Isoxazoline compound for use in treatment of animals and a composition comprising an isoxazoline compound

Abstract

The invention relates to an isoxazoline compound of formula (I) or a salt or solvate thereof for use in the treatment and / or prophylaxis of animals, characterized in that it is used in the treatment and / or prophylaxis of bee diseases. The invention also relates to a composition comprising at least one isoxazoline compound of formula (I) or a salt or solvate thereof, characterized in that it is in a form and dose acceptable for beekeeping.

Description

[0001] Isoxazoline compound for use in treatment of animals and

[0002] a composition comprising an isoxazoline compound

[0003] The invention relates to an isoxazoline compound for use in treatment of animals and a composition comprising the isoxazoline compound. The invention will find application in particular in beekeeping and the protection and treatment of bees against parasitic infestations, particularly those of the Varroidae family.

[0004] The bee family (collectively known as Apidae) is a key element of the ecosystem for vegetation and human nutrition. The bee family comprises of honeybees, eastern honeybees, giant honeybees and dwarf honeybees, and stingless honeybees (meliponines). In the early 1960s, cases of infestations of external parasitic arthropods from the Varroidae family were identified in bees, spreading across Japan and the then-Soviet Union, leading to diseases called varroosis. Since then, cases of varroa parasite infestations have been reported on all continents, as well as in ecosystems as remote and difficult to access as Hawaii and New Zealand. The two main representatives of the parasitic Varroidae family are Varroa destructor and Varroa jacobsoni. Varroas are therefore a threat to vegetation and human nutrition worldwide. They also pose a particularly serious threat to commercial beekeeping, where beehive infestations with as few as 10 parasites - if left untreated - lead to the extinction of the entire affected beehive within a few months. The pupae of this arachnid feed on the bees' fat bodies. The fat body is responsible for key vital functions of the bee, so parasitizing it leads to significant weakening of the bee and eventually death. Finally, these parasitic arthropods are vectors for transmitting numerous diseases (e.g. viruses), further weakening the hive.

[0005] In addition to previously ineffective natural methods of preventing varroa (e.g., attempts to breed resistant bees), chemical agents are primarily used to treat varroa. In Poland and the United States, dedicated preparations are available comprising, among others, thymol, amitraz, formic acid, oxalic acid, lactic acid, organic acids, coumaphos, menthol, essential oils, propolis, camphor, eucalyptol, acrinathrin, bromfenvinphos, chlorfenvinphos, cymiazole, fluvalinate, flumethrin, and cypermethrin. However, the parasite can develop resistance to acaricides within a few years, while the use of organic acids islabor-intensive and dependent on the stage of development of the bee colony. Furthermore, in the case of chemical preparations that are not approved or have untested effects on humans, there is a risk to humans through potential contamination of honey and wax. Therefore, the use of varroacides during honey production is prohibited and the waiting periods required for individual preparations must be strictly observed.

[0006] Accordingly, the main object of the present invention was to provide an improved and / or alternative formulation for use in treatment of bees, particularly in cases of parasitic infestations, especially of the Varroidae family. Surprisingly, it has been found that this and other objects can be achieved by the isoxazoline compound and a composition comprising an isoxazoline compound according to the invention.

[0007] It is worth mentioning that isoxazoline compounds are known in the art, and compounds of this class are known for their activity against external parasitic arthropods such as lice, fleas, and mites. However, despite extensive research, to date, these compounds and compositions comprising them have been administered exclusively to dogs and cats, in form of oral tablets, topical spot-on solutions and collars, as well as in treatment and control of red mite infestations in domestic chickens, in form of drinkable preparations. Furthermore, and equally importantly, established scientific consensus directly discourages the use of isoxazoline compounds in bees as unsafe and toxic (e.g., Sheng, C.-W. et al., Neurotoxicity and mode of action of fluralaner on honeybee Apis mellifera L., Pest. Manag. Sci. , 2019, 75:2901-2909). And recent scientific reports go even further, proposing isoxazoline compounds as an effective means of eradicating and controlling wasps, which are related to bees (e.g. Rust MK et. al, The Potential of Fluralaner as a Bait Toxicant to Control Pest Yellowjackets in California. Insects. 2023 Mar 23;14(4):311).

[0008] The subject of the invention is the isoxazoline compound of formula (I)

[0009]

[0010] wherein

[0011] R1= halogen, CF3, OCF3, CN,n = an integer from 0 to 3, preferably 1 , 2 or 3,

[0012] R2= Ci -Cs-haloalkyl, preferably CF3 or CF2CI,

[0013] T = a 5- or 6-membered ring which is optionally substituted with one or more radicals Y,

[0014] Y = methyl, halomethyl, halogen, CN, NO2, NH2-C=S or two adjacent Y radicals forming together a chain, especially a three- or four-membered chain;

[0015] Q= X-NR3R4or 5-membered N-heteroaryl ring which is optionally substituted with one or more radicals;

[0016] X = CH2, CH(CH3), CH(CN), CO, CS,

[0017] R3= hydrogen, methyl, haloethyl, halopropyl, halobutyl, methoxy methyl, methoxyethyl, halomethoxymethyl, ethoxymethyl, haloethoxymethyl, propoxymethyl, ethylaminocarbonylmethyl, ethylaminocarbonylethyl, dimethoxyethyl, propynylaminocarbonylmethyl, N-phenyl-N-methyl-amino, haloethylaminocarbonylmethyl, haloethylaminocarbonylethyl, tetrahydrofuryl, methylaminocarbonylmethyl, (N,N-dimethylamino)-carbonylmethyl, propylaminocarbonylmethyl, cyclopropylaminocarbonylmethyl, propenylaminocarbonylmethyl, haloethylaminocarbonylcyclopropyl,

[0018]

[0019] whereinZA= hydrogen, halogen, cyano, halomethyl (CF3);

[0020] R4= hydrogen, ethyl, methoxymethyl, halomethoxymethyl, ethoxymethyl, haloethoxymethyl, propoxymethyl, methylcarbonyl, ethylcarbonyl, propylcarbonyl, cyclopropylcarbonyl, methoxycarbonyl, methoxymethylcarbonyl, aminocarbonyl, ethylaminocarbonylmethyl, ethylaminocarbonylethyl, dimethoxyethyl, propynylaminocarbonylmethyl, haloethylaminocarbonylmethyl, cyanomethylaminocarbonylmethyl or haloethylaminocarbonylethyl;

[0021] or R3and R4together form a substituent selected from the group consisting of:

[0022] NH> SHj

[0023]

[0024] sO— CB?and ' O 'CH.

[0025] or a salt or solvate thereof for use in the treatment and / or prophylaxis of animals characterized in that it is used in the treatment and / or prophylaxis of bee diseases.

[0026] Preferably, the isoxazoline compound for use according to the invention is characterized in that the isoxazoline compound is fluralaner.

[0027] Preferably, the isoxazoline compound for use according to the invention is characterized in that the bees are selected from honeybees, eastern honeybees, giant honeybees, dwarf honeybees and stingless honeybees (meliponines).

[0028] More preferably, the isoxazoline compound for use according to the invention is characterized in that the bees are honeybees.

[0029] Preferably, the isoxazoline compound for use according to the invention is characterized in that the compound is used in the treatment and / or prophylaxis of parasitic arthropod infestations.

[0030] More preferably, the isoxazoline compound for use according to the invention is characterized in that the parasitic arthropod infestation is an ectoparasitic arthropod infestation from the Varroidae family, especially Varroa destructor and / or Varroa jacobsoni.

[0031] Preferably, the isoxazoline compound for use according to the invention is characterized in that the compound is used to combat and / or control parasitic arthropods in the bees’ environment.

[0032] Preferably, the isoxazoline compound for use according to the invention is characterized in that it is used in the prophylaxis of diseases transmitted by parasitic arthropods of bees.Preferably, the isoxazoline compound for use according to the invention is characterized in that it is administered in combination with another beekeeping acceptable active substance, preferably an antiparasitic compound.

[0033] Preferably, the isoxazoline compound for use according to the invention is characterized in that the isoxazoline compound of formula (I) is used inside a beehive in an amount from 3.75 mg to 48 mg, preferably from 6 mg to 30 mg.

[0034] Preferably, the isoxazoline compound for use according to the invention is characterized in that the isoxazoline compound of formula (I) is used outside a beehive in an amount from 200 mg to 2000 mg, preferably from 400 mg to 1000 mg.

[0035] Preferably, the isoxazoline compound for use according to the invention is characterized in that the isoxazoline compound of formula (I) is used in feeding a beehive in an amount from 8 mg to 24 mg, preferably 16 mg.

[0036] The subject of the invention is also a composition comprising at least one isoxazoline compound of formula (I)

[0037]

[0038] wherein

[0039] R1= halogen, CF3, OCF3, CN,

[0040] n = an integer from 0 to 3, preferably 1 , 2 or 3,

[0041] R2= Ci -C3-haloalkyl, preferably CF3or CF2 Cl,

[0042] T = a 5- or 6-membered ring which is optionally substituted with one or more radicals Y,

[0043] Y = methyl, halomethyl, halogen, CN, NO2, NH2-C=S or two adjacent radicals Y forming together a chain, especially a three- or four-membered chain;

[0044] Q= X-NR3R4or 5-membered N-heteroaryl ring which is optionally substituted with one or more radicals;

[0045] X = CH2, CH(CH3), CH(CN), CO, CS,

[0046] R3= hydrogen, methyl, haloethyl, halopropyl, halobutyl, methoxy methyl, methoxyethyl, halomethoxymethyl, ethoxymethyl, haloethoxymethyl, propoxymethyl,ethylaminocarbonylmethyl, ethylaminocarbonylethyl, dimethoxyethyl, propynylaminocarbonylmethyl, N-phenyl-N-methyl-amino, haloethylaminocarbonylmethyl, haloethylaminocarbonylethyl, tetrahydrofuryl, methylaminocarbonylmethyl, (N,N-dimethylamino)-carbonylmethyl, propylaminocarbonylmethyl, cyclopropylaminocarbonylmethyl, propenylaminocarbonylmethyl, haloethylaminocarbonylcyclopropyl,

[0047]

[0048] wherein

[0049] ZA= hydrogen, halogen, cyano, halomethyl (CF3);

[0050] R4= hydrogen, ethyl, methoxymethyl, halomethoxymethyl, ethoxymethyl, haloethoxymethyl, propoxymethyl, methylcarbonyl, ethylcarbonyl, propylcarbonyl, cyclopropylcarbonyl, methoxycarbonyl, methoxymethylcarbonyl, aminocarbonyl, ethylaminocarbonylmethyl, ethylaminocarbonylethyl, dimethoxyethyl, propynylaminocarbonylmethyl, haloethylaminocarbonylmethyl, cyanomethylaminocarbonylmethyl or haloethylaminocarbonylethyl;

[0051] or R3and R4together form a substituent selected from the group consisting of:

[0052] NH,

[0053]

[0054] * 'O'"' ''•CH,

[0055] or a salt or solvate thereof characterized in that it is in a form and dose acceptable for beekeeping.Preferably, the composition according to the invention is characterized in that the isoxazoline compound of formula (I) is fluralaner.

[0056] Preferably, the composition according to the invention is characterized in that it is in solid form or liquid form.

[0057] More preferably, the composition according to the invention is characterized in that the solid form is selected from a strip impregnated or coated with an active substance, an insert impregnated or coated with an active substance, a fumigation tablet, a powder, a granulate, a food and a food additive.

[0058] Even more preferably, the composition according to the invention is characterized in that the solid form for administration inside a beehive comprises from 0.1 mg to 200 mg of the isoxazoline compound of formula (I) and the solid form for administration outside a beehive comprises from 0.1 mg to 3000 mg of the isoxazoline compound of formula (I).

[0059] More preferably, the composition according to the invention is characterized in that the liquid form is selected from a concentrate, a liquid, a food and a food additive.

[0060] Even more preferably, the composition according to the invention is characterized in that the liquid form comprises from 0.1 mg / ml to 500 mg / ml of the isoxazoline compound of formula (I).

[0061] Preferably, the composition according to the invention is, characterized in that it comprises at least one additional active substance selected from thymol, amitraz, formic acid, oxalic acid, lactic acid, organic acids, coumaphos, menthol, essential oils, propolis, camphor, eucalyptol, acrinathrin, bromfenvinphos, chlorfenvinphos, cymiazole, fluvalinate, flumethrin, cypermethrin, hop beta acids, oxytetracycline, tylosin, lincomycin, permethrin, paradichlorobenzene and bacillus thuringiensis.

[0062] The use of this invention comprises to use racemic mixtures, for example, equal amounts of the enantiomers of such isoxazoline compounds as described above. In addition, the use of this invention includes isoxazoline compounds that are enriched compared to the racemic mixture in an enantiomer of formula 1. Also included are the essentially pure enantiomers of such isoxazoline compounds. When enantiomerically enriched, one enantiomer is present in greater amounts than the other, and the extent of enrichment can be defined by an expression of enantiomeric excess ("ee"), which is defined as (2x-l)-100 %, where x is the mole fraction of the dominant enantiomer in the mixture (e.g., an ee of 20 % corresponds to a 60:40 ratio of enantiomers). Preferably the compositions for use in the current invention have at least a 50 % enantiomeric excess; more preferably at least a 75 %enantiomeric excess; still more preferably at least a 90 % enantiomeric excess; and the most preferably at least a 94 % enantiomeric excess of the more active isomer. Of particular note are enantiomerically pure embodiments of the more active isomer. Isoxazoline compounds as described above can comprise additional chiral centers. The use of this invention comprises racemic mixtures as well as enriched and essentially pure stereo configurations at these additional chiral centers. The reference to isoxazoline compound in this specification includes enantiomers, salts and solvates as well as N-oxides thereof that can be produced by conventional methods.

[0063] "Treatment" means the use and / or administration of the composition according to the invention to an animal suffering from a parasitic infestation in order to combat the parasite and / or reduce the number of parasites infesting such an animal. The effect of the composition according to the invention may, for example, be lethal to eggs, larvae, adults or combinations thereof. The effect may be direct, i.e. , killing the parasites immediately and / or over time, for example, upon the onset of moulting and / or by destroying their eggs, or indirect, for example, by reducing the number of eggs laid and / or the hatching rate.

[0064] "Prophylaxis" or "prevention" means that new parasite infestations in an animal are prevented by killing adult parasites and any developmental stage / larvae that are capable of invading the host, either before or immediately after infestations of the host to be protected, and / or by preventing the production of parasite offspring, for example by reducing the number of eggs laid and / or hatching rate. "Prophylaxis" or "prevention" also includes the eradication and control of parasites in the animal's environment by, among other things, interrupting the parasite life cycle and / or reducing their numbers on other animals in the population.

[0065] An "effective amount" is the amount and / or dose of the isoxazoline compound that is required to limit and / or reduce parasite numbers in an animal and / or in the animal's environment, e.g., in a beehive and / or to inhibit the development of parasitic infections in the animal, in whole or in part. This amount can be readily determined by observing and / or detecting the number of parasites either on the animal and / or in the animal's environment (e.g., by means of a trap) both before and after administration of the isoxazoline compound in the drinking water of such animals, e.g. parasite numbers are reduced by 5% to 100% after the first administration.

[0066] "Systemic administration" means an administration at a site remote from a site wherein at least a portion of the target parasites reside.

[0067] "Infestation" means a state wherein at least one member of a parasitic arthropod (an adult parasite and / or one member of a developmental / larval stage) is permanently and / ortemporarily present on an animal. Parasitic arthropods are generally ectoparasites that live on and / or in bees and — according to their biological life cycle — may parasitize on or in the bee or its brood cells.

[0068] The term "bees" refers to all representatives of the bee (Apidae) family, especially the honeybee and the eastern (Asian) bee, regardless of the stage of development of the bee -larva, young, adult - gender and function in the hive (population).

[0069] Bee diseases are also known in the medical field as apipathologies or apidiseases. Similar medical terminology is created by combining the prefix "api," meaning bees (Latin: apis), with another word, such as therapy, giving apitherapy.

[0070] A "beehive" is defined as a natural or human-made structure used by bees to develop a bee colony and gather food supplies. In particular, a beehive is defined as a beehive and / or wild beehive — regardless of the type of structure and material used — for breeding and rearing bees. These terms are also used interchangeably in this document.

[0071] The terms "combination" and "co-administration" refer to the administration of an active substance or a composition comprising it with at least one additional active substance or composition comprising it. Such administration may occur sequentially or simultaneously by administering two or more active substances or compositions comprising them at the same time. In such a case, the active substances or compositions comprising them may be part of a single formulation that is a combination or mix of these active substances or compositions comprising them, or they may be separate formulations (co-administered at the same time).

[0072] The present invention provides a new, unexpected and long-undiscovered beekeeping use of the isoxazoline compound of formula (I). The isoxazoline compound of formula (I) or a composition comprising the isoxazoline compound of formula (I) for use according to the invention is particularly advantageous in the treatment and / or prophylaxis of parasitic arthropod infestations. Such parasitic infestations may be ectoparasitic and / or endoparasitic infestations. In one embodiment, the parasitic infestation being prevented and / or treated is an ectoparasitic infestation, especially of the family Varroidae. The composition of the invention is preferably administered by systemic routes of administration. Equally advantageously, the isoxazoline compound of formula (I) or a composition comprising the isoxazoline compound of formula (I) for use according to the invention is effective and safe in various subgroups of bees, including the particularly nutritionally and economically important honeybees and eastern honeybees.The Applicant's research has shown that the isoxazoline compound of formula (I) is effective and safe in bees. Parasitic arthropods of bees against which isoxazoline compounds have not yet been used have not developed resistance. Fluralaner - an example of the isoxazoline compound of formula (I) - has already been effectively used on a large scale in the treatment and prophylaxis of laying hens. Therefore, unlike previously used chemicals to control infestations with parasites from the Varroidae family, the isoxazoline compounds of formula (I) do not pose a nutritional threat to humans. Meanwhile, the compositions of the invention allow for dosages that avoid dangerous concentrations of the isoxazoline compound of formula (I) in honey and its disposal. The compositions of the invention are easy to use and their form allows for effective administration to multiple individuals, or even an entire hive. Equally important, the invention can be administered in occupied beehives (i.e., without the need for emptying).

[0073] The invention does not relate to and excludes known uses and compositions of the isoxazoline compounds of formula (I), in particular among dogs, cats and poultry, and compositions suitable for administration to these animals (e.g. oral tablets, rubs, collars and medicated drinking water).

[0074] The invention will be described in more detail in the following preferred and non-limiting embodiments, with reference to the attached drawings, wherein:

[0075] Fig. 1 shows Ostrowska-type beehives used in research work on the invention,

[0076] Fig. 2 shows the hygienic bottom board of the beehive covered with parasites after using the invention.

[0077] Each experiment was conducted on 10 colonies of the Carniolan bee (Apis mellifera carnica L.), Kortowka line. All colonies comprised the previous year's queens and sisters. The colonies were housed in Ostrowska's beehives on two bodies (Fig. 1). At the beginning of each experiment, the bees were of equal strength and occupied an average of 11.5 intercomb alleys.

[0078] The beehives comprising the bee colonies were equipped with hygienic bottom boards, allowing for the control of the number of parasites fallen from the beehive onto the hygienic bottom boards. To prevent any live Varroa destructor individuals from escaping, Varroa-sticky adhesive inserts were placed on the hygienic bottom boards.The apiary was located near the town of Lutomia Gorna (Lower Silesian Voivodeship, in southwestern Poland).

[0079] The parasite load was measured by the amount of parasite fall onto the hygienic bottom boards. Fig. 2 shows - for illustration purposes - one of the hygienic bottom boards with fallen arthropods after the end of the experiment within one embodiment of the invention. The efficacy of the isoxazoline compound of formula (I) for use according to the invention and the composition according to the invention was calculated in accordance with the standard methods of the European Medicines Agency EMA (2103) "Guidelines for veterinary medicinal products for the control of Varroa destructor" and in Beebook (2013) "Standard methods for varroa research".

[0080] Example 1

[0081] A number of compositions have been developed comprising at least one isoxazoline compound of formula (I)

[0082]

[0083] wherein

[0084] R1= halogen, CF3, OCF3, CN,

[0085] n = an integer from 0 to 3, preferably 1 , 2 or 3,

[0086] R2= Ci -C3-haloalkyl, preferably CF3or CF2CI,

[0087] T = a 5- or 6-membered ring which is optionally substituted with one or more radicals Y,

[0088] Y = methyl, halomethyl, halogen, CN, NO2, NH2-C=S or two adjacent radicals Y forming together a chain, especially a three- or four-membered chain;

[0089] Q= X-NR3R4or 5-membered N-heteroaryl ring which is optionally substituted with one or more radicals;

[0090] X = CH2, CH(CH3), CH(CN), CO, CS,

[0091] R3= hydrogen, methyl, haloethyl, halopropyl, halobutyl, methoxy methyl, methoxyethyl, halomethoxymethyl, ethoxymethyl, haloethoxymethyl, propoxymethyl, ethylaminocarbonylmethyl, ethylaminocarbonylethyl, dimethoxyethyl,propynylaminocarbonylmethyl, N-phenyl-N-methyl-amino, haloethylaminocarbonylmethyl, haloethylaminocarbonylethyl, tetrahydrofuryl, methylaminocarbonylmethyl, (N,N-dimethylamino)-carbonylmethyl, propylaminocarbonylmethyl, cyclopropylaminocarbonylmethyl, propenylaminocarbonylmethyl, haloethylaminocarbonylcyclopropyl,

[0092]

[0093] R*-.l3

[0094] wherein

[0095] ZA= hydrogen, halogen, cyano, halomethyl (CF3);

[0096] R4= hydrogen, ethyl, methoxymethyl, halomethoxymethyl, ethoxymethyl, haloethoxymethyl, propoxymethyl, methylcarbonyl, ethylcarbonyl, propylcarbonyl, cyclopropylcarbonyl, methoxycarbonyl, methoxymethylcarbonyl, aminocarbonyl, ethylaminocarbonylmethyl, ethylaminocarbonylethyl, dimethoxyethyl, propynylaminocarbonylmethyl, haloethylaminocarbonylmethyl, cyanomethylaminocarbonylmethyl or haloethylaminocarbonylethyl;

[0097] or R3and R4together form a substituent selected from the group consisting of:

[0098]

[0099] and "CH ;

[0100] or a salt or solvate thereof as embodiments of the invention. In preferred embodiments, the isoxazoline compound of formula (I) was fluralaner. All compositions of the invention were developed to be acceptable for beekeeping purposes. Compositions of the invention were in both liquid and solid forms. The choice of liquid or solid form depends on the beekeeper's preference and the individual situation.Without departing from the scope of the invention, the solid form may be selected from a strip impregnated or coated with an active substance, an insert impregnated or coated with an active substance, a fumigation tablet, a powder, a granulate, a food, or a food additive. Below, as non-limiting examples illustrating the broad range of forms of the composition according to the invention, a strip impregnated or coated with an active substance (Tables 1 and 2) and a fumigation tablet (Table 3) are described - in these cases, the active substance was fluralaner.

[0101] The solid form of the composition according to the invention may be used both inside and outside a beehive. In a preferred embodiment, the solid form of the composition according to the invention for administration outside a beehive comprises an excess of active substance -e.g., the isoxazoline compound of formula (I) - compared to the corresponding solid form for administration inside the beehive to account for losses during external administration. In one embodiment, the solid form of the composition according to the invention for administration inside the beehive comprised 0.1 mg, in another embodiment, the solid form of the composition according to the invention for administration inside the beehive comprises 200 mg of the isoxazoline compound of formula (I). Meanwhile, in one embodiment, the solid form of the composition according to the invention for administration outside the beehive comprised 0.1 mg, and in another embodiment, the solid form of the composition according to the invention for administration outside the beehive comprised 3000 mg of the isoxazoline of formula (I).

[0102] Table 1 A composition of a strip impregnated or coated with at least one compound of formula (I) according to the invention for administration inside a beehive.

[0103] a) Based on plywood:

[0104] Ingredient Quantity [mg] Concentration [pg / cm3] Function Fluralaner 3.6 500 Active substance Birch plywood 7000 carrier

[0105]

[0106] b) Based on polyethylene:

[0107] Ingredient Quantity [mg] Concentration [pg / cm3] Function Fluralaner 11.6 500 Active substance Polyethylene 999.46 carrier

[0108]

[0109] Table 2. Composition of a strip impregnated or coated with at least one compound of formula (I) according to the invention for administration outside of the beehive.

[0110] a) Based on plywood:

[0111] Ingredient Quantity [mg] Concentration [mg / cms] Function Fluralaner 275 38 Active substance Birch plywood 7000 carrier

[0112]

[0113] b) Based on PVC:

[0114] ingredient Quantityfmg] Cencentration[mg / cm3] Function Fluralaner 275 38 Active substance Ethylene glycol 2464.0 carrier Soybean oil 297 carrier Magnesium stearate 258.5 carrier PVC 6580,0 carrier soot 015.0 carrier

[0115]

[0116] Table 3. Composition of a fumigation tablet with at least one compound of formula (I) according to the invention.

[0117] Ingredient Quantity [mg] Function Fluralaner 15 mg Active substance Potassium carbonate 25 mg carrier Charcoal 390 mg carrier

[0118] Starch 40 mg carrier Potassium citrate 140 mg carrier Carboxymethylcelluiose 62.5 mg carrier

[0119] White spirit 10000 mg carrier

[0120]

[0121] Without departing from the scope of the invention, the liquid form may be selected from a concentrate, a liquid, a food, or a food additive. In one embodiment, the liquid form of the composition of the invention comprised 0.1 mg; in another embodiment, the solid form of the composition of the invention comprised 200 mg of the isoxazoline compound of formula (I). Table 4 below, as a non-limiting example illustrating the broad utility of the composition of theinvention, describes a liquid which, when diluted in an amount sufficient to achieve an effective therapeutic concentration, can be used in feeding or adding to the wax foundation. In this embodiment, the active substance was fluralaner.

[0122] Table 4. Composition of the liquid with at least one compound of formula (I) according to the invention.

[0123] Ingredient Quantity [mg] Function Fluralaner 15 mg Active substance Rokacet R 36 1525 mg carrier

[0124] Ethyl acetate 90 mg carrier

[0125]

[0126] The above Tables 1-4 refer to fluralaner, but the tests were successfully repeated with other isoxazolines, all results of which are not presented here for clarity. Other particularly preferred isoxazolines include afoxolaner, sarolaner, fluralaner and lotilaner. The composition of the invention comprises fluralaner or other isoxazolines alone or in combination without departing from the scope of the invention.

[0127] The compositions of the invention are prepared by conventional methods known in the art. These methods include, without departing from the scope of the invention: mixing, dissolving, emulsifying, granulating, calibrating, drying, tableting, coating, micropelletizing, encapsulating, dissolving, suspending, etc. In the case of fumigation tablet formulations, the composition of the invention was prepared by mixing all ingredients until a visually homogeneous blend was obtained, as confirmed by analytical homogeneity testing. The blend was then calibrated on 0.5 mesh sieves and finally tableted on a rotary tablet press (I KA Pressima, oval punches) using feeding and tableting parameters to obtain pharmacopeial-acceptable tablets (i.e., appropriate hardness, friability and solubility). Examples of excipients preferred for the production of tablet forms for fumigation of the composition according to the invention are: liquid paraffin, gelatin, potassium nitrate, modified starch, charcoal, ethyl potassium carbonate, other suitable excipients or any carriers that cause slow combustion and produce smoke. In the case of a liquid form for feeding, the composition according to the invention is prepared by mixing the active substance (isoxazoline compound) with a carrier and excipients by deflocculation with stirring using a spatula until a visually homogeneous mixture is obtained (dissolution). Examples of excipients preferred for the production of liquid form compositions for feeding and for wax foundation according to the invention are: ester, surfactant, ethoxylated castor oil(macrogolglycerol ricinolate), excipients or any carriers. In the case of strips impregnated or coated with an active substance, the composition according to the invention is prepared by impregnating or covering the strip with a liquid solution comprising an isoxazoline compound of formula (I). Examples of excipients preferred for the preparation of strips impregnated or coated with an active substance of the composition according to the invention for use inside a beehive are: polyethylene, any plastic, wood and / or its derivatives, paper and / or its derivatives and / or other suitable excipients and / or any carriers. Examples of excipients preferred for the preparation of strips impregnated or coated with an active substance of the composition according to the invention for administration outside a beehive are: polyvinyl chloride, any plastic, wood and / or its derivatives, paper and / or its derivatives and / or other suitable excipients, and / or any carriers.

[0128] Example 2

[0129] The efficacy and safety of the isoxazoline compound of formula (I) in the treatment and prophylaxis of bees was tested. In the embodiments, the isoxazoline compound of formula (I) was used for the treatment and prophylaxis of bees.

[0130] * .C'N

[0131] V \ ' 'f Q

[0132]

[0133] (I)

[0134] wherein

[0135] R1= halogen, CF3, OCF3, CN,

[0136] n = an integer from 0 to 3, preferably 1 , 2 or 3,

[0137] R2= Ci -C3-haloalkyl, preferably CF3or CF2 Cl,

[0138] T = a 5- or 6-membered ring which is optionally substituted with one or more radicals Y,

[0139] Y = methyl, halomethyl, halogen, CN, NO2, NH2 -C=S or two adjacent radicals Y forming together a chain, especially a three- or four-membered chain;

[0140] Q= X-NR3R4or 5-membered N-heteroaryl ring which is optionally substituted with one or more radicals;

[0141] X = CH2, CH(CH3), CH(CN), CO, CS,R3= hydrogen, methyl, haloethyl, halopropyl, halobutyl, methoxy methyl, methoxyethyl, halomethoxymethyl, ethoxymethyl, haloethoxymethyl, propoxymethyl, ethylaminocarbonylmethyl, ethylaminocarbonylethyl, dimethoxyethyl, propynylaminocarbonylmethyl, N-phenyl-N-methyl-amino, haloethylaminocarbonylmethyl, haloethylaminocarbonylethyl, tetrahydrofuryl, methylaminocarbonylmethyl, (N,N-dimethylamino)-carbonylmethyl, propylaminocarbonylmethyl, cyclopropylaminocarbonylmethyl, propenylaminocarbonylmethyl, haloethylaminocarbonylcyclopropyl,

[0142] RM. RM

[0143] R’-S RM RM RM

[0144] / \ ,A / /

[0145] w. / '* c. <

[0146] N--y

[0147] fV-7 RM NH._ 0 0

[0148] •s V s

[0149]

[0150] O-CH< CH.

[0151] R' ii R--52 ft-'- R:-1S

[0152] wherein

[0153] ZA= hydrogen, halogen, cyano, halomethyl (CF3);

[0154] R4= hydrogen, ethyl, methoxymethyl, halomethoxymethyl, ethoxymethyl, haloethoxymethyl, propoxymethyl, methylcarbonyl, ethylcarbonyl, propylcarbonyl, cyclopropylcarbonyl, methoxycarbonyl, methoxymethylcarbonyl, aminocarbonyl, ethylaminocarbonylmethyl, ethylaminocarbonylethyl, dimethoxyethyl, propynylaminocarbonylmethyl, haloethylaminocarbonylmethyl, cyanomethylaminocarbonylmethyl or haloethylaminocarbonylethyl;

[0155] or R3and R4together form a substituent selected from the group consisting of:

[0156]

[0157] andor a salt or solvate thereof. Various isoxazoline compounds of formula (I) have been tested - including afoxolaner, sarolaner, fluralaner and / or lotilaner . However, for clarity and as a preferred embodiment, the present embodiments depict fluralaner as the active substances. To further demonstrate the efficacy and safety of the isoxazoline compound of formula (I) in treatment and prophylaxis of bees, various embodiments, methods and compositions comprising at least one isoxazoline compound of formula (I) were tested.

[0158] In one embodiment, the isoxazoline compound of formula (I) was used in treatment and prophylaxis of bees in the form of strips impregnated or coated with the active substance, placed inside the beehive. Two to ten such strips were hung on frames, depending on the size of the beehive and the development of the bee colony. In colonies without brood, the strips were left in the beehive for 15 days. In colonies with brood present, the strips were left in the beehive for 42-49 days. The strips were placed so as to provide bees with access to both sides of the strips. The composition of the invention was applied during the period of full bee activity, after honey harvesting (from late July to mid-September). Application at low temperatures reduced the treatment effect.

[0159] For example, in one embodiment, the composition described in Table 1 in the form of a strip impregnated or coated with the active substance was administered to control infestations of the parasitic arthropod Varroa destructor in bees. Group A (n = 10 bee colonies) was treated with a composition according to the invention comprising 3.6 mg of at least one isoxazoline compound of formula (I) in an amount of 4 strips per beehive interior for 49 days. The colonies had brood. Group B was treated with a composition according to the invention comprising 3.6 mg of at least one isoxazoline compound of formula (I) in an amount of 6 strips per beehive interior for 49 days. The colonies had brood.

[0160] In other embodiments, 2 to 6 strips were used per beehive, depending on the needs. Advantageously, the unit doses (per strip) could be adjusted - as needed - by shortening them accordingly. In one embodiment, the strip was 6 mg, and cutting it in half would yield 3 mg. In this case, for example, 2 strips with a unit dose of 3 mg each could be used per beehive. In another embodiment, 4 strips with a unit dose of 6 mg each were successfully used per beehive. Most often, 4 strips of 3 to 6 mg each were used per beehive.

[0161] After the treatment period, fumigation with an amitraz preparation was performed to determine the drug's efficacy. The efficacy results (%) are tabulated in Table 5 below. In terms of safety, the study demonstrated good tolerability of the isoxazoline (I). Bee mortality was 800-1100 per colony, and brood mortality was 6.47% to 12.75%.Table 5. Biocidal efficacy of a strip impregnated or coated with the compound of formula (I) according to the invention used inside the beehive.

[0162] Group Varroacidal efficacy (%)

[0163] based on parasite fall on the based on the intensity of bee infection Varroa- sticky insert during and before and after treatment after treatment

[0164] A 74.80 70.72

[0165] B 97.82 98.03

[0166]

[0167] In another embodiment, the isoxazoline compound of formula (I) was used for the treatment and prophylaxis of bees in the form of strips impregnated or coated with the active substance on the outside of the beehive. Such strips were placed outside, at the entrance to the beehive, so that bees could only enter the beehive by passing through the holes in the strip. The total surface area and holes of the strip should not be covered to ensure contact between the bees and the strip and adequate ventilation of the beehive. Such strips were designed to not interfere with the removal of dead individuals. Treatment was continued for at least 9 weeks until the end of flight activity, but no longer than 4 months. In the event of continuous mite fall for 9 weeks, treatment was continued and progress was monitored. This was intended to cover the key phase of potential horizontal mite transfer, e.g., through theft.

[0168] For example, in one embodiment, the composition described in Table 2 in the form of a strip impregnated or coated with the active substance was administered to control infestations of the parasitic arthropod Varroa destructor in bees. A group of 10 bee colonies was treated with a composition according to the invention comprising 275 mg of at least one isoxazoline compound of formula (I) in an amount of 2 strips outside the beehive for 60 days. The colonies had brood.

[0169] In other embodiments, from 1 to 4 strips were used per beehive, depending on the needs. Advantageously, the unit doses (per strip) could be adjusted - as needed - by shortening them accordingly. In one embodiment, the strip was 500 mg, and cutting it into 5 equal parts would give 100 mg per strip. In this case, for example, 2 strips could be used, giving a total dose of 200 mg per beehive. In another embodiment, 4 strips with a unit dose of 500 mg each were successfully used per beehive. Most often, 2 strips per beehive, from 200 to 500 mg each, were used.

[0170] After the treatment period, fumigation with an amitraz preparation was performed to determine the drug's efficacy. The efficacy results (%) are tabulated in Table 6 below. Interms of safety, the study demonstrated good tolerability of the isoxazoline (I). Bee mortality was 450-1230 per colony, and brood mortality was 3.48% to 6.23%.

[0171] Table 6. Biocidal efficacy of a strip impregnated or coated with the compound of formula (I) according to the invention applied outside the beehive.

[0172] Varroacidal efficacy (%)

[0173] based on parasite fall on the Varroa- sticky based on the intensity of bee infection insert during and after treatment before and after treatment

[0174] 99.2 99.6

[0175]

[0176] In yet another embodiment, the isoxazoline compound of formula (I) was used in bee treatment and prophylaxis in the form of fumigation tablets comprising the active substance. These tablets were applied at a rate of 1-2 tablets per beehive - depending on the size of the beehive, the size, and the maturity of the bee colony - by ignition and blowing out the flame. The smoldering tablet was placed on a narrow (e.g., 3-4 cm) strip of dense metal mesh or a specially curved wire feeder to facilitate air access to the tablet. The smoldering tablet was then inserted through the beehive entrance, placing it on the bottom plate under the frames, and the entrance was closed for approximately 20 minutes. The entrance was then reopened and checked for combustion. If the tablet did not burn, the procedure was repeated. Fumigation is performed in the evening, after the bees have completed their flight.

[0177] For example, in one embodiment, the composition described in Table 3 in the form of fumigation tablets with the active substance was administered to control infestations of the parasitic arthropod Varroa destructor in bees. A group of 10 bee colonies was treated with a composition according to the invention comprising 15 mg of at least one isoxazoline compound of formula (I) per tablet. In an amount of 0.5 - 1 tablet per beehive. The colonies had brood.

[0178] In other embodiments, depending on the needs, 14 to 2 tablets per beehive were used. Most often, 0.5-1 15 mg tablet per beehive was used.

[0179] 48 hours after fumigation with the composition according to the invention, fumigation with the amitraz preparation was performed to determine the drug's efficacy. The efficacy results (%) are tabulated in Table 7 below. In terms of safety, the study showed average tolerability of the isoxazoline of formula (I) administered in this form of the composition according to the invention. Bee mortality was 1300-2400 per colony, and brood mortality was 12.75% to 21.50%.Table 7. Biocidal efficacy of fumigation tablets with the compound of formula (I).

[0180] Varroacidal efficacy (%)

[0181] based on parasite fall on the Varroa- sticky based on the intensity of bee infection insert during and after treatment before and after treatment

[0182] 95.6 95.8

[0183]

[0184] In yet another embodiment, the isoxazoline compound of formula (I) is used in treatment and prophylaxis of bees in the form of a liquid with the active substance. Such liquids, after dissolving (e.g., in syrup) in an amount appropriate to obtain a treatment concentration, are used in feeding or added to wax foundation.

[0185] For example, in one embodiment, the composition described in Table 4 in the form of a liquid with the active substance in feeding was administered to control infestations of the parasitic arthropod Varroa destructor in bees. A group of 10 bee colonies was treated with the composition of the invention dissolved in sugar syrup and a volume equivalent to 16 mg of the isoxazoline compound of formula (I) was administered at one time per beehive. The colonies had brood. In other embodiments, depending on the needs, from 8 to 24 mg was used per beehive.

[0186] Five months after administration of the composition according to the invention, fumigation with an amitraz preparation was performed to determine the efficacy of the drug. The results regarding efficacy (%) are tabulated in Table 8 below. In terms of safety, the study showed average tolerability of the isoxazoline of formula (I) administered in this form of the composition according to the invention. Bee mortality was 1150-1570 per colony, and brood mortality was 25.8% to 32.5%.

[0187] Table 8. Biocidal efficacy of the liquid with the compound of formula (I).

[0188] Varroacidal efficacy (%)

[0189] based on parasite fall on the Varroa- sticky based on the intensity of bee infection insert during and after treatment before and after treatment

[0190] 64.5 72.1

[0191]

[0192] The above results demonstrate the efficacy and safety of using the isoxazoline compound of formula (I) according to the invention and the composition according to the invention in treatment and / or prophylaxis of bees. Additionally, bees subjected to treatment and cured from parasites showed better immunity and the queen bees produced better brood (egglaying). In a preferred embodiment, the invention was used in honeybees. The results of other embodiments - not reported here for space reasons - confirm a similar level of efficacy and safety among other representatives of the bee family, which include eastern honeybees, giant honeybees, dwarf honeybees, or stingless bees (meliponines). In a preferred embodiment, the isoxazoline compound of formula (I) and the composition of the invention were used in treatment and / or prophylaxis of infestations by parasitic arthropods. In a particularly preferred embodiment, the isoxazoline compound of formula (I) and the composition of the invention were used in the treatment and / or prophylaxis of infestations by parasitic arthropods of the Varroidae family. In addition to the above-mentioned examples of the efficacy and safety of the isoxazoline compound of formula (I) and the composition of the invention against the bee parasite (Varroa destructor), the results not reported here for space reasons confirm the efficacy also against Varroa jacobsoni.

[0193] As part of the research, a reduction in the number of parasites in the apiary was observed. Therefore, the isoxazoline compound of formula (I) is effective in eliminating or controlling parasites in the bees’ environment, reducing the chance of the infestation spreading, including controlling the apiary's infestation. Furthermore, due to its prophylactic effect — directly through the bees being treated and indirectly by eliminating or controlling parasites in the bees’ environment the isoxazoline compound of formula (I) for use in the present invention is also effective in prophylaxis of diseases (e.g., viral diseases) transmitted by parasites.

[0194] Example 3

[0195] In yet other embodiments, the possibility of administering the isoxazoline compound of formula (I) in combination with other beekeeping acceptable active substances was investigated. In a preferred embodiment, such beekeeping acceptable active substances were antiparasitic compounds. Examples of such active substances are active substances known in the art against Varroa mite such as, among others, thymol, amitraz, formic acid, oxalic acid, lactic acid, organic acids, coumaphos, menthol, essential oils, propolis, camphor, eucalyptol, acrinathrin, bromfenvinphos, chlorfenvinphos, cymiazole, fluvalinate, flumethrin, cypermethrin , as well as active substances against other parasites such as, among others, hop beta acids, oxytetracycline, tylosin, lincomycin, permethrin, paradichlorobenzene and / or bacillus thuringiensis. In another preferred embodiment, the possibility of administering the isoxazoline compound of formula (I) in combination with other isoxazolines of formula (I) such as afoxolaner, sarolaner, fluralaner and / or lotilaner has been successfully tested. Suchadministrations have proven to be advantageous in terms of the possibility of reducing the dose of at least one of the active substances administered in combination. This is particularly preferred when the active substances are administered in combination with the isoxazoline of formula (I) for use according to the invention or the composition according to the invention simultaneously or when the additional active substance is from the same class as or has the same indication.

Claims

Claims:

1. The isoxazoline compound of formula (I)whereinR1= halogen, CF3, OCF3, CN,n = an integer from 0 to 3, preferably 1 , 2 or 3,R2= Ci -C3-haloalkyl, preferably CF3or CF2 Cl,T = a 5- or 6-membered ring which is optionally substituted with one or more radicals Y,Y = methyl, halomethyl, halogen, CN, NO2, NH2-C=S or two adjacent radicals Y forming together a chain, especially a three- or four-membered chain;Q= X-NR3R4or 5-membered N-heteroaryl ring which is optionally substituted with one or more radicals;X= CH2, CH(CH3), CH(CN), CO, CS,R3= hydrogen, methyl, haloethyl, halopropyl, halobutyl, methoxymethyl, methoxyethyl, halomethoxymethyl, ethoxymethyl, haloethoxymethyl, propoxymethyl, ethylaminocarbonylmethyl, ethylaminocarbonylethyl, dimethoxyethyl, propynylaminocarbonylmethyl, N-phenyl-N-methyl-amino, haloethylaminocarbonylmethyl, haloethylaminocarbonylethyl, tetrahydrofuryl, methylaminocarbonylmethyl, (N,N-dimethylamino)-carbonylmethyl, propylaminocarbonylmethyl, cyclopropylaminocarbonylmethyl, propenylaminocarbonylmethyl, haloethylaminocarbonylcyclopropyl,CH.(O-CH, O '"p -N V -NO > C > H c Y -YS' \— / V:::: / 5N"whereinZA= hydrogen, halogen, cyano, halomethyl (CF3);R4= hydrogen, ethyl, methoxymethyl, halomethoxymethyl, ethoxymethyl, haloethoxymethyl, propoxymethyl, methylcarbonyl, ethylcarbonyl, propylcarbonyl, cyclopropylcarbonyl, methoxycarbonyl, methoxymethylcarbonyl, aminocarbonyl, ethylaminocarbonylmethyl, ethylaminocarbonylethyl, dimethoxyethyl, propynylaminocarbonylmethyl, haloethylaminocarbonylmethyl, cyanomethylaminocarbonylmethyl or haloethylaminocarbonylethyl;or R3and R4together form a substituent selected from the group consisting of:CH: andor a salt or solvate thereof for use in the treatment and / or prophylaxis of animals, characterized in that it is used in the treatment and / or prophylaxis of bee diseases.

2. The isoxazoline compound for use according to claim 1, characterized in that the isoxazoline compound is fluralaner.

3. The isoxazoline compound for use according to any of claims 1 - 2, characterized in that the bees are selected from honeybees, eastern honeybees, giant honeybees, dwarf honeybees and stingless honeybees (meliponines).

4. The isoxazoline compound for use according to claim 3, characterized in that the bees are honeybees.

5. The isoxazoline compound for use according to any of claims 1 - 4, characterized in that the compound is used in the treatment and / or prophylaxis of parasitic arthropod infestations.

6. The isoxazoline compound for use according to claim 5, characterized in that the parasitic arthropod infestation is an ectoparasitic arthropod infestation from the Varroidae family, especially Varroa destructor and / or Varroa jacobsoni.

7. The isoxazoline compound for use according to any of claims 5 - 6, characterized in that the compound is used to combat and / or control parasitic arthropods in the bees’ environment.

8. The isoxazoline compound for use according to any of claims 5 - 7, characterized in that it is used in the prophylaxis of diseases transmitted by parasitic arthropods of bees.

9. The isoxazoline compound for use according to any of claims 1 - 8, characterized in that it is administered in combination with another beekeeping acceptable active substance, preferably an antiparasitic compound.

10. The isoxazoline compound for use according to any of claims 1 - 9, characterized in that the isoxazoline compound of formula (I) is used inside a beehive in an amount from 3.75 mg to 48 mg, preferably from 6 mg to 30 mg.

11. The isoxazoline compound for use according to any of claims 1 - 9, characterized in that the isoxazoline compound of formula (I) is used outside a beehive in an amount from 200 mg to 2000 mg, preferably from 400 mg to 1000 mg.

12. The isoxazoline compound for use according to any of claims 1 to 9, characterized in that the isoxazoline compound of formula (I) is used in feeding a beehive in an amount from 8 mg to 24 mg, preferably 16 mg.

13. A composition comprising at least one isoxazoline compound of formula (I)whereinR1= halogen, CF3, OCF3, CN,n = an integer from 0 to 3, preferably 1 , 2 or 3,R2= Ci -C3-haloalkyl, preferably CF3or CF2 Cl,T = a 5- or 6-membered ring which is optionally substituted with one or more radicals Y,T1Y = methyl, halomethyl, halogen, CN, NO 2 , NH2-C=S or two adjacent radicals Y forming together a chain, especially a three- or four-membered chain;Q= X-NR3R4or 5-membered N-heteroaryl ring which is optionally substituted with one or more radicals;X= CH2, CH(CH3), CH(CN), CO, OS,R3= hydrogen, methyl, haloethyl, halopropyl, halobutyl, methoxymethyl, methoxyethyl, halomethoxymethyl, ethoxymethyl, haloethoxymethyl, propoxymethyl, ethylaminocarbonylmethyl, ethylaminocarbonylethyl, dimethoxyethyl, propynylaminocarbonylmethyl, N-phenyl-N-methyl-amino, haloethylaminocarbonylmethyl, haloethylaminocarbonylethyl, tetrahydrofuryl, methylaminocarbonylmethyl, (N,N-dimethylamino)-carbonylmethyl, propylaminocarbonylmethyl, cyclopropylaminocarbonylmethyl, propenylaminocarbonylmethyl, haloethylaminocarbonylcyclopropyl,CH,ONr-NS''"o3- 0o -swhereinZA= hydrogen, halogen, cyano, halomethyl (CF3);R4= hydrogen, ethyl, methoxymethyl, halomethoxymethyl, ethoxymethyl, haloethoxymethyl, propoxymethyl, methylcarbonyl, ethylcarbonyl, propylcarbonyl, cyclopropylcarbonyl, methoxycarbonyl, methoxymethylcarbonyl, aminocarbonyl, ethylaminocarbonylmethyl, ethylaminocarbonylethyl,dimethoxyethyl, propynylaminocarbonylmethyl, haloethylaminocarbonylmethyl, cyanomethylaminocarbonylmethyl or haloethylaminocarbonylethyl;or R3and R4together form a substituent selected from the group consisting of:t ij <,sO — CI?! and * 0 Cihor a salt or solvate thereof, characterized in that it is in a form and dose acceptable for beekeeping.

14. The composition according to claim 13, characterized in that the isoxazoline compound of formula (I) is fluralaner.

15. The composition according to any of claims 13 - 14, characterized in that it is in solid form or liquid form.

16. The composition according to claim 15, characterized in that the solid form is selected from a strip impregnated or coated with an active substance, an insert impregnated or coated with an active substance, a fumigation tablet, a powder, a granulate, a food and a food additive.

17. The composition according to claim 16, characterized in that the solid form for administration inside a beehive comprises from 0.1 mg to 200 mg of the isoxazoline compound of formula (I) and the solid form for administration outside a beehive comprises from 0.1 mg to 3000 mg of the isoxazoline compound of formula (I).

18. The composition according to claim 15, characterized in that the liquid form is selected from a concentrate, a liquid, a food, and a food additive.

19. The composition according to claim 18, characterized in that the liquid form comprises from 0.1 mg / ml to 500 mg / ml of the isoxazoline compound of formula (I).

20. The composition according to any of claims 13 - 18, characterized in that it comprises at least one additional active substance selected from thymol, amitraz, formic acid, oxalic acid, lactic acid, organic acids, coumaphos, menthol, essential oils, propolis, camphor, eucalyptol, acrinathrin, bromfenvinphos, chlorfenvinphos, cymiazole, fluvalinate, flumethrin, cypermethrin, hop beta acids, oxytetracycline, tylosin, lincomycin, permethrin, paradichlorobenzene and bacillus thuringiensis.

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