(12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (10) International Publication Number (43) International Publication Date WO 2015/067959 Al 14 May 2015 (14.05.2015) P O P C T

(51) International Patent Classification: (81) Designated States (unless otherwise indicated, for every A61K 31/341 (2006.01) A61K 31/7052 (2006.01) kind of national protection available): AE, AG, AL, AM, A61K 31/405 (2006.01) A61K 45/06 (2006.01) AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, A61K 31/44 (2006.01) A61P 11/00 (2006.01) BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DK, DM, A61K 31/5415 (2006.01) DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IR, IS, JP, KE, KG, KN, KP, KR, (21) International Application Number: KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, PCT/GB2014/053321 MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, (22) International Filing Date: PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, 7 November 2014 (07.1 1.2014) SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW. (25) Filing Language: English (84) Designated States (unless otherwise indicated, for every (26) Publication Language: English kind of regional protection available): ARIPO (BW, GH, (30) Priority Data: GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, 13 19762.' 8 November 2013 (08. 11.2013) GB TZ, UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, TJ, TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, (71) Applicant: NORBROOK LABORATORIES LIMITED DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, [GB/GB]; 105 Armagh Road, Newry, Co. Down, Northern LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, Ireland, BT35 6PU (GB). SM, TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, KM, ML, MR, NE, SN, TD, TG). (72) Inventor: HAUGHEY, James; c/o Norbrook Laboratories Limited, 105 Armagh Road, Newry, Co. Down, Northern Published: Ireland BT35 6PU (GB). — with international search report (Art. 21(3)) (74) Agent: HGF LIMITED; 140 London Wall, Greater Lon don, EC2Y 5DN (GB).

© o (54) Title: TULATHROMYCIN AND NONSTEROIDAL ANTI-INFLAMMATORY DRUG COMPOSITIONS (57) Abstract: The present invention relates to novel compositions containing tulathromycin and nonsteroidal anti-inflammatory drugs, pharmaceutical formulations comprising said compositions and their use for treatment and prevention of respiratory diseases in mammals. Tulathromycin and Nonsteroidal Anti-Inflammatory Drug Compositions

TechnicalField

The present invention relates to novel compositions containing tulathromycin and nonsteroidal anti-inflammatory drugs, pharmaceutical formulations comprising said compositions and their use for treatment and prevention of respiratory diseases in mammals.

Background Art

Non-steroidal anti-inflammatory drugs (NSAID) are a group of medicaments with , anti-inflammatory and effects. NSAID are widely administered for the treatment of mild to severe . Among the most commonly used members of the NSAID class of drugs are , fluxinin, , and . See Peterson K, et ah, "Drug Class Review: Nonsteroidal Antiinflammatory Drugs (NSAIDs)" (Oregon Health and Science University, Portland, Oregon, US, 2010). Macrolides are broad spectrum antibiotics formed by a large lactone ring that varies in size from 12 to 16 atoms. Macrolide antibiotics exhibit good activity against Gram-positive aerobes such as Streptococcus pneumoniae and Haemophilus influenzae. See Zhanel G, et ah, Drugs 2001; 61(4):443-498. Their principal mechanism of action against pathogens involves the direct inhibition of essential protein biosynthesis by selective binding to bacterial 50S ribosomal subunits. In addition to their antimicrobial function, macrolide antibiotics are known to modulate immune cell functions. This class of antibiotics is known to influence the recruitment and infiltration of neutrophils and to alter their ability to secrete histotoxic compounds and proinflammatory cytokines. See Ichikawa Y, et ah, Am. Rev. Respir. Dis. 1992; 146: 196-203, Lee W, et ah, Vet. Res. 2004; 35:213-224, Mikasa K, et ah, J. Antimicrob. Chemother. 1992; 30:339-348, Takeshita K, et ah, Drugs Exp. Clin. Res. 1989; 15:527-533 and Tsuchihashi Y, et ah, Antimicrob. Agents Chemother. 2002; 46:1101-1104. Recent findings also indicate that some macrolides may induce cellular death by apoptosis and block NF-κΒ signaling in T lymphocytes, macrophages or epithelial cells. See Aoshiba K, et al, Antimicrob. Agents Chemother. 1995; 39:872-877, Buret A, et al, Can. J. Vet. Res. 2010; 74:1-10, Chin A, et al, Antimicrob. Agents Chemother. 2000; 44:2465-2470, Chin A, et al, Am. J. Vet. Res. 1998; 59:765-771, Lee W, et al, Vet. Res. 2004; 35:213-224, Leiva M, etal, FEMS Immunol. Med. Microbiol. 2008; 53:343-350, Ou X, et al, Respirology 2008; 13:63-72 and Wu L, et al, Int. Immunopharmacol. 2009; 9:1105-1109. However, the precise mechanisms underlying the anti-inflammatory and proapoptotic activities of macrolides in neutrophils remain unclear. Tulathromycin is a triamilide macrolide characterized by a 15-membered ring composed of 3 polar amine groups. This particular structure distinguishes it from other macrolides. See Evans N, Vet. Ther. 2005; 6:83-95. The drug is used in the treatment and prevention of respiratory diseases in cattle and swine. The therapeutic success of tulathromycin is partially attributed to its pharmacodynamic concentration in appropriate tissues and low inhibitory concentrations against various bacterial pathogens. Tulathromycin also has a very high affinity for uptake within neutrophils and, to a lesser extent, macrophages, which helps target the delivery of the drug to infected tissues. Compared to other antibiotics used in the treatment and prevention of bovine respiratory disease, tulathromycin has shown superior clinical efficacy and clears the infection and inflammation for reasons that remain not completely understood. See Nutsch R, et al , Vet. Ther. 2005; 6:167-179, Mount J, et al, Can. J. Vet. Res. 2009; 73:49-57, Skogerboe T, et al, Vet. Ther. 2005; 6:180-196.

Summary of the Invention

The present invention relates to a composition comprising tulathromycin and at least one nonsteroidal anti-inflammatory drug ("NSAID"). Surprisingly, the same antibiotic efficacy is attained when a lower than recommended amount of tulathromycin is used in the presence of a NSAID. In addition to the obvious economic benefits of this composition, a lower tulathromycin have the further advantage of reducing the risk of developing antibiotic-resistant bacteria. In a first aspect, the present invention is directed to a composition comprising tulathromycin and at least one NSAID. In a second aspect, the present invention refers to a pharmaceutical formulation comprising the composition of the first aspect and at least one pharmaceutically acceptable carrier, additive, adjuvant or vehicle. In a third aspect, the present invention relates to a process for preparing the pharmaceutical formulation of the invention, which comprises mixing the composition of the first aspect and at least one pharmaceutically acceptable carrier, additive, adjuvant or vehicle. In a fourth aspect, the present invention refers to a composition according to the first aspect or a pharmaceutical formulation according to the second aspect for its use as a medicament. In a fifth aspect, the present invention is directed to a composition according to the first aspect or a pharmaceutical formulation according to the second aspect for its use in the treatment or prevention of respiratory diseases in mammals. Alternatively, the present invention is directed to the use of the composition according to the first aspect in the manufacture of a medicament for the treatment or prevention of a respiratory disease in mammals. In a sixth aspect, the present invention refers to a method of treating or preventing the inception of a respiratory disease in a subject, which comprises administering to the subject a therapeutically effective amount of the composition of the first aspect or the pharmaceutical formulation of the third aspect of the invention.

Description of the Embodiments

This invention relates to a novel composition that enables the administration of a lower yet effective dosage of tulathromycin to subjects suffering, or in risk of suffering, respiratory diseases. The components of the composition are tulathromycin and at least one NSAID. The NSAID can be an derivative, an enolic acid derivative, a derivative, a derivative, a derivative, a selective cyclooxigenase-2 inhibitor, a sulphoanilide compound or a pharmaceutically acceptable salt, isomer, enantiomer or tautomers thereof.

1. Definitions The term "auxiliary" as used herein, means any component, other than the active substance(s) intentionally added to the formulation of a dosage form. Exemplary auxiliaries are acids (e.g. citric or tartaric acid), alcohols, a bicarbonate (e.g. sodium or potassium), binders, colorants, excipients, disintegrants, fillers, flavoring agents, flavor masking agents, foaming agents, lubricants, plastizicers, pore-forming agents, sodium chloride, stabilizers, sugars and sweeteners. Some auxiliaries can serve multiple purposes (e.g. filler and disintegrant). See US Pharmacopoeia. The term "pharmaceutically acceptable salt" as used herein, refers o any salt of acidic or basic nature which may be formed with any of the compounds of the invention. Examples of acidic salts include, but are not limited to, the hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, acid citrate, tartrate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucaronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate salts. Examples of basic salts include, but are not limited, to the calcium, magnesium, sodium and potassium salts. The terms "prevent", "preventing" and "prevention" as used herein, refer to inhibiting the inception, or decreasing the occurrence or recurrence, of a disease in a subject. Prevention may be complete (e.g. the total absence of pathological cells in a subject) or partial. Prevention also refers to a reduced susceptibility to a clinical condition. The term "subject" as used herein, refers to a mammalian, such as a human being, a non-human primate (e.g. chimpanzees and other apes and monkey species), a farm animal (e.g. cattle, sheep, pigs, goats and horses), a domestic mammal (e.g. dogs and cats), or a laboratory animal (e.g. rodents, such as mice, rats and guinea pigs. The term does not denote any particular age or sex. The term "therapeutically effective amount" as used herein, refers to any amount of a composition or pharmaceutical formulation which, when administered to a subject: i) prevents the inception or recurrence or ii) causes the reduction or remission of the disease or codition against which the composition or formulation is effective. The term "treat" or "treatment" as used herein, refers to the control of the progression of a disease after its clinical signs have appeared. Control of the disease progression is understood to mean the beneficial or desired clinical results that include, but are not limited to, reduction of the symptoms, reduction of the duration of the disease, stabilization of pathological states (specifically to avoid additional deterioration), delaying the progression of the disease, improving the pathological state, and remission (both partial and total). The control of progression of the disease also involves an extension of survival, compared with the expected survival if treatment is not applied. The term "tulathromycin" as used herein, refers to ( ,3 ,4 ,5 ,8 ,

1OR,11R,12S,13S,14R)-13-[[2,6-dideoxy-3 -C-methy 1-3 -O-methyl-4-C- [(propylamino)meth yl]-a-L-ribo-hexopyrano-syl]oxy]-2-ethyl-3,4,10-trihydroxy-3,5,8,10,12,14-hexamethyl-ll -[[3,4,6-trideoxy-3-(dimethylamino )-P-D-xylo-hexopyranosyl]-oxy]-l-oxa-6-azacyclopenta decan-15-one, CAS [217500-96-4], of molecular formula C41H79N3O12 and structural formula:

Tulathromycin exists in two isomer forms: A and B . The product has been proved effective for the treatment and prevention of respiratory diseases caused by Mannheimia haemolytica, Pasteurella multocida, Histophilus somni and Mycoplasma bovis in cattle and Actinobacillus pleuropneumoniae, Pasteurella multocida, Bordetella bronchiseptica, Haemophilus parasuis and Mycoplasma hyopneumoniae in pigs.

Tulathromycin and nonsteroidal anti-inflammatory drugs compositions In a first aspect, the present invention is directed to a composition comprising tulathromycin and at least one NSAID. Preferably, tulathromycin is about 95% (w/w) or less, and the NSAID is about 5% (w/w) or more, of the total composition weight. More preferably, tulathromycin is about 95% to about 50% w/w, and the NSAID is from about 5% to about 50% w/w, of the total composition weight. A composition wherein tulathromycin is from about 80% to about 60% w/w, and the NSAID is from about 20% to about 40% w/w, of the total composition weight is further preferred. In one embodiment of the first aspect of the invention, the composition comprises a mixture of the tulathromycin A and B isomers. More preferably, the isomers are present in the mixture in a ratio of about 9 to 1 of the tulathromycin isomer A form to the tulathromycin isomer B form. The tulathromycin compounds of the invention comprise the pharmaceutically acceptable salts, isomers, enantiomers and tautomers of the tulathromycin isomer A and B forms and the mixtures thereof. In one embodiment, the nonsteroidal anti-inflammatory drug of the invention is an acetic acid derivative, an enolic acid derivative, a fenamic acid derivative, a propionic acid derivative, a salicylic acid derivative, a selective cyclooxigenase-2 inhibitor, a sulphoanilide or a pharmaceutically acceptable salt, isomer, enantiomer or tautomer thereof. Preferably, the acetic acid derivative is , , benzofenac, , cinmetacin, clidenac, clopirac, , , , fenclorac, , fluxinin, furafenac, ibufenac, indomethacin, isoxepac, , , , , oxepinac, pirozolac, , , , or a pharmaceutically acceptable salt, isomer, enantiomer or tautomer thereof. Preferably, the fenamic acid derivative is bucloxic acid, etofenamic acid, , , , piromidic acid, protizinic acid, , or a pharmaceutically acceptable salt, isomer, enantiomer or tautomer thereof. Preferably, the propionic acid derivative is , butibufen, carprofen, cicloprofen, , , , , , , furaprofen, furobufen, , , , , , , oxaprozi, , or a pharmaceutically acceptable salt, isomer, enantiomer or tautomer thereof. Preferably, the salicylic acid derivative is acetylsalicylic acid, choline salicylate, , , methylsalicylate, mesalamine, , , , sodium thiosalicylate, sulfasalazine or a pharmaceutically acceptable salt, isomer, enantiomer or tautomer thereof. Preferably, the selective cyclooxigenase-2 inhibitor is , , , firocoxib, , , , , , , valecoxib or a pharmaceutically acceptable salt, isomer, enantiomer or tautomer thereof. Preferably, the sulphoanilide is . Preferably, the enolic acid derivative is , , , meloxicam, , tecnoxicam or a pharmaceutically acceptable salt, isomer, enantiomer or tautomer thereof. More preferably, the enolic acid derivative is meloxicam or a pharmaceutically acceptable salt, isomer, enantiomer or tautomer thereof. Any compound referred to herein is intended to represent such specific compound as well as certain variations or forms of it. In particular, compounds referred to herein may have asymmetric centers and therefore exist in different enantiomeric or diastereomeric forms. Thus, any given compound referred to herein is intended to represent any one of a racemate, one or more enantiomeric forms, one or more diastereomeric forms, and mixtures thereof. Likewise, stereoisomerism or geometric isomerism about the double bond is also possible, therefore in some cases the molecule could exist as trans and cis isomers. If the molecule contains several double bonds, each double bond will have its own stereoisomerism, that could be the same as, or different to, the stereoisomerism of the other double bonds of the molecule. Furthermore, compounds referred to herein may exist as atropisomers. All the stereoisomers including enantiomers, diastereoisomers, geometric isomers and atropisomers of the compounds referred to herein, and mixtures thereof, are considered within the scope of the present invention. Furthermore, any compound referred to herein may exist as tautomers. Specifically, the term tautomer refers to one of two or more structural isomers of a compound that exist in equilibrium and are readily converted from one isomeric form to another. Common tautomeric pairs include, but are not limited to, amine-imine, amide-imidic acid, keto-enol, and lactam-lactim.

3. Pharmaceuticalformulations

In another aspect, the present invention relates to a pharmaceutical formulation comprising a composition of tulathromycin and at least one NSAID, a pharmaceutically acceptable salt, isomer, enantiomer or tautomer thereof, and at least a pharmaceutically acceptable carrier, additive, adjuvant or vehicle. The tulathromycin and NSAID compositions of the invention may be formulated for their simultaneous, separate or sequential administration, with at least one pharmaceutically acceptable carrier, additive, adjuvant or vehicle. Therefore, these compositions may be administered:

i) as part of the same pharmaceutical formulation, the two active ingredients being then administered always simultaneously or

ii) as two units, each unit with one of the active ingredients and making possible their simultaneous, sequential or separate administration. Preferably, tulathromycin and the NSAID are administered simultaneously in the same pharmaceutical formulation. In another embodiment, tulathromycin is independently administered from the NSAID (i.e. in two units) but at the same time. An example of this dosage regime would be the administration of the tulathromycin and firocoxib composition. The auxiliary materials of the pharmaceutical formulations can be carriers, excipients, support materials, lubricants, fillers, solvents, diluents, colorants, and flavor conditioners such as sugars, antioxidants or agglutinants. In the case of suppositories, this may imply waxes or fatty acid esters or preservatives, emulsifiers or carriers for parenteral application. The selection of these auxiliary materials and the amounts to be used will depend on the form of application of the pharmaceutical formulation. Examples of auxiliaries that may be used according to the invention include, but are not limited to, glycerol triacetate, monothioglycerol, polyethylene glycol, propylene glycol and water. The pharmaceutical formulations according to the invention can be adapted to any form of administration, such as the subcutaneous, intramuscular, intra-articular, intraperitoneal, intravenous, pulmonary, nasal, percutaneous or oral administration. Suitable formulations for oral administration include, but are not limited to, tablets, capsules, chewing gums, granules, drops, syrups, pastes, feed premixes, gels or boluses. Suitable preparations for parenteral administration are solutions, suspensions, reconstitutable dry preparations or sprays. Preferably, the pharmaceutical formulation is suitable for intramuscular or subcutaneous injection. The formulations of the invention may be produced following methods known in the art. See Gunnar A, Ed., "Remington: The Science and Practice of Pharmacy" 20th ed. (Lippincott Williams & Wilkins, Philadelphia, PA, US, 2003).

4. Use of compositions andformulations for the treatment or prevention of respiratory diseases

In another aspect, the present invention refers to the use of a composition or a pharmaceutical formulation according to the invention for the treatment and prevention of respiratory diseases. In a particular embodiment, said respiratory diseases are mediated by bacterial pathogens of the Mannheimia spp, Pasteurella spp, Histophilus spp, Mycoplasma spp, Actinobacillus spp, Bordetella spp, and Haemophilus spp. Alternatively, the present invention refers to a method for treating or preventing a respiratory disease in a subject, which comprises administering to the subject a therapeutically effective amount of a composition or pharmaceutical formulation of the invention. Preferably, the subject is cattle or a pig. Preferably, the composition or formulation of the invention is administered once by injection. The subcutaneous route of administration is preferred for cattle. The intramuscular route is the preferred way of delivery for pigs. This invention is further illustrated by the following examples which should not be construed as limiting. The contents of all documents cited throughout this application are incorporated herein in their entirety by reference.

Generalprocedures

1. Tulathromycin, carprofen, fluxinin meglumine and meloxicam compositions

Liquid formulations containing the composition of tulathromycin with carprofen, fluxinin meglumine and meloxicam are prepared by adding the active ingredients to a monothioglycerol, propylene glycol and water vehicle as described in the art to achieve the concentrations of the examples.

2. Tulathromycin andfirocoxib composition An aqueous formulation of tulathromycin is prepared by adding the active ingredient to a monothioglycerol, propylene glycol and water vehicle as described in the art. A non aqueous formulation of firocoxib is prepared by adding the active ingredient to a glycerol triacetate and polyethylene glycol vehicle as described in the art. The aqueous and non aqueous formulations are administered separately.

3. Statistical analysis

P values are determined based on a chi-square analysis and by comparing the results obtained for each tulathromycin and tulathromycin-NSAID compositions and their respective placebo vehicle.

Example! Effectiveness of the tulathromycin and NSAID compositionsfor the treatment bovine respiratory diseases

Nasopharyngeal swabs were collected from calves aged 4 to 11 months old and weighing 150 to 320 kg. The samples were analyzed for the presence of Mannheimia haemolytica, Pasteurella multocida, Histophilus somni and Mycoplasma spp pathogens. Calves positive for any of the pathogens and having a rectal temperature of > 40 °C were enrolled in the assay. Tulathromycin and the tulathromycin and NSAID compositions were administered as a single dosage at a ratio of 2.4 mL of solution per 100 kg of body weight. Saline solution at an equivalent volume of 0.025 mL/kg of body weight was used as placebo. The solutions were administered subcutaneously in the neck at day 0. See Table 1. The treatment success was determined on day 14. A treatment was classified as successful if it the subject survived through day 14 without being classified as a treatment failure. A calf was classified as a treatment failure if it had a rectal temperature of > 40 °C on any of days 3-14.

Table 1 Percentage Assay Active ingredients Treated Cured cured p value 1 Placebo 33 8 24.2% ... 2 Tulathromycin 100 mg/mL 35 27 77.1% p< 0 1

3 Tulathromycin 90 mg/mL 38 28 72.5% p< 0 1 Carprofen 50 mg/mL 4 Tulathromycin 80 mg/mL 34 23 67.6% p< 0 1 Carprofen 55 mg/mL 5 Tulathromycin 90 mg/mL 36 22 61.1% p< 0 1 Firocoxib 20 mg/mL 6 Tulathromycin 80 mg/mL 32 18 56.2% p< 0 1 Firocoxib 22.5 mg/mL 7 Tulathromycin 90 mg/mL 33 24 72.7% p< 0 1 Fluxinin meglumine 50 mg/mL 8 Tulathromycin 80 mg/mL 35 23 65.7% p< 0 1 Fluxinin meglumine 55 mg/mL 9 Tulathromycin 90 mg/mL 38 28 76.3% p< 0 1 Meloxicam 20 mg/mL 10 Tulathromycin 80 mg/mL 35 24 68.6% p< 0 1 Meloxicam 22.5 mg/mL

Example 2 Effectiveness of the tulathromycin and NSAID compositionsfor the treatmentporcine respiratory diseases

Nasopharyngeal swabs were collected from pigs aged 6 to 24 weeks of age and weighing 10 to 62.7 kg. The samples were analyzed for the presence of Actinobacillus pleuropneumoniae, Pasteurella multocida, Bordetella bronchiseptica and Haemophilus parasuis pathogens. Pigs positive for any of the pathogens and having a rectal temperature of > 40 °C were enrolled in the assay. Tulathromycin and the tulathromycin and NSAID compositions were administered as a single dosage at a ratio of 0.25 mL of solution per 10 kg of body weight. Saline solution at an equivalent volume of 0.025 mL/kg of body weight was used as placebo. The solutions were administered by intramuscular injection at day 0. See Table 2. The treatment success was determined on day 7. A treatment was classified as successful if it the subject survived through day 7 and had a rectal temperature of < 40 °C.

Table 2 Percentage Assay Active ingredients Treated Cured cured p value 1 1 Placebo 49 23 46.9% ... 12 Tulathromycin 100 mg/mL 4 1 29 70.7% p< 0 1

13 Tulathromycin 90 mg/mL 42 28 66.7% p< 0 1 Carprofen 50 mg/mL 14 Tulathromycin 80 mg/mL 46 28 60.8% p< 0 1 Carprofen 55 mg/mL 15 Tulathromycin 90 mg/mL 47 28 59.6% p< 0 1 Firocoxib 20 mg/mL 16 Tulathromycin 80 mg/mL 44 23 52.3% p< 0 1 Firocoxib 22.5 mg/mL 17 Tulathromycin 90 mg/mL 45 30 66.7% p< 0 1 Fluxinin meglumine 50 mg/mL 18 Tulathromycin 80 mg/mL 44 26 59.1% p< 0 1 Fluxinin meglumine 55 mg/mL 19 Tulathromycin 90 mg/mL 43 30 69.7% p< 0 1 Meloxicam 20 mg/mL 20 Tulathromycin 80 mg/mL 49 30 61.2% p< 0 1 Meloxicam 22.5 mg/mL

Example 3 Effectiveness of the tulathromycin and NSAID compositions in the prevention of bovine respiratory diseases

Calves exposed to pathogens associated to bovine respiratory diseases were recruited and distributed randomly among the different treatment groups. The subjects aged 4 to 1 1 months old and weighed 150 to 320 kg. Tulathromycin and the tulathromycin and meloxicam compositions were administered as a single dosage at a ratio of 2.4 mL of solution per 100 kg of body weight. Saline solution at an equivalent volume of 0.025 mL/kg of body weight was used as placebo. The solutions were administered subcutaneously in the neck at day 0. See Table 3. The treatment failure was determined on days 1-14. A treatment was classified as a failure if it the subject developed a rectal temperature of > 40 °C on any of days 1-14.

Table 3 Assay Active ingredients Treated Failures failures p value 2 1 Placebo 104 60 58.2% — 22 Tulathromycin 100 mg/mL 139 15 10.8% p< 0 1

23 Tulathromycin 90 mg/mL 134 15 11.1% p< 0 1 Meloxicam 20 mg/mL 24 Tulathromycin 80 mg/mL 141 17 12.1% p< 0 1 Meloxicam 22.5 mg/mL

The results above suggest that a when a NSAID is co-administered with tulathromycin, a lower antibiotic dosage can be used and still be efficacious in the treatment and prevention of respiratory diseases caused by of Mannheimia spp, Pasteurella spp, Histophilus spp, Mycoplasma spp, Actinobacillus spp, Bordetella spp and Haemophilus spp pathogens in cattle and pigs. Claims

1. A composition comprising tulathromycin and at least one nonsteroidal anti inflammatory drug. 2. The composition according to claim 1 wherein tulathromycin is from about 95% to about 50% w/w or the nonsteroidal anti-inflammatory drug is from about 5% to about 50% w/w based on the total weight of the composition. 3. The composition according to claim 1 wherein tulathromycin is from about 80% to about 60% w/w or the nonsteroidal anti-inflammatory drug is from about 20% to about 40% w/w based on the total weight of the composition. 4. The composition according to any preceding claim, wherein tulathromycin is a mixture of the tulathromycin A and B isomers.

5. The composition according to claim 4, wherein the isomers are present in the mixture in a ratio of about 9 to 1 of the tulathromycin isomer A form to the tulathromycin isomer B form.

6. The composition according to any preceding claim wherein the nonsteroidal anti inflammatory drug is an acetic acid derivative, an enolic acid derivative, a fenamic acid derivative, a propionic acid derivative, a salicylic acid derivative, a selective cyclooxigenase-2 inhibitor and a sulphoanilide.

7. The composition according to any one of claims 1 to 6 wherein the acetic acid derivative is aceclofenac, acemetacin, aclofenac, benzofenac, bromfenac, cinmetacin, clidenac, clopirac, diclofenac, etodolac, fenclofenac, fenclorac, fentiazac, fluxinin, furafenac, ibufenac, indomethacin, isoxepac, ketorolac, lonazolac, nabumetone, oxaprozin, oxepinac, pirozolac, sulindac, tenclofenac, tepoxalin, tiopinac, tolmetin, zidometacin, zomepirac or a pharmaceutically acceptable salt, isomer, enantiomer or tautomer thereof

8. The composition according to any one of claims 1 to 6 wherein the fenamic acid derivative is bucloxic acid, etofenamic acid, flufenamic acid, meclofenamic acid, mefenamic acid, niflumic, piromidic acid, protizinic acid, tiaprofenic acid, tolfenamic acid or a pharmaceutically acceptable salt, isomer, enantiomer or tautomer thereof. 9. The composition according to any one of claims 1 to 6 wherein the propionic acid derivative is , benoxaprofen, butibufen, carprofen, cicloprofen, dexibuprofen, dexketoprofen, fenbufen, fenoprofen, flunoxaprofen, fluprofen, flurbiprofen, furaprofen, furobufen, ibuprofen, indoprofen, ketoprofen, loxoprofen, miroprofen, naproxen, oxaprozin, pirprofen, , surprofen, tioxaprofen or a pharmaceutically acceptable salt, isomer, enantiomer or tautomer thereof. 10. The composition according to any one of claims 1 to 6 wherein the salicylic acid derivative is acetylsalicylic acid, choline salicylate, diflunisal, flufenisal, magnesium salicylate, methylsalicylate, mesalamine, salicylamide, salsalate, sodium salicylate, sodium thiosalicylate, sulfasalazine or a pharmaceutically acceptable salt, isomer, enantiomer or tautomer thereof 11. The composition according to any one of claims 1 to 6 wherein the selective cyclooxigenase-2 inhibitor is celecoxib, deracoxib, etoricoxib, firocoxib, lumiracoxib, paracetamol, parecoxib, phenylbutazone, robenacoxib, rofecoxib, valecoxib or a pharmaceutically acceptable salt, isomer, enantiomer or tautomer thereof. 12. The composition according to any one of claims 1 to 6 wherein the sulphoanilide is nimesulide.

13. The composition according to any one of claims 1 to 6 wherein the enolic acid derivative is droxicam, isoxicam, lornoxicam, meloxicam, piroxicam, sudoxicam, tecnoxicam a pharmaceutically acceptable salt, isomer, enantiomer or tautomer thereof . 14. A pharmaceutical formulation comprising the composition according to any one of

claims 1 to 13 or a pharmaceutically acceptable salt, isomer, enantiomer or tautomer thereof, and at least a pharmaceutically acceptable carrier, additive, adjuvant or vehicle.

15. The composition according to any one of claims 1 to 13 or the pharmaceutical formulation according to claim 14 for use as a medicament.

16. The composition according to any one of claims 1 to 13 or the pharmaceutical formulation according to claim 14 for use in the treatment or prevention of a respiratory disease in mammals. a medicament for the treatment or prevention of a respiratory disease in a mammal. 18. A method for treating or preventing a respiratory disease in a subject, which comprises administering to the subject a therapeutically effective amount of the

composition according to any one of claims 1 to 13 or the pharmaceutical formulation according to claim 14.

19. The method of claim 18 wherein the subject is cattle. 20. The method of claim 18 wherein the subject is a pig. A . CLASSIFICATION O F SUBJECT MATTER INV. A61K31/341 A61K31/405 A61K31/44 A61K31/5415 A61K31/7052 A61K45/06 A61P11/00 ADD.

According to International Patent Classification (IPC) o r t o both national classification and IPC

B . FIELDS SEARCHED Minimum documentation searched (classification system followed by classification symbols) A61K

Documentation searched other than minimum documentation to the extent that such documents are included in the fields searched

Electronic data base consulted during the international search (name of data base and, where practicable, search terms used)

EPO-Internal , WPI Data, BEI LSTEIN Data, EMBASE, BIOSIS, CHEM ABS Data

C . DOCUMENTS CONSIDERED T O B E RELEVANT

Category* Citation of document, with indication, where appropriate, of the relevant passages Relevant to claim No.

CN 101 461 821 A (QINGDAO CONTINENT 1-20 PHARMACEUTIC [CN] QINGDAO CONTINENT PHARM CO LTD) 24 June 2009 (2009-06-24) the whol e document 1-20

GUZEL M ET AL: "Cl i ni cal effi cacy of 1-7 , di cl ofenac sodi um and f l uni x i n megl umi ne 14-19 as adjuncts t o anti bacteri al treatment of respi ratory di sease of cal ves" , AUSTRALIAN VETERINARY JOURNAL, vol . 88, no. 6, June 2010 (2010-06) , pages 236-239 , XP002733513 , abstract 1-20 page 236 page 239 -/-

X| Further documents are listed in the continuation of Box C . See patent family annex.

* Special categories of cited documents : "T" later document published after the international filing date o r priority date and not in conflict with the application but cited to understand "A" document defining the general state of the art which is not considered the principle o r theory underlying the invention to be of particular relevance "E" earlier application o r patent but published o n o r after the international "X" document of particular relevance; the claimed invention cannot be filing date considered novel o r cannot b e considered to involve a n inventive "L" documentwhich may throw doubts o n priority claim(s) orwhich is step when the document is taken alone cited to establish the publication date of another citation o r other "Y" document of particular relevance; the claimed invention cannot be special reason (as specified) considered to involve a n inventive step when the document is "O" document referring to a n oral disclosure, use, exhibition o r other combined with one o r more other such documents, such combination means being obvious to a person skilled in the art "P" document published prior to the international filing date but later than the priority date claimed "&" document member of the same patent family

Date of the actual completion of the international search Date of mailing of the international search report

15 December 2014 16/01/2015

Name and mailing address of the ISA/ Authorized officer European Patent Office, P.B. 5818 Patentlaan 2 N L - 2280 HV Rijswijk Tel. (+31-70) 340-2040, Fax: (+31-70) 340-3016 Opravz , Petra C(Continuation). DOCUMENTS CONSIDERED TO BE RELEVANT

Category* Citation of document, with indication, where appropriate, of the relevant passages Relevant to claim No.

Y Anonymous : "Nonsteroi dal 1-20 anti - i nfl ammatory drugs" , The Merck Veteri nary Manual

5 October 2013 (2013-10-05) , XP002733514, Retri eved from the Internet: URL : http : //web . arch i ve . org/web/2013 1005055 326/http : //www. merckmanual s. com/vet/pharma col ogy/anti - i nf 1ammatory_agents/nonsteroi d al anti - i nfl amrnatory drugs . html [retri eved on 2014-12-09] the whol e document

T Anonymous : "Tul athromyci n" , 4,5 DrugFuture

XP002733512 , Retri eved from the Internet: URL: http : //www. drugf uture . com/chemdata/tul athromyci n. html [retri eved on 2014-12-09] the whol e document Patent document Publication Patent family Publication cited in search report date member(s) date

CN 101461821 24-06-2009 NONE