RU2578036C2 - New injectable compositions of eprinomectin - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to veterinary medicine and concerns an injectable parasiticide composition applicable for controlling ecto- and endoparasites contaminating warm-blood domestic animals and containing a therapeutically effective amount of eprinomectin, polyethylene glycol, benzyl alcohol and N-methylpyrrolidone.

EFFECT: invention provides improved bioavailability of the active substance and provides prolonging the time of parasiticide residence in the mammals different from a human.

14 cl, 4 ex, 10 tbl

Description

FIELD OF THE INVENTION

The present invention relates to new compositions suitable in the field of veterinary medicine, and, more specifically, in relation to antiparasitic compositions containing macrocyclic lactones, like eprinomectin, with improved bioavailability and long-term therapeutic effects.

BACKGROUND OF THE INVENTION

Warm-blooded animals are attacked by parasites, and humans have been trying for a long time to combat such parasites that infect companion animals, farm animals, and exotic animals to alleviate suffering as well as commercial gain. Internal and external parasites, such as gastrointestinal worms and lice, are organisms that feed on host animal tissues, blood and tissue fluids, and they can adversely affect performance. Internal parasites can cause appetite suppression, reduced feed digestibility and absorption of nutrients, blood loss and anemia, which, in turn, can lead to reduced weight gain and milk production, weakening of the immune system, and damage to tissues and organs. External parasites can cause hair loss and scabs, blood loss and anemia, skin irritation, and can also act as carriers of diseases. Impacts of external parasites on productivity may include reduced weight gain and milk production, skin damage and damage to facilities and fences due to friction and combing.

Macrocyclic lactones, such as, for example, avermectins and milbemycins, are very potent antiparasitic agents at a low dose and are applicable against a wide range of mammalian endoparasites and ectoparasites, and also have agricultural applications against various parasites found in and on crops and soils. They are active against many immature nematodes (including hypobiont larvae) and arthropods. In addition, a single therapeutic dose may be maintained in concentrations sufficient to be effective against existing nematode infections for extended periods after treatment.

Commercial avermectins are ivermectin, abamectin, doramectin, eprinomectin and selamectin. Commercially available milbemycins are milbemycin oxime and moxidectin. The basic avermectin compounds are isolated from the enzymatic broth of the soil microorganism Streptomyces avermitilis, and these compounds are described in US patent No. 4310519. In addition, derivatives of these basic compounds of avermectin were obtained using a number of chemical agents.

In particular, eprinomectin, also called 4'-epi-acetylamino-4'-deoxyavermectin B1, is a new semi-synthetic derivative of the avermectin family and is characterized by the following structure:

Eprinomectin is a mixture of two homologs, Eprinomectin B1a (90%) and Eprinomectin B1b (10%), the difference between which is the presence of a methylene group at position C-25. These structures have a wide spectrum of activity against nematodes and arthropods and, due to their effectiveness against both endo- and ectoparasites, they are called endectocides.

The pharmaceutical activity of these molecules increases the permeability of the muscles and nerve cells of parasites to chlorine ions, thereby causing parasite paralysis and death. These molecules bind to the glutamate-dependent channels for chlorine ions of invertebrate cells. They can also bind to other GABA-dependent channels for chlorine ions. Since mammals do not possess this type of glutamate-dependent channel for chlorine ions, these molecules provide a high degree of safety, even if used at high doses.

Macrocyclic lactones are generally lipophilic compounds with sufficiently high distribution coefficients or log P values; for example, these values are about 6 for moxidectin, about 5.6 for doramectin and about 4.8 for ivermectin, with the exception of eprinomectin, which has the lowest log P value of about 4.4 (see example 2 and Kaliszan R. and al., Pure Appl. Chem., Vol. 73, No. 9, p. 1465-1475, IUPAC 2001). Thus, they are generally well absorbed when administered orally, parenterally or in the form of formulations for the application of a limited volume of preparation. They are widely distributed throughout the body, including the gastrointestinal tract, lungs and skin, and are especially concentrated in adipose tissue, having a long residence time and half-life (T _1/2 ) in animal tissues.

The residence time and half-life of these macrocyclic lactones, however, can be affected by the state of the animal’s body, the route of administration, and also by composition and / or solvents. For example, it is known that oil formulations have a longer residence time than aqueous formulations. Also parenteral and topical formulations, as a rule, have a better residence time than oral formulations. For example, the residence time of macrocyclic lactones when administered as formulations for applying a limited volume of the preparation may be approximately 12.8 days for moxidectin and doramectin, approximately 8.4 days for ivermectin, and approximately 4.16 days for eprinomectin.

Eprinomectin-based products currently available on the market include formulations for external use, such as formulations for applying a limited volume of preparation containing a 0.5% w / v solution of eprinomectin that can be applied to the skin of an animal. Permission has been obtained to enter the market of eprinomectin for topical administration by applying a limited volume of the drug, and is currently on the market by Merial under the trademark Eprinex ^® for the treatment and / or prevention of damage by roundworms, pulmonary nematodes, vermiform larvae, lice, ticks and small cow livers in cattle. A limited volume formulation of Eprinex ^® currently comprises 5 mg of eprinomectin, 0.10 mg of butylhydroxytoluene, 0.06 mg of vitamin E and excipients.

Although methods for applying a limited volume of the preparation have several advantages in terms of the safety of the specialist in applying pesticides, they are also significantly affected by several factors that can reduce their effectiveness due to inaccurate dosage.

The effectiveness of the absorption or adsorption of the agent is usually affected by the skin conditions of the animal such as the presence of fur, cuts, wounds, burns, as well as the cleanliness of the skin of animals (dust, dirt, etc.). Also, environmental factors, including climatic conditions like rains, solar radiation, etc., can affect the efficiency of adsorption.

All these adverse conditions cause a forced more than twofold excess of the required amount of funds in order to combat the parasite. The requirement for this additional addition of eprinomectin has economic implications regarding the cost of the treated animal, it can have a negative impact on the environment, and additionally can lead to potential side effects resulting from the use of high doses of the drug.

Eprinomectin has been proposed for use as subcutaneous or intramuscular injection formulations. In fact, in addition to its endectocidal activity, eprinomectin has not been shown to be excreted with milk fat in productive animals. In studies, they came to the conclusion that only 0.1% of the applied product is excreted in milk, that is, 50 times less in comparison with other well-known macrocyclic lactones, such as, for example, ivermectin or moxidectin.

Although eprinomectin could theoretically be the preferred macrocyclic lactone, especially for treating dairy cattle, it has weak lipophilic properties, a short residence time and a shorter duration of effect or shorter duration of action compared to other macrocyclic lactones.

Thus, there is a clear need to improve bioavailability and increase the residence time or half-life of an eprinomectin-based agent in the animal’s body, as well as providing methods for administering agents with low toxicity and high tolerance. Also, when choosing media for the composition, the specialist should consider a number of issues, including the solubility of eprinomectin, the affinity of the agent for certain media, whether it will affect the main excipients, pH, stability over time, viscosity and, naturally, the risk of any toxic effect on the animals being treated but still keep a very low content of residues in milk. Thus, the preparation of this parasiticide is a complex task, and not ordinary experimentation.

In this regard, US patent No. 5773422 describes compositions for parenteral or topical administration of avermectin based on the use of N-methylpyrrolidone or 2-pyrrolidone or mixtures thereof to dissolve avermectin, such as ivermectin. Also, European application No. EP 873127 describes macrocyclic lactone-based injectable anthelmintic compositions containing macrocyclic lactone, vegetable oil and aromatic alcohol having four or more carbon atoms as a solvent. Preferred aromatic solvent alcohols include benzyl alcohol, ethylbenzyl alcohol, phenethyl alcohol and other aromatic monohydric alcohols. Also preferred vegetable oils are soybean oil, sesame oil and corn oil. It is disclosed that the use of these specific formulations enables the antihelminthic agent to remain unchanged on external and internal parasites. However, none of these previously known formulations has solved the problem of increasing the residence time and bioavailability of the agent in treated non-human mammals.

Although polyethylene glycol (PEG) has never been described as a substance for increasing the bioavailability and residence time of an agent, it was unexpectedly discovered in the present invention that incorporating an effective amount of PEG had a significant effect on the bioavailability of eprinomectin in parasiticidal formulations. In injectable formulations, therefore, a lower proportion of eprinomectin was required. The new eprinomectin compositions of the present invention also unexpectedly showed a longer lasting effect and greater efficacy, especially with respect to the strongilides of the gastrointestinal tract and ectoparasites in ruminants and dairy cattle. In addition, the compositions of eprinomectin gave a stable plasma concentration after administration, which led to greater overall effectiveness throughout the herd of cattle, combined with a lower risk of inducing resistance to the active agent.

SUMMARY OF THE INVENTION

Accordingly, it is an object of the present invention to provide an improved long-acting veterinary injection composition comprising a therapeutically effective amount of eprinomectin in a solvent system comprising polyethylene glycol (PEG) and optionally other solvents and excipients.

In particular, it is an object of the invention to provide a veterinary injection composition containing a therapeutically effective amount of eprinomectin and PEG as a solvent to control a wide range of endoparasites and ectoparasites in non-human mammals.

An additional objective of the invention is the provision of a veterinary injection composition containing a therapeutically effective amount of eprinomectin and PEG as a solvent for the treatment and / or prevention of infections caused by endoparasites and / or ectoparasites in non-human mammals, especially dairy cattle. The compositions of the present invention are particularly effective for controlling the strongilides of the digestive tract and lungs.

Another additional objective of the invention is the provision of a veterinary injection composition containing a therapeutically effective amount of eprinomectin and PEG, where the active eprinomectin compound has improved bioavailability and a longer half-life (T _1/2 ) compared to known macrocyclic lactones, such as ivermectin, doramectin and moxidectin, and compared with the compositions of eprinomectin for applying a limited volume of the drug.

The main objective of the invention is to provide a veterinary composition containing a therapeutically effective amount of eprinomectin and PEG as a solvent for parenteral administration to bovine, more specifically lactating cows in such a way that there are less residues of macrocyclic lactone, especially eprinomectin, in milk or milk, compared to using eprinomectin to apply a limited amount of the drug.

An alternative embodiment of the present invention provides an injectable parasiticidal composition prepared for treating animals containing a weakly lipophilic macrocyclic lactone, i.e. having a distribution coefficient or log P of less than 5, preferably from about 4 to about 5, and most preferably about 4.4 or about 4.5 in a physiologically acceptable injectable solvent system, wherein the composition comprises polyethylene glycol that is effective To improve bioavailability, and provides long-lasting effect or longer duration of action less lipophilic macrocyclic lactone.

BRIEF DESCRIPTION OF GRAPHIC MATERIALS

Figure 1 is a graph showing on a linear scale the plasma concentration profiles of eprinomectin (ng / ml) based on the mean and standard deviation obtained after a single subcutaneous injection of 200 μg / kg EPRINOJECT and a single application of a limited volume of the drug to cows 500 mcg / kg EPRINEX.

Figure 2 is a graph showing, on a logarithmic scale, the plasma concentration profiles of eprinomectin (ng / ml) based on the mean and standard deviation obtained after a single subcutaneous injection of 200 μg / kg EPRINOJECT and a single application of a limited volume of the drug to cows 500 mcg / kg EPRINEX.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to an improved long-acting veterinary injection composition comprising a therapeutically effective amount of eprinomectin in a solvent system containing polyethylene glycol (PEG) and optionally other solvents and excipients.

It was unexpectedly found that although eprinomectin is weakly lipophilic compared to other macrocyclic compounds (i.e., moxidectin, doramectin or ivermectin), injectable eprinomectin-based formulations can be prepared with an effective amount of PEG solvent, and these formulations possess what was previously not known reported improved properties with respect to residence time and, therefore, a longer effect / longer duration of action compared to other macrocyclic compounds. Additionally, the eprinomectin compositions of the present invention showed excellent bioavailability, improved dissolution properties of eprinomectin prior to administration, as well as faster delivery of eprinomectin to target sites.

The new compositions of the present invention contain from about 0.1% to about 10%, preferably from about 1% to about 5%, most preferably about 1% of eprinomectin. Eprinomectin will usually be completely dissolved in the composition. The composition according to the invention may contain at least 20% PEG by weight. The composition according to the present invention may also include from 20 to 98% PEG by weight, preferably from about 40% to 95% and most preferably from about 93% PEG by weight.

A number of PEG solvents are commercially available in accordance with the molecular weight, and any or others that may still be available may be selected for convenience provided that the PEG is present or is available as a liquid at the time of preparation. Typically, PEG 200-1500 can be used for the purposes described herein, preferably a low molecular weight PEG 200-600 can be effectively used in the present invention. A preferred solvent system includes PEG-200. PEG for use in the present invention is commercially available from various sources, such as, for example, Sigma Chemical Co. or Fisher Scientific Co.

In a most preferred embodiment, the injectable veterinary composition of eprinomectin according to the present invention comprises eprinomectin, PEG, N-methylpyrrolidone and benzyl alcohol.

Additionally, a pH adjuster can be used to adjust the pH in the range of from about 5.5 to about 9.5. Examples of pH adjusters for use in the present invention include citric acid, acetate, phosphoric acid, ascorbic acid, gluconic acid, succinic acid, tartaric acid, lactic acid and the like, their salts and mixtures thereof.

The invention further relates to a veterinary injection composition containing a therapeutically effective amount of eprinomectin and PEG as a solvent, optionally together with other solvents and excipients, for treating and / or preventing infections caused by endoparasites and / or ectoparasites in non-human mammals.

In a preferred embodiment, the present invention relates to a veterinary injection composition comprising a therapeutically effective amount of eprinomectin and a solvent system comprising PEG, N-methylpyrrolidone and benzyl alcohol for the treatment and / or prevention of infections caused by endoparasites and / or ectoparasites in non-human mammals .

The injection veterinary composition according to the present invention may contain a therapeutically effective amount of eprinomectin from about 0.5% to about 5%, benzyl alcohol in an amount from about 0.5% to about 5%, N-methylpyrrolidone in an amount from about 0.5% to approximately 5% and is composed in polyethylene glycol. Preferably, the injectable veterinary composition contains eprinomectin in an amount of from about 0.1% to about 1%, benzyl alcohol in an amount of from about 0.1% to about 1%, N-methylpyrrolidone in an amount of from about 0.5% to about 5% and is composed in polyethylene glycol.

Injectable veterinary eprinomectin may contain, in addition to PEG, N-methylpyrrolidone and benzyl alcohol, one or more pharmaceutically acceptable components that are suitable for use in non-human mammals without undue side effects (such as toxicity, irritation and allergic reaction), as appropriate benefit / risk ratio.

For example, the composition may include a liquid excipient, or a pharmaceutically acceptable solvent, or a combination of solvents or cosolvents selected from the group consisting of pyrrolidone, propylene glycol, formalglycerol, isosorbide dimethyl ether, ethanol, dimethyl sulfoxide, glycerol, triethylene glycol, tetrahydrofurfurfuryl, .

The standard parameter of lipophilicity, namely the distribution coefficient or log P, provides a thermodynamic measure of the hydrophilic-lipophilic balance of a chemical compound in a system containing two immiscible liquids, including an aqueous phase and an organic phase. Once the compound reaches complete equilibrium with the two solvents, the distribution coefficient can be calculated by dividing the amount of the compound present in the organic phase by the amount of the compound present in the aqueous phase and taking logarithms. As discussed above, eprinomectin has a low log P value of approximately 4.4 compared to other known macrocyclic lactones and, therefore, has a short residence time or half-life after injection to non-human mammals.

As will become apparent from the following examples, an unexpectedly longer half-life of approximately 12 days was obtained by subcutaneous injection of the eprinomectin composition of the present invention.

By “injection composition” or “injection composition” is meant a composition or composition that can be drawn into a syringe and injected subcutaneously, intravenously, intraperitoneally or intramuscularly to a non-human mammal without causing side effects. According to a preferred embodiment of the present invention, the veterinary composition of eprinomectin is administered to non-human mammals by subcutaneous injection.

As used herein, “bioavailability” means the degree or rate at which an active substance is absorbed into a living system or becomes available in a site with physiological activity. As for the active substances that must be absorbed into the bloodstream, the bioavailability data for a given composition can provide an estimate of the relative fraction of the administered dose that is absorbed into the systemic circulation. “Bioavailability” can be characterized by one or more pharmacokinetic parameters.

"Pharmacokinetic parameters" describe the in vivo characteristics of the active substance over time, such as plasma concentration (C), C _max , C _n , C ₂₄ , T _max and AUC. “C _max ” is the measured plasma concentration of the active substance at the point of maximum or peak concentration. "C _min " is the measured concentration of the active substance in plasma at the point of minimum concentration. "AUC" is the area under the curve of the graph measured by the concentration of the active substance in the plasma, measured from one point in time to another. For example, AUC _0-t is the area under the curve of plasma concentration versus time in the time range from time 0 to time t, where t can be the last time with a measurable plasma concentration for a single composition. AUC _0-∞ or AUC _0-INF is the estimated area under the curve of plasma concentration versus time in the time range from time 0 to infinity. In inpatient studies, AUC _0-τ represents the area under the plasma concentration curve in the dosing interval (i.e., in the time range from time 0 to time τ (tau), where tau represents the length of the dosing interval). Other pharmacokinetic parameters are the parameter K _e or K _el , the constant of the final elimination rate, which is calculated from the semilogarithmic graph of the curve of plasma concentration versus time; T _1/2 represents the final elimination half-life, calculated as 0.693 / K _el ; CL / F stands for apparent total clearance after administration, calculated as the ratio of total dose / total AUC _∞ ; and V _area / F means the apparent total distribution after administration, calculated as the ratio of total dose / (total AUC _∞ × K _el ).

As used herein, “T _max ” refers to the observed time (day) to reach the maximum concentration of the eprinomectin in the animal plasma after injection to non-human mammals.

As used herein, “peak plasma concentration (C _max ) of eprinomectin,” “area under the curve (AUC) of eprinomectin in blood plasma,” “half-life of eprinomectin (T _1/2 ) in plasma” are pharmacokinetic parameters, known to those skilled in the art (Pharmaceutical Research, Volume 8, Number 4 / April, 1991 Laursen et al., Eur. J. Endocrinology, 135: 309-315, 1996).

"Area under the curve (AUC)" measures the concentration of eprinomectin in the serum of a subject over time after administering a dosage of eprinomectin to a subject by any injection route. "C _max " represents the maximum concentration of eprinomectin in the serum of a subject after a single dosage of eprinomectin.

As used herein, “approximately” will be construed by one of skill in the art and will vary to some extent depending on the context in which it is used. If there are applications of this value that are not clear to those skilled in the art, taking into account the context in which it is applied, then “approximately” will mean up to plus or minus 10% of the specific value.

In the formula according to the present invention, the PEG-associated epinomectin agent is characterized by improved bioavailability and extended residence time, which results in slow release and long-term effect of the agent. Thus, the antiparasitic effect of the drug lasts for a longer period due to its presence in the blood. Additionally, it was found that there is less variation in plasma concentrations of eprinomectin, which translates into better efficacy in the herd, as well as a lower risk of resistance induction.

The veterinary composition of eprinomectin for parenteral administration to non-human mammals delivers less than about 0.5 mg / kg of eprinomectin and preferably less than about 0.4 mg / kg or from about 0.1 to about 0.4 mg / kg. Preferably, the dose of eprinomectin in the composition is in the range from about 0.1-0.3 mg / kg to about 0.3 mg / kg and is about 0.2 mg / kg. The required dosage of the injectable veterinary composition is thus lower than the dosage for existing formulations for applying a limited volume of preparation. Indeed, as indicated above, the recommended subcutaneous dose of eprinomectin is approximately 0.2 mg / kg. In one embodiment, such dosages are administered to the animal only once (single administration) for a period of 40 days, without any other doses of eprinomectin being given during this period. This is less compared to days 14, 21, 28 for formulations for applying a limited volume of the preparation or subcutaneous formulations of eprinomectin and ivermectin.

The injectable veterinary compositions of eprinomectin according to the present invention show an increase in peak plasma concentration of eprinomectin (C _max ) and an increase in the area under the plasma concentration curve (AUC) over time and an additionally excellent half-life (T _1/2 ) compared to existing formulations basis of macrocyclic lactones. Increasing C _max and AUC leads to greater efficacy of the macrocyclic lactone composition of the present invention and makes it possible to use lower administration doses, which may still be effective in removing parasites.

In a preferred embodiment, the methods and compositions of the present invention are characterized by (i) a peak concentration (C _max ) of eprinomectin in the blood plasma of said non-human mammals from about 45 to about 75 ng / ml or from about 45 to about 60 ng / ml and preferably approximately 50 ng / ml; (ii) the area under the concentration curve (AUC) of eprinomectin in the blood plasma of said non-human mammals from about 7000 to about 9000 (ng / ml) × h or from about 8000 to about 8500 (ng / ml) × h, preferably about 8300 (ng / ml) × h; (iii) a half-life (T _1/2 ) of eprinomectin in blood plasma of approximately 10-15 days or 11-13 days and preferably at least 12 days; and / or (iv) T _max more than 1.5 days or from about 2 to about 3 days and preferably about 2.5 or about 2.3 days. Methods for measuring the concentration of eprinomectin in a blood plasma are well known to a person skilled in the art and are described in the examples below using a previously published method (Danaher et al., “Development and optimization of an improved derivatization procedure for the determination of avermectins and mylbemycins in bovine liver ”, The Analyst, 126, 576-580).

These increased speeds and delivery levels correlate with the increased therapeutic efficacy of the methods and compositions of the present invention for the prevention and / or treatment of these parasitic diseases and conditions in non-human mammals.

The present invention thus provides a veterinary injection composition comprising a therapeutically effective amount of eprinomectin, PEG solvent and optionally other solvents and excipients, where the active eprinomectin compound has increased bioavailability and a longer half-life (T _1/2 ), as shown in the examples, and / or in comparison with known macrocyclic lactones, such as ivermectin, doramectin and moxidectin.

The formulations of the present invention may be sterile and / or non-sterile injectable formulations. For example, the composition may be an aqueous or oily suspension. Suspensions may be formulated in accordance with methods known in the art using suitable dispersing agents or wetting agents and suspending agents. The injection composition may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent. Suitable diluents and solvents for injectable formulations include 1,3-butanediol, water, Ringer's solution and isotonic sodium chloride solution. Sterile fixed oils are conventionally used as a solvent or suspending medium. Suitable fixed oils include, but are not limited to, synthetic mono- or diglycerides; fatty acids, such as oleic acid and its glyceride derivatives, as well as natural pharmaceutically acceptable oils, such as olive oil, castor oil and their polyoxyethylene derivatives (Sigma Chemical Co., Fisher Scientific).

Additionally, the injection composition may further include one or more additional antiparasitic agents, such, without any restrictions, such as compounds of salicylanilides, avermectins and / or milbemycins.

In addition, the injection composition may further include any of the following many other ingredients in a pharmaceutically acceptable amount, such as, for example, one or more protective light stabilizers, crystallization inhibitors, colloids, adhesives, thickeners, thixotropic agents, wetting agents, stabilizers, sequestering antioxidants, solubilizers, fluidizing agents, complexing agents, vitamins, minerals, preservatives, buffers, antiseptic agents, or combinations thereof. More generally, the active ingredients can be combined with any solid or liquid additives in accordance with conventional formulation formulation techniques. Examples of antioxidants include, but are not limited to, sulfate compounds, L-cysteine, thiodipropionic acid, thiolactic acid, monothioglycerol, propyl gallate sodium metabisulfite, sodium formaldehyde, sulfoxylate acetate, butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT, vitamin A, vitamin C), vitamin B12, or a combination of these agents. Illustrative thickeners include methyl cellulose, hydroxyethyl cellulose, sodium carboxymethyl cellulose, hydroxypropyl cellulose, polyvinylpyrrolidone, and mixtures thereof. Illustrative complexing agents include EDTA and its salts, phosphates, nitrates, acetates, citrates, and mixtures thereof. Illustrative antiseptics include methyl p-hydroxybenzoate, propyl p-hydroxybenzoate, PHB ester, chlorobutanol, benzyl alcohol, butanol, butane-1,3-diol, chlorhexidine salts, benzoic acid and its salts, sorbic acid and mixtures thereof.

The compositions of the present invention can be administered to non-human mammals, especially mammals, including, without limitation, ruminant mammals, such as bovine or sheep, in a more general sense, cows, sheep, goats, buffaloes, oxen, bulls, horses, pigs, cats, dogs, hamsters, mice, rats, monkeys, rabbits, deer, llamas and yaks and, more specifically, dairy and / or pedigree cattle, such as cows and goats. In a preferred embodiment, non-human mammals are bovine. In most preferred embodiments, non-human mammals are dairy cows, lactating cows or dairy cows.

The present invention further relates to a veterinary composition comprising a therapeutically effective amount of eprinomectin and PEG as a solvent, optionally together with other solvents and excipients, for parenteral administration by bovine, more particularly dairy cows, such that in milk or meat and offal of cows fewer residues of macrocyclic lactone, especially eprinomectin, compared with applications for applying a limited volume of the drug.

In fact, an amazing feature of the present invention is that after the injection of the injectable veterinary composition of the eprinomectin according to the present invention, the cows continued to produce milk without delay with fewer macrocyclic lactone residues in the milk.

The present invention also relates to a veterinary injection composition containing a therapeutically effective amount of eprinomectin and PEG as a solvent, optionally together with other solvents and excipients, for controlling a wide range of endoparasites and ectoparasites in non-human mammals. Such eprinomectin formulations are thus particularly useful for the treatment and / or prevention of infections caused by endoparasites and / or ectoparasites in non-human mammals, especially dairy cattle. Thus, the composition can be given to a healthy animal in order to prevent parasitic infection. The composition can be given to an already infected animal, such as an animal suffering from weight loss due to infection.

In various embodiments, the compositions used in the methods of the present invention may further comprise a second pharmaceutically active compound. Typical classes of second pharmaceutically active compounds include, but are not limited to, other antibacterial, antifungal, antiparasitic, antiviral, and anti-inflammatory agents.

The compositions and methods of the present invention are particularly effective for controlling the strongilides of the digestive tract and lungs and ectoparasites.

In an additional aspect, the present invention relates to a method for treating and / or preventing infections caused by endoparasites and / or ectoparasites, in which an effective amount of an antiparasitic composition described herein is administered to non-human mammals.

According to this aspect, mammals that can be processed include, without limitation, ruminant mammals, such as bovine or sheep, in a more general sense, cows, sheep, goats, buffaloes, oxen, bulls, horses, pigs, cats, dogs, hamsters, mice, rats , monkeys, rabbits, deer, llamas and yaks, and more particularly dairy and / or breeding cattle. In most preferred embodiments, non-human mammals are a dairy cow or goat.

In another embodiment of an anti-ectoparasite treatment, the ectoparasite may be one or more insects or arachnids, including those of the genera Ctenocephalides, Rhipicephalus, Dermacentor, Ixodes, Ambylomma, Haemaphysalis, Hyalomma, Sarcoptes, Psoroptes, Otodetesus Chathora, Chorosa, Linus , Solenoptes, Trichodectes and Felicola.

In yet another embodiment, an anti-ectoparasite treatment, the ectoparasite may be from the genera Ctenocephalides, Rhipicephalus, Dermacentor, Ixodes and / or Boophilus. Treated ectoparasites include, but are not limited to, fleas, ticks, ticks, mosquitoes, flies, lice, meat flies, and combinations thereof. Specific examples include, but are not limited to, feline and canine fleas (Ctenocephalides felis, Ctenocephalides sp., Etc.), ticks (Boophilus sp., Ixodes sp., Dermacentor sp., Amblyoma sp., Etc.) and pruritus ( Demodex sp., Sarcoptes sp., Otodectes sp., Etc.), lice (Trichodectes sp., Cheyletiella sp., Lignonathus sp., Etc.), mosquitoes (Aedes sp., Culex sp., Anopheles sp. ., etc.) and flies (Hematobia sp., Musca sp., Stomoxys sp., Dermatobia sp., Coclyomia sp., etc.). In yet another embodiment of the anti-ectoparasite treatment, the ectoparasite is a flea and / or ixodid tick.

Additional examples of ectoparasites include, without limitation, ixodid ticks of the genus Boophilus, especially those of the microplus species (tick boofilus), decoloratus, and anulatus; miazy flies such as Dermatobia hominis (known as Berne in Brazil) and Cochlyomia hominivorax (green carrion); sheep miase flies, such as Lucilia sericata, Lucilia cuprina (known as meat bee fly in Australia, New Zealand and South Africa); adult parasite flies, Haematobia irritans (small cow lance); lice, such as Linognathus vitulorum, etc., and pruritus, such as Sarcoptes scabici and Psoroptes ovis. The above list is not exhaustive, and other ectoparasites are well known in the art as harmful to animals and humans. These include, for example, migrating dipterous larvae.

The composition can also be used for treatment against endoparasites, such as helminths from the group of genera Anaplocepheda, Ancylostoma, Anecator, Ascaris, Capillaria, Cooperia, Dipyllidinum, Dirofilaria, Echinococcus, Enterobius, Fasciola, Haemonchus, Oesophocagag, Oesophagag , Trichinella, Trichuris, and Trichostrongylus. Specifically, gadfly larvae, strongilides, roundworms and pinworms, roundworms like wireworms (Haemonchus placei), ostertagia (Ostertagia spp.), Nodular worm (Oesophagostomum spp.), Hookworm (Bunostomum spp.), Cooperium (Cooperium). paraphilia (Parafilaria bovicola); tapeworms similar to moniesia (Moniezia spp.). Trematodes, a group of trematodes include hepatic trematode (Fasciola spp. - hepatic trematode and giant hepatic trematode) and cone-shaped trematode (Calicophoron - former Paramphistomum), the latter of which is found in the gastrointestinal tract.

Additionally, with or without the addition of additional pesticidal agents, the composition according to the present invention can also be used for treatment against other pests, which include without limitation the following pests:

from the order Isopoda, for example, Oniscus asellus, Armadillidium vulgare and Porcellio scaber;

from the order of Diplopoda, for example Blaniulus guttulatus;

from the order Chilopoda, for example, Geophilus carpophagus and Scutigera spp .;

from the order Symphyla, for example Scutigerella immaculata;

from the order Thysanura, for example Lepisma saccharina;

from the order Collembola, for example Onychiurus armatus;

from the order Orthoptera, for example, Acheta domesticus, Gryllotalpa spp., Locusta migratoria migratorioides, Melanoplus spp. and Schistocerca gregaria;

from the order Blattaria, for example, Blatta orientalis, Periplaneta americana, Leucophaea maderae and Blattella germanica;

from the order Dermaptera, for example Forficula auricularia;

from the order Isoptera, for example Reticulitermes spp .;

from the order Phthiraptera, e.g. Pediculus humanus corporis, Haematopinus spp., Linognathus spp., Trichodectes spp. and Damalinia spp .;

from the order Thysanoptera, for example, Hercinothrips femoralis, Thrips tabaci, Thrips palmi and Frankliniella accidentalis;

from the Heteroptera order, for example, Eurygaster spp., Dysdercus intermedins, Piesma quadrata, Cimex lectularius, Rhodnius prolixus and Triatoma spp .;

from the order Homoptera, for example, Aleurodes brassicae, Bemisia tabaci, Trialeurodes vaporariorum, Aphis gossypii, Brevicoryne brassicae, Cryptomyzus ribis, Aphis fabae, Aphis pomi, Eriosoma lanigerum, Hyalopterus arundxepphemphylphisphaephi phi phylphis phylumphriphysphyphphis, Phalipidae, Macaurus Phorodon humuli, Rhopalosiphum padi, Empoasca spp., Euscelis bilobatus, Nephotettix cincticeps, Lecanium corni, Saissetia oleae, Laodelphax striatellus, Nilaparvata lugens, Aonidiella aurantii, Aspidiotus hedera. and Psylla spp .;

from the order Lepidoptera, for example, Pectinophora gossypiella, Bupalus piniarius, Cheimatobia brumata, Lithocolletis blancardella, Hyponomeuta padella, Plutella xylostella, Malacosoma neustria, Euproctis sppisppri, Buppl. ., Earias insulana, Heliothis spp., Helicoverpa spp., Mamestra brassicae, Panolis flammea, Spodoptera spp., Trichoplusia ni, Carpocapsa pomonella, Pieris spp., Chilo spp., Pyrausta nubilalis, Epellaia tella giella khella pellionella, Hofmannophila pseudospretella, Cacoecia podana, Capua reticulana, Choristoneura fumiferana, Clysia ambiguella, Homona magnanima, Tortrix viridana and Cnaphalocerus spp .;

from the order Coleoptera, for example, Anobiur punctatum, Rhizopertha dominica, Bruchidius obtectus, Acanthoscelides obtectus, Hylotrupes bajulus, Agelastica alni, Leptinotarsa decerlineata, Phaedon cochleariae, Diabrotica sphida pppippsa ppp. , Sitophilus spp., Otiorrhynchus sulcatus, Cosmopolites sordidus, Ceuthorrhynchus assimilis, Hypera postica, Dermestes spp., Trogoderma spp., Anthrenus spp., Attagenus spp., Lyctus spp., Meligethes aeneus, holtibulus polentibus, p. Tribolium spp., Tenebrio molitor, Agriotes spp., Conoderus spp., Melolontha melolontha, Amphimallon solstitialis and Costelytra zealandica;

from the Hymenoptera order, for example, Diprion spp., Hoplocampa spp., Lasius spp., Monomorium pharaonis and Vespa spp .;

from the order Diptera, for example, Aedes spp., Anopheles spp., Culex spp., Drosophila melanogaster, Musca spp., Fannia spp., Calliphora erythrocephala, Lucilia spp., Chrysomyia spp., Cuterebra spp., Gastrophilus sppa. Hy ., Stomoxys spp., Oestrus spp., Hypoderma spp., Tabanus spp., Tannia spp., Bibio hortulanus, Oscinella frit, Phorbia spp., Pegomyia hyoscyami, Ceratitis capitata, Dacus oleae, Tipula paludosa, Hylemyiapp. and Liriomyza spp .;

from the order Siphonaptera, for example, Xenopsylla cheopis and Ceratophyllus spp .;

from the arachnid class, for example, Scorpio maurus, Latrodectus mactans, Acarus siro, Argas spp., Ornithodoros spp., Dermanyssus gallinae, Eriophyes ribis, Phyllocoptruta oleivora, Boophilus spp., Rhipicephalus sompp., Amb. spppp. Amb. ., Psoroptes spp., Chorioptes spp., Sarcoptes spp., Tarsonemus spp., Bryobia praetiosa, Panonychus spp., Tetranychus spp., Hemitarsonemus spp. and Brevipalpus spp.

EXAMPLES

Example 1. Obtaining an injectable veterinary composition of eprinomectin

An injectable veterinary composition of eprinomectin was obtained by mixing the following components, as shown in table 1 below:

Table 1

Ingredients Amount per 100 ml Eprinomectin 1.0 g Benzyl alcohol 1.0 g N-methylpyrrolidone 5.0 g Peg 200 how much is needed up to 100 ml

Example 2. Comparison of the pharmacokinetic properties of the composition of the present invention with existing compositions

table 2

  Moxidectin Doramectin Ivermectin Eprinomectin (EPRINOJECT) Route of administration Inj SC Inj SC Inj SC Inj SC Dosage (0.2 mg / kg) (0.2 mg / kg) (0.2 mg / kg) (0.2 mg / kg) Route of administration application of a limited volume of the drug application of a limited volume of the drug application of a limited volume of the drug Dosage (0.5 mg / kg) (0.5 mg / kg) (0.5 mg / kg) Lipophilicity (log P) 6.0 5,6 4.8 4.4 Structure:       As provided in example 1 Inj water oil organic solvent PO oil alcoholic oil Tmax (day)         Inj 1,5 6 2.12 2,3 PO 4.3 4.3 3.4  - Cmax (ng / ml)         Inj 18.3 32.6 46.4 47.8 PO 2,33 12,2 12,2 - T _{1 / 2el} (day)         Inj 14.5 5,4 5 12,2 PO - 9.82 5.3 - MRT (average residence time) (day)         Inj 25.7 11.8 5,4 12 PO 12.8 12.8 8.4 - AUC 0-inf ((ng / ml) × hour) 6719 / 549.6 12264/4334 6384/2919 8326 / - Sources Alvinerie et al, J Vet Pharmacol Ther. 1997 Apr; 20 (2): 91-9. Sallovitz et al, 2000. J. Vet. Pharmacol Ther, 23, Supp. 1, abstract H17 Toutain et al, Vet Parasitol. 1997 Sep; 72 (1): 3-8. Gayrard et al, 1999. Vet Parasitol, 81:47 - 55 Lifschitz et al, Vet Parasitol. 2007 Jul 20; 147 (3-4): 303-10. Gayrard et al, 1999. Vet Parasitol 81: 47-55 Example 1

* Inj SC - subcutaneous injection; Inj - injection; PO: application of a limited volume of the drug.

The results show that the use of a half-dose subcutaneous composition of eprinomectin compared with the corresponding topical formulation for applying a limited volume of the drug resulted in improved pharmacokinetic performance, including a large C _max and / or greater bioavailability (AUC) of eprinomectin.

Example 3. The definition of pharmacokinetic data

The injection composition of eprinomectin, referred to as EPRINOJECT, was administered to 5 lactating cows. 10 ml blood samples were collected by puncture of the anterior vena cava into heparinized tubes prior to treatment, and also after 6, 12, 24, 40, 48, 60 hours, 3, 5, 7, 11, 16, 21, 25, 35 and 43 days after treatment. Samples were immediately centrifuged at 10,000 rpm. within 10 minutes

Plasma eprinomectin concentrations were determined by the validated HPLC method after derivatization and fluorescence detection.

The results are presented in the following table 3. In particular, the ratio in table 3: these values mean the ratio of the application of a limited volume of the drug / injection. The t _max values are equivalent (ratio = 0.8), the C _max value is 2.4 times higher during injection, the AUC value is 3 times higher during injection.

Table 3 Pharmacokinetic Parameters

Options Unit EPRINOJECT Mean ± SD Attitude t _max (h) 55 ± 11 0.8 C _poppy (ng / ml) 47.8 ± 9.58 2,4 AUC _0-t (h × ng / ml) 7783 ± 1703 3 AUC _0-∞ (h × ng / ml) 8309 ± 1775 3,1 Stay time (h) 300 ± 72 -

Also, the time until the concentration exceeded 1 ng / ml for eprinomectin, calculated after a single subcutaneous injection of 200 mg / kg of the injectable composition of eprinomectin according to the present invention, is presented in the following tables 4.1 and 4.2.

Table 4.1 Individual Values

Animal The time until the concentration exceeded 1 ng / ml (hours) An_261 858 An_296 1015 An_4022 795 An_768 1016 An_9305 1031

Table 4.2. Statistical data

N (amount) 5 Average 943 SD (standard deviation) 109 SEM (standard error of the mean) 49 Min (minimum value) 795 Median 1015 Max (maximum value) 1031 CV% (coefficient of variation) 12 Geometric mean 938 Harmonic mean 932

Example 4. Comparative study with the composition of eprinomectin for applying a limited volume of the drug (Eprinex® = EPRINEX)

The veterinary composition of the eprinomectin according to the present invention (EPRINOJECT) was obtained as shown in example 1 above in this document.

Lactating dairy cows (10 in total, 5 in each group) were divided into 2 treatment groups. Group 1 received EPRINEX (Merial) by applying a limited volume of the drug at 500 μg / kg body weight, and group 2 received EPRINOJECT subcutaneously at 200 μg / kg body weight.

Blood samples (10 ml) were collected by puncture of the anterior vena cava into heparinized tubes before treatment, and also after 6, 12, 24, 40, 48, 60 hours, 3, 5, 7, 11, 16, 21, 25, 35 and 43 days after treatment. Samples were immediately centrifuged at 10,000 rpm for 10 minutes.

Plasma eprinomectin concentrations were determined by the validated HPLC method after derivatization and fluorescence detection. (Danaher et al., ”Development and optimization of an improved derivatization procedure for the determination of avermectins and mylbemycins in bovine liver”, The Analyst, 126, 576-580).

The results are presented in table 5 and in figures 1 and 2.

Table 5 Pharmacokinetic Parameters

Options Unit Mean ± SD Attitude EPRINOJECT Eprinex t _max (h) 55 ± 11 70 ± 30 0.8 C _max (ng / ml) 50.3 ± 12.2 20.33 ± 10.34 2,4 AUC _0-t (h × ng / ml) 7799 ± 1698 2538 ± 1211 2.07 AUC _0-∞ (h × ng / ml) 8326 ± 1772 2677 ± 1177 3,1 Stay time (h) 299 ± 72 299 ± 85 -

Also, the time until the concentration exceeded 1 ng / ml for eprinomectin, calculated after a single subcutaneous injection of 200 mg / kg EPRINOJECT to the cows, the composition according to the present invention, is presented in the following tables 6.1 and 6.2.

Table 6.1. Individual Values

Animal Time until concentration exceeded 1 ng / ml (hours) An_261 858 An_296 1015 An_4022 795 An_768 1016 An_9305 1031

Table 6.2 Statistics

N (amount) 5 Average 943 SD (standard deviation) 109 SEM (standard error of the mean) 49 Min (minimum value) 795 Median 1015 Max (maximum value) 1031 CV% (coefficient of variation) 12 Geometric mean 938 Harmonic mean 932

Also, the time until the concentration exceeded 1 ng / ml or 2 ng / ml for eprinomectin, calculated after topical application of a single dose of 500 mg / kg of eprinomectin in the form of EPRINEX for applying a limited volume of the drug, is presented in the following tables 7.1 and 7.2.

Table 7.1 Individual Values

Animal The time until the concentration exceeded 1 ng / ml (hours) ID3337 333 ID4524 343 ID7176 502 ID8087 484 ID8138 479

Table 7.2 Statistics

N (amount) 5 Average 428 SD (standard deviation) 83 SEM (standard error of the mean) 37 Min (minimum value) 333 Median 479 Max (maximum value) 502 CV% (coefficient of variation) 19 Geometric mean 421 Harmonic mean 414

Claims (14)

1. Injectable parasiticidal veterinary composition containing a therapeutically effective amount of eprinomectin, polyethylene glycol, benzyl alcohol and N-methylpyrrolidone. 2. The composition according to claim 1, containing at least about 20% polyethylene glycol by weight. 3. The composition according to p. 2, containing from 20 to 98% polyethylene glycol by weight or from 40 to 95% polyethylene glycol by weight. 4. The composition according to p. 3, comprising approximately 93% polyethylene glycol by weight. 5. The composition according to claim 1, containing from about 0.5% to about 5% of eprinomectin, from about 0.5% to about 5% gasoline alcohol, from about 0.5% to about 5% N-methylpyrrolidone and formulated in polyethylene glycol. 6. The composition according to claim 5, containing approximately 1% eprinomectin, approximately 1% benzyl alcohol, approximately 5% N-methylpyrrolidone, and formulated in polyethylene glycol. 7. The composition according to p. 1 for the treatment and / or prevention of infections caused by endoparasites and / or ectoparasites in non-human mammals. 8. The composition according to p. 7 for the treatment and / or prevention of damage to the strongilides of the digestive tract and / or lungs in non-human mammals. 9. The composition of claim 7 or 8, wherein said non-human mammals are bovine or sheep. 10. The composition of claim 7, wherein said non-human mammals are cows, sheep, goats, buffaloes, oxen, bulls, horses,
pigs, cats, dogs, hamsters, mice, rats, monkeys, rabbits, deer, llamas or yaks. 11. The composition of claim 7, wherein said non-human mammals are cattle or breeding cattle. 12. The composition of claim 7, wherein said non-human mammals are a dairy cow or dairy goat. 13. The composition of claim 7, wherein said composition is administered to said non-human animal in a single administration over a period of 40 days. 14. The composition of claim 7, wherein said composition is administered by said non-human mammal by subcutaneous injection.

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