MX2013008407A - Injectable compositions for mastitis comprising an nsaid and an antibiotic in a non-aqueous solvent. - Google Patents

Abstract

An injectable composition when used for the treatment of a microbial infection in a mammary gland of an animal, wherein the composition includes a) a non-steroidal anti-inflammatory drug (NSAID); b) an antibiotic selected from the group consisting of a beta lactam antibiotic and macrolide antibiotic; characterised in that the composition includes a non-aqueous solvent, and the NSAID and antibiotic in the composition are dissolved in the non-aqueous solvent.

Description

METHOD FOR THE TREATMENT OF AN INFECTION Field of the invention The invention relates to a compound for the treatment of an infection and more preferably, although not in a specific form for the treatment of mastitis.

BACKGROUND OF THE INVENTION The present invention will be discussed in relation to a means by which the infection can be treated in an animal.

While reference is made throughout the specification to the treatment of farm animals, such as dairy cattle, it should be kept in mind that the principles of the present invention can be applied to others, including humans. Throughout this specification, specific examples are provided in relation to the treatment of mastitis. It should also be understood that the scope of the invention is not limited to mastitis and can be substantially related to any type of bacterial infection in an animal.

Consequently, the entire specification refers to an infection of a bacterial nature, and in particular related to the mastitis caused by this bacterial infection.

It is common to use antibiotics to treat bacterial infections in animals. This can be done by means of several routes of administration, including oral or topical administration. A preferred application is by an injection close to the infected site (such as the udder).

The administration of antibiotics is very expensive.

First, because the medication itself is expensive.

Second, in cases such as mastitis, which adversely affects milk production, there is a time of milk suspension in which the milk produced by the animal is wasted because the contaminated milk must be discarded. If the entire herd is considered, this can result in annual losses of thousands of dollars. Likewise, there are regulations that regulate the introduction of animal products in the food chain (whether eggs or meat) that come from animals recently treated with antibiotics.

Therefore, it would be desirable to have an antibiotic treatment that would be effective against bacterial infections with minimal interruption as regards the milk discarded.

Mastitis is a costly disease, which affects the production of animals, whether immediate (milk) and long-term: (shorter productive life with the risk of premature slaughter), and it also compromises the welfare of the animal and increases stress for people caring for the animals (milker), especially during the peak of lactation.

There is a need to find new treatments that are more convenient to administer and still be effective. This can result in better compliance and better treatment outcomes (cure rates).

Injectable antibiotics have the advantage of treating all quarters of the udder, (in comparison to intramammary antibiotics, which only treat a quarter) as long as the concentrations of the active principle reached are high enough to be effective against the causative bacterium. .

The minimum inhibitory concentrations (MIC) of the antibiotic necessary for the inhibition of the specific bacterium are the target concentrations of the products for the treatment of mastitis. It is likely that products that do not reach the desired MICs are not effective in curing mastitis.

For example, the bacterium Staphylococcus aureus produces a difficult-to-cure mastitis and for the treatment of such infections the indication of tylosin-containing antibiotics is frequent. However, there are not many international studies to determine the MIC of tylosin for S. aureus. Most of the MIC values have been determined for erythromycin, less frequently used in the treatment of mastitis.

MIC values of erythromycin for S. aureus of 0.5 pg / ml, and 2 pg / ml of tylosin for S. aureus have been reported. This means that the concentration of tylosin required to effectively treat S. aureus is approximately four times greater than the concentration of erythromycin required.

In fact, there are pharmacokinetic studies that show that an injectable preparation of tylosin at 20% only reaches peaks of concentration of 1.4 pg / ml cf during treatment, so that they do not reach the required therapeutic concentrations (MIC). ~ Therefore, it is necessary to use either a larger dose and / or a higher concentration of active ingredient (> 20%) to achieve milk concentrations higher than the MIC.

Therefore, it is necessary to formulate improved antibiotic treatments (for other antibiotics in addition to tylosin) to avoid the need to use larger doses or higher concentrations of active substance (> 20%) to achieve the required MIC and achieve an effect on S. aureus and other causes of bacterial infections.

In addition, it is necessary to devise compounds in which other excipients or active ingredients are included to allow the antibiotic to be more effective in the treatment of bacterial infections.

It is known that when an NSAID is administered together with the antibiotic, the response of the antibiotic is more effective. However, it is difficult that these active principles can be combined in a stable compound, so they are generally administered separately. This is a drawback, and it would be preferable if it were possible to administer a single compound that included both the antibiotic and the NSAIDs.

Also, it is important to consider animal welfare. Infections such as mastitis cause pain. Therefore, it is usually not only the infection, but also the pain associated with infection with active agents such as NSAIDs. It is a relatively new combined treatment for the industry.

The patent 2005/0277634 describes a combination of NSAID (meloxicam) and an antibiotic (penetamate iohydrate) in a suspension that is injected for the treatment of mastitis. There, the synergic reaction between the NSAID and the antibiotic is discussed, which allows reaching concentrations higher than the MIC with lower concentrations of antibiotic. Preferably, the higher the concentration of NSAIDs in the compound, the better the function of the antibiotic appears to be.

However, suspension formulations have some disadvantages. They include the possibility of sedimentation compact, problems of dispersion before its application, difficulties in its administration, pain or reaction in the site after its injection and a poor absorption of the active principles. Therefore, it is preferable to administer the NSAID and a combination of antibiotics in the form of stable compound in solution. In addition, it would be advantageous to administer a compound solution in which the NSAID is charged to improve the effectiveness of the antibiotic. However, the above problems have not allowed this objective to be successfully achieved.

In general, one of the significant problems associated with many injectable compounds is that a reaction or pain appears at the site in the animal to which the compound has been administered intramuscularly or subcutaneously.

Frequently, this reaction or pain at the injection site can be produced by the active agent, such as the antibiotic tylosin. However, it is possible that these problems are exacerbated by other excipients of the compounds, generally the solvents that are vehicles or solubilizers of the active agents. The article by Strickley et al., Pharmaceutical Research, Vol., 21 No.2 February 2004 entitled "Solubilizing Excipient in Oral and Injectable Formulations" fully explains this problem.

For this reason, veterinary chemists generally attempt to make compounds in the form of solutions with the lowest concentration and / or volume of solvent possible in the injectable compounds. Consequently, the ability to reach higher concentrations of active agents may decrease, and the compound as a whole may lose stability, and this ultimately decreases the chances of successful treatment for the animal.

Therefore, veterinary chemists prefer to use water-based compounds to avoid reactions and pain at the site of injection.

While this may be effective in decreasing side effects such as reaction and pain at the injection site, water-based compounds may confer instability to the active agents, which shortens the preservation time of the drug. compound (and ultimately its bioavailability). It can also limit the ability to retain higher concentrations of active ingredients. It may then be necessary to reuse the suspensions discussed in US 2005/0277634.

WO 02/41899 (WO'899) describes injectable or topical compositions which include an antibiotic and an analgesic, both dissolved in the solvent. However, there is a distinction between the types of antibiotics exemplified (florfenicol, gentamicin, and oxytetracycline) and those that are they are involved with the compounds of the present invention, namely beta-lactam antibiotics and macrolides.

In fact, it is known that beta-lactam antibiotics and macrolides are particularly unstable and probably react with NSAIDs such as flunixin. Although the difficulties associated with the combination of beta-lactam antibiotics and macrolides with NSAIDs, these antibiotics are very useful for treating microbial infections related to mastitis.

WO'899 does not provide stability data suggesting that many compounds, particularly those with a higher concentration of active agent (such as that described in Example 4) would not be stable in storage. In fact, the studies carried out by the inventors have shown the instability of similar compounds.

It is the object of the present invention to address the above problems or at least to offer the public a useful option.

All references, including patents or patent applications cited in this specification are included herein by reference. The references are not taken as previous inventions. The discussion of the references establishes what the authors affirm, and the applicants reserve the right to question the accuracy and relevance of the documents cited. It is clearly understood that, despite the fact that reference may be made to publications of previous inventions, this reference does not constitute an acknowledgment that any of these documents forms part of the general knowledge of the technique, in New Zealand or in any other nation.

In the present specification, the term "includes" or its variants, as "include" or "including" implies the inclusion of the element, the integer, the stage, or the group of elements, integers or steps, but not implies the exclusion of any other element, integer or stage number, or groups of elements, integers or stages.

There will be other aspects and advantages of the present invention that will be apparent after revealing the description, which is presented as an example only.

Brief Description of the Invention According to a first aspect of the present invention, an injectable compound is described when it is used for the treatment of a microbial infection in the mammary gland of an animal.

The compound includes to. A non-steroidal anti-inflammatory drug (NSAID); b. An antibiotic selected from the group that includes a beta-lactam antibiotic and a macrolide antibiotic; With the characteristic that the compound includes a non-aqueous solvent, and The NSAID and the antibiotic in the compound dissolve in the non-aqueous solvent.

According to another aspect of the present invention, a method is described for treating an animal with an internal microbial infection, which is characterized by the stages of administering a compound substantially as described above by injection.

According to another aspect of the present invention, the use of a compound as described above for the preparation of a medicament for the treatment of a microbial infection is described.

A method for preparing a compound, substantially as described, which is characterized by a step in which at least one non-aqueous base solvent is added to the compound.

Brief Description of the Figures Figure 1 is a graph where the solid lines represent the tylosin serum concentrations when treated with the study compound of Example 4, which is discussed in detail below. Dotted lines represent serum tylosin concentrations when treated with Tylan.

Figure 2 the solid lines represent the serum concentrations of ketoprofen when treated with the study compound of example 4, which is discussed in detail below. The dashed lines represent the serum concentrations of ketoprofen when treated with Ketofen.

Detailed description of the invention The characteristics and advantages of the present invention are set forth below with greater specificity Discussion of the advantages associated with the invention It is known to the inventors that significant inflammation at the site of infection and in its vicinity can interfere with the effectiveness of the antibiotic, because inflammation decreases the contact and effect of antibiotics on bacteria.

In the past, most NSAIDs were used to treat inflammation and / or pain. However, the inventors have found that the combination in a single injectable compound of an NSAID and an antibiotic produce an important synergistic effect. Thanks to this synergistic effect, it is possible to increase the effectiveness of the antibiotic over the microbial infection if we compare it with the antibiotic administered alone. Similarly, the inventors have found that the combination of the NSAID and the antibiotic in a single formulation increases the synergistic effect that will not be present when the animal is treated with NSAIDs and antibiotics separately in two formulations. This may be due in part to the possibility of using two active ingredients together in a stable compound.

The inventors consider that, without limiting the proposed mode of action, the synergistic interaction may be caused by the fact that the NSAID blunts inflammation at the site of infection and in its vicinity (in the udder in the case of mastitis) , and this allows the effective distribution of the antibiotic in the site that without the NSAID would be inflamed.

In addition, it is possible that the synergistic effect is a consequence of the binding of NSAIDs to endotoxins (endotoxemia). The inventors found that if this does not occur, the effectiveness of the antibiotic decreases.

For example, mastitis is associated with the presence of gram-negative bacteria, which produce endotoxins. These bacteria can cause systemic diseases (fever and endotoxemia). Although currently in New Zealand it is relatively rare (incidence less than 1%), the increased use of feeding tracks and sheds may produce a higher incidence of mastitis caused by gram-negative pathogens. Therefore, it is predicted that in the future, the present formulation will be an important factor of the regimes for the treatment of infections, for example, of mastitis.

In addition, it is possible to perform even more synergistic effect, since NSAID decreases fever (pyrexia) which increases the effectiveness of the antibiotic and improves the health status and comfort of the animal.

During the experimental use of certain NSAIDs and antibiotics, the inventors recorded a synergistic effect, as mentioned above. Based on the experimental uses mentioned, and the identification of beneficial results, the expert of the subject may consider that it is to be expected that there is a synergistic result not only in the specific examples that have been studied but in the most frequent types of antibiotics. For example, it is expected that all beta-lactam antibiotics and macrolides have the same synergistic effect when combined with an NSAID. The mode of action of both types of antibiotics is similar (inhibition of cellular protein synthesis).

Another important advantage of the combination identified by the inventors may be the ability of NSAIDs to improve animal welfare, because it reduces the pain and inflammation produced by internal infections such as mastitis.

In addition, the ability to produce two different active ingredients in a combined formulation allows avoiding the use of multiple compounds, storing compounds, reducing costs, dispensing with packaging, among other benefits. The inventors identified a particular type of solvent that allowed the combination of two active ingredients, whose combination in a stable solution previously had been very complicated. In the past, these active ingredients were combined into a single compound by suspensions.

The inventors found another unexpected synergistic effect between the two active ingredients as the NSAID concentration was increased, as discussed below. The objective in increasing the concentration of NSAIDs was to improve the synergistic effect initially identified by the inventors to allow improving the effectiveness of the antibiotic at the site of infection.

The recommended doses for active agents are based on what is defined as effective doses. Therefore, when combined two agents, this combination should include the appropriate doses of each of the active ingredients. For example, for the recommended dose for a particular antibiotic (for example, in the case of tylosin, 10 mg / kg), a 20% concentration of tylosin is generally used in the compound. It is common that this high concentration of tylosin is the limiting factor in said compound, because it can produce significant irritation at the injection site. To apply a concomitant dose of NSAID (eg, ketoprofen), preferably, a concentration of 6% w / v is used. In fact, the degree of compatibility of the dosing regimes among the multiple active principles used can be an important aspect to determine the success of the active compound in the administration of the correct amount of active principle.

The inventors tried to use a compound with 6% ketoprofen, but it lost stability in an aqueous solvent. With the other NSAIDs studied, similar results were obtained, with low stability in water-based compounds, when NSAIDs were included at the concentrations required to obtain the recommended doses.

To avoid using suspensions, the inventors had to try other ways.

To compensate for the higher concentrations of NSAIDs, the inventors used a non-aqueous solvent system, in order to obtain a stable compound in solution at different conditions (for example, at temperatures above 4 ° C). However, it is expected that the use of non-aqueous solvents, produce greater reaction or pain at the administration site compared to that produced by the aqueous compounds, as is known in the art.

However, in the inventors' studies it was unexpectedly found that said reaction or pain at the site did not increase with respect to what can be seen for commercially available compounds (eg, Tylan and Ketofen) which are aqueous compounds.

Without limiting itself to a particular mode of action, it is believed that the highest concentrations of NSAIDs provide synergy that can alleviate inflammation and local pain at the injection site. Therefore, it is possible that these higher concentrations of NSAIDs counteract the effect of the more severe solvents that would normally increase the reaction and pain at the injection site.

Therefore, unexpectedly, the inventors found that it is possible to include an antibiotic and an NSAID in a stable compound in solution, with higher concentrations of NSAIDs than they would use when it is combined in a water-based system. The important thing is that this use does not produce a greater reaction or pain at the administration site compared to the reference industrial compounds (Tylan and Ketofen).

Another effect of this finding was that the concentration (and ultimately the dose) of the NSAID and even the antibiotic may increase without producing a greater reaction and / or pain at the site of administration. It is then possible that this synergistic effect at the site of infection is further enhanced, which improves the overall outcome of the treatment.

Preferred modalities of the compound The compound of the present invention is presented as an injectable liquid formulation. In the specification, the term liquid formulation is equivalent to any therapeutic formulation whose consistency allows it to be placed in a syringe.

This is the preferred route of administration by many breeders of farm animals and veterinarians, particularly, the local application of an NSAID, which may cause a greater effect to treat the infection for example of the mammary glands in mastitis. Preferably, the viscosity of the liquid formulation is suitable for syringes at low temperatures.

The liquid formulation is a solution. This means that in the compound all the excipients and active agents are solubilized in the solution, and not in the suspension. As mentioned above, there are many benefits associated with the use of solutions with respect to the use of suspensions. For example, the problems of the formation of compact deposits, the resuspension, or the difficulties in the injection are avoided.

It can be a solution with one or more phases.

Nonsteroidal anti-inflammatory drugs In this specification, the term non-steroidal anti-inflammatory drug (NSAID) is equivalent to any drug or active ingredient with anti-inflammatory anti-inflammatory effects without the use of glucocorticoid steroids.

There are many different types of known NSAIDs. The invention includes the use of any combination of NSAIDs already known or to be known.

Preferably, an NSAID is selected from the group of carprofen, naproxen, ibuprofen, ketoprofen, piroxicam, diclofenac, etodolac, flunixin, deracoxib, meloxicam, celecoxib, rofecoxib, and combinations thereof.

It is preferable to use an NSAID from the flunixin group, carprofen, ketoprofen and combinations thereof.

The inventors found that these NSAIDs have the greatest synergistic effect when combined with antibiotics as claimed. In addition, it has been shown that all of these NSAIDs are effective as anti-inflammatory agents in medicines for cattle, from the point of view of their efficacy and toxicity.

Preferably, the NSAID is present in liquid formulations at concentrations of 1 to 15% w / v. As previously discussed, this may be to increase the concentration of NSAIDs and thus improve the synergistic effect between the two active ingredients. Although the benefits associated with the use of this combination are valued, it has not been possible to produce a stable compound in solution, let alone a compound that does not produce a greater reaction or pain at the site when administered, compared to the effects produced by water-based compounds.

Preferably, the NSAID flunixin meglumine is found in the compound at concentrations of about 7.3% w / v. This means about 4.4% flunixin in the compound. This is per dose when combined with tylosin.

The approximate concentration that is preferred for each NSAID may also depend on its anti-inflammatory efficacy and its toxicity profiles, as well as the concentration of antibiotic used in the compound.

It is preferable that the NSAID meloxicam be present in the compounds at concentrations of about 1% w / v.

It is preferable that the NSAID caprofen be present in the compounds at concentrations of about 3% w / v.

Preferably, the NSAID ketoprofen is present in the compound at concentrations of about 4% to 10% w / v. For example, when the concentration of the antibiotic tylosin is in the compound by 20% w / v, the corresponding concentration of ketoprofen is preferably 6% w / v. If we increase the concentration of tylosin to 30% w / v, the preferred concentration of ketoprofen is 9% w / v.

Alternatively, the NSAID is selected from the natural forms of anti-inflammatory agents. For example, they may include : green mussel extract, omega 3, and the like. .

Antibiotics In the specification the term antibiotic is equivalent to a substance or compound, whether natural, synthetic or semi-synthetic, with bactericidal and / or bacteriostatic effect.

The antibiotic in the compound is selected from the group of macrolide and beta lactam antibiotics.

In this specification, the term macrolide is equivalent to any antibiotic whose activity arises from the presence of a macrolide ring to which one or more deoxy sugars may be attached, generally cladinose cladinose and desosamine desosamine. Generally, lactone rings are 14, 15 or 16 member rings.

Generally in veterinary medicine antibiotics of the macrolide class are used, which are considered antibiotics with the same mode of action. The consideration is that macrolide antibiotics act by interfering with the synthesis of proteins, for example, they block translation at the ribosome level.

Preferably, antibiotics of the macrolide class are selected from azithromycin Azithromycin, clarithromycin Clarithromycin, Dirithromycin dirithromycin, erythromycin Erythromycin, roxithromycin Roxithromycin, telithromycin Telithromycin, carbomycin A Carbomycin A, Josamycin josamycin, kitasamycin itasamycin, midecamycin / midecamycin acetate Midecamicine / midecamicine acétate, oleandomycin. Oleandomycin, Spiramycin spiramycin, Troleandomycin troleandomycin, Tylosin tylosin / tylocine.

More preferably, the macrolide antibiotic is tylosin.

Tylosin is a class of antibiotic of the macrolide type used in veterinary medicine to treat bacterial infections in a wide range of species and has a high safety margin. Another advantage associated with tylosin is that it is the antibiotic authorized for treatment in New Zealand.

Tylosin has a spectrum of activity against gram-positive bacteria including Staphylococci, Streptococci, Corynebacteria, and Erysipelothrix. This antibiotic is also active against Gram-negative bacteria Campylobacter coli, and some Spirochaete. It is also active against the Mycoplasma species.

Injectable tylosin formulations can cause pain, swelling, and itching at the injection site. For 16 ', it is important to moderate the above-mentioned side effects when tylosin is injected. Another advantage associated with the use of an antibiotic such as tylosin with an NSAID is that the side effects of the antibiotic are not observed. The synergistic effect between the NSAID and the antibiotic administered in a single formulation allows to moderate the side effects. In this specification, the term beta-lactam antibiotic is equivalent to any antibiotic that includes a beta-lactam ring in its structure. For example, this may include derivatives of the penicillin (penam), cephalosporins (cephems), monobactams and carbapenems.

The beta-lactam antibiotics act by attacking bacterial cell walls. Beta-lactam antibiotics can be added with beta-lactam inhibitors such as a clavulanic acid.

Preferably, the beta-lactam antibiotic is the penetamate. In a recent study, it was found that the efficacy of penetamate and tylosin are similar (the latter being the antibiotic particularly preferred for the present invention).

With respect to other types of antibiotics, beta-lactam antibiotics and macrolides both have the advantage that they are weak bases and are lipophilic. This may allow higher concentrations of the antibiotic in body fluids (such as milk) than in plasma. Then, they have 'greater possibilities than less lipophilic compounds to achieve higher concentrations at minimum inhibitory concentrations (CIM) Unexpectedly, these types of antibiotics appear to have a synergistic effect with NSAIDs. However, it is a synergistic effect that is not observed with other types of antibiotics. Preferably, the antibiotic is in the liquid formulation at concentrations of 10 to 35% w / v, and more preferably about 20.0 to 30.0 w / v.

Generally, when the concentration of antibiotic is greater, the volume of antibiotic administered per dose is lower, which may be more convenient for breeders of farm animals and more comfortable for small animals. If the concentration of NSAIDs in the compound is increased, it will be possible to use higher concentrations of antibiotic in the compound without aggravating the reaction at the site of administration (similar to the effect that can be observed when the inventors switched to a non-aqueous base solvent, for which also expected greater reaction or pain at the site of administration).

For example, it is known that if we talk about a treatment against Staph. aureus, it is possible to increase the concentration of antibiotic (for example, tylosin) up to 60% w / v to reach the minimum inhibitory concentration (MIC).

Preferably, the ratio of NSAID and antibiotic (w / v) in the liquid formulation is approximately 1: 3, respectively. This may be a particularly applicable ratio if the NSAID is flunixin. However, if said NSAID is a different type of NSAID, such as meloxicam or carprofen, this Quotient may vary.

Solvents In this specification the term solvent is equivalent to any solvent or combination of different solvents that are present in the compound.

Preferably, the solvent has one or more of the following characteristics: Not watery; - Dielectric point above 30 to 25 ° C; A boiling point greater than 100 ° C; I Classified as a bipolar aprotic solvent.

More preferably, the entire compound is non-aqueous. The inventors have studied a range of different water-based compounds, which are known to be relatively unstable at elevated temperatures and even at room temperature. For example, tylosin was found to be particularly unstable when combined with an NSAID in a water-based formulation. However, if the NSAID and the antibiotic are combined in a non-aqueous solvent based system, the compound preserves its stability, especially at lower temperatures (eg, 2 to 15 ° C).

It was found that if the aqueous solvents were replaced by non-aqueous solvents, more stability in the solution was observed.

In addition, with a non-aqueous solvent compound, the inventors can increase the concentration of NSAIDs to values that could not be achieved in aqueous systems. Although an increase in the reaction at the administration site was expected, the inventors did not perceive this in their studies.

Apparently, if the non-aqueous solvent has a dielectric point greater than 30 at 25 ° C, it improves the stability of the compound. Non-aqueous solvents such as ethanol (with a dielectric point of 25) turned out to be poor solvents for the present invention and did not allow to reach solubilities and / or stabilities equivalent to solvents with dielectric points greater than 30. It would be expected that the solvents of the same classes will produce the same positive effects.

More preferably, the non-aqueous solvent has a dielectric point between 30 and 50. It was found that propylene glycol (32), 1), NMP (32.2), DMSO (46.7) and DMA (38) improve the stability of the active principle. It could be seen that the reaction and / or pain at the administration site did not increase, as expected, in relation to what is seen in the aqueous compounds (possibly, due to the secondary synergistic effect mentioned above). Solvents of similar kinds are expected to produce the same positive effects.

In the same way, it was found that a non-aqueous solvent with a boiling point above 100 ° C allows the stability to improve while not increasing the reaction at the site of administration. Once again, ethanol and water turned out to be poor solvents. On the other hand, it was observed that other solvents such as propylene glycol (PE = 188 ° C), glycerin formaldehyde (PE = 190-195 ° C), NMP (PE = 202-204 ° C), DMSO (PE = 189 ° C) C), DMA (PE = 164-166 ° C) all present the positive advantages discussed above. Solvents of similar kinds are expected to produce the same positive effects.

In addition, the inclusion of at least one non-aqueous solvent, broadly classified as a bipolar aprotic solvent, proved to be particularly beneficial. For example, it was found that NMP is a very useful solvent for improving stability without increasing the reaction at the site of application, although it is considered an irritant solvent and a solvent that produces a reaction at the injection site. Preferably, the non-aqueous solvent is selected from N-methyl-2-pyrrolidone (NMP), glycerin formaldehyde, propylene glycol, and benzyl alcohol.

It is well known that solvents such as NMP and propylene glycol, which are particularly effective in stabilizing the compound and allowing the use of higher concentrations of NSAIDs, are highly irritating when injected. Thus, it was not expected that these solvents would not produce a greater reaction at the site when the compound was injected, as was the case.

Preferably, the liquid formulation includes at least two different non-aqueous solvents.

More preferably, the liquid formulation includes a first and second. nonaqueous base solvent, wherein the first solvent acts as a solubilizing agent and wherein the second solvent acts as a carrier solvent.

For example, the inventors have identified that the combined NMP and propylene glycol provide particularly stable active ingredients in the formulation. The inclusion of NMP, glycerin formaldehyde and / or benzyl alcohol can serve as a solubilizing agent for the active, while a solvent such as propylene glycol can be used as a vehicle in the compound to improve stability.

Preferably, only a minimum concentration of solubilizing agent is used, such as NMP. The concentration of solubilizing agent may vary according to the type of active agent and the solubility of the active agent that is present in the compound.

Preferably, the concentration of solubilizing agent in the compound is less than 40% w / v.

More preferably, the concentration of solubilizing agent in the compound is about 30% w / v.

In the present invention, it is preferable to use NMP at minimal concentrations and only as a solubilizing agent.

This differs from what is described in Examples 1 to 4 of WO 02/41899 (O'899), in which the main solvent is NMP, and the compound is prepared with respect to NMP (in most of the cases this is equivalent to approximately 60% p / or more in the final compound). Simply using the NMP as the main solvent (described on page 4 of WO '899 as 95% w / v of the final compound) allows to see that betalactam antibiotics or macrolides were not used.

Preferably, the concentration of carrier solvent in the compound is at least 30% w / v. More preferably, the concentration of vehicle solvent in the formulation is at least 40% w / v.

Preferably, the ratio of the first and second solvent is between 1:10 and 1: 1. More preferably, the ratio of the first solvent and the second solvent is about 1: 2 to 1: 1.

Vehicle solvents such as propylene glycol are much more severe solvents and are considered to produce a greater reaction at the site when injected compared to NMP. Based on As a general concept of the technique, it is possible that this is another reason why descriptions such as WO'899 do not incline the reader to use a vehicle solvent such as propylene glycol, on the contrary it is mainly based on the use of large concentrations of the NMP.

It is believed that the carrier solvent in the present invention is what provides good stability to the beta-lactam antibiotic and / or macrolide and NSAID in the compound. WO'899 does not explain any use of a vehicle solvent, let alone the types of antibiotics used in the present invention.

Furthermore, the solvent combinations described in WO'899 would not achieve the same results in terms of stability as those of the present invention, even if macrolide and / or beta-lactam antibiotics were used in place of the antibiotic and NSAID combinations more stable which are examples in WO'899.

It is known to the inventors that beta-lactam antibiotics and macrolides are very unstable with flunixin, the WO'899 NSAID. In the studies carried out by the inventors, the stability observed in compounds similar to those described in WO'899 was poor.

On the other hand it is demonstrated that the stability of the compounds of the present invention, the macrolide antibiotics and / or beta-lactams, is satisfactory under a range of conditions (see in Example 6 of the Best Practices of the Invention).

Preferably, the non-aqueous solvent is more than 10% w / v of the compound.

More preferably, the non-aqueous solvent comprises between 30% w / v and 80% of the compound. As discussed above, these high concentrations of the non-aqueous solvent (and particularly a carrier solvent as compared to the solubilizing agent) allowed to obtain more stable compounds and higher concentrations of NSAIDs and antibiotics, and it was expected that under these circumstances, the reaction and the pain at the site were more severe compared to the water-based compounds, however, this was not the case.

Antioxidants Preferably, the liquid formulation includes at least one antioxidant.

Preferably, the antioxidant is selected from sodium formaldehyde sulfoxylate or hydroxybutyl toluene (BHT). However, the person skilled in the art will know that it is possible to use any antioxidant in the present invention and that the Antioxidant should not be considered outside the scope of the invention.

Preferably, the antioxidant is in the liquid formulation at concentrations of 0.01-0.3% w / v, and more preferably about 0.05% w / v.

Particularly preferred examples of the formulations are described in detail in The Best Practices of the Invention of the specification.

The preferred modalities of the treatment method In this specification, the term "administration" is equivalent to the administration of active principles of the compound at the site of infection, either by local or systemic application (ie, by the bloodstream and by the body of the animal).

Preferably, the compound serves to treat an internal infection. In this specification, the term "internal infection" is equivalent to a site inside the body, including the cavities of the body which can be accessed from the outside. These cavities include for example the mouth, the teat canal, the udder, the anus , the vagina, among others.

Preferably, the compound is administered systemically. Examples of systemic administration include administration by injection, bolus, or spray, and subsequently the active ingredients are distributed through the bloodstream of the animal to the site of infection and / or inflammation.

More preferably, the compound is administered by injection. It should be noted that the injection can be performed at or near the site of infection and yet the active ingredient enters the system optimally.

It is possible to inject the compound subcutaneously or intramuscularly.

Preferably, the compound is used to treat clinical mastitis. However, in certain embodiments, it is possible to use the compound to prevent the early stages of mastitis. The use of NSAIDs in a compound to treat mastitis is a new concept, which as discussed in this specification has numerous associated advantages.

However, the inventors recognized that it is possible to use the compound to treat any type of internal microbial infection and / or inflammation and / or associated pain.

Preferably, the compound is used to treat animals such as cattle, sheep, goats and other frequent farm animals.

The preferred modalities for dose regimens Preferably, the compound is administered at doses of 5 to 20 mg / kg / day (by weight of the drug / animal) of antibiotic and 0.2 mg to 4 mg / kg / day of NSAIDs.

In preliminary studies, this dose seems to reach the MIC required for tylosin (2 g / ml), and thus the antibiotic reaches the therapeutic concentration. However, due to the synergistic effect between the antibiotic and the NSAID, it is possible that the antibiotic is more effective than normal, with which with lower concentrations of antibiotic it is still possible to treat the bacterial infection effectively.

The inventors found that the concentration of NSAIDs can vary depending on the type of NSAID. For example, the preferred approximate dose of each NSAID may depend on its anti-inflammatory efficacy and its toxicity profiles.

Preferably, the dose of flunixin is from 1 to 3 mg / kg / day. More preferably, the dose of flunixin is about 2.2 mg / kg / day.

Preferably, the dose of meloxicam is 0.1 to 1 mg / kg / day. More preferably, the dose of meloxicam is about 0.5 mg / kg / day.

Preferably, the dose of ketoprofen is from 2 to 4 mg / kg / day. More preferably, the dose of ketoprofen is approximately 3 mg / kg / day.

Preferably, the dose of carprofen is from 0.5 to 2.5 mg / kg / day. More preferably, the dose of carprofen is approximately 1.4 mg / kg / day.

The treatment method can use a flexible dose approach to allow doses to be regulated when diseases are treated based on the type of disease and its severity For example, the inventors noted that a flexipack may be particularly useful for this approach to treatment. use flexible doses.The flexipack is generally a bottle that includes a mixture of high and low density polyethylene.This is a type of container offered by different manufacturers.

The preferred modalities of the preparation method As discussed in this specification, one of the main advantages associated with the use of non-aqueous solvents is that it is possible to increase the concentration of active principle in the compound. This is associated with many benefits, including the ability to administer a greater amount of active ingredient to the animal in smaller volumes of compound. This allows to shorten the time of administration, and discomfort to the animal that occurs during the injection.

In addition, the dose at lower volumes is an advantage in terms of storage as it takes up less space.

Preferably, it would increase the concentration (% w / v) of NSAIDs in the compound which would remove storage stability at room temperature to the corresponding aqueous-based compound.

Preferably, the NSAID is ketoprofen and the concentration of ketoprofen is greater than 4% w / v.

Some of the preferred aspects of the elaboration process are discussed below.

Preferably, a first and second non-aqueous solvent is added to the mixing vessel before adding either the NSAID or the antibiotic.

The first and second non-aqueous solvent chosen may depend on the type, number or concentration of NSAIDs and the antibiotic used in the formulations. The examples given of the first and second non-aqueous solvent are presented in The Best Practices of the Invention. However, they are non-limiting examples.

In certain embodiments, it is possible to add other non-aqueous solvents. Alternatively, it may be suitable to include only a non-aqueous solvent.

Preferably, non-aqueous solvents are heated to approximately 55 to 80 ° C before adding the NSAID or antibiotic. The temperature used may depend on the specific active ingredients used in the compound.

It is preferable to add an antibiotic to the non-aqueous solvents and dissolve it before adding the NSAID. Subsequently, the NSAID can be added and dissolved in a mixture of dissolved antibiotic and non-aqueous solvent. The inventors concluded that adding in stages the solvents, the antibiotic and finally the NSAID improved the stability and the solubility of the compound.

Preferably, the temperature is maintained at 55 to 80 ° C during the stepwise addition of the NSAID and the antibiotic to the formulation.

The compound containing the NSAID and the antibiotic is then cooled to room temperature (for example, between 20 and 30 ° C) before completing a final volume (s) with the non-aqueous solvent and mixing for about 10 to 20 minutes. minutes The compound can then be stored at approximately 4 ° C before administration.

Some of the advantages of the present invention include: The synergistic relationship between the NSAID and the antibiotic (NSAID improves the effectiveness of the antibiotic).

The use of a non-aqueous solvent that allows to overcome the problems of incompatibility between the active ingredients, in particular the instability in many antibiotics.

The use of a non-aqueous solvent allows to have a stable compound in solution (not a suspension) in a range of storage conditions.

The solubilization avoids the formation of compact deposits, the need to resuspend the compound before administering it, and the problems associated with the injection (obstruction of the needle, and possible pain to the animal as a result of particulate matter).

The use of non-aqueous solvents allows the concentration of NSAIDs to be increased to values higher than those obtainable in an aqueous compound. Although the reaction and pain at the site of administration are the expected characteristics of the non-aqueous solvents, an aggravation of these side effects was not observed.

It is possible that the higher concentrations of NSAIDs facilitate the moderation of the reaction at the site, produced by solvents and also potentially by the antibiotic (the irritant nature of tylosin is known). It is possible to increase the concentration of tylosin in the compound because the reaction at the injection site is less severe.

It is believed that if higher concentrations of NSAIDs are used in the compound, the synergy between the two active ingredients at the site of infection will increase. A more effective antibiotic allows decreasing the concentrations of antibiotic used, obtaining the same results.

As the water in the compound is dispensed with, it is not necessary to regulate the pH of the compound to the desired value, thus simplifying the manufacturing process.

The pain and inflammation associated with internal infections diminish.

The efficiency of the administration improves (one and not two compounds are administered). For example, the combination of antibiotic (for example tylosin) and NSAID in a single product avoids the need to apply many injections and simplifies the administration, which improves treatment compliance and improves results.

Versatility in administration options The preferred administration is the injection that allows to improve the distribution of the drug NSAID improves the welfare of the animal while the antibiotic is acting In addition to relieving pain and fever, NSAIDs bind to endoxins (of gram-negative bacteria), which improves treatment outcomes, and this attribute makes them an important element of the mastitis treatment regimens that will come .

It is a formulation that is possibly particularly effective for the treatment of mastitis caused by gram-negative bacteria, which is expected to increase its frequency in New Zealand in the years to come.

- BEST WAYS TO PRACTICE THE INVENTION Example 1 The following table 1 describes a number of initial formulations studied by the inventors in line with the present invention.

Next, the suitability of the formulations studied is discussed.

Table 1: Tylosin / flunixin combinations Fl F2 F3 F4 F5 F6 F7 F8 %%%%%%% Ingredients p / v p / v p / v p / v p / v p / v p / v p / v 1. Tylosin base ** 20.4 20.4 20.4 20.4 20.4 20.4 20.4 flunix na meglundna Í7.3% provides 4, 4% of J. flunLxine) - 8.3 7.3 7.3 7.3 7.3. 7.3 7.3 Sulfur oxyl ato 3. sodium fQDüBldehyde - 0.05 0.05 0.05 - - - Hydroxybutyl toluene < . (?? G) - - - - 0.05 0.05 0.05 0.05 S. Disodium edetate - 0.1 0.1 0.1-0.1 0.1 -s. -nenyl-2-pyrrole dana < NKE > > - - - - - - - 20 says formaldehyde quarrel 7. is abil zada - - - - 40 - 4,18 - 18% v to. Propi lengli col 52 / v 52 52 is 52 52 qs 9. Phenol liquid - 0,25 - - - - - - 10 Benzyl alcohol co 4, 18 0 4.18 4.18 - 4, 18 1 1 Monoe tanolami na *** eleven (to regulate the pK) - es es - - - - - sodium füdróxido *** 12 < to regulate the pH) - - - C3 - es es Hydrochloric acid 13 (to regulate the pH) - es es CS - es es - 1ß Water injection is es es C3 - es es - added 2% of authorized surpluses anger regulate the pH to values between 8 and 9, 5 A tylosin base injection formulation registered in the Law on Agricultural Compounds and Veterinary Drugs (ACVM).

An existing injection formulation of flunixin (Trade name: Fluxamine injection) registered in ACVM.

At 48 hours the product turned brown at room temperature.

The pH was adjusted with 20% sodium hydroxide. The color of the product remained stable at 4 and 40 ° C.

To avoid problems with pH, the product was formulated using glycerin formaldehyde with propylene glycol. The color of the product remained stable at 4 and 40 ° C.

The antioxidant was changed from sodium formaldehyde sulfoxylate to BHT. The color of the product remained stable at 4 and 40 ° C.

Stabilized glycerin formaldehyde was added to stabilize the formulation. The color of the product remained stable at 4 and 40 ° C.

The color of the product remained stable at 4 and 40 ° C.

Example summary It seems that the most stable formulation of the tylosin and flunixin formulations is the F5 formulation. Next, a method for preparing an F5 formulation is described.

Procedure for preparing the F5 formulation In a clean and dry production container: 1. Load the required amount of glycerin Stabilized formaldehyde 2. Add and mix 25% propylene glycol 3. Heat the mixture from 70 to 80 ° C Add and dissolve the tylosin base and mix correctly (keep at temperatures of 70 to 80 ° C) Add and dissolve flunixin meglumine and mix correctly (keep at temperatures of 70 to 80 ° C) Cool the dough to less than 30 ° C Complete the final volume with propylene glycol and Mix correctly for 10 minutes Example 2 Two other formulations (F9 and FIO) were studied, with tylosin and caprofen or meloxicam. These formulations and processing procedures are described below.

Table 2 Combinations of tylosin / carprofen and tylosin / meloxicaro.

F9 FIO ** 5% of authorized surpluses were added Production procedure for F9 and FIO 1. In a container of clean and dry processing 2. Load the required amount of NMP 3. Add and mix benzyl alcohol 4. Add and mix (35%) of propylene glycol 5. Heat the mixture from 70 to 80 ° C 6. Add and dissolve the tylosin base and mix correctly (maintain at temperatures of 70 to 80 ° C) 7. Add and dissolve meloxicam / carprofen and mix correctly (maintain at temperatures of 70 to 80 ° C) 8. Cool the dough to less than 30 ° C 9. Complete the final volume with propylene glycol and mix correctly for 10 minutes.

The inventors studied a combination that contains tylosin / ketoprofen 5% of aggregate authorized surpluses (20% according to prospectus) '* 2% of added authorized surpluses (4% according to prospectus) In this water-based compound, tylosin / ketoprofen was stable. However, when the inventors increased the concentration of ketoprofen to more than 4% w / v, the compound lost its stability in an aqueous-based compound.

Production procedure for Fll 1. In a processing vessel, load the required amount of propylene glycol, benzyl alcohol and mix well 2. Heat the mixture at 60 to 70 ° C 3. Add and dissolve the tylosin base and mix correctly (maintain at temperatures of 60 to 70 ° C) Corroborate the clarity of the solution 4. Add and dissolve ketoprofen and mix correctly Cool the dough at room temperature 5. Control the pH Should be between 8.8 to 9.5 If necessary, regulate the pH between 8.8 to 9.5 using 10% w / v sodium hydroxide solution or 5% w / v hydrochloric acid 6. Complete the final volume with water for injection and mix correctly for 10 to 15 minutes 7. Control the pH. It should be between 8.8 to 9.5 Example 4 The objective of the inventors is to achieve a compound that. contains higher concentrations of NSAID ketoprofen than the one described in Example 3, together with tylosin. The reasoning was that increasing the temperature of NSAIDs would synergistically improve the effectiveness of the antibiotic at the site of infection, while also allowing for more pain relief.

The inventors did not wish to return to a suspension, and then studied more severe non-aqueous base solvents to stabilize the NSAID at higher loads. A stable compound in solution could be successfully achieved. However, it was expected that the compound obtained would produce a more severe reaction and pain at the injection site when non-aqueous solvents were used. ** 5% of added authorized surpluses (20% according to prospectus) Production procedure for F12 1. In a clean and dry production vessel, load the required amount of N-methyl-2-pyrrolidone (NMP) 2. Load the required amount of benzyl alcohol into the container and mix correctly 3. Load propylene glycol (35% of the batch size) into the container and mix correctly 4. Heat the dough from 50 to 65 ° C 5. Add and dissolve the hydroxybutyl toluene (BHT) and mix 6. Add and dissolve the tylosin base and mix (keep at temperatures of 55 to 65 ° C) I. Add and dissolve ketoprofen and mix 8. Corroborate the clarity of the solution 9. Complete the volume to 98% of the batch size with propylene glycol and mix correctly 10. Cool the dough to 25 to 30 ° C and mix II. Complete the final volume with propylene glycol and mix correctly Example 5: Preliminary studies in animals to study the reaction at the injection site A study was conducted to compare therapeutic equivalence and side effects (site reaction and pain during administration) between the study compound containing 20% tylosin and 6% ketoprofen (as in Example 4) and the compounds reference industrial The reference industrial compounds were Tylan (Elanco Animal Health, Auckland, New Zealand) and Ketofen (Merial New Zealand, Auckland, New Zealand) when they are administered intramuscularly to dairy cattle. Tylan contains 20% w / v tylosin; with a prescribed dose of 10 mg / kg of body weight. Ketofen contains 10% w / v of ketoprofen with the prescribed dose of 3 mg / kg. Both Tylan and Ketofen are water-based compounds.

Twenty individuals were selected at various stages of lactation, in a range of 4 to 6 years of age from a herd of approximately 1200 individuals.

They were separated in pairs and placed randomly in group 1 or group 2. The study was cross-linking in both directions, in which each group was treated with each of the two treatments during the course of the study phase in animals, obtaining 20 treated animals in each treatment period. The constant dose was maintained between both groups, namely 10 mg / kg for tylosin, 3 mg / kg ketoprofen.

After milking on day 0, the dairy cattle were treated based on the standing weight of the animal. Blood samples were collected in 11 instances and recorded in real time for a period of 24 hours.

As indicated below, the studies revealed that the study compound and the reference industrial compounds are bioequivalent.

These results can be more easily understood by consulting the graph of Figure 1. Where the solid lines represent the tylosin serum concentrations when treated with the study compound of Example 4. The dotted lines represent serum tylosin concentrations when treated with Tylan.

In the graph of Figure 2, the solid lines represent the serum concentrations of ketoprofen when treated with the study compound of Example 4. The dotted lines represent the serum concentrations of ketoprofen when treated with Ketofen.

In summary, according to the study, the bioequivalence of both compounds was similar for constant doses. This indicates that the bioavailability of the combined compound is good, relative to the reference industrial compounds that are administered separately. This in itself is a great advantage.

Another advantage that was observed in this study is that contrary to what was expected, the study compound did not show more Adverse reactions (pain or reaction at the site) compared to the reference industrial compounds (Tylan and Ketofen).

This was unexpected because it was thought that the non-aqueous solvent used in the study compound in Example 4 would produce adverse side effects upon injection. If one takes into account that the dose was the same in the study of group 1 and group 2, it could be expected that a dose of NSAID (for example ketoprofen) greater than 3 mg / kg would avoid a greater reaction and / or pain in the site (and even its moderation) compared to what results from the use of the compounds currently available. It is possible to use higher doses with higher concentrations of NSAIDs because a non-aqueous solvent is used to stabilize the active ingredient in the compound.

Example 6: Stability studies A stability study was carried out in three batches of 4.5 L of the compounds of Example 4.

The batches were placed in 100 ml glass flasks and stored under the assigned storage conditions of 25 ° C / 60% relative humidity, 30 ° C / 65% relative humidity and 40 ° C / 75% relative humidity.

The physical and chemical characteristics of the product were recorded at regular intervals as recommended by the ACV.

As shown in the following tables, the conditions were stable for at least six months at 25 ° C / 60% relative humidity and 30 ° C / 65% relative humidity.

Storage conditions: 25 ° C / 60% relative humidity Storage conditions: 30 ° C / 65% relative humidity Storage conditions: 40 ° C / 75% relative humidity Storage conditions: 25 ° C / 60% relative humidity Storage conditions: 30 ° C / 65% relative humidity Storage conditions: 40 ° C / 75% relative humidity The aspects of the present invention have been described by way of example only and it should be understood that it is possible to apply modifications and additions thereto without departing from the spirit thereof as defined in the appended claims.

Claims (1)

1. Claims An injectable compound that is used for the treatment of a microbial infection in the mammary gland of an animal. The compound includes A non-steroidal anti-inflammatory drug (NSAID); An antibiotic selected from the group that includes a beta-lactam antibiotic and a macrolide antibiotic; A vehicle With the characteristic that The compound includes at least two non-aqueous solvents, At least one of the non-aqueous solvents is a solubilizing agent; Y At least one non-aqueous solvent is a vehicle, The concentration of vehicle in the compound is greater than the concentration of solubilizing agent in the compound; The solubilizing agent is characterized by the property that it dissolves the NSAID and / or the antibiotic more rapidly than a vehicle; Y The NSAID and the antibiotic dissolve in the at least two non-aqueous solvents. The compound claimed in claim 1, wherein the compound is non-aqueous. The compound as claimed in claim 1 or claim 2, wherein the non-aqueous solvent has one or more of the following characteristics: a) Dielectric point above 30 to 25 ° C; b) A boiling point greater than 100 ° C; I c) It is a bipolar aprotic solvent, d) The compound as claimed in any of the preceding claims wherein at least two non-aqueous solvents are selected from the group of N-methyl-2-pyrrolidone (NMP), dimethyl sulfoxide (DMSO), dimethylacetamide (DMA), glycerin formaldehyde, propylene glycol, benzyl alcohol, and combinations thereof. The compound as claimed in claim 4, wherein the first non-aqueous solvent is glycerin formaldehyde or NMP and wherein the second non-aqueous solvent is propylene glycol. The compound as claimed in the preceding claims, wherein an NSAID of the group of carprofen, naproxen, ibuprofen, ketoprofen, piroxicam, diclofenac, etodolac, flunixin, deracoxib, meloxicam, celecoxib, rofecoxib, and combinations thereof. The compound as claimed in the preceding claims, wherein the NSAID of flunixin, meloxicam, carprofen, ketoprofen and combinations thereof is selected. The compound as claimed in any of the preceding claims, wherein NSAID is present at concentrations of 1 to 15% w / v. The compound as claimed in claim 7, wherein the flunixin concentration is about 7.3% w / v, the concentration of meloxicam is about 1.0% w / v, the concentration of carprofen is about 3%. , 0% w / v the concentration of ketoprofen is approximately 4 to 10% w / v. The compound as claimed in any of the preceding claims wherein the macrolide class antibiotic is selected from azithromycin A zithromycin, clarithromycin Clarithromycin, dirithromycin dirithromycin, erythromycin Erythromycin, roxithromycin Roxithromycin, telithromycin Telithromycin, carbomycin A Carbomycin A, josamycin Josamycin, kitasamycin Kitasamycin, midecamycin / raidecamycin acetate Midecamicine / midecamicine acétate, oleandomycin Oleandomycin, spiramycin Spiramycin, troleandomycin Troleandomycin, and tylosin Tylosin / tylocine, and combinations thereof. The compound as claimed in any of the preceding claims, wherein the macrolide antibiotic is tylosin. The compound as claimed in any of the preceding claims, wherein the beta-lactam antibiotic is a penetamate. The compound as claimed in any of the preceding claims, wherein the antibiotic is present at concentrations of 10 to 35% w / v. The compound as claimed in any of the preceding claims, wherein the antibiotic is present at concentrations of about 20.0% w / v. The compound as claimed in any of claims 1 to 14, wherein the quotient of NSAID and antibiotic in the compound is about 1: 3 w / v. The compound as claimed in any of the preceding claims, the compound includes at least one antioxidant. The compound as claimed in claim 16, wherein the antioxidant is selected from sodium formaldehyde sulfoxylate or hydroxybutyl toluene (BHT) and combinations thereof. The compound as claimed in any of claims 16 or 17, wherein the antioxidant is present in the compound at concentrations between 0.01 - 0.3% w / v. A method to treat an animal against a microbial infection wherein the method for treating an animal against a microbial infection is a method for treating an animal against an internal microbial infection which is characterized by the stages of administering a compound as claimed in claims 1 to 20 by injection to an animal. The method as claimed in claim 19 wherein the compound is injected subcutaneously. The method as claimed in claim 19 or 20 wherein the animal is treated with a dose of 5 to 10 milligrams of antibiotic per kg of animal per day (mg / kg / day) and 0.2 to 4 mg / kg / day of NSAIDs. The method as claimed in any of claims 19 to 21 wherein the animal is treated with 5 to 10 mg / kg / day of antibiotic and any of the following doses of NSAIDs: a2.2 mg / kg / day of flunixin b.0,5 mg / kg / day of meloxicam c.3 mg / kg / day of ketoprofen d. 1.4 mg / kg / day of carprofen The method as claimed in any of claims 19 to 22 wherein the compound is used to treat mastitis and / or early stages of clinical mastitis. The use of a compound as claimed in any of claims 1 to 18 in the production of a medicament for the treatment of a microbial infection. A method for preparing a compound when used for the treatment of a microbial injection in the mammary gland containing NSAIDs and an antibiotic selected from the group including a macrolide and beta-lactam antibiotic which is characterized by the stages of a) Adding at least two non-aqueous solvents to the compound, wherein at least one non-aqueous solvent is a solubilizing agent and at least one non-aqueous solvent is a vehicle, wherein the vehicle concentration in the compound is greater than the concentration of solubilizing agent in the compound; b) Dissolve the NSAID and the antibiotic in the at least two non-aqueous solvents. The method as claimed in claim 25, wherein the NSAID in the compound increases to a concentration (% w / v) which would remove storage stability at room temperature in the corresponding aqueous-based compound. The method as claimed in claim 25 or claim 26 wherein NSAID is ketoprofen and wherein the concentration of ketoprofen is greater than 4% w / v. The method as claimed in any of claims 25 to 27, wherein at least two non-aqueous solvents are added to a mixing vessel before adding NSAID or antibiotic. A method as claimed in any of claims 25 to 28 wherein the vehicle is added to a solubilizing agent before adding either NSAID or antibiotic. A method as claimed in any of claims 25 to 29 wherein the compound is maintained between 60 and 80 ° C until NSAIDs and antibiotics are substantially dissolved in at least two non-aqueous solvents. A method as claimed in any. ' of claims 25 to 30, wherein the antibiotic is added to at least two non-aqueous solvents and dissolved before adding the NSAID. A method as claimed in any of claims 25 to 31, wherein the NSAID is added to the dissolved antibiotic mixture and to at least two non-aqueous solvents. A method as claimed in any of claims 25 to 31 wherein the compound containing dissolved NSAID and antibiotic is cooled to about 30 ° C before reaching the final volume (s) with a non-aqueous solvent and mixing for about 10 minutes. A compound as is substantially described herein, with reference to formulations F3-F10 which are depicted in Examples 1-3 in the Best Ways to Practice the Invention. A method for preparing a compound that includes an NSAID and an antibiotic substantially as described herein with reference to the production procedure detailed in Examples 1 to 3 in the Best Ways to Practice the Invention.