阅读说明：本技术 可注射药物组合物及其用途 (Injectable pharmaceutical composition and use thereof ) 是由 B·L·瓦尔·科伦 K·弗雷豪夫 F·格里诺 C·D·库尔科扎 B·卡里略 于 2020-05-01 设计创作，主要内容包括：包含式(I)的异恶唑啉化合物或其盐或其N-氧化物和莫西菌素微球的可注射药物组合物以及使用其预防或治疗寄生虫感染的方法。(Injectable pharmaceutical compositions comprising an isoxazoline compound of formula (I) or a salt thereof or an N-oxide thereof and moxidectin microspheres and methods of preventing or treating parasitic infections using the same.)

1. An injectable veterinary composition comprising

a) Moxidectin microspheres comprising from about 50% to about 99% by weight of a fat, wax or mixture thereof, and 0.01-10% by weight of an antioxidant; and

b) isoxazoline particles represented by the formula (I);

wherein

R¹Halogen, CF₃，OCF₃，CN，

n is an integer from 0 to 3, preferably 1,2 or 3,

R²＝C₁-C₃haloalkyl, preferably CF₃Or CF₂Cl，

T ═ 5 to 12 membered monocyclic or bicyclic ring systems, optionally substituted with one or more groups Y,

y ═ methyl, halomethyl, halogen, CN, NO₂、NH₂-C ═ S, or two adjacent groups Y are chained, in particular ternary or quaternary;

Q＝X-NR³R⁴or a 5-membered N-heteroaryl ring, optionally substituted with one or more groups;

X＝CH₂，CH(CH₃)，CH(CN)，CO，CS，

R³hydrogen, methyl, haloethyl, halopropyl, halobutyl, methoxymethyl, methoxyethyl, halomethoxymethyl, ethoxymethyl, haloethoxymethyl, propoxymethyl, ethylaminocarbonylmethyl, ethylaminocarbonylethyl, dimethoxyethyl, propynylaminocarbonylmethyl, N-phenyl-N-methyl-amino, haloethylaminocarbonylmethyl, haloethylaminocarbonylethyl, tetrahydrofuranyl, methylaminocarbonylmethyl, (N, N-dimethylamino) -carbonylmethyl, propylaminocarbonylmethyl, cyclopropylaminocarbonylmethyl, propenylaminocarbonylmethyl, haloethylaminocarbonylcyclopropyl,

wherein Z^AHydrogen, halogen, cyano, halomethyl (CF)₃)；

R⁴Hydrogen, 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:

wherein the moxidectin microspheres and isoxazoline compound particles are suspended in an aqueous carrier comprising one or more suspending agents, one or more wetting agents and/or one or more preservatives and water.

2. The injectable veterinary composition according to claim 1, wherein the fat, wax or mixture thereof has a melting point higher than about 40 ℃.

3. The injectable veterinary composition according to claim 1 or 2, wherein the moxidectin microspheres comprise about 75 to about 95 wt.% of fat, wax or a mixture thereof.

4. The injectable veterinary composition according to any one of claims 1 to 3, wherein the fat, wax or mixture thereof is a fatty acid ester.

5. The injectable veterinary composition according to claim 4, wherein the fatty acid ester is a triglyceride of fatty acids each containing 12-22 carbon atoms.

6. The injectable veterinary composition according to claim 5, wherein the fatty acid ester is glyceryl tristearate.

7. The injectable veterinary composition according to any one of claims 1 to 6, wherein the moxidectin microspheres comprise moxidectin in an amount of about 9-12% by weight and glyceryl tristearate in an amount of 87-92% by weight.

8. The injectable veterinary composition according to any one of claims 1 to 7, wherein the moxidectin microsphere and/or isoxazoline compound particle has a volume weighted particle size distribution D50 of about 25 μm to about 250 μm measured by static light scattering instrument.

9. The injectable veterinary composition according to claim 8, wherein the moxidectin microsphere and/or the isoxazoline compound has a particle size distribution D50 of about 75 μm to about 150 μm.

10. An injectable veterinary composition according to any of claims 1-9 wherein the isoxazoline compound particles have a thickness of more than 10 μm but less than 100 μm, preferably more than 20 μm but less than 90 μm, as measured by scanning electron microscopy.

11. The injectable veterinary composition according to claim 10, wherein the isoxazoline compound particles have a thickness of more than 30 μ ι η but less than 80 μ ι η.

12. The injectable veterinary composition according to any one of claims 1 to 11, wherein the volume weighted particle size of the moxidectin microsphere and/or isoxazoline compound particle has a D10 of about 20 to 35 μ ι η, D50 of about 90 to 105 μ ι η and D90 of about 155 to 175 μ ι η measured by static light scattering.

13. The injectable veterinary composition according to any one of claims 1 to 12, wherein the suspending agent is selected from sodium carboxymethylcellulose, polyvinylpyrrolidone and methylcellulose.

14. The injectable veterinary composition according to any one of claims 1 to 13, wherein the composition comprises from about 0.5 to about 15 wt% of a suspending agent, by weight of the composition.

15. The injectable veterinary composition according to any one of claims 1 to 14, wherein the wetting agent is a poloxamer.

16. The injectable veterinary composition according to any one of claims 1 to 15, wherein the composition comprises from about 0.01% to about 1.0% of a wetting agent by weight of the composition.

17. The injectable veterinary composition according to any one of claims 1 to 16, wherein the isoxazoline compound is selected from the group consisting of flurarana, alfurana, sarorana or loratadine.

18. The injectable veterinary composition according to any one of claims 1 to 17, wherein the isoxazoline compound of formula (I) is flurarana.

19. A kit, wherein the kit comprises:

a) a first container comprising a solid mixture of particles of an isoxazoline compound of formula (I) as described in claims 1, 8, 9, 10, 11, 12, 17 and 18 and moxidectin microspheres as described in claims 1 to 12, and;

b) a second container having an aqueous carrier comprising one or more suspending agents, wetting agents and/or preservatives and water; and

c) instructions for reconstituting the moxidectin microspheres and isoxazoline compound particles with an aqueous vehicle prior to administration to an animal by subcutaneous or intramuscular injection.

20. The kit of claim 19, wherein the first container comprises an effective amount of moxidectin and an isoxazoline compound of formula (I) as described in claim 1, 17, or 18 sufficient to treat or prevent a parasitic infestation in an animal.

21. The kit of claim 19 or 20, wherein the kit further comprises a device for reconstituting the mixture of compositions from the first container and the second container and parenterally administering the mixture of compositions to an animal.

22. The kit of claim 21, wherein the device comprises a syringe.

23. The kit according to any one of claims 19 to 22, wherein in the first container the isoxazoline compound of formula (I) and/or the moxidectin microspheres have a volume weighted particle size distribution D50 of about 25 microns to about 250 microns as measured by static light scattering instrumentation.

24. The kit according to any one of claims 19-23 wherein in said first container the isoxazoline compound has a particle size D10 of about 20-35 μm, a particle size D50 of about 90-105 μm, and a particle size D90 of about 155-175 μm.

25. A method of treating or preventing a parasitic infection in an animal comprising administering to an animal in need thereof an injectable veterinary composition according to claims 1 to 18.

26. A method of preparing an injectable veterinary composition according to any one of claims 1 to 18, comprising the steps of:

a) preparing isoxazoline particles;

b) moxidectin microspheres are prepared by melting fats, waxes or mixtures thereof, adding moxidectin and optionally antioxidants, and preparing the microspheres by rotary disk atomization and sieving;

c) filling the moxidectin microspheres obtained by the step b) together with the isoxazoline particles obtained by the step a) into a first container;

d) preparing the aqueous carrier by dissolving excipients including a suspending agent, a wetting agent and/or a preservative in water and filling into a second container;

e) reconstituting the solid by transferring the aqueous carrier from the second container d) to the first container c) and shaking to form the ready-to-use suspension.

27. A method of treating or preventing a parasitic infection in an animal using the kit of any one of claims 19 to 24, the method comprising administering to the animal by subcutaneous or intramuscular injection the reconstituted suspension obtained by the method of claim 26.

Background

Isoxazoline compounds are known in the art, and these compounds and their use as antiparasitic agents are described, for example, in U.S. patent application No. US 2007/0066617 and international patent applications WO 2005/085216, WO2007/079162, WO 2009/002809, WO 2009/024541, WO 2009/003075, WO2010/070068 and WO 2010/079077, the disclosures of which and references cited herein are incorporated by reference. Such compounds are known to have excellent activity against ectoparasitic arthropods.

WO2015/048371 discloses long acting injectable compositions comprising a spirocyclic isoxazoline (spirocy isoxazoline) compound, one biopolymer and at least one carrier, solvent or excipient.

WO2016/138339 discloses long acting injectable formulations comprising at least one isoxazoline active agent, a poloxamer and a co-solvent.

WO2016/164487 discloses an extended release injectable veterinary formulation for combating parasites comprising at least one isoxazoline active agent, a pharmaceutically acceptable polymer and a solvent.

Us patent No.9,609,869 discloses pesticidal compounds based on isoxazoline derivatives for controlling pests associated with agriculture, horticulture, animal husbandry and companion animals.

U.S. patent application publication No. 2017/0239218 discloses long acting injectable compositions for combating parasites comprising at least one isoxazoline active agent, liquid PEG and/or a neutral oil.

Another ectoparasite compound has recently been described: 2-chloro-N- (1-cyanocyclopropyl) -5- [1 '-methyl-3' - (1,1,2,2, 2-pentafluoroethyl) -4'- (trifluoromethyl) [1,5' -bis-1H-pyrazol ] -4-yl ] benzamide; tegolana (Tigolaner) (CAS RN 1621436-41-6) which is disclosed in WO 2019/012377.

Moxidectin (Moxidectin) is an active ingredient which is useful in the prevention and treatment of infections and infestations caused by helminths, nematodes, mites and endoparasitic and ectoparasitic arthropods, particularly when administered to animals without parenteral administration.

Moxidectin is disclosed in U.S. patent No. 4,916,154. EP0525307 and EP1197207 disclose moxidectin microspheres and injectable compositions, their preparation and use.

An advantageous injectable pharmaceutical composition for veterinary use would be one that enables a single injection to provide effective concentration levels of the two active compounds (moxidectin and isoxazoline compounds) in the plasma of treated animals over an extended period of time.

In addition to the duration of release of such compositions, technical features of injectable veterinary formulations, such as ease of administration (injectability and resuspension) and absence of side effects (local injection site reactions and systemic side effects after administration) as well as the possibility of sterilizing the formulation, are important features.

It is therefore desirable to have a technically feasible injectable formulation that allows for the effective and safe release of an effective amount of the isoxazoline compound and moxidectin as described above in a combined formulation over an extended period of time. This would allow these modern compounds to be used under single injection conditions without the need for repeated dosing. The composition should also ensure that the excipients do not interfere with the moxidectin microspheres and provide a stable moxidectin content.

Accordingly, there is a need for injectable pharmaceutical compositions for the prolonged release of isoxazoline compounds and moxidectin that overcome one or more limitations of the prior art.

Disclosure of Invention

Accordingly, the present invention provides injectable compositions comprising isoxazoline compounds and moxidectin microspheres with long term efficacy against parasites, safety, physical and chemical stability and reduced risk of injection site irritation.

One embodiment of the invention is an injectable veterinary composition comprising

a) Moxidectin microspheres comprising from about 50% to about 99% by weight of a fat, wax or mixture thereof, and 0.01-10% by weight of an antioxidant; and

b) particles of isoxazoline compounds of formula (I)

Wherein

R¹Halogen, CF₃、OCF₃、CN，

n is an integer from 0 to 3, preferably 1,2 or 3,

R²＝C₁-C₃haloalkyl, preferably CF₃Or CF₂Cl，

T ═ 5 to 12 membered monocyclic or bicyclic ring systems, optionally substituted with one or more groups Y,

y ═ methyl, halomethyl, halogen, CN, NO₂、NH₂-C ═ S, or two adjacent groups Y together form a chain, in particular a ternary or quaternary chain;

Q＝X-NR³R⁴or a 5-membered N-heteroaryl ring, optionally substituted with one or more groups;

X＝CH₂、CH(CH₃)、CH(CN)、CO、CS，

R³hydrogen, methyl, haloethyl, halopropyl, halobutyl, methoxymethyl, methoxyethyl, halomethoxymethyl, ethoxymethyl, haloethoxymethyl, propoxymethyl, ethylaminocarbonylmethyl, ethylaminocarbonylethyl, dimethoxyethyl, propynylaminocarbonylmethyl, N-phenyl-N-methyl-amino, haloethylaminocarbonylmethyl, haloethylaminocarbonylethyl, tetrahydrofuranyl, methylaminocarbonylmethyl, (N, N-dimethylamino) -carbonylmethyl, propylaminocarbonylmethyl, cyclopropylaminocarbonylmethyl, propenylaminocarbonylmethyl, haloethylaminocarbonylcyclopropyl,

wherein Z^AHydrogen, halogen, cyano, halomethyl (CF)₃)；

R⁴Hydrogen, ethyl, methoxymethyl, halomethoxymethyl, ethoxymethyl, haloethoxymethyl, propoxymethyl, methylcarbonyl, ethylcarbonyl, propylcarbonyl, cyclopropylcarbonyl, ethoxycarbonyl, methoxyethylcarbonyl, aminocarbonyl, ethylaminocarbonylmethyl, ethylaminocarbonylethyl, dimethoxyethyl, propynylaminocarbonylmethyl, haloethylaminocarbonylmethyl, cyanomethylaminocarbonylmethyl, or haloethylaminocarbonylethyl;

or R³And R⁴Together form a substituent selected from:

or a salt or solvate thereof;

wherein the moxidectin microspheres and isoxazoline compound particles are suspended in an aqueous carrier comprising one or more suspending agents, one or more wetting agents and/or one or more preservatives and water.

Another embodiment is a method of treating or preventing a parasitic infection in an animal comprising administering to an animal in need thereof such an injectable veterinary composition.

Another embodiment is a method for producing such an injectable veterinary composition comprising the steps of:

a) preparing isoxazoline particles, preferably by crystallization;

b) moxidectin microspheres are prepared by melting fats, waxes or mixtures thereof and adding moxidectin and optionally antioxidants and preparing the microspheres, preferably by rotary disk atomization and optionally sieving;

c) filling the moxidectin microspheres obtained by the step b) together with the isoxazoline particles obtained by the step a) into a first container;

d) preparing an aqueous carrier by dissolving excipients including a suspending agent, a wetting agent and/or a preservative in water and filling into a second container;

e) reconstituting the solid by transferring the aqueous carrier from the second container d) to the first container c) and shaking to form the ready-to-use suspension.

Another embodiment is a kit, wherein the kit comprises:

a) a first container comprising a solid mixture of particles of an isoxazoline compound of formula (I) above and moxidectin microspheres as described above;

b) a second container having an aqueous carrier comprising one or more suspending agents, wetting agents and/or preservatives and water; and

c) instructions for reconstituting the moxidectin microspheres and the isoxazoline compound particles with an aqueous vehicle prior to administration to an animal by subcutaneous or intramuscular injection.

Another embodiment is a method of treating or preventing a parasitic infection in an animal by administering the reconstituted suspension to the animal by subcutaneous or intramuscular injection using such a kit.

Drawings

FIG. 1: scanning electron micrographs of 10% moxidectin in glyceryl tristearate microspheres (GTS)

FIGS. 2 and 3: plasma levels following subcutaneous administration of moxidectin (2) and fluranide (3) to dogs

Detailed Description

The present invention provides injectable compositions comprising isoxazoline compounds and moxidectin microspheres with long-term efficacy against parasites, safety, physical and chemical stability and reduced risk of injection site irritation.

The physical stability of an injectable suspension is particularly important to allow proper administration by injection of a homogeneous suspension containing both the isoxazoline compound and moxidectin in the correct amounts in one common formulation. The present inventors had to overcome the problem of different densities of the two different solid components, isoxazoline (crystalline) particles and moxidectin microspheres-difficulty in providing a homogeneous suspension. Chemical stability in the formulation is particularly challenging for moxidectin.

A stable suspension should be formed that can be easily resuspended with an aqueous vehicle by gentle shaking without causing foaming or floating or settling of the suspended particles, which would affect the accuracy of administration.

Furthermore, it is important that the final composition to be injected remains physically (and chemically) stable throughout use after resuspension/reconstitution.

In addition, it is advantageous that the injection is safe for the animal after injection and does not cause side effects, particularly without unacceptable injection site irritation.

More importantly, the composition can be sterilized by known and acceptable procedures as it will be administered by injection.

The present invention provides such advantageous compositions.

Isoxazoline compounds are known in the art, and such compounds are known to have excellent activity against parasitic infections (e.g., ticks and fleas). Embodiments of various isoxazoline compounds useful in the present invention are provided below.

Injection site irritation is the damage that occurs at the injection site and surrounding tissues when an animal receives an injection of a pharmaceutical composition. Such damage may be swelling, skin discoloration and tissue necrosis. Although some injection site irritation is inevitable in some animals, veterinarians and animal owners generally consider injection site swelling of more than 2 x 2cm lasting for more than 2 to 3 days to be unacceptable. By minimal injection site stimulation is meant injection site stimulation of less than 2 x 2cm, which lasts for less than 2 to 3 days. In the case of animals receiving injections such as rabies vaccine, veterinarians and their customers generally receive this criterion.

As used herein, reported particle size data is volume weighted, such as static light scattering (also known as laser diffraction), image analysis, or sieving, as measured by conventional particle techniques well known to those skilled in the art. More discussion of particle size measurement is provided below.

Injectable means that the suspension can be readily removed from the ampoule/vial/container into a syringe having a needle, and subsequently injected from such a syringe through the needle intramuscularly (im) or subcutaneously (sc) (e.g. 18 gauge needle).

The particle size of the active ingredient in a suspension can affect resuspendability and injectability, i.e., it should be small enough to prevent compaction or clumping and facilitate resuspension.

A pharmaceutically acceptable excipient is an inert substance that forms the vehicle or medium of the drug product.

By "infestation" of a parasite is meant the presence of an amount of the parasite that poses a risk to a human or animal. Presence may be in the environment, for example in animal bedding, on the skin or fur of an animal, etc. When reference is made to an infestation in an animal, for example in blood or other internal tissue, unless otherwise indicated, the term infestation is also intended to be synonymous with the term "infection", as that term is generally understood in the art.

By aqueous suspension is meant a composition comprising particles that are mixed with, but not dissolved in, an aqueous liquid comprising water or a water-miscible liquid.

Liquid aqueous vehicles are aqueous carriers or inert media used as solvents (or diluents) in which the active agent is formulated and/or administered.

A reconstitutable (or resuspendable) formulation is one in which the liquid vehicle is one container, one or more active ingredient solids are in another container, and the contents of the two containers are combined at some point prior to administration to an animal to form a liquid solution or suspension final formulation.

Reconstitution is the process of adding a liquid/diluent to dry ingredients to make a solution or suspension.

Aqueous liquid vehicles contain some excipients for formulation, such as one or more aqueous diluents, suspending agents, one or more wetting agents, one or more preservatives, and the like.

The pharmaceutical compositions of the present invention are of particular value in the control of ectoparasites, i.e., arthropods that are harmful to humans and livestock and companion animals or that transmit diseases in humans and livestock and pets or that act as carriers of diseases.

Important arthropod parasites-ectoparasites (insect and acarine pests) are described in more detail below.

Biting insects include, for example, migrating diperous lavae, such as dermomycops (Hypoderma sp.), gastrophilis sp of horses and flavoviridae (currebra sp.) in rodents, as well as various types of biting flies and mosquitoes (mosquitoes spp.), for example, blood sucking adults including, for example, horn or western horn flies (haemantobia irritans), horse flies or gadflies (talus spp.), fly or stable stox flies, black flies or gnats (Simulium spp.), horseflies or horseflies (chrysomyiids spp.), Meloxys flies or sheds flies (melegus inus), tongue or tongue flies (Glossina spp.). Parasitic fly maggots include, for example, skin flies (ovine flies (Oestris ovis) and Flavobrussias (Cuterebra spp.)), blowflies (blow flies) or Chlorobia (Phaenicia spp.), gyrus or homo saprophaga (Cochliomyia hominivorax), bovine grubs or dermomycosis (Hypoderma spp.), and ovine caterpillars (fleecem). Mosquitoes include, for example, Culex spp, Anopheles spp and Aedes spp.

The mite includes chicken mite, Dermanyssus gallinae (Dermanyssus gallinae); itch or scab mites or scabies (altigmaa), such as sarcoptedae (Sarcoptidae spp.), e.g., Sarcoptes scabiei; scabies mites, such as the pediculosis (Psopotidae spp.), including the bovine Demodex mites (Choroptes bovis), the ovine Demodex ovis and the canine Demodex Canis; otomites octodes cyanides; trombiculidae (Trombiculidae spp.), such as Trombiculidae, such as trombiculid mite, Trombicula aifreddugesi.

Ticks include, for example, soft-bodied ticks, including the genus Argasidae spp, such as the genus rhynchophorus (Argas spp.) and the genus iruthoros (ornithoroos spp.); hard tick including hard tick (Ixodidae spp.), such as hard tick ricini (ixoders ricinus), hard tick scapula (ixoders scapularis), Rhipicephalus sanguineus (Rhipicephalus sanguineus), Haemaphysalis (Haemaphysis spp.), and cicada (German pigment reptilis); dermacentor variabilis (Dermacentor variabilis), Amblyomma americanum (Amblyomma americanum) and Boophilus spp.

Lice include, for example, lice sucking, such as Menopon spp, and bovine feather (Bovicola spp); biting lice, for example, the genera diaphorina (haemantopinus spp.), hemizygium (linoganthus spp.) and louse bovines (solenoptes spp.).

Fleas include, for example, the genus Ctenocephalides spp, such as canine fleas (Ctenocephalides canis) and feline fleas (Ctenocephalides fells); daphnia species (Xenopsylla spp.) such as daphnia magna (Xenopsylla cheopis); and fleas (Pulex spp.) such as human fleas (Pulex irritans).

Real bed bugs include, for example, the family of bed bugs (Cimicidae) or, for example, the common bed bugs (Cimex lectularius); triatomine spp includes lygus, also known as lygus; for example, long red plant bugs (Rhodnius prolixus) and Triatoma (Triatoma spp.).

The inventive compositions are valuable for treating and controlling various life cycle stages of parasites including egg, nymph and larva, juvenile and adult stages.

For the avoidance of doubt, "treatment" as used herein includes both therapeutic and palliative treatment and "controlling ectoparasites" includes killing, repelling, incapacitating, arresting, eliminating, mitigating, minimizing, eradicating pests on and in the animal's environment.

Prevention is the establishment of a new or incoming infestation or infection.

By "controlling an ectoparasite infestation" is meant preventing the infestation, or reducing or diminishing the number of parasites in and/or on the animal body, in whole or in part, and/or inhibiting the development of a parasitic infestation in or on the animal body.

By "controlling endoparasite infestation" is meant wholly or partially reducing or diminishing the number of endoparasites, such as nematode and cestode parasites, in and/or on animals, and/or inhibiting the development of parasite infestation in or on animals.

Efficacy against endoparasites can be assessed by directly counting the endoparasites (particularly worms) after necropsy of the host animal.

The reduction in parasite numbers, particularly gastrointestinal helminth parasites, may alternatively be measured indirectly by fecal egg or differential larval counts. In this case, the effective amount of the composition is determined by treating the animal feces before and after treatment for a reduction in the number of worm eggs or larvae excreted.

The compositions of the present invention are administered by parenteral injection. The concentration of the active ingredient in the composition needs to be sufficient to provide the desired therapeutically or prophylactically effective amount, the volume of which is acceptable for injectable administration by the animal to be treated.

The compositions according to the invention are useful for the treatment of helminth infections, for example infections caused by one or more helminths selected from the group consisting of:

ancylostomus (Ancylostoma spp.); analyzer spp; chicken ascaris (ascaris spp.); ascaris (Ascaris spp.); brugia spp; meloidogyne spp (bunostom spp.); capillariella spp; caenorhabditis spp; cupressus spp (Cooperia spp.); genus california (Cyathostomum spp.); genus cycocyclus spp; genus dicorophorus (cyelodinophorus spp.); calixas (cyclostephanus spp.); stomatochia (guillerostomum spp.); dictyocaulus spp (Dictyocaulus spp.); nematodiasis (Dipetalonema spp.); heartworm (Dirofilaria spp.); the genus Loxothrix (Dracruplus spp.); enterobiasis (Enterobius spp.); filariasis (filiaroides spp.); littertagia (Habronema spp.); haemonchus spp (Haemonchus spp.); heterodera (Heterakis spp.); porcine circinella (Hyostrongylus spp.); metastrongylus spp); meullerius spp.; ostertagia (Necator spp.); nematoda spp (Nematodirus spp.); a hookworm (nippotungylus spp.); nodorhodopsomum spp; genus Onchocerca spp; ostertagia spp); ostertagia spp (Oxyuris spp.); genus Parascaris (Parascaris spp.); coronarium (stephanuus spp.); strongyloides (strongylous spp.); winged nematode (Syngamus spp.); toxocara spp; strongyloides spp; suspensa nematodes (Teladorsagia spp.); toxocara (Toxascaris spp.); trichinella spp; whipworm (Trichuris spp.); strongyloides (trichostrongylous spp.); tridentate nematode (trichontophous spp.); hookworm (Uncinaria spp.) and Wuchereria spp.

Another endoparasite that severely damages animals is Dirofilaria immitis, also known as heartworm. The most common hosts are dogs and cats, but other animals such as ferrets and raccoons may also be infected. The parasites are transmitted by mosquito bites that carry heartworm larvae. Adults live in the major blood vessels of the lungs, causing vascular inflammation and possibly causing cardiac damage and early death. In late stage infections, worms also enter the heart.

In a particularly preferred embodiment of the invention, the composition of the invention is used for the treatment or prevention of heartworm (Dirofilaria immitis) infection. In another embodiment, the compounds and compositions of the invention are used for the treatment or prevention of white spot heartworm infection (Dirofilaria repens).

Control or "efficacy" of a compound refers to the amount of reduction in parasite count by 5% to about 100% after the first administration. Control of arthropods (e.g., insects, mites) can be insecticidal and/or acaricidal. The effect of the compounds of the invention may be, for example, ovicidal, larvicidal, nymphs and/or adults or combinations thereof.

The effect may be manifested directly, i.e. killing the parasites immediately or after a period of time, for example when molting occurs, or by destroying their eggs, or indirectly, for example by reducing the number of eggs laid and/or the hatchability.

For in vivo administration of a compound according to the invention, an effective amount is synonymous with "pharmaceutically effective amount", which is a dose or amount that wholly or partially treats or ameliorates symptoms and/or signs of a parasitic infection or infestation in an animal being treated, or reduces the number of parasites in and/or on the animal body, and/or inhibits the development of a parasitic infestation in or on the animal body. The latter amount can also be readily determined by one of ordinary skill in the art, for example, by observing or detecting changes in the clinical condition or behavior of the treated animal, as well as by observing or detecting relative changes in the number of parasites following such treatment.

Systemic administration of a drug means reaching the target (organ or parasite) by the bloodstream.

Animals refer to mammals, including companion animals. Companion animal (or pet) refers to a dog, cat or horse, in particular a dog or cat.

In one embodiment, the isoxazoline compounds useful in the present invention also include pharmaceutically acceptable salts, esters, and/or N-oxides thereof. Furthermore, reference to isoxazoline compounds also refers to any of its polymorphic forms or stereoisomers.

In one embodiment, the pharmaceutical composition according to the present invention may use a racemic mixture of isoxazolines used in the present invention, which contains equal amounts of the enantiomers of such isoxazoline compounds as described above.

Alternatively, the pharmaceutical compositions may employ isoxazoline compounds that contain an enriched stereoisomer compared to the racemic mixture, which is one of the enantiomers of the isoxazoline as defined herein.

In addition, the pharmaceutical compositions may employ substantially pure stereoisomers of such isoxazoline compounds. Such enriched or purified stereoisomeric formulations of isoxazolines for use in the present invention can be prepared by methods known in the art.

Examples are chemical methods using catalytic asymmetric synthesis, or separation of diastereomeric salts (see, for example, WO 2009/063910 and JP 2011/051977, respectively).

The S enantiomer is particularly preferred.

In one embodiment of the isoxazolines useful in the present invention, T is selected from

Wherein in T-1, T-3 and T-4 the group Y ═ hydrogen, halogen, methyl, halomethyl, ethyl or haloethyl.

In one embodiment of the isoxazolines useful in the present invention, Q is selected from

Wherein R is³、R⁴X and Z^AAs defined above, and

in one embodiment, the isoxazolines useful in the present invention are as shown in table 1.

Table 1:

table 1 (next):

in one embodiment, the isoxazolines useful in the present invention are as shown in table 2.

Table 2:

in one embodiment, the isoxazolines useful in the present invention are the following compounds:

wherein R is^1a、R^1b、R^1cIndependently of one another: hydrogen, Cl or CF₃

Preferably, R^1aAnd R^1cIs Cl or CF₃And R is^1bIs a hydrogen atom, and is,

t is

Wherein Y is methyl, bromo, Cl, F, CN or C (S) NH₂(ii) a n is 1 or 2; and Q is as described above.

In one embodiment of the isoxazolines as defined herein, R is³Is H, and R⁴The method comprises the following steps: -CH₂-C(O)-NH-CH₂-CF₃，-CH₂-C(O)-NH-CH₂-CH₃，-CH₂-CH₂-CF₃or-CH₂-CF₃。

In one embodiment of the pharmaceutical composition according to the invention, the isoxazoline is one or more selected from the group consisting of fluralan (fluralaner), afalorane (afoxolaner), loratadine (lotilaner) or sarolaner (sarolaner). In another embodiment of the pharmaceutical composition according to the invention, the isoxazoline compound is one or more selected from the group consisting of fluralan (fluralaner), afalorane (afoxolaner), tegoramide (tigolaner), lotilaner (lotilaner) or sarolaner (sarolaner).

In one embodiment, the compound of formula (I) is 4- [5- (3, 5-dichlorophenyl) -5-trifluoromethyl-4, 5-dihydroisoxazol-3-yl ] -2-methyl-N- [ (2,2, 2-trifluoro-ethylcarbamoyl) -methyl ] -benzamide (CAS RN 864731-61-3-USAN fluralaner).

In another embodiment, the compound of formula (I) is 4- [5- [ 3-chloro-5- (trifluoromethyl) phenyl ] -4, 5-dihydro-5- (trifluoromethyl) -3-isoxazolyl ] -N- [ 2-oxo-2- [ (2,2, 2-trifluoroethyl) amino ] ethyl ] -1-naphthamide as disclosed in WO2007/079162 (CAS RN 1093861-60-9, USAN-aflavoner).

In one embodiment of the pharmaceutical composition according to the invention, the isoxazoline is lotilaner (CAS RN: 1369852-71-0; 3-methyl-N- [ 2-oxo-2- (2,2, 2-trifluoroethylamino) ethyl ] -5- [ (5S) -5- (3,4, 5-trichlorophenyl) -5- (trifluoromethyl) -4H-1, 2-oxazol-3-yl ] thiophene-2-carboxamide).

In one embodiment of the pharmaceutical composition according to the invention, the isoxazoline is sarolaner (CAS RN: 1398609-39-6; 1- (5' - ((5S) -5- (3, 5-dichloro-4-fluorophenyl) -5- (trifluoromethyl) -4, 5-dihydroisoxazol-3-yl) -3' -H-spiro (azetidine-3, 1' - (2) benzofuran) -1-yl) -2- (methylsulfonyl) ethanone).

In another embodiment, the compound of formula (I) is (Z) -4- [5- (3, 5-dichlorophenyl) -5-trifluoromethyl-4, 5-dihydroisoxazol-3-yl ] -N- [ (methoxyimino) methyl ] -2-methylbenzamide (CAS RN 928789-76-8).

In another embodiment, the compound of formula (I) is 4- [5- (3, 5-dichlorophenyl) -5- (trifluoromethyl) -4H-isoxazol-3-yl ] -2-methyl-N- (thietane-3-yl) benzamide (CAS RN 1164267-94-0) disclosed in WO 2009/0080250.

In another embodiment, the compound of formula (I) is 4- [5- [ 3-chloro-5- (trifluoromethyl) phenyl ] -4, 5-dihydro-5- (trifluoromethyl) -3-isoxazolyl ] -N- [ 2-oxo-2- [ (2,2, 2-trifluoroethyl) amino ] ethyl ] -1-naphthamide as disclosed in WO2007/079162 (CAS RN 1093861-60-9, USAN-aflavoner).

In another embodiment, the compound of formula (I) is 5- [5- (3, 5-dichlorophenyl) -4, 5-dihydro-5- (trifluoromethyl) -3-isoxazolyl ] -3-methyl-N- [ 2-oxo-2- [ (2,2, 2-trifluoroethyl) amino ] ethyl ] -2-thiophenecarboxamide disclosed in WO2010/070068 (CAS RN 1231754-09-8).

In an alternative embodiment, the isoxazoline compound is 2-chloro-N- (1-cyanocyclopropyl) -5- [1 '-methyl-3' - (1,1,2,2, 2-pentafluoroethyl) -4'- (trifluoromethyl) [1,5' -di-1H-pyrazol ] -4-yl ] benzamide; tigolalan (CAS RN 1621436-41-6), which is disclosed in WO 2019/012377.

Isoxazoline compounds can exist in various isomeric forms. References to compounds for use in the present invention always include all possible isomeric forms of such compounds.

In one embodiment, the racemic form of the isoxazoline compound is present in the composition according to the invention. In another embodiment, the S-enantiomer is present.

In a particularly preferred embodiment, the S-enantiomer of frataxin is present.

In another preferred embodiment, the isoxazoline compound according to formula (I) is the (S) -enantiomer of alfafoxolaner (also known as esafoxolaner).

An important aspect of the present invention is the combination of particles of an isoxazoline compound with stabilizing microspheres comprising moxidectin dissolved in a fat, wax or mixture thereof having a melting point above about 40 ℃, preferably above about 50 ℃ in an aqueous suspension.

These moxidectin microspheres can be sterilized by gamma radiation or electron beam without significant degradation.

The microspheres are straightSmall spherical particles of 1-1000mm diameter, consisting of polymerized wax or other protective material.

Preferred stable moxidectin microspheres for injectable formulations according to the present invention comprise about 75 to 95% by weight of fat, wax or mixtures thereof on a weight basis. Preferably, the microspheres comprise about 1-25% moxidectin and about 0.01-1% antioxidant.

In an alternative embodiment, the composition comprises microspheres of different macrolide compounds as described herein, including but not limited to avermectins or milbemycins. In some embodiments, such avermectins or milbemycins are eprinomectin, avermectin, ivermectin, selamectin, milbemectin, milbemycin D, or milbemycin oxime.

In one embodiment, the composition comprises a combination of flurandrine and eprinomectin, or a combination of flurandrine and milbemycin oxime, selamectin, or moxidectin.

In one embodiment, the composition comprises a combination of afoxolaner and eprinomectin, or a combination of afoxolaner and milbemycin oxime, selamectin, or moxidectin.

In one embodiment, the composition comprises a combination of sarolaner and eprinomectin, or a combination of sarolaner and milbemycin oxime, selamectin, or moxidectin.

In one embodiment, the composition comprises lotilaner in combination with eprinomectin or lotilaner in combination with milbemycin oxime, selamectin or moxidectin.

Injectable compositions typically require sterilization prior to administration to an animal. Gamma radiation or electron beam radiation is an effective sterilization method for eliminating microbial contaminants.

However, moxidectin is susceptible to degradation upon irradiation and loses most of its biological activity. This destructive and degradative response to radiation precludes the use of gamma radiation or electron beams as a means of sterilizing certain moxidectin containing compositions.

The moxidectin microspheres can be radiation sterilized for injection as shown in example 2 without adversely affecting the stability of the active ingredient. The moxidectin microsphere comprises or consists essentially of: from about 50% to 99% by weight of a fat, wax or mixture thereof having a melting point above about 40 ℃, from about 1% to 50% of moxidectin, and from about 0.01-10% of an antioxidant.

The injectable pharmaceutical composition achieves an effective prolonged release of moxidectin and isoxazoline compounds.

The present invention also provides a method for introducing and maintaining blood levels of moxidectin and isoxazoline compounds, particularly moxidectin and loratidine, in an animal for an extended period of time; and methods for preventing or treating infections and infestations in animals caused by helminths, nematodes, mites and endoparasitic and ectoparasitic arthropods.

In one embodiment, moxidectin is present in the injectable veterinary composition in an amount from about 0.01% to about 1.0% by weight.

In a specific embodiment, the injectable composition of the invention comprises 15% fluranide and 0.17% moxidectin.

Waxes and fats suitable for use in the ingredients of the present invention typically have melting points above 40 c, preferably above 50 c.

The term "wax" as used herein is defined as a low melting organic mixture or compound of high molecular weight, as described in Hawley's The Condensed Chemical Dictionary, 11 th edition, is solid at room temperature, and is generally similar in composition to fats and oils except that it does not contain glycerides.

Some are hydrocarbons; others are esters of fatty acids and alcohols. These compounds include saturated or unsaturated long-chain C₁₀-C₂₄Fatty acids, alcohols, esters, salts, ethers, or mixtures thereof. They are classified as lipids. Waxes are thermoplastic, but since they are not high polymers, they are not considered a family of plastics.

Common properties of these waxes include water resistance; the texture is smooth; no toxicity; and no unpleasant odor and color. They are flammable and have good dielectric properties. They are soluble in most organic solvents and insoluble in water. The main types are as follows:

A. natural substance (such as natural gas)

1. Animal wax (beeswax, lanolin, shellac wax, Chinese insect wax)

2. Vegetable (Brazil palm, candelilla, waxberry, sugarcane)

B. Mineral substance

1. Paraffin wax or earth wax (ceresin, montan wax)

2. Petroleum waxes (Paraffins, microcrystals) (slack wax or flake wax)

D. Synthesized

1. Olefinic polymers and polyol ether-esters ("carbowax")

2. Chlorinated naphthalenes ("photowaxes").

The term "fat" as used herein is defined as glycerides of higher fatty acids such as stearic acid and palmitic acid. Such esters and mixtures thereof are solid at room temperature and exhibit a crystalline structure. Lard and tallow are examples.

The term "fat" generally refers specifically to triglycerides, while "lipid" is inclusive.

The fat preferably consists of triglycerides of long chain C12-C22 fatty acids, such as stearates, palmitates, laurates, myristates, arachidates and behenates, and mixtures thereof; those having melting points greater than 50 ℃ are most preferred.

In the practice of the present invention, glyceryl tristearate is the most preferred fat.

Antioxidants suitable for use in the practice of the present invention include any antioxidant known in the art suitable for stabilizing moxidectin compounds.

The antioxidant of the present invention may be defined as an organic compound added to rubber, natural fats and oils, food, gasoline and lubricating oil to delay oxidation, deterioration, rancidity and gum formation, respectively. Rubber antioxidants are generally of the aromatic amine type, such as di-, 8-naphthyl-p-phenylenediamine and phenyl-, 8-naphthylamine.

Many antioxidants are substituted phenolic compounds (butylated hydroxyanisole, di-tert-butyl-p-cresol and propyl gallate). Food antioxidants are effective at very low concentrations and not only delay rancidity, but also preserve nutritional value by minimizing the breakdown of vitamins and essential fatty acids.

The moxidectin microspheres of the invention can be sterilized by gamma radiation or electron beam and maintain shelf life without significant loss of biological activity. Moxidectin is generally susceptible to degradation and loss of most of its biological activity, especially upon irradiation.

Antioxidants suitable for use in the microsphere compositions of the present invention include tocopherol, ascorbic acid, ascorbyl palmitate, fumaric acid, malic acid, sodium ascorbate, sodium metabisulfite, n-propyl gallate, BHA (butylated hydroxyanisole), BHT (butylated hydroxytoluene) monothioglycerol, t-butyl hydroxyquinone, 6-ethoxy-1, 2-dihydro-2, 2, 4-trimethylquinoline, and the like, with butylated hydroxytoluene being a preferred antioxidant.

In certain embodiments, antioxidants are generally added to the formulation in amounts of about 0.01 to about 2.0%, with about 0.05 to about 1.0% being particularly preferred, based on the total weight of the formulation.

The microsphere compositions of the invention can be sterilized by gamma radiation or electron beam and maintained for shelf life without significant loss of biological activity.

Microspheres for use in the compositions of the present invention may be prepared by incorporating moxidectin, an antioxidant and optionally other excipients with molten fat, wax or mixtures thereof and then forming the microspheres of the resulting mixture by various techniques, such as emulsifying or atomizing the mixture, or by machining the ingredients and molten fat, wax or mixtures thereof and cooling, such as with a centrifuge disk.

Alternatively, the mixture of active ingredient, antioxidant, excipient and fats, waxes and mixtures thereof and oils may be cooled to obtain a solid which may then be processed by procedures such as milling, grinding and the like.

The stabilized microspheres of the invention are dispersed in a pharmaceutically acceptable carrier to obtain a sustained release composition for parenteral administration.

Excipients such as surfactants, salts, buffers or mixtures thereof may be included in the vehicles of the present invention.

The amount of the excipient suitable for use in the present invention ranges from about 0.1 to 20% by weight.

Preferably, the cellulose derivative, such as carboxymethyl cellulose, comprises about 1-5% by weight of the carrier and the inorganic salt, such as NaCl, comprises about 0.1-2% by weight of the carrier.

Maintenance of blood levels of the active compounds is associated with the protection or treatment of warm-blooded animals against infection and infestation by helminths, nematodes, mites and endoparasitic and ectoparasitic arthropods.

Maintaining blood levels is an indication of slow release of the active ingredient.

The present invention includes the use of the compositions herein to introduce and maintain the levels of moxidectin and isoxazoline compounds, particularly flurarana, in the bloodstream of an animal.

It has been found that the injectable compositions of the present invention comprising particles of isoxazoline compounds and moxidectin microspheres of defined particle size exhibit desirable bioavailability and duration of efficacy while causing minimal irritation at the injection site.

The compositions also provide the desired safety profile for warm-blooded and avian recipients.

Furthermore, it has been found that a single administration of such compositions typically provides effective activity against one or more parasites (e.g., ectoparasites, such as fleas, ticks, or mites), while also tending to provide a rapid onset of activity, a long duration of activity, and/or a desirable safety profile.

The present invention also provides a method for treating or preventing parasitic infections and infestations in an animal comprising administering an effective amount of an injectable composition comprising an antiparasitic effective amount of at least one isoxazoline compound and moxidectin microspheres of defined particle size and a pharmaceutically acceptable excipient.

Surprisingly, it has been found that the compositions of the invention described herein exhibit excellent broad spectrum efficacy against harmful parasites (e.g., ectoparasites such as fleas and ticks) more rapidly and for a long duration than other injectable compositions known in the art, while exhibiting minimal irritation at the site of injection.

The pharmaceutical composition of the present invention may be administered by subcutaneous or intramuscular injection.

The long-acting injectable composition of the present invention includes a pharmaceutically acceptable excipient.

Pharmaceutically acceptable excipients include, but are not limited to, surfactants, antioxidants, preservatives, pH stabilizers (e.g., buffers), and other inactive excipients.

In another embodiment, the compositions of the present invention may comprise from about 0.01% to about 20% (w/v) of a pharmaceutically acceptable excipient.

In other embodiments, the composition may comprise from about 0.01% to about 5% (w/v), from about 0.1% to about 10% (w/v), or from about 0.1% to about 5% (w/v) of a pharmaceutically acceptable excipient. In other embodiments, the composition may comprise from about 5 to about 15% (w/v), or from about 5 to about 10% (w/v), of a pharmaceutically acceptable excipient.

In another embodiment, the composition may comprise from about 7 to about 10% of a pharmaceutically acceptable excipient.

The surfactant may be present in the compositions of the present invention at a concentration of about 0.1% to about 10% (w/w), about 1% to about 10% (w/w), or about 5% to about 10% (w/w). More typically, the surfactant may be present at a concentration of about 0.1% to about 5% (w/w), or about 1 to about 5% (w/w).

Examples of surfactants that may be used in the composition include, but are not limited to, glycerol monooleate, polyoxyethylene sorbitan fatty acid esters, sorbitan esters (including sorbitan monooleate)) Polyvinyl alcohol, polysorbate (including polysorbate 20 and polysorbate 80), D-alpha-tocopherol polyethylene glycol 1000 succinatePericates (TPGS), sodium lauryl sulfate, copolymers of ethylene oxide and propylene oxide (e.g., poloxamers, e.g.F87, etc.), polyethylene glycol castor oil derivatives (including polyoxyethylene 35 castor oil)Polyoxyethylene 40 hydrogenated castor oil (40) Polyoxyethylene 60 hydrogenated castor oil) (ii) a Propylene glycol monolaurateGlycerides, including glyceryl caprylate/capratePolyglycolyzed glycerides (A), (B), (C)PEG 300 caprylic/capric glyceridesPEG 400 caprylic/capric glyceridesPEG 300 glyceryl oleatePEG 300 glyceryl linoleatePolyethylene glycol stearate and polyethylene glycol hydroxystearate, including polyethylene glycol 8 stearate (PEG 400 monostearate), polyethylene glycol 40 stearate (PEG 1750 monostearate)Fatty acid esters, etc.).

Polyethylene glycol stearates (synonyms including macrogolstearates), polyoxyethylene stearates, ethoxylated stearates; CAS nos. 9004-99-3, 9005-08-7) are mixtures of mono and distearates of mixed polyoxyethylene polymers. Polyethylene glycol hydroxystearate is a mixture of mono-and diesters of hydroxystearic acid with polyethylene glycol. One polyethylene glycol hydroxystearate that may be used in the composition is polyethylene glycol 12-hydroxystearate. In another embodiment, the compositions of the present invention may comprise the surfactant polyethylene glycol 1512-hydroxystearate (from BASF)15) Mixtures of monoesters and diesters of 12-hydroxystearic acid with 15 moles of ethylene oxide.

Again, these compounds and their amounts are well known in the art.

In another embodiment of the invention, the composition of the invention may comprise a polyoxyethylene 35 castor oilAs a surfactant. In other embodiments, the compositions of the present invention may comprise polyoxyethylene 40 hydrogenated castor oil(s) ((s))40) Or polyoxyethylene 60 hydrogenated castor oil as a surfactant. The compositions of the present invention may also include a combination of surfactants.

The compositions of the invention may contain other inert ingredients such as antioxidants, preservatives or pH stabilizers. These compounds are well known in the composition art.

Antioxidants such as vitamin E, alpha-tocopherol, ascorbic acid, ascorbyl palmitate, citric acid, fumaric acid, malic acid, sodium ascorbate, sodium metabisulfite, n-propyl gallate, BHA (butylated hydroxyanisole), BHT (butylated hydroxytoluene), BHA and citric acid, monothioglycerol, tert-butylhydroquinone (TBHQ), benzyl alcohol, and the like may be added to the compositions of the present invention.

Antioxidants are generally included in the compositions of the present invention in amounts of from about 0.01% to about 3%, or from about 0.01% to about 2% (w/v), based on the total weight (w/w) of the composition. In another embodiment, the composition comprises from about 0.05% to about 1.0% (w/w) of an antioxidant or a mixture thereof.

Preservatives such as benzyl alcohol are suitably employed in the composition in an amount of from about 0.01 to about 10.0%, with from about 0.05 to about 5.0% being particularly preferred. Other preservatives include parabens (methyl and/or propyl paraben), benzalkonium chloride, benzethonium chloride, benzoic acid, benzyl alcohol, bronopol, butyl paraben, cetyltrimethylammonium bromide, chlorhexidine, chlorobutanol, chlorocresol, cresol, ethyl paraben, imidurea, methyl paraben, phenol, phenoxyethanol, phenylethyl alcohol, phenylmercuric acetate, phenylmercuric borate, phenylmercuric nitrate, potassium sorbate, sodium benzoate, sodium propionate, sorbic acid, thimerosal, and the like.

Preferred ranges for these compounds include from about 0.01 to about 5%.

Benzyl alcohol is preferred.

Compounds that stabilize the pH of the composition may also be present. Likewise, these compounds and how to use them are well known to those skilled in the art. The buffer system comprises, for example, a system selected from: acetic acid/acetate, malic acid/malate, citric acid/citrate, tartaric acid/tartrate, lactic acid/lactate, phosphoric acid/phosphate, glycine/glycidate, tris, glutamic acid/glutamate and sodium carbonate, especially sodium phosphate or citrate.

The aqueous suspension may comprise particles of the isoxazoline compound and moxidectin microspheres as described herein mixed with excipients suitable for the manufacture of aqueous suspensions.

Such excipients include suspending agents, for example, sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia; dispersing or wetting agents include naturally-occurring phosphatides, for example lecithin, or condensation products of an alkylene oxide with fatty acids, for example polyoxyethylene stearate, or condensation products of ethylene oxide with long chain aliphatic alcohols, for example heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example polyethylene sorbitan monooleate.

The aqueous suspensions may also contain one or more preservatives, for example ethyl or n-propyl p-hydroxybenzoate.

Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water can provide isoxazoline compounds and moxidectin microspheres in admixture with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified by those already mentioned above.

In one embodiment, the isoxazoline compound is suspended in an aqueous suspension wherein the liquid carrier (diluent) is water.

In another embodiment, the liquid carrier (diluent) of the aqueous suspension comprises water and a cosolvent.

The co-solvent that can be used in the injectable composition of the present invention comprising an isoxazoline compound and a moxidectin microsphere can be a single co-solvent or a blend of co-solvents.

In one embodiment, the co-solvent used in the aqueous injectable composition of the present invention comprises a polar solvent miscible with water.

Non-limiting examples of such co-solvents include ethanol, isopropanol, benzyl alcohol, glycol ethers (e.g., including, but not limited to, diethylene glycol monoethyl ether (DGME, and DGME),Butyl diglycol, dipropylene glycol n-butyl ether, ethylene glycol monoethyl ether, ethylene glycol monomethyl ether,Dipropylene glycol monomethyl ether, propylene glycol monoethyl ether, etc.), liquid polyethylene glycols (PEGs) (e.g., PEG 400), propylene glycol, carbonates (e.g., propylene carbonate), 2-pyrrolidone, N-methylpyrrolidone, dimethyl isosorbide (DMI), dimethylacetamide, dimethylsulfoxide, glycerol formal, or a mixture of at least two of these solvents.

In one embodiment, the composition of the present invention comprises a polar protic solvent, including but not limited to an alcohol such as ethanol, isopropanol or a glycol or glycol ether. In yet another embodiment, the long acting injectable composition of the present invention comprises a polar aprotic solvent, such as N-methylpyrrolidone, dimethyl isosorbide, dimethylacetamide, dimethylsulfoxide, or propylene carbonate.

In one embodiment, the isoxazoline compound can exist in various isomeric forms. Reference to isoxazoline compounds always includes all possible isomeric forms of such compounds.

Unless otherwise indicated, a compound structure that does not indicate a particular conformation is intended to encompass compositions of all possible conformers of the compound, as well as compositions comprising less than all possible conformers. In some embodiments, the compound is a chiral compound.

The (S) enantiomer is particularly preferred. In some embodiments, the compound is an achiral compound.

In one embodiment, the isoxazoline compounds of formula (I) can be prepared according to, for example, patent applications US 2007/0066617, WO2007/079162, WO 2009/002809, WO 2009/080250, WO2010/070068, WO 2010/079077, 2011/075591 and WO 2011/124998 or any other method within the capabilities of the person skilled in the art as experts in chemical synthesis.

For the chemical preparation of the products of the invention, the person skilled in the art is to be considered as having, inter alia, the "chemical abstracts" and the literature cited therein in their entirety.

In one embodiment, the isoxazoline compound is suspended in the composition. In one embodiment, the suspension is aqueous. In an alternative embodiment, the suspension is non-aqueous.

In one embodiment, the pharmaceutical composition is substantially free of organic solvents.

In one embodiment, the pharmaceutical composition comprises a surfactant/wetting agent. In another embodiment, the surfactant/wetting agent is a poloxamer.

Alternatives to poloxamers are other water-soluble/miscible nonionic surfactants including sorbitan fatty acid esters (Spans), polyoxyethylene sorbitan fatty acid esters (polysorbates/Tweens), polyoxyethylene castor oil derivatives (Cremaphors), polyoxyethylene stearates, lecithin and TPGS (d- α -tocopheryl polyethylene glycol 1000 succinate).

The surfactant/wetting agent is present in the composition in an amount of about 0.01% w/v to about 0.5% w/v or about 0.05% w/v to about 0.1% w/v.

Poloxamers are nonionic triblock copolymers of polyoxyethylene (poly (ethylene oxide)) consisting of a central hydrophobic chain of polyoxypropylene (polypropylene oxide) flanked by two hydrophilic chains (see U.S. patent No. 3,740,421).

Poloxamer 124 is poly (ethylene glycol) -block-poly (propylene glycol) -block-poly (ethylene glycol), CAS No. 9003-11-6, also known as Lutrol L44 or Kollisolv P124.

Lutrol F68 is another poly (ethylene glycol) -block-poly (propylene glycol) -block-poly (ethylene glycol), also known as poloxamer 188 or Kolliphor P188.

In one embodiment, the pharmaceutical composition comprises a suspending agent. In one embodiment, the suspending agent is carboxymethylcellulose, especially sodium carboxymethylcellulose (NaCMC).

In an alternative embodiment, the suspending agent is methylcellulose or polyvinylpyrrolidone.

In one embodiment, the pharmaceutical composition comprises a preservative. In one embodiment, the preservative is benzyl alcohol.

In an alternative embodiment, the preservative is m-cresol, benzalkonium chloride, methyl paraben, or propyl paraben.

Injectable pharmaceutical compositions may be prepared by combining and mixing solid components and then suspending the solid mixture in a diluent.

A method of preparing an injectable pharmaceutical composition comprises combining isoxazoline particles with moxidectin microspheres to form a solid mixture, which is reconstituted with an aqueous liquid carrier in a subsequent step to form an aqueous suspension ready for injection.

In one embodiment, the aqueous liquid carrier is water.

In an alternative embodiment, the diluent is an oil or solvent having little or no solubility for the isoxazoline compound and moxidectin microsphere component.

The pharmaceutical composition further comprises a surfactant/wetting agent.

Specific surfactants/wetting agents and alternatives to surfactants/wetting agents are discussed in the specification and examples.

The pharmaceutical composition may further comprise additional excipients, such as suspending agents or preservatives.

Specific examples of suitable excipients and alternative agents are set forth in the description and examples discussed below in this application.

Particle size and measurement of isoxazoline compound particles and moxidectin microspheres

It has been found that the injectable compositions of the present invention comprising particles of isoxazoline compounds having a defined particle size have particularly beneficial properties.

In one embodiment, the moxidectin microsphere and the isoxazoline compound particle have the same particle size.

Thus, in this specification, reference to "isoxazoline compound particle size" includes reference to compositions in which the moxidectin microspheres have the same particle size and are measured by the same method.

In one embodiment, the isoxazoline compound and/or moxidectin microspheres have a D50 particle size distribution of about 25 microns to about 250 microns, a particle size of about 11 microns to about 250 microns, a particle size of about 50 microns to about 150 microns, a particle size of about 75 microns to about 130 microns, a particle size of about 90 microns to about 110 microns, a particle size of about 30 microns to about 100 microns as measured by static light scattering.

The particle size distribution describes the relative amount of particles present according to size.

D10 is the particle size distribution representing the size below which 10% of the particles are smaller.

D50 is the granulometry distribution representing the size below which 50% of the particles are smaller.

D90 is the granulometry distribution representing the size below which 90% of the particles are smaller.

In another embodiment, the particle sizes are different.

In one embodiment, the particle size of the isoxazoline compound and moxidectin microspheres are in similar ranges.

In another embodiment, the particle sizes are not in a similar range.

In a specific embodiment, the isoxazoline compound has a particle size D10 of greater than 10 μm, a particle size D50 of 80-120 μm, and a particle size D90 of less than 210 μm.

In a particular embodiment, for moxidectin microspheres, the D10 particle size is greater than 50 μm, the D50 particle size is from 100 to 150 μm, and the D90 particle size is less than 200 μm.

In a particular embodiment, the D10 for the particle size is about 10 μm, about 20 μm, about 30 μm, about 40 μm, about 50 μm, about 60 μm, or about 80 μm.

In a specific embodiment, the D10 for the particle size of the moxidectin microsphere is about 80 μm.

In a particular embodiment, the D50 for the particle size is about 50 μm, about 75 μm, about 80 μm, about 90 μm, about 100 μm, about 110 μm, about 120 μm, about 130 μm, about 140 μm, or about 150 μm.

In a specific embodiment, the D50 for the particle size of the moxidectin microsphere is about 110 μm.

In a particular embodiment, the D90 for the particle size is about 100 μm, about 130 μm, about 150 μm, about 175 μm, about 200 μm, or about 250 μm.

In one embodiment, the D10 for the particle size of the isoxazoline compound is about 20-35 μm, the D50 for the particle size is about 90-105 μm and the D90 for the particle size is about 155-175 μm.

In a specific embodiment, for moxidectin microspheres, D10 of particle size is about 60 to 85 μm, D50 of particle size is about 90 to 115 μm, and D90 of particle size is about 145 to 165 μm.

In a specific embodiment, for moxidectin microspheres, the particle size D10 is about 80 μm, the particle size D50 is about 110 μm, and the particle size D90 is about 150 μm.

In a specific embodiment, D10 for particle size is about 25 to 30 μm, D50 for particle size is about 95 to 100 μm, and D90 for particle size is about 160 to 170 μm.

In a specific embodiment, D10 for particle size is about 10 to 20 μm, D50 for particle size is about 85 to 110 μm, and D90 for particle size is about 170 to 185 μm.

In a specific embodiment, D10 for particle size is about 10 to 15 μm, D50 for particle size is about 95 to 105 μm, and D90 for particle size is about 175 to 180 μm.

In a specific embodiment, D10 for particle size is about 10 to 25 μm, D50 for particle size is about 40 to 60 μm, and D90 for particle size is about 95 to 100 μm.

In a specific embodiment, D10 for particle size is about 15 to 20 μm, D50 for particle size is about 45 to 55 μm, and D90 for particle size is about 90 to 95 μm.

In a specific embodiment, the D10 of the particle size is about 30 to 50 μm and the D50 of the particle size is about 70 to 130 μm.

In one embodiment, D10 for particle size is about 35 to 45 μm and D50 for particle size is about 90 to 110 μm.

In a specific embodiment, the D10 for the particle size is about 40 μm and the D50 for the particle size is about 100 μm.

Volume weighted particle size can be measured by sieving, microscopy or laser diffraction (Malvern or Sympatec).

Volume weighted particle size measurements can be made with a Malvern Mastersizer 2000 with Hydro 2000G measuring cell or with a Horiba LA-910 laser scattering particle size distribution analyzer. Volume weighted particle size can be measured by Sympatec Helos instrument.

For use in the present invention, the isoxazoline compound is present in the pharmaceutical composition according to the invention in an amount of from about 0.1 to about 50% w/v of the final pharmaceutical composition according to the invention.

The isoxazoline is present in an amount of about 10 to about 45% w/v; about 20 and about 45% w/v; about 15 and 35% w/v or about 25% w/v and about 35% w/v or about 1% w/v and about 12% w/v or about 3% w/v and about 9% w/v of the pharmaceutical composition of the present invention.

In an embodiment of the invention and/or embodiments thereof, the composition comprises eprinomectin as (a1) a physiologically active macrolide and flurarana, preferably (S) -flurarana, as (b) an isoxazoline compound of formula (I).

In an embodiment of the invention and/or embodiments thereof, the composition comprises milbemycin oxime as (a) a physiologically active macrolide and flurarana, preferably (S) -flurarana, as (b) an isoxazoline compound of formula (I).

In an embodiment of the invention and/or embodiments thereof, the composition comprises selamectin as (a) a physiologically active macrolide and frataxin, preferably (S) -frataxin, as (b) an isoxazoline compound of formula (I).

In an embodiment of the invention and/or embodiments thereof, the composition comprises moxidectin as (a) a physiologically active macrolide and afuravir, preferably (S) -fluroradine, as (b) an isoxazoline compound of formula (I).

In an embodiment of the invention and/or embodiments thereof, the composition comprises eprinomectin as (a) a physiologically active macrolide and afuram, preferably (S) -afuram, as (b) an isoxazoline compound of formula (I).

In an embodiment of the invention and/or embodiments thereof, the composition comprises milbemycin oxime as (a) a physiologically active macrolide and afuram, preferably (S) -afuram, as (b) an isoxazoline compound of formula (I).

In an embodiment of the invention and/or embodiments thereof, the composition comprises selamectin as (a) a physiologically active macrolide and afuravir, preferably (S) -afuravir, as (b) an isoxazoline compound of formula (I).

In an embodiment of the invention and/or embodiments thereof, the composition comprises moxidectin as (a) a physiologically active macrolide and afuravir, preferably (S) -afuravir, as (b) an isoxazoline compound of formula (I).

In an embodiment of the invention and/or embodiments thereof, the composition comprises eprinomectin as (a) a physiologically active macrolide and saraladine as (b) an isoxazoline compound of formula (I).

In an embodiment of the invention and/or embodiments thereof, the composition comprises milbemycin oxime as the physiologically active macrolide and saraladine as (b) the isoxazoline compound of formula (I).

In an embodiment of the invention and/or embodiments thereof, the composition comprises selamectin as a physiologically active macrolide and saraladine as (b) an isoxazoline compound of formula (I).

In an embodiment of the invention and/or embodiments thereof, the composition comprises moxidectin as the physiologically active macrolide and saraladine as (b) the isoxazoline compound of formula (I).

In an embodiment of the invention and/or embodiments thereof, the composition comprises eprinomectin as the physiologically active macrolide and loratadine as (b) the isoxazoline compound of formula (I).

In an embodiment of the invention and/or embodiments thereof, the composition comprises milbemycin oxime as the physiologically active macrolide and loratadine as (b) the isoxazoline compound of formula (I).

In an embodiment of the invention and/or embodiments thereof, the composition comprises selamectin as a physiologically active macrolide and loratadine as (b) an isoxazoline compound of formula (I).

In an embodiment of the invention and/or embodiments thereof, the composition comprises moxidectin as the physiologically active macrolide and loratadine as (b) the isoxazoline compound of formula (I).

Injectable compositions typically require sterilization prior to administration to an animal. In a preferred embodiment of the invention and/or embodiments thereof, the microspheres are sterilized, for example by irradiation with gamma radiation or electron beam.

Although it has been reported that physiologically active macrolides degrade upon irradiation and lose most of their biological activity, microspheres (a) can be sterilized by irradiation for injection without negatively affecting the stability of the active ingredient.

In one embodiment, the amount of isoxazoline compound in the pharmaceutical composition according to the invention is about 30% w/v of the pharmaceutical composition according to the invention.

In one embodiment, the amount of isoxazoline compound in the pharmaceutical composition according to the invention is about 7.5% w/v of the pharmaceutical composition according to the invention.

In one embodiment, the pharmaceutical composition must be reconstituted prior to injection. For example, the pharmaceutical compositions are reconstituted in an aqueous liquid carrier prior to injection.

In another embodiment, the pharmaceutical composition is a ready-to-use composition ready for injection.

In one embodiment, the pharmaceutical composition is administered in combination with an additional therapeutic agent.

The administration of the additional therapeutic agent may be in the same composition or in a separate composition.

The additional therapeutic agent may be a parasiticide or a vaccine.

In another embodiment, the additional therapeutic agent is another parasiticide. The other active ingredient is selected from isoxazoline compounds, macrolides, avermectins (e.g., ivermectin, selamectin, doramectin, avermectin, and eprinomectin); milbemycins (milbemycin oximes); benzimidazole precursors (pro-benzimidazoles) (e.g., febantel, netobimin, and thiophanate), benzimidazole derivatives (e.g., thiazole benzimidazole derivatives (e.g., thiabendazole and canandazole), carbamate benzimidazole derivatives (e.g., fenbendazole, albendazole (oxide), mebendazole, oxfendazole, parbendazole, oxibendazole, flubendazole, and triclabendazole), imidazothiazoles (e.g., levamisole and tetramisole), tetrahydropyrimidines (e.g., morantel and pyrantel), salicylanilides (e.g., closantel, hydroxychlorozamide, rafenide, and niclosamide), nitrophenolic compounds (e.g., nifenitrile and nitrothiocyanamide), benzenesulfonamides (e.g., clorsulone), pyrazinoisonols (e.g., praziquantel, and epothilones), heterocyclic compounds (e.g., piperazine) Diethylcarbamazine and phenothiazine); bischlorophenols, arsenides (e.g., thioacetamol, meroxamine and arsenamide); cyclooctadepsipeptides (e.g., emodepside); para-herquamides (e.g., derquantel); and aminoacetonitrile compounds (e.g., monatomic, AAD 1566); amidine compounds (e.g., amitalr and triphenidine), including all pharmaceutically acceptable forms, such as salts, solvates or N-oxides.

One embodiment of the present invention is a method of treating or preventing a parasitic infection in an animal comprising administering to an animal in need thereof an effective amount of the injectable pharmaceutical composition described above.

In one embodiment, the animal suffers minimal injection site irritation.

As noted above, minimal injection site stimulation means injection site stimulation of less than 2 x 2cm, which lasts for less than 2 to 3 days.

In one embodiment, the animal is a companion animal. In one embodiment, the companion animal is a dog or cat.

The optimum effective amount employed to achieve optimum results will, of course, depend upon the particular isoxazoline compound employed, the species of animal being treated and the type and severity of the parasitic infection or infestation.

When using the isoxazoline compounds of formula (I), good results are generally obtained when about 0.01 to 200mg/kg, in one embodiment 0.1 to 100mg/kg, or 0.5 to 50mg/kg or 1 to 30mg/kg of animal body weight is administered, such total dose being given once or in divided doses.

Good results are generally obtained when moxidectin is administered at about 0.01 to 10mg/kg, and in one embodiment 0.1 to 5mg/kg, of animal body weight, such total doses being administered once or in divided doses.

The injectable pharmaceutical composition may be administered daily, weekly, monthly, semi-annually or annually.

The injectable pharmaceutical composition may be administered monthly, bimonthly, every three months, every four months, every five months, every 6 months, every seven months, every eight months, every nine months, every ten months, every eleven months, every twelve months, every 13 months, every 14 months, every 15 months, every 16 months, every 17 months, or every 18 months.

Administration every 6 months is particularly preferred.

It is also preferred to administer once every 12 months. This provides long-term protection of the animal from ectoparasites (especially fleas and ticks) and endoparasites (especially heartworms and/or gastrointestinal worms). Particularly preferred is long-term protection against heartworm infestation.

The injectable composition of the invention can advantageously be used together with annual vaccination against infectious diseases such as distemper, influenza, rabies and other vaccines with conventional antigens.

In one embodiment of the injectable pharmaceutical composition, the isoxazoline compound and/or moxidectin microsphere has a particle size D50 of about 75 microns to about 130 microns and a particle size D10 of about 30 microns to about 100 microns.

One embodiment of the invention is a kit for treating or preventing parasite infestation in an animal, the kit comprising two or more containers:

a) solid crystalline isoxazoline compounds and moxidectin microspheres;

b) a vehicle comprising a pharmaceutically acceptable excipient capable of forming a suspension with the compound of a); and

c) instructions for combining solid crystalline isoxazoline compounds and moxidectin microspheres with an aqueous liquid carrier prior to injection.

Wherein for solid crystalline isoxazoline compounds and moxidectin microspheres, the D50 particle size is from about 75 microns to about 130 microns and the D10 particle size is from about 30 microns to about 50 microns.

In another embodiment, the isoxazoline compound is flurandrine.

One embodiment of the present invention is a kit, wherein the kit comprises:

a) a first container comprising a solid mixture of particles of an isoxazoline compound of formula (I) as described in claims 1, 8, 9, 10, 11, 12, 17 and 18 and moxidectin microspheres as described in claims 1 to 12, and;

b) a second container having an aqueous carrier comprising one or more suspending agents, wetting agents and/or preservatives and water; and

c) instructions for reconstituting the moxidectin microspheres and isoxazoline compound particles with an aqueous vehicle prior to subcutaneous or intramuscular injection to an animal.

In one embodiment, the first container comprises an effective amount of moxidectin and an isoxazoline compound of formula (I) as described above sufficient to treat and/or prevent a parasitic infestation of an animal.

In one embodiment, the kit further comprises means for reconstituting and administering parenterally (by injection) a mixture of the compositions from the first and second containers to the animal, particularly using a syringe (e.g., No. 18).

In one embodiment, in the first container, the isoxazoline compound of formula (I) and the moxidectin microspheres have a volume weighted particle size distribution D50 of about 25 microns to about 250 microns as measured by static light scattering instrumentation.

In another embodiment, in the first vessel, the isoxazoline compound has a particle size D10 of about 20-35mm, a particle size D50 of about 90-105mm and a particle size D90 of about 155-175 mm.

Another aspect of the invention is a method of treating or preventing a parasitic infection in an animal comprising administering to the animal in need thereof an injectable veterinary composition.

Another aspect of the invention is a method for preparing an injectable veterinary composition according to the invention, comprising the following steps:

a) preparing particles of the isoxazoline compound by crystallization;

b) preparing moxidectin microspheres by melting a fat, wax or mixture thereof and adding moxidectin and optionally an antioxidant, and preparing the microspheres by rotary disk atomization and sieving;

c) filling the moxidectin microspheres obtained by the step b) together with the isoxazoline particles obtained by the step a) into a first container;

d) preparing an aqueous carrier by dissolving excipients including a suspending agent, a wetting agent and/or a preservative in water and filling into a second container;

e) reconstituting the solid by transferring the aqueous carrier from the second container d) to the first container c) and shaking to form a.

Rotating discs are considered a production technique that produces uniform spherical particles with a low span of particle size distribution by controlling key process parameters such as melt temperature, flow rate and disc velocity.

The rotating disk method is a packaging technique that uses mechanical energy to pressurize a liquid film or increase its kinetic energy to break up, possibly in the form of droplets.

Alternatively, the moxidectin may be uniformly incorporated into the fat, wax or mixture thereof using other methods such as hot melt extrusion, hot melt granulation, film evaporation, and the like. The resulting mixture may be milled or sprayed to achieve the desired particle size.

If desired, sieving of the material can be performed to produce batches with defined particle sizes.

In other words, the microspheres (a) can be considered as microspheres prepared by the steps of: the physiologically active macrolide is incorporated and microspheres of the resulting mixture are then formed by various techniques as described above.

Alternatively, the mixture of physiologically active macrolide and optionally other excipients may be cooled to give a solid, which may then be processed by procedures such as milling, grinding and the like.

In general, solvent evaporation, rotary disc atomization, spray drying and sieving are methods known to those skilled in the art.

In one embodiment of the present invention and/or embodiments thereof, the mixture in the first container comprises an effective amount of microspheres as defined in any one of claims 1 to 14 and particles of an isoxazoline compound of formula (I) as defined in any one of claims 1 to 14 and/or a compound of formula (II) as defined in any one of claims 1 to 14 sufficient to treat or prevent a parasitic infestation of an animal.

In one embodiment of the invention and/or embodiments thereof, the kit further comprises a device, in particular a syringe, for reconstituting and parenterally administering the mixture of compositions from the first and second containers to the animal.

Another embodiment is a method of treating and/or preventing a parasitic infestation in an animal over an extended period of 6 months, or alternatively 12 months, the method comprising administering to an animal in need thereof a reconstituted liquid prepared when using a kit as described above and administering it to the animal by injection according to instructions.

The parasites are ectoparasites and endoparasites as described previously.

Preferred target animals are companion animals, such as cats or dogs, in particular dogs.

The optimum effective amount employed to achieve optimum results will, of course, depend upon the particular isoxazoline compound employed as well as the physiologically active macrolide, the animal species to be treated and the type and severity of the parasitic infection or infestation.

In a preferred embodiment of the present invention and/or embodiments thereof, the isoxazoline compound of formula (I), preferably fluridone, is administered at about 0.01 to about 200mg/kg animal body weight, preferably about 0.1 to about 100mg/kg animal body weight, more preferably about 0.5 to about 50mg/kg animal body weight, especially about 1 to about 30mg/kg animal body weight. The total dose may be administered in a single dose or in divided doses.

In a preferred embodiment of the present invention and/or embodiments thereof, the physiologically active macrolide, preferably moxidectin, is administered at about 0.01 to about 10mg/kg, preferably about 0.1 to about 5mg/kg, of animal body weight. The total dose may be administered in a single dose or in divided doses.

The results show that when the injectable veterinary composition of the invention is used for the treatment and/or prevention of parasitic infestation in animals, the treated animals are subjected to minimal injection site irritation.

As noted above, minimal injection site stimulation means injection site stimulation of less than 2 x 2cm, which lasts for less than 2 to 3 days.

Another aspect of the invention is a method for the treatment and/or prevention of a parasitic infection in an animal comprising administering to a subject in need thereof a therapeutically effective amount of an injectable veterinary composition according to the invention or a kit according to the invention.

Likewise, the same applies with respect to injectable veterinary compositions and kits. The same applies to parasites and parasite infestations.

The features of the present invention have been described in the embodiments of the present application; however, for the sake of brevity, not all combinations of features are literally described.

However, combinations of features as described above are explicitly considered to be part of the present invention.

Examples

Example 1: preparation of 10% moxidectin microsphere

10% moxidectin tristearate (GTS) microspheres were prepared by spinning the discs and formulated as follows:

table 1: example formulation of 10% moxidectin in Glycerol tristearate microspheres

Component (A)	％w/w
		Moxidectin	10.00
Tristearin	89.97
		Butylated hydroxytoluene	0.03

Briefly, 180g of glyceryl tristearate was melted in a vessel and heated to a temperature of 180 ℃ with stirring. 20g of moxidectin and 0.06g of butylated hydroxymethylbenzene were added and stirred until dissolved. The resulting molten solution was cooled to-80 ℃ and pumped onto a 4 "pan heated to-90 ℃ at 3000 rpm. The resulting microspheres were screened and less than 150 μm of material was collected for further characterization and study.

Example 2

Sterilization of 10% moxidectin in GTS microspheres-stability of microspheres to irradiation

The microsphere compositions listed below were placed in 20mL serum vials, two of which were flushed with dry nitrogen to remove oxygen. The vial was then closed with an elastomeric septum and crimped aluminum cap. Next, the microspheres were irradiated to 15, 20, and 25kGy by both gamma irradiation and electron beam to be sterilized. The microspheres were extracted into acetonitrile/water (1: 1) and analyzed by high performance liquid chromatography for 23- (O-methyloxime) -F28249 a. The results of this experiment are summarized in table 2 below. GTS microspheres were sterilized at low or room temperature with or without a nitrogen blanket.

The samples were evaluated for assay variation. The% was determined as% of non-irradiated measurements.

Table 2: effect of irradiation type, irradiation dose, temperature and Nitrogen blanketing on Moxidectin determination during Sterilization

Example 3: preparation of liquid aqueous vehicle

About 50% of the water for injection is filled into a container and heated to about 70-80 ℃, and the suspending agent NaCMC, hypromellose E50 or PVP is added and homogenized until dissolved. The other ingredients were added slowly and mixed with stirring to achieve dispersion. The heat was removed and cold water for injection was added to bring the volume to 10 liters. In examples 3B and 3D, the pH was adjusted to 4.5-5.5 by addition of HCl.

Each vehicle was sterilized by autoclave and the vehicle solution was stored in a sterile container.

Dose uniformity of ready-to-use injectable suspension products

Although the moxidectin microspheres and the frataxin particles exhibit similar particle sizes, they differ in density. Thus, many different vehicles were tested to find a vehicle suitable for uniform resuspension and administration.

Samples containing 15% frataxin and 1.7% moxidectin GTS microspheres were shaken by hand until visually dispersed in each vehicle for-30 seconds.

The following vehicles were studied:

example formulations of viscous aqueous vehicles for reconstitution/resuspension:

example 3A

Example 3B

Example 3C

Example 3D

Example 3E

Then 61 mL samples were withdrawn and assayed for moxidectin and frataxin by experiment, and the results are shown in table 4 below:

TABLE 4

Example 4: preparation and use of the final formulation

In use, the vehicle prepared in example 3 was added to the moxidectin microspheres and crystalline frataxin particles prepared in example 1, and the container was shaken to disperse the microspheres and frataxin particles in the liquid vehicle. The formulation was then drawn into a syringe and injected subcutaneously in the dose volume specified for the body weight of the dog to be treated.

Example 5

Pharmacokinetic evaluation of injectable formulations comprising GTS moxidectin microspheres and flurarana particles

Preparation:

a.15% freirana + 0.17% moxidectin microspheres (unscreened, GTS, 25kGy)

B.15% Fluralazine + 0.17% moxidectin microspheres (unscreened, GTS, 15kGy)

C.15% Fluraniana + 0.17% moxidectin microsphere (D50 ═ 75 μm, GTS, 25kGy)

D.15% Flurania sodium + 0.17% moxidectin microsphere (D50 ═ 150 μm, GTS, 25kGy)

The formulation was prepared using the following method:

A. media

1. Injecting water accounting for 80 percent of the total volume.

2. The suspending agent (sodium carboxymethylcellulose (CMCNa)) was added and mixed for about 5 minutes with an overhead mixer.

3. The mixture was further mixed with a homogenizer until free of agglomerates.

4. Add the wetting agent (poloxamer 124) and mix with an overhead mixer until homogeneous.

5. Add the preservative (benzyl alcohol (BA)) and mix with an overhead mixer until homogeneous.

6. Sodium phosphate was added and mixed with an overhead mixer until homogeneous.

7. The antifoam (dimethicone) was gently mixed with an overhead mixer until homogeneous (5 minutes).

8. The pH of the mixture was adjusted to pH 7.0-7.4 by addition of HCl. Mix gently with an overhead mixer until homogeneous (5 min).

9. Water was added to the final weight for injection and gently mixed with an overhead mixer until homogeneous (5 minutes).

10. The resulting formulation was packaged into injectable vials and sealed with stoppers.

11. The vials were autoclaved at 121 ℃ for 15 minute cycles.

B. Active ingredient

1. Solid fluranide and moxidectin GTS microspheres (prepared as described above) were added to the vials and sealed.

2. The vials were terminally sterilized by gamma radiation.

C. Formation of reconstituted injectable formulations

1. The vial of a vehicle was added to the vial of B active ingredient and shaken.

2. The resulting suspension is ready for injection.

Similar procedures were used to prepare the formulations of examples 5B-H. Batch sizes ranged from 50mL to 1000 mL.

Vehicle for all formulations:

a sample of formulation A, B, C or D was administered subcutaneously in a single dose to different groups of eight beagle dogs each at 0.1mL/kg BW (i.e., 15mg fluranide/kg BW, 0.17mg moxidectin/kg BW).

The formulations were evaluated for local tolerability for at least 21 days after administration.

Blood samples for determination of moxidectin and total fluoride rana plasma concentrations were collected before treatment, 8 hours after treatment and 1, 3,5, 7, 10, 14, 21, 28, 35, 42, 49, 56, 70, 84, 98, 112, 126, 140, 154, 168 and 182 days.

For all formulations tested, favorable pharmacokinetic profiles were obtained showing prolonged plasma levels of moxidectin and loratidine in dogs after sc administration.

There was no significant injection reaction during the evaluation of the formulation of example 5.