalosporins) or cell membrane (polymixins), or interfere with essential bacterial enzymes (rifamycins, lipiarmycins, quinolones, and sulfonamides) have bactericidal activities. Those that target protein synthesis (aminoglycosides, macrolides, and tetracyclines) are usually bacteriostatic. Further categorization is based on their target specificity. "Narrow- spectrum" antibacterial antibiotics target specific types of bacteria, such as Gram- negative or Gram-positive bacteria, whereas broad-spectrum antibiotics affect a wide range of bacteria. Following a 40-year hiatus in discovering new classes of antibacterial compounds, four new classes of antibacterial antibiotics have been brought into clinical use: cyclic lipopeptides (such as daptomycin), glycylcyclines (such as tigecycline), oxazolidinones (such as linezolid) and lipiarmycins (such as fidaxomicin). Preferred antibiotics are aminoglycosides, such as neomycin, beta-lactam antibiotics such as ampicillin and amoxicillin, macrolides such as spiramycin and tylosin, polymixins, such as colistin and family of chloramphenicol, such as chloramphenicol, thiamphenicol and florfenicol.

Anti- inflammatory agents refers to the property of a substance or treatment that reduces inflammation and for example but not limited to steroids (e.g. glucocorticoids, corticosteroids), non-steroidal anti-inflammatory drugs (NSAIDs) or immune selective anti- inflammatory derivatives (ImSAIDs).

Anthelmintics are drugs that are used to treat infections with parasitic worms. This includes both flat worms, e.g., flukes and tapeworms and round worms, i.e., nematodes. They are separated into classes on the basis of similar chemical structure and mode of action such as for example but not limited to piperazine, flubendazole, benzimidazole, oxybendazole, nicotinic receptor agonists (levamisole, pyrantel and morantel), oxindole alkaloid (paraherquamide A and marcfortine A), macrocylic lactones and milbemycins, cyclodepsipeptide molecule (emodepside) and pyruvate ferredoxin oxidoreductase inhibitor (nitazoxanid). Endectocides are used in the treatment and control of internal (endo-) and external (ecto-) parasites in certain animals. Endectocides comprise two groups of closely related compounds, the avermectins such as but not limited to doramectin, eprinomectin, and ivermectin and the milbemycin such as but not limited to moxidectin.

Vitamins are organic compounds required by an animal as a vital nutrient in limited amounts such as but not limited to Vitamin A, Vitamin Bl to 12, vitamin C, etc.

Minerals are trace elements, micronutrients or ions such as but not limited to calcium, copper, magnesium, selenium or iron. Generally minerals are elements known by a skilled person in the veterinary field according to the needs of each non- human animal treated. A preferred mineral for use in the invention is an iron complex. Iron complexes

Because iron salts may be toxic at high concentrations, the iron is preferably complexed to a macromolecule, such as carbohydrate or polymer. For example we can cite iron dextran, glucoheptonic acid of iron dextran, iron isomaltoside, iron carboxymaltose iron-sucrose, iron-oligosaccharide and iron-gluconate compounds. Examples of other preferred iron complexes include, without limitation, an aqueous colloidal solution of beta-ferric oxyhydroxide (or ferric hydroxide) and dextran glucoheptonic acid (Gleptoferron commercialized under the trademark Gleptosil® or Ursoferran®) or a ferric hydroxide with a low molecular weight dextran (commercialized under the trademark Uniferon® or Dexafer®) or a ferric hydroxide with macromolecular dextran (commercialized under the trademark Ferroforte®) or a ferric compound of type I.

The iron (as complex) represents preferably from 10% to 25% by weight of the total weight of the composition. Preferentially, the compositions of the invention contain from 10% to 20% of iron (as complex(es)), and even more preferentially from 12% to 20% of iron (as complex(es)). The most preferred iron complex for use in the invention is a colloidal solution of beta- ferric oxyhydroxide (or ferric hydroxide) and dextran glucoheptonic acid (Gleptoferron), the percentage of iron (as complex) in the compositions of the invention may be comprised between 12% and 20% by weight. Advantageously, the iron complex used may contain an effective amount of promoters of non-heme iron absorption, such as but not limited to vitamin C (ascorbic acid), derivatives of ascorbic acid, such as ascorbyl palmitate, an ester synthetic compound of palmitic acid with L ascorbic acid (vitamin C palmitate, or erythorbic acid).

Surfactants

The term "surfactant" designates an agent which modifies the liquid surface tension, in particular that of water.

Preferred surfactants of the invention are surfactants having a low HLB (Hydrophilic- Lipophilic Balance), more preferably a HLB ranging from 1 to 8. The invention shows that such particular class of surfactants allows the formation of suspensions of a triazine and another active ingredient which do not form foam upon stirring or in which foam (if formed) disappears very quickly.

A surfactant molecule can be described as a hydrophobic chain (affinity for nonpolar solvents) linked to a hydrophilic chain (affinity for polar solvents). Consequently, surfactants present antagonistic properties and, once formulated in water, the hydrophobic part rejects the molecule on the surface and the hydrophilic part tends to plunge within the liquid.

The HLB is used to determine the hydrophilic or hydrophobic dominant character of a surfactant. HLB values have been proposed in 1949 by Griffin (Griffin WC, Surface- Active Classification of Agents by HLB, Newspaper of the Society of Cosmetic Chemists 1 (1949): 31). This method allows the determination of reference points which quantify existing balance between the hydrophilic part and the lipophilic part of the molecule of surfactant, and which are related to its solubility in water. The scale varies from 0 to 40: the higher the value of HLB, the bigger is the solubility in water. In 1957, Davies suggested a method based on calculating a value based on the chemical groups of the molecule. The advantage of this method is that it takes into account the effect of stronger and weaker hydrophilic groups. According to the method of Davies:

H LB H LBgrom es hydrophiles H LBg _rml p _es hydrophobes ~T~

Preferred surfactants of the invention are hydrophobic with a low HLB, more preferably ranging from 1 to 8, typically of 1, 2, 3, 4, 5, 6, 7, or 8. The surfactants for use in the compositions of the invention are selected but not limited to soybean lecithins, sorbitan esters, polyol esters, silicone emulsion, simethicone emulsion, propylene glycol monolaurate, propylene glycol monocaprylate, glyceryl monooleate, phospholipids, lauroyl polyoxylglycerides, linoleoyl polyoxylglycerides, oleoyl polyoxylglycerides, or polyoxyethylene alkyl ether. Most preferred surfactants are sorbitan monooleate, propylene glycol monolaurate and simethicone emulsion.

In a particular embodiment, the preferred surfactant is a simethicone emulsion, which typically comprises about 25 to 35% of simethicone USP by weight. In a specific example, the simethicone emulsion contains the following constituents: polydimethylsiloxane, octamethylcyclotetrasiloxane, methylcellulose, decamethylcyclopentasiloxane, methylated silica and sorbic acid.

The surfactant(s) are preferably present in the compositions to a total amount of 0.01 to 5% by weight of the total weight of the composition. Preferentially, the compositions contain from 0.05 to 3% of surfactant with a low HLB, and even more preferentially from 0.05 to 1%.

The surfactants of the invention are efficient to prevent foaming by lowering surface tension and cohesive binding of the liquid phase. They are also compatible with triazines and another active ingredient, and can for stable aqueous suspensions which are particularly adapted. In this regard, another object of the invention resides in the use of a surfactant having a HLB between 1 and 8 to prevent foaming in a triazine and iron complex aqueous suspension.

Another object of the invention resides in a method for reducing foaming in an aqueous triazine suspension with another active ingredient, preferably iron complex, comprising adding to the suspension an effective amount of a surfactant having a HLB between 1 and 8.

The term "prevent foaming" and "reduce foaming" designates aqueous suspension which do not form foam upon stirring or in which foam (if formed) disappears very quickly.

Optional high HLB surfactants

In particular embodiments, in addition to the low HLB surfactant as defined above, the compositions of the invention also contain a further surfactant, particularly a high HLB surfactant. The addition of such a high HLB surfactant in the suspension can further improve triazine incorporation into the final formulation. A high HLB surfactant is a surfactant having a HLB strictly superior to 8.

Preferred high HLB surfactants are selected but not limited to polyethylene castor oil derivatives, polyoxyethylene alkyl ether, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearate, sodium lauryl sulfate, sodium docusate, cetrimide, phospholipids, or cethylpyridinium chloride.

Usually, high HLB surfactant(s) constitute(s) from 0% to 5% by weight, with respect to the total weight of the composition. Preferentially, when present, the high HLB surfactant(s) represent(s) from 0.05% to 5% of the composition, preferably from 0.05 to 1%. Solvents

The composition according to the invention may optionally comprise one or more additional hydrophilic solvents. The hydrophilic co-solvents are used e.g., in order to reduce the time of wetting and the time of dispersal of the aqueous suspension. Preferred solvents are hydrophilic semi polar to polar solvents. Preferentially, the solvents are selected but not limited to diethylene glycol monoethyl ether, glycerol, polyethlene glycol and propylene glycol.

Usually, the hydrophilic solvent(s) constitute(s) constitute from 0% to 30% by weight, with respect to the total weight of the composition. Preferentially, when present, the solvent(s) represent(s) from 1% to 30%> of the composition, preferably from 1 to 20%>, even more preferentially from 1% to 15%.

Salts

The composition according to the invention may optionally comprise one or several salts. Salts are used in order to complete surfactant efficiency. Preferred salts are selected but not limited to calcium gluconate, calcium phosphate, calcium chloride, calcium sulfate, calcium carbonate, magnesium gluconate, magnesium phosphate, magnesium chloride, magnesium sulfate, magnesium carbonate, potassium phosphate, potassium gluconate, potassium chloride, potassium sulfate, potassium carbonate, sodium gluconate, sodium chloride, sodium carbonate, sodium phosphate, sodium citrate, sodium sulfate, ammonium chloride, or ammonium carbonate.

Usually, salt(s) represent from 0%> to 10%> by weight, with respect to the total weight of the composition. Preferentially, when present, the salt(s) represent(s) from 0.1% to 10% of the composition, preferably from 0.5 to 4%, even more preferentially from 0.5% to 2%.

Preservatives

The composition according to the invention may optionally comprise one or several preservatives. Preferred preservatives are selected but not limited to ethanol, benzoic acid and the sodium or potassium salts thereof, chlorobutanol, benzyl alcohol, phenylethanol, the methyl, ethyl, propyl or butyl p-hydroxybenzoates, phenol, m-cresol, p-chloro-m-cresol or benzalkonium chloride.

Usually, preservatives represent from 0 % to 10 % by weight of the composition. Preferentially, when present, the preservative(s) represent(s) from 0.05 % to 5 %, preferably, from 0.1 % to 3 %.

Thickeners

The composition according to the invention may optionally comprise one or several thickeners. Preferred thickeners are selected but not limited to octyldecanol, aluminium monostearate, calcium phosphate, sodium phosphate, chitosan, dextran, glyceryl behenate, glyceryl monoleate, glyceryl monostearate, colloidal silicon dioxide, cellulose and its derivatives, starch, propylene carbonate, hydroxypropyl methylcellulose, polyvinylpyrrolidone, sorbitol, trehalose or sucrose.

Usually, thickeners represent from 0 % to 10 % by weight of the composition. Preferentially, when present, the thickener(s) represent(s) from 0.05 % to 5 %, preferably, from 0.05 % to 2 %.

Preferred compositions of the invention

The present invention provides optimized combined compositions comprising a triazine with another active ingredient, both suspended in an aqueous medium containing one or more specific surfactants. Furthermore, the invention provides optimal dosages and administration routes for all the compounds, in order to confer best protective effect during the treatment procedures.

Preferred compositions of the invention comprise one triazine and one iron complex, which may be combined with one or several, preferably one, surfactant having a low HLB. Most preferred compositions comprise a further hydrophilic solvent.

A particular object of the invention resides in an oral or injectable composition comprising: from 1 % to 10 % by weight of a triazine, preferably a triazinetrione, more preferably toltrazuril;

from 10 % to 25 % by weight of an iron (as a complex), preferably Gleptoferron;

- from 0.01 % to 5 % by weight of one or several surfactant(s), preferably sorbitan monooleate, propylene glycol monolaurate or a simethicone emulsion;

from 0 to 30 % by weight of one or several hydrophilic solvent(s),;

from 0 to 10% by weight of one or several salt(s); and

from 0 to 5% by weight of one or several high HLB surfactant(s).

In a particular embodiment, the composition further comprises:

from 0% to 10% by weight of one or several thickening agent(s), and

from 0%) to 10%) by weight of one or several preservative(s).

The above weight% are provided by reference to the total weight of the composition.

Another particular object of the invention resides in an oral or injectable composition comprising:

from 1 % to 6 % by weight of a triazine, preferably a triazinetrione, more preferably toltrazuril;

from 10 % to 20 % by weight of an iron (as complex, preferably Gleptoferron); from 0.05 % to 3 % by weight of one or several surfactant(s), preferably selected sorbitan monooleate, propylene glycol monolaurate or simethicone emulsion; from 0%o to 20 % by weight of one or several hydrophilic solvent(s), typically from 1% to 20% ;

from 0%) to 4%o by weight of one or several salt(s), typically from 0.5%> to 4%; - from 0% to 4% by weight of high HLB surfactant, typically from 0.05% to 4%, - from 0% to 5 % by weight of one or several thickening agent(s), typically from

0.05% to 5%, and

from 0%) to 5%> by weight of one or several preservative(s).

The above weight% are provided by reference to the total weight of the composition. In another further preferred embodiment, the composition according to the invention is an aqueous suspension comprising: from 2 % to 5 % by weight of toltrazuril,

from 12 % to 20 % by weight of an iron complex (as Gleptoferron),

from 0.05 % to 1 % by weight of one or several surfactant(s), preferably selected from sorbitan monooleate, propylene glycol monolaurate or simethicone emulsion,

optionally from 1 to 15 % by weight of one or several hydrophilic solvent(s), from 0% to 2% by weight of one or several salt(s),

from 0% to 2% by weight of high HLB surfactant,

from 0% to 2% by weight of one or several thickening agent(s), and

- from 0% to 3% by weight of one or several preservative(s).

The above weight% are provided by reference to the total weight of the composition.

A specific example of a composition of the invention comprises 2.9 % toltrazuril, 18.8 % of iron (as Gleptoferron), 0.3 % of sorbitan monooleate, 0.1 % of copolymer Polyoxyl 35 - hydrogenated castor oil, 1.5% of sodium chloride and water.

Another specific example of a composition of the invention comprises 4.2 % toltrazuril, 13.5 % of iron (as Gleptoferron), 0.3 % of sorbitan monooleate, 0.1 % of copolymer Polyoxyl 35 - hydrogenated castor oil, 1.5% of sodium chloride and water.

Another more specific example of a composition of the invention comprises 4.2 % toltrazuril, 13.5 % of iron (as Gleptoferron), 1% of sodium chloride, 1% of polyvinylpyrrolidone, 0.6% of phenol, 0.5% of colloidal silicon dioxide, 0.1 % of sodium docusate, 0.1% of simethicon emulsion and water.

Further specific examples of preferred compositions of the invention are formulations 2- 14 as disclosed in the examples. Such compositions contain the amount of triazine and other active ingredient which are required to obtain the minimal effective dose to ensure efficient protection.

The compositions of the invention generally comprise water, qsp 100%. They may also contain additional excipients or vehicles, as will be disclosed below. Preferred administration route and excipients

The compositions of the invention are suitable for oral or parenteral administration.

In this regard, the compositions may further comprise additional veterinary acceptable excipient(s) such as surfactant with low HLB, diluents, solvents, salts, solubilizers, adjuvants, stabilizers, surfactants such as high HLB, antioxidants, preservatives, thickeners.

Powders, dispersible powders, or granules suitable for preparation of an aqueous suspension by addition of water are convenient dosage forms for oral or parenteral administration. Formulation as a suspension provides the active ingredient in a mixture with a surfactant, and, optionally, with a dispersing or wetting agent, suspending agent, and, one or more preservatives.

The compositions of the invention may also be administered parenterally by injection (e.g., intramuscular, subcutaneous, intravenous, or the like), infusion or implantation in dosage forms, formulations, or via suitable delivery devices or implants containing conventional, non-toxic pharmaceutically acceptable carriers and adjuvants.

The preferred administration route of the compositions of the invention is by injection. The intramuscular administration route is most preferred.

The compositions of the invention may be administered by injection(s) using techniques and/or devices known per se in the art. In this regard, injection, such as intramuscular injection can be performed with a syringe, a gun, a micro-needle injection device, a needle-free injection device, a pulse device, etc. In a preferred embodiment, injection is performed with a needle injector or a syringe. In another particular embodiment, injection is performed with a needle-free injection device such as a pulse needle-free system, more particularly a spring-powered, battery-powered, or compressed-gas- powered device. Specific examples of needle free technologies are described e.g., in WO2006/058426, WO2007/140610, or WO2009/111794. A preferred needle-free injection device for use in the present invention is AcuShot™ needle free technology described in the international patents WO2006/058426 and WO2007/140610. Intramuscular injection may be made in any muscle. For livestock, such as cattle, intramuscular injection is preferably made in the area of the neck, or behind the ear, rather than in the hind limb/ham muscle or in the inguinal fold.

In this regard, a particular object of the invention resides in a use of the composition as described above, in the treatment of coccidiosis in animal, wherein said composition is administered by intramuscular injection in the neck or behind the ear for non-human mammals and in muscles adjacent to the breast-bone of adult avian.

Another particular object of the invention resides in the use of a composition as described above, in the treatment of coccidiosis in avians, particularly in young avians, wherein said composition is administered by subcutaneous injection.

The compositions of the invention are more preferably administered by a single injection to the non-human animal

The dose of triazine may vary depending on the animal species and nature of the triazine. Conventional dosage rates from 1 to 60 mg of triazine per kg bodyweight (mg/kg) of the non-human mammals may be used, preferably 5 to 50 mg/kg, and more preferably from 10 to 30 mg/kg. Illustrative dosages in the compositions of the invention are 10 mg, 20 mg, 30 mg, 40 mg, 50 mg or 60 mg toltrazuril/dose.

Within the context of the invention, the term "effective amount of designates, preferably, a dose of the active agent which produces a clinical benefit in the treated non- human animals. Particularly, an effective amount is an amount sufficient to reduce infection, disease development, or to improve the symptoms.

Preferred dosages for intramuscular toltrazuril are disclosed below, for some non- human animals 1 species example:

. pigs: 20 mg/kg bodyweight/treatment (preferably one single administration);

. cattle: 15 mg/kg bodyweight/treatment (preferably one single administration);

. sheep: 20 mg/kg bodyweight/treatment (preferably one single administration);

-poultry: 25 mg/kg bodyweight/treatment (preferably one single administration). Uses of compositions of the invention

The compositions of the invention may be used for the treatment of any non-human animals. The term treatment includes, particularly, the preventive treatment of non- human animal against a disease. The preventive treatment of a non-human animal against a disease designates a treatment made before the non-human animal has been exposed to or in contact with the causative agent of the disease (e.g., a pathogen, virus, protozoan, cell, etc.), or after said exposure/contact but before development of the symptoms or at an early stage of development of the disease. Also, the term preventive treatment, in relation to a population of non-human animals, designates the treatment of all members of the population even after the disease has been detected in certain members, to limit or avoid spreading of and contamination to the others. The term treatment or preventive treatment also includes the alleviation of the symptoms, as well as a delay, reduction or cure of an existing infection.

The term treatment also includes an increase in the welfare of the treated non-human animals, for example in increasing the production of meat, milk, wool, etc.

The invention may be used to treat (e.g., protect, prevent, delay, reduce or cure) an infectious disease in non-human animals, preferably a protozoan or microbial disease. The invention is particularly suited to treat protozoan diseases of the blood and tissues (such as babesiosis, sarcocystosis, or toxoplasmosis), or of the digestive tract (such as coccidiosis).

The invention may be used, in particular, to treat coccidiosis, particularly to prevent or protect non-human animals against coccidiosis.

In a particular embodiment, diseases which may be efficiently treated with compositions of the invention, are coccidiosis and protozoal diseases, which are very frequent parasitic infectious diseases caused by coccidian protozoa such as, e.g. Eimeria, Isospora, Neospora, Sarcosporidia, Cryptosporidium, Hammondia, besnoitia, hepatozoon and Toxoplasma. Coccidiosis is frequent. Thus, for example, protozoans from the genera cause coccidioses and protozoal diseases all over the world. The predominant sign of cocci diosis is diarrhea, which usually persists for 4 to 6 days. The faeces may vary from white to yellow in color and from a fluid to a pasty consistency, usually without the presence of blood. Coccidiosis predisposes the non- human animal to the incidence of secondary bacterial infections and severely affected non-human animal may die.

Although the morbidity is usually high, mortality is variable, probably due to the difference in the number of oocysts ingested, differences in the environment and the presence of other co-existing disease problems. Although the disease has only a slight influence on mortality, it does influence the presence of concomitant infections and the amount of antibiotics needed to control them. The disease also drastically affects the development of non-human animals, where the daily weight gain of the infected non- human animals decreases compared to uninfected non-human animals, causing herds to have poor uniformity at weaning age. There is notable emaciation and stunting, coccidiosis reduces growth by about 15% on average, that is, minimum 500g at weaning age, and this contributes to weaning herds that are highly heterogenous.

The compositions of the invention may be used, in particular, to treat coccidiosis and protozoal diseases caused by various protozoans such as, more particularly, Mastigophora (Flagellata), Sarcomastigophora (Rhizopoda), Myxospora, Microspora, or Pneumocystis carinii. Specific examples of Mastigophora include Trypanosomatidae such as, without limitation, Trypanosoma brucei, T.gambiense, T. rhodesiense, T. congolense, T. cruzi, T. evansi, or T. equinum. Specific examples of Sarcomastigophora (Rhizopoda) include Entamoebidae and Apicomplexa (Sporozoa) such as Eimeridae, for example E. acervulina, E. adenoides, E. alabahmensis, E. anatis, E. anseris, E. arloingi, E. ashata, E. auburnensis, E. bovis, E. brunetti, E. canis, E. chinchillae, E. clupearum, E.columbae, E. contorta, E. crandalis, E. debliecki, E. dispersa, E. ellipsoidales, E. falciformis, E. faurei, E. flavescens, E. gallopavonis, E. hagani, E. intestinalis, E. iroquoina, E. irresidua, E. labbeana, E. leucarti, E. magna, E. maxima, E. media, E. meleagridis, E. meleagrimitis, E. mitis, E. necatrix, E ninakohlyakimovae, E. ovis, E. parva, E. pavonis, E. perforans, E. phasani, E. piriformis, E. praecox, E. residua, E. scabra, E. spec, E. stiedai, E. suis, E. tenella, E. truncata, E. truttae, E. zuernii, Globidium spec, Isospora belli, L canis, L felis, L ohioensis, L rivolta, L spec, L suis, Neospora caninum, N. hugesi, Cystisospora spec, Toxoplasma gondii, Sarcocystis bovicanis, S. bovihominis, Leucozytozoon simondi, Plasmodium berghei, P. falciparum, P. malariae, P. ovate, P. vivax, P. spec, Babesia argentina, B. bovis, B. canis, B. spec, Theileria parva, Theileria spec, such as Adeleina, for example Hepatozoon canis, H. spec

The protozoan genera and species which in pigs lead to subclinical or clinical infections must be very especially emphasized, in particular: Eimeria debliecki, E. suis, E. scabra, E. perminuta, E. spinosa, E. polita, E. porci, E. neodebtiecki, Isospora suis, Cryptosporidium, Toxoplasma gondii, Sarcocystis miescheriana, S. suihominis, Babesia trautmanni, B. perroncitoi, Balantidium coli. Particular examples of economically important coccidioses are: infections of pigs with coccidia of the genus Isospora or of cattle with coccidia of the genus Eimeria.

The compositions according to the invention are effective against all stages of development of the pathogen. A specific object of the invention resides in a use of the compositions as described above, in treating any disease in a non-human animal, preferably a microbial disease.

Another particular object of the invention resides in a use of the compositions as described above to protect a non-human animal against coccidiosis, wherein said compositions are administered orally. A particular object of the invention resides in a use of the compositions as described above to protect a non-human animal against coccidiosis via parenteral administration, and preferably by intramuscular or subcutaneous injection.

Storage of the compositions of the invention

The compositions of the invention may be provided in unit dosage forms (e.g., in single- dose ampoules), in vials containing several doses or in various containers such as bottles (in which a suitable preservative may be added). The containers (e.g., bottles) may be made of glass or of plastic, e.g., polyethylene, and preferably high density polyethylene (HDPE). Such HDPE bottles may be, for instance, of nominal capacity of 100 ml, 250 ml, 500 ml, 750 ml or 1000 ml, and closed respectively with polypropylene screw caps with a tamper-evident seal. Other materials and containers can also be used for storing compositions of the invention. Such containers should comply with the strict regulations and requirements for storing pharmaceutical compositions.

For example, the compositions of the invention are preferably stored in multi-layered polymeric stable containers as described in a patent application n° FR2917381. Such containers may be sterilized by irradiation with or without the composition and allow a stable conservation of said composition for a long period of time in sterile conditions. The above containers are in general formed by the association of several material layers (sheets), thereby improving the properties of the packagings, which is then particularly less rigid, less breakable, and more resistant to heat, to light, to gas and chemical treatments. These multi-layered containers are made of materials such as polyamides, polyolefm (PO) (polypropylene (PP), polyethylene (PE)), polyethylene terephtalate (PET), polyvinyl chloride (PVC), poly(ethylene vinyl acetate) which are associated with gas barrier layer made of ethylene vinyl acetate copolymer (copolymer EVOH), ethyl vinyl acetate (EVA), and polyamides, in order to yield an increased barrier protection against oxygen and water vapor and limit any possible alterations of the pharmaceutical composition. In a specific embodiment, the compositions of the invention are stored in a multi-layered plastic polymeric container comprising at least one polyolefm as described in patent application n° FR2917381. Some compositions, such as injectable compositions, may require to be sterilized before administration. These compositions are thus in general manufactured and stored in sterile conditions. The container may be sterilized either empty or filled with a composition. Alternatively, the container and the composition may be sterilized separately, and the container may then be filled in sterile conditions according to well known methods in the art.

The container for storing compositions of the invention preferably maintain properties of resistance after sterilization, as well as stability, mechanical resistance, transparency, and impermeability towards environmental factors, chemical products or various treatments; the reference being the glass material. The material for the packaging or container should be pharmaceutically/veterinary acceptable and should therefore not alter the quality of the pharmaceutical compositions. Also, preferably, the compositions should not alter the nature and composition of the container or packaging in contact therewith. Such alterations may result in the migration of chemicals from and to the packaging or container and the composition. Such chemicals may be impurities, degradation products that appear over time under the action of oxygen, light and temperature, or due to processing of the container or packaging, such as sterilization process by irradiation. These interactions may alter over time the chemical properties of the composition, such as the stability of the active ingredient and/or of the container or packaging, transparency and/or color of the composition or container, thereby reducing the lifetime of the container or packaging. Furthermore, such interactions may modify the sterility, safety and efficacy of the composition.

The container may contain all of the ingredients of the compositions in admixture. In particular, the container (e.g., flask, bottle, etc.) can comprise, in suspension, the triazine compound, the further active agent(s,) the surfactant(s) and, when present, the solvent(s) and/or excipient(s).

Alternatively, the ingredients may be formulated in separate containers, or in separate compartments of a same container. In this regard, in a particular embodiment, the invention uses two containers, a first container comprising the triazine compound, for instance as a powder, and a second container comprising the other ingredient(s) in an aqueous suspension with one or more surfactant(s) having an HLB comprised between 1 and 8. The content of the two containers are typically mixed prior to administration.

In another particular embodiment, the invention uses one container comprising a first and a second compartments, said compartments being separated from each other, for instance, by a membrane or wall, a first compartment comprising the triazine compound, for instance as a powder, and a second compartment comprising the other ingredient(s) in an aqueous suspension with one or more surfactant(s) having an HLB comprised between 1 and 8. The content of the two compartments are typically mixed prior to administration, for instance by breaking or removing the separating membrane or wall. Another object of the invention relates to a method for preparing a composition as defined above, comprising providing a triazine, a further active ingredient, and a surfactant having an HLB comprised between 1 and 8, and mixing said ingredients, generally in an appropriate vehicle or carrier. Typically, the method comprises (i) providing a triazine compound, preferably in the form of a powder, (ii) providing the further active ingredient in an aqueous suspension with one or more surfactant(s) having an HLB comprised between 1 and 8, and (iii) mixing said ingredients in an appropriate container. The compositions of the invention may be prepared in advanced and stored in any appropriate container (flask, bottle, etc.). Alternatively, the compositions may be prepared extemporaneously, e.g., by mixing the ingredients shortly before administration.

A further object of the invention is a kit comprising one or several containers as defined above.

Preferred non-human animals treated by compositions of the invention

The compositions of the present invention may be used for the treatment of any non- human animals, including porcine, ovine, bovine, canine, feline or avian and preferably livestock, breeding animals, avian animals, companion animals, and laboratory animals.

Livestock and breeding animals include non-human mammals such as, for example, cattle, horses, sheep, pigs, goats, camels, water buffalos, donkeys, rabbits, fallow deer, reindeer, fur bearers such as, for example, mink, chinchilla, or raccoon.

Avian animals include, for example, chickens, hens, ducks, turkeys, guinea fowl, quail, geese, pigeons or parrots ostriches, and bird species for keeping in domestic premises and in zoos.

Companion animals include such as, for example, horses, dogs and cats.

Laboratory animals and experimental animals include such as, for example, mice, rats, guinea pigs, or golden hamsters.

The compositions of the invention are particularly suited for treating pigs, ovine, bovine, horses, sheeps, cattle, dogs, rabbits, or cats. The compositions of the invention may be used in adults or young non-human animals, such as newborn to 10 days old non- human animals

Further aspects and advantages of the invention will be disclosed in the following illustrative experimental section.

Examples

A - Preparation of compositions

The following compositions are prepared:

Formula 1

Ingredient g/lOOmL

Toltrazuril 3.5

Iron (as Gleptoferron) 18.3

Water Qs

Formula 2

Ingredient g/lOOmL

Sorbitan Monooleate 0.3

Toltrazuril 3.5

Iron (as Gleptoferron) 19.4

Water Qs

Formula 3

Ingredient g/lOOmL

Sorbitan Monooleate 0.3

diethylene glycol 10

monoethyl ether

Toltrazuril 3.5

Iron (as Gleptoferron) 17.8

Water Qs Formula 4

Ingredient g/lOOmL

Sorbitan Monooleate 0.3 diethylene glycol 10 monoethyl ether

Copolymer Polyoxyl 35 - 0.1 hydrogenated castor oil

Toltrazuril 3.5

Iron (as Gleptoferron) 17.8

Water Qs

Formula 5

Ingredient g/lOOmL

Sorbitan Monooleate 0.3

Copolymer Polyoxyl 35 - 0.1 hydrogenated castor oil

Toltrazuril 3.5

Iron (as Gleptoferron) 19.4

Water Qs

Formula 6:

Ingredient g/lOOmL

Sorbitan Monooleate 0.1

Sodium chloride 1.0

Diethylene glycol 10 monoethyl ether

Toltrazuril 3.5

Iron (as Gleptoferron) 17.8

Water Qs Formula 7:

Formula 8:

Formula 9 :

Ingredient g/100 niL

Sodium docusate 0.1

Silicone emulsion 0.1

Sodium chloride 1.5

Iron ( as gleptoferron) 13.5

Toltrazuril 4.2

Water qs Formula 10 :

Formula 11 : Formula 12:

Formula 13:

Ingredient g/100 niL

Sodium docusate 0.1

Simethicone emulsion 0.1

Sodium chloride 1.0

Povidone 1.0

Colloidal silicon dioxide 0.5

Phenol 0.6

Iron (as gleptoferron) 13.5

Toltrazuril 4.2

Water QS Formula 14:

B- Evaluation of the foaming properties of the compositions

The foaming aspect of the formulations is estimated according to the following procedure:

- Filling of graduated test-tube with 7 g of product.

- Vigorous agitation of the test-tube during 45 seconds.

- Measuring of the foam formed at the following time intervals:

• right after agitation,

• 1 minute after agitation,

• 3 minutes after agitation,

• 5 minutes after agitation,

· 10 minutes after agitation The results are presented in the following Table.

Formula 1, which does not contain a low HLB surfactant according to the invention, exhibits high foaming 10 minutes after agitation. In contrast, Formula 2 to 8 have a foam height below 3 mm after 10 minutes, and have therefore remarkable antifoaming properties.

Formula 3, 4, 7 and 11 to 14 are remarkable since they are characterized by a rapid decrease of the foam thickness, which almost disappeared 10 minutes post agitation. Formula 9 and 10 are particularly remarkable since they are characterized by a very fast decrease of the foam thickness, which almost disappeared 1 to 3 minutes post agitation

The formation of foams when reconstituting liquid dosage formulations is undesirable and disruptive. The remarkable capacity with the formulations of the invention to avoid and/or quickly stop formation of foam in an aqueous suspension after reconstitution is therefore particularly advantageous since it allows the user to quickly syringe the suspension in optimal condition, to secure the toltrazuril and iron dose taken, and to minimize the phenomenon of re-suspension observed after the agitation.