US 20110281920A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2011/0281920 A1 Holmes et al. (43) Pub. Date: Nov. 17, 2011
(54) INJECTABLE PARASITICIDAL Publication Classi?cation FORMULATIONS OF LEVAMISOLE AND MACROCYCLIC LACTONES (51) Int. Cl. A61K 31/429 (2006.01) (76) Inventors: Robert Holmes, Biirkenhead (NZ); A61P 33/10 (2006.01) Majid Razzak, The Palms (NZ); Alan Johnson, Papatoetoe (NZ); (52) US. Cl...... 514/368 Jitendra GosWami, Miyapur (IN); Atul AWasthi, Manukau City (NZ) (57) ABSTRACT (21) Appl. No.: 13/106,677 The present invention provides levamisole/macrocyclic lac tone-containing inj ectable formulations that are effective (22) Filed: May 12, 2011 against animal pests, including endoparasites. The formula tions may be used for combating parasites in or on birds and Related US. Application Data mammals. The invention also provides for an improved (60) Provisional application No. 61/333,882, ?led on May method for eradicating, controlling and preventing parasite 12, 2010. infestation of birds and mammals. Patent Application Publication Nov. 17, 2011 Sheet 1 of 3 US 2011/0281920 A1
Dimethylacetamide (DMA)
Add ‘ 4 . . Eprinomectin V ‘ Mix until clear
Add BHT Mix until clear
Methylparaben 2 Mix until clear
polysorbate 80 Mix until clear / Cremophor EL v DMA/Eprinomecrin/B HTAl/lelhylparaben/Pol 80mm,“ A ysorbate80 or Cremophor EL
Water for Injection
Add Levamisole > < Mix until clear Phosphate
V
Add Sol u t'onl A w“; Bulksohmon ‘H Mix~ with- silverson'
1831mm Wm] --——--~—->- Bulk Salim-On 4- Mix With Silverson
V Aseptic ?ltration through 0.22 micron
FIG. I Aseptic Filling Patent Application Publication Nov. 17, 2011 Sheet 2 0f 3 US 2011/0281920 A1
Levamisoile Conc in Plasma of Cattle Treated with Eprin/Levinj 1.2
E‘ 1 "" " i‘ u U3 " E 5 E16 7' rrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrrr '7 .E WWBNZDS '3 £14 “?n-‘SW09
5’ £12 * "main
0 i 0 '5 l0 15 20 25 30 Time?ws}
FIG. 2 Patent Application Publication Nov. 17, 2011 Sheet 3 0f3 US 2011/0281920 A1
Eprinomectin Conc in Plasma of Cattle ‘treated with Eprin/Lev inj 0.06
0.05
{mg/kg)Eprinomectinplasmacane 0.03
56 10E! 159 20G 25G 300
FIG. 3 US 2011/0281920 A1 Nov. 17, 2011
INJECTABLE PARASITICIDAL cins are described in Aoki et al., US. Pat. No. 3,950,360 as FORMULATIONS OF LEVAMISOLE AND Well as in the various references cited in “The Merck Index” MACROCYCLIC LACTONES 12”’ ed., S. Budavari, Ed., Merck & Co., Inc. Whitehouse Station, N]. (1996). Semi-synthetic derivatives of these INCORPORATION BY REFERENCE classes of compounds are Well knoWn in the art and are [0001] This application claims priority to US. provisional described, for example, in US. Pat. No. 5,077,308, US. Pat. patent application No. 61/333,882, ?led May 12, 2010. All No. 4,859,657, US. Pat. No. 4,963,582, US. Pat. No. 4,855, documents cited or referenced in the applicant cited docu 317, US. Pat. No. 4,871,719, US. Pat. No. 4,874,749, US. Pat. No. 4,427,663, US. Pat. No. 4,310,519, US. Pat. No. ments, and all documents cited or referenced herein (“herein 4,199,569, US. Pat. No. 5,055,596, US. Pat. No. 4,973,711, cited documents”), and all documents cited or referenced in US. Pat. No. 4,978,677, and US. Pat. No. 4,920,148, all herein cited documents, together With any manufacturer’s incorporated herein by reference in their entirety. A particu instructions, descriptions, product speci?cations, and product larly useful macrocyclic lactone is eprinomectin, Which is sheets for any products mentioned herein or in any document incorporated by reference herein, are hereby incorporated described in US. Pat. No. 6,174,540, US. Pat. No. 6,733,767, herein by reference, and may be employed in the practice of US. Pat. No. 5,602,107 to Merck (all incorporated by refer ence). This compound provides for a very short milk With the invention. draWal period due to it tendency to partition aWay from the milk of a lactating mammal. FIELD OF THE INVENTION [0006] To overcome the problem of resistance, various [0002] The present invention is directed to neW injectable combinations of anthelmintics have been extensively parasiticidal formulations comprising levamisole and macro researched. Unfortunately, combinations of macrocyclic lac cyclic lactones. The present invention also provides methods tone anthelmintics With anthelmintics of other families have for eradicating, controlling, and preventing parasite infesta proven di?icult to formulate at high concentration and With tion and/or infection in birds, ?sh and mammals. The formu an acceptable viscosity for injection. For example, Closamec lations of the invention may be administered to animals, tin (manufactured by Norbrook) contains iverrnectin at 0.5% particularly mammals, ?sh and birds, to prevent or treat para and closantel at 12.5% and is so viscous that the product is sitic infestation and/ or infection. supplied in a special pack that has a Warming device. The Warming device is activated 15 minutes before administration BACKGROUND OF THE INVENTION and the pack requires periodic shaking throughout this time to [0003] Animals, such as mammals, ?sh and birds are often ensure temperature reduces the viscosity suf?ciently for susceptible to parasite infestation and/or infection. These acceptable administration. parasites may be ectoparasites, such as insects and acarine [0007] Another example, Which has been the subject of species, and endoparasites such as ?lariae and other Worms. considerable research, is combining a macrocyclic lactone Endoparasites pose a signi?cant problem to those Who raise (ML) With levamisole. The di?iculties of combining these production animals such as bovines, ovines, and porcines. tWo actives lies in the stability of each being incompatible in Thus, there is an ongoing need to develop active pesticidal the environment in Which the other is stable. and parasiticidal compounds to protect animals against attack [0008] Levamisole is stable in acid conditions usually a pH or infestation/infection by pests. of about 4 or beloW, While macrocyclic lactones are unstable [0004] Many endoparasiticides or anthelmintics exist in the in conditions Where the pH is acidic. Both levamisole and art for treating internal parasites and other animal pests. macrocyclic lactones are Well knoWn in the art. In particular, These endoparasiticides vary in cost as Well as their effective levamisole has been used as an anthelmintic to treat nema ness to a particular parasite. HoWever, the results of treatment todes and respiratory Worm infestations in both humans and With these pesticides are not alWays satisfactory because of animals. Levamisole may be combined With other actives, continued development of resistance by the parasite to the although often With signi?cant dif?culty, as is described in available therapeutic agents. Thus, there is a need in the art for various patents and applications. For example, US 2009/ more effective parasiticidal agents for treatment and protec 0075918 A1 to Neto et al. appears to contemplate adding tion of animals, eg mammals, ?sh and birds from infestation various organic salts of levamisole to macrocyclic lactones, by animal parasites. but provides no Working examples. Other applications that [0005] The averrnectin and milbemycin series of com mention levamisole in combination With macrocyclic lac pounds are potent anthelmintic and antiparasitic agents tones include WO 00/061068, Which describes non-aqueous against a Wide range of internal and external parasites. The pour-on formulations that further contain triclabendaZole, compounds Which belong to this series are either natural and WO 04/009080, Which describes stable pyrrolidone products or are semi-synthetic derivatives thereof. The struc based formulations of averrnectins or milbemycins combined ture of these tWo series of compounds are closely related and With levamisole. Even more recently, WO2010021555 (to they both share a complex 16-membered macrocyclic lactone Intervet) has described oral drenches containing levamisole ring; hoWever, the milbemycins do not contain the disaccha and a macrocyclic lactone, Where the macrocyclic lactone is ride substituent in the 13-position of the lactone ring. The apparently stabiliZed via the inclusion of a protective agent, natural product averrnectins are disclosed in US. Pat. No. namely hydroxypropyl starch phosphate. Since this type of 4,310,519 to Albers-Schonberg, et al., and the 22,23-dihydro “protective agent” also serves as a thickener and emulsion averrnectin compounds are disclosed in Chabala, et al., US. stabiliZer, this particular approach to stabiliZing the macro Pat. No. 4,199,569. For a general discussion of averrnectins, cyclic lactone component may be ill-suited to developing Which include a discussion of their uses in humans and ani injectable formulations. Hydroxypropyl starch phosphate mals, see “Ivermectin and Abamectin,” W. C. Campbell, ed., (HP starch) is typically used for creams & lotions, founda Springer-Verlag, NY. (1989). Naturally occurring milbemy tions, mascara, color cosmetics, liquid make-up, liquid talc, US 2011/0281920 A1 Nov. 17, 2011
conditioners, shampoos, sunscreen products, and antiperspi that the Applicants reserve the right to this invention and rants. Finally, AU200310102 (to Nufarm and Novar‘tis) hereby disclose a disclaimer of any previously knoWn prod describes formulations having levamisole, avermectins, and uct, process, or method. benZimidaZole, said formulations appearing to make use of a [0014] It is noted that in this disclosure and particularly in biphasic solvent system, in an attempt to overcome the prob the claims and/or paragraphs, terms such as “comprises”, lematic instability of formulations Which comprise levami “comprised”, “comprising” and the like can have the meaning sole and macrocyclic lactones. Several patents describe long acting injectable combination formulations comprising attributed to it in US. patent laW; e.g., they can mean macrocyclic lactones and other actives, for example US “includes”, “included”, “including”, and the like; and that patent numbers US. Pat. No. 6,174,540 and US. Pat. No. terms such as “consisting essentially of’ and “consists essen 6,733,767 (to Merck), Which use castor oil and liquid poly tially of’ have the meaning ascribed to them in US. patent mers, respectively, to achieve the sustained release of the laW; e.g., they alloW for elements not explicitly recited, but macrocyclic lactones and the other actives. After revieWing exclude elements that are found in the prior art or that affect the current literature, applicants are unaWare of any previ a basic or novel characteristic of the invention. ously described, effective combinations of levamisole and [0015] These and other embodiments are disclosed or are eprinomectin, inj ectable or otherWise. obvious from, and encompassed by, the folloWing Detailed [0009] It is expressly noted that citation or identi?cation of Description. any document in this application is not an admission that such document is available as prior art to the present invention. Any foregoing applications, and all documents cited therein BRIEF DESCRIPTION OF DRAWINGS or during their prosecution (“application cited documents”) and all documents cited or referenced in the application cited [0016] The folloWing Detailed Description, given by Way documents, and all documents cited or referenced herein of example, and not intended to limit the invention to speci?c (“herein cited documents”), and all documents cited or refer embodiments described, may be understood in conjunction enced in herein cited documents, together With any manufac With the accompanying Figures, incorporated herein by ref turer’s instructions, descriptions, product speci?cations, and erence, in Which: product sheets for any products mentioned herein or in any [0017] FIG. 1 is a How diagram outlining the preparation of document incorporated by reference herein, are hereby incor formulations according to the instant invention porated herein by reference, and may be employed in the [0018] FIG. 2 depicts levamisole plasma concentration at practice of the invention. time points after administration of formulations according to the instant invention SUMMARY OF THE INVENTION [0019] FIG. 3 depicts eprinomectin plasma concentration [0010] The invention provides novel parasiticidal formula at time points after administration of formulations according tions comprising levamisole and macrocyclic lactones, par to the instant invention ticularly avermectins such as ivermectin and eprinomectin, and milbemycins, such as moxidectin, milbemectin, and mil DETAILED DESCRIPTION bemycin oxime. The formulations are storage-stable and pro vided in a single solvent system that is miscible in Water. [0020] The present invention provides novel injectable Therefore, the formulations according to the present inven levamisole/macrocyclic lactone formulations With parasiti tion may be classi?ed as micellar solutions as opposed to cidal activity, for the treatment or prevention of parasitic emulsions or biphasic emulsions. The invention also provides infestations and/or infection in an animal. Also provided are methods for the treatment and prevention of parasitic infes methods for the treatment or prevention of parasitic infesta tation and/or infection of animals. tions and/or infection in animals, comprising administering [0011] The inventive formulations comprising the levami an effective amount of the formulation of the invention to the sole and macrocyclic lactones are highly effective for the animal. treatment or prophylaxis of parasites in or on mammals, ?sh [0021] An important aspect of formulations containing and birds, and in particular, cats, dogs, horses, chickens, pigs, eprinomectin as the macrocyclic lactone component is that sheep and cattle With the aim of ridding these hosts of all the said formulations may have shortened WithdraWal times parasites commonly encountered by mammals, ?sh and birds. before it is appropriate to obtain milk from lactating mam The invention also provides for effective and long-lasting mals. This is because of the macrocyclic lactones, eprinomec defense against endoparasites, such as helminths, nematodes, tin has relatively the loWest milk to plasma partition ratio ?lariae, hookWorms, WhipWorms and roundWorms of the (~0.2). For a point of reference and de?nition, a milk to digestive tract of animals and humans. plasma partition ratio of 1 Would indicate the active tends to [0012] Accordingly, the present invention provides meth distribute evenly betWeen milk and plasma. ods for preventing and treating parasites in or on animals, [0022] Formulations described herein are particularly comprising administering a parasiticidally effective amount effective for controlling endoparasites. Endoparasites of a storage-stable injectable formulation comprising levami include, but are not limited to, nematodes (such as round sole and macrocyclic lactone to the animal. The invention Worms, hookWorms, WhipWorms and heartWorms) and ces also provides a method for combating or controlling animal todes (tapeWorms) and trematodes (?ukes). Therefore, the pests and parasites. inventive formulations have substantial utility in preventing [0013] It is an object of the invention to not encompass damage to crops, plants, plant propagation material and Within the invention any previously knoWn product, process Wood-containing property, and in controlling and preventing of making the product, or method of using the product such the infestation and/ or infection of animals by parasites. US 2011/0281920 A1 Nov. 17, 2011
[0023] The invention includes at least the following fea ated With the molecule in the crystalline form. The composi tures: tions of the invention may include hydrates and solvates of the [0024] (a) In one embodiment, the invention provides novel active agents. inj ectable formulations comprising levamisole and macrocy clic lactones, Which are active against animal pests, including De?nitions: insects and parasites; [0033] Terms used herein Will have their customary mean [0025] (b) methods for treating a parasitic infestation/infec ings in the art unless speci?ed. As used herein the term tion in or on an animal are provided, Which methods comprise “anthelmintic” and variations thereof encompasses one or administering a parasiticidally effective amount of a levami more nematocidal, trematocidal and cestocidal active com sole/macrocyclic lactone-containing formulation to the ani pounds. The term “pesticidal” and variations thereof includes mal in need thereof; any said anthelmintic, any endectoparasiticidal, and/or any ectoparasiticidal compound. Where the context alloWs the [0026] (c) methods for the prevention of a parasitic infes term “ectoparasiticidal” includes compounds effective tation/ infection of an animal, Which comprise administering a against any one or more ectoparasites including ticks, lice, parasiticidally effective amount the levamisole/macrocyclic ?eas, mites and the like. Further, as used herein the “endec lactone-containing formulation to the animal in need thereof; toparasiticidal” and variants thereof includes compounds [0027] (d) use of the formulations as a veterinary medica and/ or formulations that are active against internal (endo) and ment for controlling pests, including parasites; and external (ecto) parasites. [0028] (e) processes for the preparation of the levamisole/ [0034] As used herein the term “solubility” refers to the macrocyclic lactone-containing formulations. ability of a compound to be dissolved in a speci?c phase; and “lipophilic” means a greater tendency to an organic, oil or the [0029] Also contemplated Within the scope of the invention like phase as opposed to another phase (for example, the are acid or base salts of the compounds in the compositions of aqueous phase). the invention, Where applicable. [0035] As referred to herein the term “injectable” in the [0030] The term “acid” contemplates all pharmaceutically context of ?uids or liquids covers viscosities ranging from a acceptable inorganic or organic acids. Inorganic acids include free ?oWing liquid to a thin gel consistency that is capable of mineral acids such as hydrohalic acids such as hydrobromic being expelled by syringe and suitable forbeing administered acid and hydrochloric acid, sulfuric acid, phosphoric acids to an animal via injection. As used herein, a suitable or appro and nitric acid. Organic acids include all pharmaceutically priate viscosity for injection may be betWeen 1-50 cp or from acceptable aliphatic, alicyclic and aromatic carboxylic acids, 1-10 cp. Routes of injection may be parenteral, for example dicarboxylic acids, tricarboxylic acids and fatty acids. In one intramuscular (IM), intraperitoneal (IP), or subcutaneous embodiment of the acids, the acids are straight chain or (SQ) branched, saturated or unsaturated C 1 -C20 aliphatic carboxy [0036] As referred to herein the term “acceptable storage lic acids, Which are optionally substituted by halogen or by stability” means stable for greater than 3 months at room hydroxyl groups, or C6-Cl2 aromatic carboxylic acids. temperature. Examples of such acids are carbonic acid, formic acid, acetic acid, propionic acid, isopropionic acid, valeric acid, ot-hy Formulations of the Invention: droxy acids such as glycolic acid and lactic acid, chloroacetic [0037] The formulations of the invention comprise at least acid, benZoic acid, methane sulfonic acid, and salicylic acid. levamisole and at least one macrocyclic lactone active com Examples of dicarboxylic acids include oxalic acid, malic pound, and are provided as micellar solutions of actives, acid, succinic acid, tartaric acid, fumaric acid, and maleic Which are miscible in Water for injection, and are particularly acid. An example of a tricarboxylic acid is citric acid. Fatty effective at controlling/ combating parasites, particularly acids include all pharmaceutically acceptable saturated or endoparasites. In certain embodiments, the formulations of unsaturated aliphatic or aromatic carboxylic acids having 4 to the invention are useful in veterinary applications, including 24 carbon atoms. Examples include butyric acid, isobutyric for controlling parasites in or on an animal. acid, sec-butyric acid, lauric acid, palmitic acid, stearic acid, [0038] In one embodiment, the at least one macrocyclic oleic acid, linoleic acid, linolenic acid, and phenylsteric acid. lactone active agent in the formulations of the invention may Other acids include gluconic acid, glycoheptonic acid and be at least one avermectin or milbemycin compound. In vari lactobionic acid. ous embodiments, the at least one macrocyclic lactone com [0031] The term “base” contemplates all pharmaceutically pound may be abamectin, avermectin, dimadectin, doramec acceptable inorganic or organic bases, including hydroxides, tin, emamectin, eprinomectin, iver'mectin, latidectin, carbonates or bicarbonates of alkali metal or alkaline earth lepimectin, selamectin, milbemectin, milbemycin D, milbe metals. Salts formed With such bases include, for example, mycin oxime, moxidectin or nemadectin. the alkali metal and alkaline earth metal salts, including, but [0039] In another embodiment, the at least one macrocyclic not limited to, as the lithium, sodium, potassium, magnesium lactone active agent may be any one of the avermectins or or calcium salts. Salts formed With organic bases include the avermectin monosaccharides modi?ed in the 4' or 4" position common hydrocarbon and heterocyclic amine salts, Which describedinU.S. Pat. No. 7,704,961,U.S. Pat. No. 7,521,429, include, for example, ammonium salts (NH4+), alkyl- and US 2006-0105970A1, US Pat. No. 7,678,740, U.S. Pat. No. dialkylammonium salts, and salts of cyclic amines such as the 7,632,820, U.S. Pat. No. 7,678,773, U.S. Pat. No. 7,605,134, morpholine and piperidine salts. and Us. Pat. No. 6,933,260 (each to Merial). It Will be [0032] In addition, the compounds Within the compositions appreciated by a skilled person that any of the above-recited of the invention may exist as hydrates or solvates, in Which a avermectins exhibiting a relatively loW milk to plasma parti certain stoichiometric amount of Water or a solvent is associ tion ratio (for example less than about 1, less than about 0.5, US 2011/0281920 A1 Nov. 17, 2011
and even more favorably, less than about 0.2) Will be a par embodiment, the macrocyclic lactone is either ivermectin or ticularly desirable component of formulations according to eprinomectin and is present in an amount su?icient to deliver the instant invention. a dose of at least 150 ug/kg animal bodyWeight. [0040] In one embodiment the invention provides formula [0083] In a particular embodiment, the present invention is tions according to the following: a stable inj ectable formulation comprising eprinomectin and [0041] (a) Levamisole; levamisole and is suitable for use in cattle. In an embodiment, [0042] (b) At least one macrocyclic lactone; the formulation is an injectable combination anthelmintic [0043] (c) At least one Water miscible organic solvent; containing about 7 g/ L eprinomectin and about 223 g/L [0044] (d) A surfactant; levamisole phosphate. [0045] (e) Water for injection (referred to hereafter as [0084] In an emodiment, the formulation requires not more “WFI”). than a 35 day Withholding period for either milk or meat. [0046] In another embodiment, the invention provides a [0085] In an embodiment, the formulation is administered formulation comprising: at a dose rate of 1 mL per 35 kg (200 pg eprinomectin and 6.4 [0047] (a) Levamisole; mg levamisole phosphate per kg, equivalent to 5 mg levami [0048] (b) At least one of ivermectin, abamectin, or epri sole HCI per kg). In another embodiment, the formulation is effective against a broad range of internal parasites including nomectin, or a mixture thereof; Oslerlagla spp., Trichoslrongylus axel, Cooperia spp., suck [0049] (c) At least one Water miscible organic solvent; ling lice, and certain strains resistant to certain macrocyclic [0050] (d) A surfactant; and lactones and/or benZamidaZoles. [0051] (e) WFI. [0086] In an embodiment, the formulations have a shelf life [0052] In still another embodiment, the invention provides and storage condition of at least 2 years at 2-6° C. In another a formulation comprising: embodiment, the formulations are prepared as pack siZes of [0053] (a) Levamisole; 100 to 1000 mL. In a particular embodiment, the pack siZes [0054] (b) At least one of dimadectin, doramectin ema are 500 mL ?exible packages, or ?exipacks. mectin, latidectin, lepimectin or selamectin, or a mixture [0087] In a preferred embodiment, the Water miscible sol thereof; vent Will be an organic solvent, including an amide, alcohol, [0055] (c) At least one Water miscible organic solvent; ester or sulfoxide. In another preferred embodiment, the for [0056] (d) A surfactant; and mulations of the invention Will comprise a pharmaceutically [0057] (e) WFI. acceptable amide including, but not limited to, dimethylfor [0058] In yet another embodiment, the invention provides a mamide, dimethylacetamide, 2-pyrrolidone, N-methylpyr formulation comprising: rolidone and the like. [0059] (a) Levamisole; [0088] Surfactants are Well knoWn in the art and may [0060] (b) At least one of ivermectin, abamectin, or epri include non-ionic surfactants, cationic surfactants and nomectin, or a mixture thereof; anionic surfactants. Anionic surfactants include, but are not [0061] (c) At least one Water miscible organic solvent; limited to, alkaline stearates (e. g. sodium, potassium or [0062] (d) A surfactant; and ammonium stearate); calcium stearate or triethanolamine [0063] (e) WFI. stearate; sodium abietate; alkyl sulfates, Which include but [0064] In another embodiment, the invention provides a are not limited to sodium lauryl sulfate and sodium cetyl formulation comprising: sulfate; sodium dodecylbenZenesulphonate or sodium dioctyl [0065] (a) Levamisole; sulphosuccinate; or fatty acids (eg coconut oil); [0066] (b) At least one of milbemectin, milbemycin D, [0089] Cationic surfactants include, but are not limited to, milbemycin oxime, moxidectin or nemadectin, or a mix any knoWn Water-soluble quaternary ammonium salts such as ture thereof; cetyltrimethylammonium bromide and the like; and knoWn [0067] (c) At least one Water miscible organic solvent; amine salts such as octadecylamine hydrochloride and the [0068] (d) A surfactant; and like. [0069] (e) WFI. [0090] Non-ionic surfactants include, but are not limited to, [0070] In a preferred embodiment, the invention provides a optionally polyoxyethylenated esters of sorbitan, e.g. formulation comprising: Polysorbate 80, or polyoxyethylenated alkyl ethers; polyeth ylene glycol stearate, polyoxyethylenated derivatives of cas [0071] (a) Levamisole; tor oil, polyglycerol esters, polyoxyethylenated fatty alco [0072] (b) At least one of ivermectin or eprinomectin, or hols, polyoxyethylenated fatty acids or copolymers of a mixture thereof; ethylene oxide and of propylene oxide. Other surfactants that [0073] (c) At least one Water miscible organic solvent; are suitable for the formulations of the present invention [0074] (d) A surfactant; and include amphoteric surfactants, such as substituted lauryl [0075] (e) WFI. compounds of betaine. The surfactant of the inventive formu [0076] In another embodiment, the invention provides a lations may also be a mixture of at least tWo different surfac formulation comprising: tants. [0077] (a) Levamisole; [0091] In another embodiment, the levamisole is levami [0078] (b) At least one of avermectin or avermectin sole phosphate and the macrocyclic lactone is either epri monosaccharide, each modi?ed at the 4' or 4" position, nomectin or ivermectin. In some embodiments, the Water or a mixture thereof; miscible organic solvent is dimethylacetamide (DMA) and [0079] (c) At least one Water miscible organic solvent; the surfactant is CREMOPHOR EL. Thus in an embodiment, [0080] (d) A surfactant; and the formulation of the present invention may comprise: [0081] (e) WFI. [0092] (a) Levamisole phosphate; [0082] In an embodiment, the levamisole is levamisole [0093] (b) Eprinomectin, ivermectin, abamectin, dor phosphate and is present in an amount suf?cient to deliver a amectin or moxidectin; dose of at least 4 mg/kg animal bodyWeight. In another [0094] (c) DMA US 2011/0281920 A1 Nov. 17, 2011
[0095] (d) CREMOPHOR EL; and [0101] In one embodiment for treatment against ectopara [0096] (e) WFI sites, the ectoparasite is one or more insect or arachnid includ [0097] The inventive formulations may contain other inert ing those of the genera Clenocephalides, Rhipicephalus, Der ingredients such as antioxidants, preservatives, or pH stabi macenlor, Ixodes, Boophilus, Ambylomma, Haemaphysalis, liZers. These compounds are Well knoWn in the formulation Hyalomma, Sarcoples, Psoroples, Olodecles, Chorioples, art. Antioxidant such as an alpha tocopherol, ascorbic acid, Hypoderma, Damalinia, Linognalhus, Haemalopinus, Sole ascrobyl palmitate, fumaric acid, malic acid, sodium ascor noples, Trichodecles, and Felicola. bate, sodium metabisulfate, n-propyl gallate, BHA (butylated [0102] In another embodiment for the treatment against hydroxy anisole), BHT (butylated hydroxy toluene) mono ectoparasites, the ectoparasite is from the genera Cleno thioglycerol and the like, may be added to the present formu cephalides, Rhipicephalus, Dermacenlor, Ixodes and/or B00 lation. In certain embodiments, the antioxidants are generally philus. The ectoparasites treated include but are not limited to added to the formulation in amounts of from about 0.01 to ?eas, ticks, mites, mosquitoes, ?ies, lice, bloW?y and combi about 2.0%, based upon total Weight of the formulation, With nations thereof. Speci?c examples include but are not limited about 0.05 to about 1.0% being especially preferred. to cat and dog ?eas (Clenocephalidesfelis, Clenocephalides [0098] In some embodiments, preservatives, such as the sp. and the like), ticks (Rhipicephalus sp., Ixodes sp., Derma parabens (methylparaben and/ or propylparaben), are suitably cenlor sp., Amblyoma sp. and the like), and mites (Demodax used in the formulation in amounts ranging from about 0.01 to sp., Sarcoples sp., Olodecles sp. and the like), lice (Tri about 2.0%, With about 0.05 to about 1.0% being especially chodecles sp., Cheyleliella sp., Lignonalhus sp., and the like), preferred. Other preservatives include benZalkonium chlo mosquitoes (Aedes sp., Culex sp., Anopheles sp., and the like) ride, benZethonium chloride, benZoic acid, benZyl alcohol, and ?ies (Hemalobia sp., Musca sp., Slomoxys sp., Demalo bronopol, butylparaben, cetrimide, chlorhexidine, chlorobu bia sp., Cochliomyia sp., and the like). In yet another embodi tanol, chlorocresol, cresol, ethylparaben, imidurea, meth ment for the treatment against ectoparasites, the ectoparasite ylparaben, phenol, phenoxyethanol, phenylethyl alcohol, is a ?ea and/or tick. Additional examples of ectoparasites phenylmercuric acetate, phenylmercuric borate, phenylmer include but are not limited to the tick genus Boophilus, espe curic nitrate, potassium sorbate, sodium benZoate, sodium cially those of the species microplus (cattle tick), decoloralus propionate, sorbic acid, thimerosal, and the like. Preferred and annulalus; myiases such as Dermalobia hominis (knoWn ranges for these compounds include from about 0.01 to about as Berne in Brazil) and Cochliomyia hominivorax (green 5%. bottle); sheep myiases such as Lucilia sericala, Lucilia cup [0099] Compounds Which stabiliZe the pH of the formula rina (knoWn as bloW?y strike in Australia, NeW Zealand and tion are also contemplated and may be used in certain South Africa). Flies proper, namely those Whose adult con embodiments of the inventive formulations. Again, such com stitutes the parasite, such as Haemalobia irrilans (horn ?y); pounds are Well knoWn to a practitioner in the art as Well as lice such as Linognalhus vilulorum, etc.; and mites such as hoW to use these compounds. Buffering systems include, for Sarcoples scabici and Psoroples ovis. The above list is not example, systems selected from the group consisting of acetic exhaustive and other ectoparasites are Well knoWn in the art to acid/acetate, malic acid/malate, citric acid/citrate, tataric be harmful to animals and humans. These include, for acid/tar‘trate, lactic acid/lactate, phosphoric acid/phosphate, example migrating dipterous larvae. glycine/glycimate, tris, glutamic acid/glutamates and sodium carbonate. [0103] When an anthelmintic agent is added to the compo sition of the invention, the composition can also be used to Parasites Controlled: treat against endoparasites such as those helminths selected from the group consisting of Anaplocephala, Ancylosloma, [0100] The injectable levamisole/macrocyclic lactone for Anecalor, Ascaris, Capillaria, Cooperia, Dipylidium, Diro? mulations are particularly effective for e?iciently controlling laria, Echinococcus, Enlerobius, Fasciola, Haemonchus, endoparasites, such as ?ukes, hookWorms, and helminths Oesophagoslumum, Oslerlagia, Toxocara, Slrongyloides, such as cestodes, nematodes, and trematodes. Endoparasites Toxascaris, Trichinella, Trichuris, and Trichoslrongylus. further include helminths such as Anaplocephala, Ancylos [0104] In another embodinient of the invention, the com Zoma, Anecalor, Ascaris, Capillaria, Cooperia, Dipylidium, pounds and compositions of the invention are suitable for Diro?laria, Echinococcus, Enlerobius, Fasciola, Haemon controlling pests such as insects selected from the group chus, Oesophagoslumum, Oslerlagia, Toxocara, Slrongy consisting of Blalella germanica, Heliolhis virescens, Lepli loides, Toxascaris, Trichinella, Trichuris, and Trichoslrongy nolarsa decemlineala, Telramorium caespilum and combina lus. Or others from the class of helminths, such as from the tions thereof. class of helminths, for example, Ancylosloma duodenale, [0105] The phytoparasitic nematodes include, for example, Ancylosloma ceylanicum, Acylosloma braziliensis, Ancylos Anguina spp., Aphelenchoides spp., Belonoaimus spp., Bur Zoma spp., Ascaris lubricoides, Ascaris spp., Brugia malayi, saphelenchus spp., Dilylenchus dipsaci, Globodera spp., Brugia Zimori, Bunoslomum spp., Chaberlia spp., Clonorchis Heliocolylenchus spp., Helerodera spp., Longidorus spp., spp., Cooperia spp., Dicrocoelium spp, Diclyocaulus?laria, Meloidogyne spp., Pralylenchus spp., Radopholus similis, Diphyllobolhrium lalum, Dracunculus medinensis, Echino Rozylenchus spp., Trichodorus spp., Zj1lench0rhynchus spp., coccus granulosus, Echinococcus mullilocularis, Enlerobius Elenchulus spp., Elenchulus semipenelrans, Xiphinema vermicularis, Faciola spp., Haemonchus spp., Helerakis spp., spp. Hymenolepis nana, Hyoslrongulus spp., Loa L011, Nemalodi rus spp., Oesophagoslomum spp., Opislhorchis spp., [0106] In addition, With or Without the other pesticidal Onchocerca volvulus, Oslerlagia spp., Paragonimus spp., agents added to the composition, the invention can also be Schislosomen spp., Strongloidesfuelleborni, Slrongyloides used to treat other pests Which include but are not limited to slercoralis, Slronyloides spp., Taenia saginala, Taenia pests: solium, Trichinella spiralis, Trichinella naliva, Trichinella [0107] (1) from the order of Isopoda, for example Oniscus brilovi, Trichinella nelsoni, Trichinella pseudopsiralis, Tri asellus, Armadillidium vulgare and Porcellio scaber; choslrongulus spp., Trichuris Zrichuria, Wuchereria ban [0108] (2) from the order of Diplopoda, for example Bla cro?i. niulus gullulalus; US 2011/0281920 A1 Nov. 17, 2011
[0109] (3) from the order of Chilopoda, for example Geo [0121] (15) from the class of Gastropoda, for example, philus carpophagus and Sculigera spp.; Anion spp., Biomphalaria spp., Bulinus spp., Deroceras spp., [0110] (4) from the order of Symphyla, for example Sculi Galba spp., Lymnaea spp., Oncomelania spp., Succinea spp.; gerella immaculala; [0122] (16) from the class of helminths, for example, Ancy [0111] (5) from the order of Thysanura, for example Lepi losloma duodenale, Ancylosloma ceylanicum, Acylosloma sma saccharina; braziliensis, Ancylosloma spp., Ascaris lubricoides, Ascaris [0112] (6) from the order of Collembola, for example Ony spp., Brugia malayi, Brugia Zimori, Bunoslomum spp., Chab chiurus armalus; erlia spp., Clonorchis spp., Cooperia spp., Dicrocoelium spp, [0113] (7) from the order of Blattaria, for example Blalla Diclyocaulus?laria, Diphyllobolhrium lalum, Dracunculus orienlalis, Periplanela americana, Leucophaea maderae and medinensis, Echinococcus granulosus, Echinococcus mul Blallella germanica; Zilocularis, Enlerobius vermicularis, Faciola spp., Haemon [0114] (8) from the order of Hymenoptera, for example chus spp., Helerakis spp., Hymenolepis nana, Hyoslrongulus Diprion spp., Hoplocampa spp., Lasius spp., Monomorium spp., Loa Loa, Nemalodirus spp., Oesophagoslomum spp., pharaonis and Vespa spp.; Opislhorchis spp., Onchocerca volvulus, Oslerlagia spp., [0115] (9) from the order of Siphonaptera, for example Paragonimus spp., Schislosomen spp., Slrongyloidesfuelle Xenopsylla cheopis and Ceralophyllus spp.; borni, Slrongyloides slercoralis, Slronyloides spp., Taenia Saginaw, Taenia solium, Trichinella spiralis, Trichinella [0116] (10) from the order of Anoplura (Phthiraptera), for naliva, Trichinella brilovi, Trichinella nelsoni, Trichinella example, Damalinia spp., Haemalopinus spp., Linognalhus pseudopsiralis, Trichoslrongulus spp., Trichuris Zrichuria, spp., Pediculus spp., Trichodecles spp.; Wuchereria bancro?i; [0117] (l 1) from the class of Arachnida, for example, [0123] (17) from the order of Heteroptera, for example, Acarus siro, Aceria sheldoni, Aculops spp., Aculus spp., Anasa Zrislis, Anlesliopsis spp., Blissus spp., Calocoris spp., Amblyomma spp., Argos spp., Boophilus spp., Brevipalpus Campylomma livida, Cavelerius spp., Cimex spp., Creonlia spp., Bryobiapraeliosa, Chorioples spp., Dermanyssus galli des dilulus, Dasynus piperis, Dichelops furcalus, Dicono nae, Eolelranychus spp., Epilrimerus pyri, Eulelranychus coris hewelli, Dysdercus spp., Euschislus spp., Eurygasler spp., Eriophyes spp., Hemilarsonemus spp., Hyalomma spp., spp., Heliopellis spp., Horcias nobilellus, Leplocorisa spp., Ixodes spp., Lalrodeclus maclans, Melalelranychus spp., Oli Leploglossus phyllopus, Lygus spp., Macropes excavalus, gonychus spp., Ornilhodoros spp., Panonychus spp., Phyllo Miridae, Nezara spp., Oebalus spp., Penlomidae, Piesma coplrula oleivora, Polyphagolarsonemus lotus, Psoroples quadrala, Piezodorus spp., Psallus serialus, Pseudacysla spp., Rhipicephalus spp., Rhizoglyphus spp., Sarcoples spp., persea, Rhodnius spp., Sahlbergella singularis, Scolino Scorpio maurus, Slenolarsonemus spp., Tarsonemus spp., Telranychus spp., Vasales lycopersici; phora spp., Slephanilis nashi, Hbraca spp., Trialoma spp.; [0124] (18) from the order of Homoptera, for example, [0118] (12) from the class of Bivalva, for example, Dreis Acyrlhosipon spp., Aeneolamia spp., Agonoscena spp., Aleu sena spp.; rodes spp., Aleurolobus barodensis, Aleurolhrixus spp., [0119] (13) from the order of Coleoptera, for example, Amrasca spp., Anuraphis cardui, Aonidiella spp., Apha Acanlhoscelides obleclus, Adorelus spp., Agelaslica alni, nosligmapini, Aphis spp.,Arboridia apicalis, Aspidiella spp., Agrioles spp., Amphimallon solslilialis, Anobiumpunclalum, Aspidiolus spp., Alanus spp., Aulacorlhum solani, Bemisia Anoplophora spp., Anlhonomus spp., Anlhrenus spp., Apogo spp., Brachycaudus helichrysii, Brachycolus spp., Brevico nia spp., Alomaria spp., Allagenus spp., Bruchidius obleclus, ryne brassicae, Calligypona marginala, Carneocephala Bruchus spp., Ceulhorhynchus spp., Cleonus mendicus, Con fulgida, Ceralovacuna lanigera, Cercopidae, Ceroplasles oderus spp., Cosmopoliles spp., Costelylra Zealandica, Cur spp., Chaelosiphonfragaefolii, Chionaspis Zegalensis, Chlo culio spp., Cryplorhynchus lapalhi, Dermesles spp., rila onukii, Chromaphisjuglandicola, Chrysomphalus?cus, Diabrolica spp., Epilachna spp., Fauslinus cubae, Gibbium Cicadulina mbila, Coccomylilus halli, Coccus spp., Crypto psylloides, Heleronychus aralor, Hylamorpha elegans, myzus ribis, Dalbulus spp., Dialeurodes spp., Diaphorina Hylolrupes bajulus, Hypera poslica, Hypolhenemus spp., spp., Diaspis spp., Doralis spp., Drosicha spp., Dysaphis Lachnoslerna consanguinea, Leplinolarsa decemlineala, spp., Dysmicoccus spp., Empoasca spp., Eriosoma spp., Lissorhoplrus oryzophilus, Lixus spp., Lyclus spp., Erylhroneura spp., Euscelis bilobalus, Geococcus co?‘eae, Meligelhes aeneus, Melolonlha melolonlha, Migdolus spp., Homalodisca coagulala, Hyaloplerus arundinis, kerya spp., Monochamus spp., Naupaclus xanlhographus, Niplus holo Idiocerus spp., Idioscopus spp., Laodelphax slrialellus, leucus, Orycles rhinoceros, Oryzaephilus surinamensis, Lecanium spp., Lepidosaphes spp., Lipaphis erysimi, Macro Oliorrhynchus sulcalus, Oxycelonia jucunda, Phaedon siphum spp., Mahanarva?mbriolala, Melanaphis sacchari, cochleariae, Phyllophaga spp., Popilliajaponica, Premnol Melcal?ella spp., Melopolophium dirhodum, Monellia cos rypes spp., Psylliodes chrysocephala, Plinus spp., Rhizobius Zalis, Monelliopsis pecanis, Myzus spp., Nasonovia ribisni venlralis, Rhizoperlha dominica, Silophilus spp., Spheno gri, Nepholellix spp., Nilaparvala lugens, Oncomelopia spp., phorus spp., Slernechus spp., Symphyleles spp., Tenebrio Orlhezia praelonga, Parabemisia myricae, Paralrioza spp., molilor, Tribolium spp., Trogoderma spp., Zjwhius spp., Parlaloria spp., Pemphigus spp., Peregrinus maidis, Phen Xylolrechus spp., Zabrus spp.; acoccus spp., Phloeomyzus passerinii, Phorodon humuli, [0120] (14) from the order of Diptera, for example, Aedes Phylloxera spp., Pinnaspis aspidislrae, Planococcus spp., spp., Anopheles spp., Bibio horlulanus, Calliphora erythro Prolopulvinaria pyrlformis, Pseudaulacaspis penlagona, cephala, Ceralilis capilala, Chrysomyia spp., Cochliomyia Pseudococcus spp., Psylla spp., Pleromalus spp., Pyrilla spp., Cordylobia anlhropophaga, Culex spp., Culerebra spp., spp., Quadraspidiolus spp., Quesada gigas, Raslrococcus Dacus oleae, Dermalobia hominis, Drosophila spp., Fannia spp., Rhopalosiphum spp., Sal'sselia spp., Scaphoides lilanus, spp., Gaslrophilus spp., Hylemyia spp., Hyppobosca spp., Schizaphis graminum, Selenaspidus arliculalus, Sogala spp., Hypoderma spp., Liriomyza spp., Lucilia spp., Musca spp., Sogalella furclfera, Sogalodes spp., Sliclocephala feslina, Nezara spp., Oeslrus spp., Oscinella fril, Pegomyia hyos Tenalaphara malayensis, ?nocallis caryaefoliae, Tomaspis cyami, Phorbia spp., Slomoxys spp., Tabanus spp., Tannia spp., Toxoplera spp., Trialeurodes vaporariorum, Trioza spp., Hpula paludosa, Wohlfahrlia spp.; spp., Zj/phlocyba spp., Unaspis spp., [?leus vilifolii; US 2011/0281920 A1 Nov. 17, 2011
[0125] (19) from the order of Isoptera, for example, Reli culilermes spp., Odonlolermes spp.; TABLE 1 [0126] (20) from the order of Lepidoptera, for example, Acronicla major, Aedia leucomelas, Agrolis spp., Alabama Formulation of Eprinomectin levamisole Iniection argillacea, Anlicarsia spp., Baralhra brassicae, Bucculalrix Formulation 1 Formulation 2 Zhurberiella, Bupalus piniarius, Cacoecia podana, Capua % W/v % W/v reliculana, Carpocapsa pomonella, Cheimalobia brumala, Ingredient Composition Composition Function Chilo spp., Charisloneura fumlferana, Clysia ambiguella, Eprinomectin 0.70* 0.70* Active Cnaphalocerus spp., Earias insulana, Epheslia kuehniella, Levamisole Phosphate 22.3 * 22.3 * Active Euproclis chrysorrhoea, Euxoa spp., Fellia spp., Galleria BHT 0.1 0.1 Antioxidant mellonella, Helicoverpa spp., Heliolhis spp., Hofmannophila Dimethylacetamide 5 .0 5 .0 Solvent pseudosprelella, Homona magnanima, Hyponomeula Methyl Paraben 0.15 0.15 Preservative padella, Laphygma spp., Lilhocollelis blancardella, Litho Polysorbate 80 5.0 i Surfactant phane anlennala, Loxagrolis albicosla, Lymanlria spp., Cremophor EL i 5 .0 Surfactant Malacosoma neuslria, Mameslra brassicae, Mocis repanda, Water For Injection q.s. to 100% Vehicle Mylhimna separala, Oria spp., Oulema oryzae, Panolis?am mea, Peclinophora gossypiella, Phyllocnislis cilrella, Pieris *Corrected for potency spp., Plulella xyloslella, Prodenia spp., Pseudalelia spp., Pseudoplusia includens, Pyrausla nubilalis, Spodoplera [0135] Stability of both formulations has been demon spp., Thermesia gemmalalis, llnea pellionella, llneola bis strated in real-time storage conditions. selliella, Torlrix viridana, Trichoplusia spp.; [0127] (21) from the order of Orthoptera, for example, TABLE 2 Achela domeslicus, Blalla orienlalis, Blallella germanica, Stability of Eprinomectin Levamisole Iniection Formulation 1 Gryllolalpa spp., Leucophaea maderae, Locusla spp., Mel anoplus spp., Periplanela americana, Schislocerca gregaria; 2-8° C. 25° C./60% RH [0128] (22) from the order of Thysanoptera, for example, Baliolhrips biformis, Enneolhrips Havens, Frankliniella Batch No. Control 1 month 2 month 3 month spp., Heliolhrips spp., Hercinolhrips femoralis, Kakolhrips Formula- Description VLYS VLYS VLYS VLYS spp., Rhipiphorolhrips cruenlalus, Scirlolhrips spp., Taenio tion 1. Assay of Eprinomectin 100.9 99.6 98.0 98.6 Zhrips cardamoni, Thrips spp.; Assay ofLevamisole 98.0 97.9 98.3 99.2 Phosphate [0129] (23) from the class of Protozoa, for example, E ime ria spp. VLYS: Very LightYelloW Solution LYS: LightYelloW Solution YS: Yellow Solution EXAMPLES
[0130] The following examples are provided to illustrate TABLE 3 certain embodiments of the invention and are not to be con strued in any Way as limiting the scope of the invention. Stability of Eprinomectin Levamisole Iniection Formulation 2 [0131] Example 1 is particularly, though not exclusively, representative of inj ectable eprinomectin/levamisole formu 2-8° C. 25° C./60% RH lations according to the present invention. Example 2 pro Batch No. Control 1 month 2 month 3 month vides evidence that iver'mectin-containing formulations according to the present invention are stable and safe in ani Formula- Description VLYS VLYS VLYS VLYS tion 2. Assay of Eprinomectin 101.1 100.0 98.0 99.6 mals. Finally, example 3 provides extensive, though not sole Assay ofLevamisole 98.9 99.3 98.8 100.3 evidence of the signi?cant amount of time and effort that Was Phosphate invested in developing the inventive formulations. VLYS: Very LightYelloW Solution LYS: LightYelloW Solution Example 1 YS: Yellow Solution Development of Stable Inj ectable Levamisole/Epri nomectin Formulations TABLE 4 [0132] Levamisole is Water soluble and eprinomectin is Admini tration of Treatments nearly Water insoluble. Eprinomectin is soluble, for example, in methanol, ethanol, ethyl acetate, and dimethyl acetate Animal ID Group ID Product/BN Weight (kg) Dose (ml) (DMA). Applicants are aWare of no existing combination 2 1 Formulation 324.0 9.3 inj ectable formulation comprising eprinomectin and levami 3 1 294.0 8.4 sole. The development Work fully disclosed and described 5 BN: 08 334.0 9.5 herein out Was carried out to provide a formulation in Which 6 315.0 9 0 9 343.0 9 8 the anthelmintics eprinomectin and levamisole (as the phos 11 297.0 8 5 phate) could be combined in a formulation having physical 12 285.0 8 1 and chemical properties suitable for injection. 13 289.0 8 3 [0133] Stress studies have identi?ed at least tWo formula 14 302.0 8 6 20 313.0 8.9 tions that are stable and suitable in terms of irritancy. 1 2 Formulation 270.0 7.7 [0134] Constituents of these formulations are provided in 4 2 303.0 8.7 Table 1. US 2011/0281920 A1 Nov. 17, 2011
TABLE 4-continued TABLE 5-continued
Admini tration of Treatments Injection Site Reactions Formulation 1 Formulation 1 Batch no: 08 Animal ID Group ID Product/BN Weight (kg) Dose (ml) Animal Weight Dose 48 96 Day Day Day 7 EN: 09 288.0 8.2 ID (kg) (mL) 24 hours hours hours 10 21 35 8 317.0 9.1 10 326.0 9.3 14 302.0 8.6 4 4 1 1 0 0 16 331.0 9.5 20 313.0 8.9 4 4 1 0 0 0 17 291.0 8.3 18 309.0 8.8 19 296.0 8.5 21 340.0 9.7 TABLE 6 15 3 Control 305.0 f Injection Site Reactions Formulation 2 [0136] Both formulations Were also tested in cattle studies Formulation 2 Batch no: 09 to determine the absorption characteristics and injection site Animal Weight Dose 48 96 Day Day Day acceptability. TWenty-one Hereford Cross 15 month old ID (kg) (mL) 24 hours hours hours 10 21 35 female cattle Were selected for the study from a larger group 1 270.0 7.7 held at the study site. On Day —1 the study animals Were 4 303.0 8.7 individually identi?ed With uniquely numbered ear tags. 7 288.0 8.2 [0137] The study animals Were Weighed and ranked 8 317.0 9.1 10 326.0 9.3 according to body Weight. The animal With the median body 16 331.0 9.5 Weight (305 kg) Was selected as the control animal. The 17 291.0 8.3 remaining animals Were assigned to tWo groups of ten by restrictive randomization based on body Weight. Treatments Were administered as per Table 4. [0138] Animals in Group 1 Were treated With Formulation 1 at a dose rate of 1 ml/35 kg body Weight. Animals in Group Blood Plasma Pro?le 2 Were treated With Formulation 2 at a dose rate of 1 ml/35 kg [0147] On Day —1, (3 Jan. 2010) ?ve animals Were ran bodyWeight. All calculated doses Were rounded up to the domly selected from each of the tWo treated groups for blood nearest 0.2 ml. The control animal received no treatment. sampling as illustrated in Table 7: [0139] Treatments Were administered by subcutaneous injection to the anterior half of the neck, using a neW sterile 18 TABLE 7 gauge 25 mm needle for each injection. The area for injection Was clipped and sWabbed With 70% isopropyl alcohol prior to Eprinomectin Levamisole Blood Sampling Schedule injection. The injection sites Were inspected and palpated at 24, 48 and 96 hours post treatment and on Days 10, 21 and 35. Blood Plasma Group 1 Blood Plasma Group 2 All observations Were recorded. 3 1 [0140] The degree of injection site swelling Was recorded 11 10 according to the following scale: 12 16 14 18 [0141] 0:no edema 20 21 [0142] lqlery slight edema (barely noticed) [0143] 2:slight edema (Well de?ned by de?nite raising) [0148] These ten animals plus the untreated control Were [0144] 3:moderate edema (raised approximately 1 mm) blood sampled via jugular venipuncture on Day —1 prior to [0145] 4:(raised more than 1 mm and extending beyond treatment. These same animals Were blood sampled at 1, 2, 6, the area of injection) 12, 24, 48 and 96 hours post treatment, and then at Day 10, [0146] The maximum length, Width and height of any vis Day 21 and Day 35 after treatment. Blood samples Were ible or palpable site reaction Was determined using calipers, collected by caudal venipuncture into green top heparinised and all ?ndings Were recorded. Vacutainer’s, and spun in a centrifuge for ten minutes at 3000 RPM. Plasma for each animal Was decanted into individually TABLE 5 labeled plasma vials and froZen Within tWo hours of collec tion. Injection Site Reactions Formulation 1 [0149] At the conclusion of the animal phase of the study Formulation 1 Batch no: 08 (Day 35) the froZen plasma samples Were submitted to an analytical laboratory for Eprinomectin and Levamisole Animal Weight Dose 48 96 Day Day Day analysis. Samples from 0, 1, 2, 6, 12, 24 and 48 hours Were ID (kg) (mL) 24 hours hours hours 10 21 35 analyZed for Levamisole concentration. Samples from 0, 48, 2 324.0 9.3 96 hours and 10, 20 and 35 days Were analyZed for Epri 3 294.0 8.4 nomectin concentration. 5 334.0 9.5 [0150] The mean eprinomectin and levamisole pharmaco 6 315.0 9.0 9 343.0 9.8 kinetic pro?les in cattle plasma after treatment With the tWo 11 297.0 8.5 formulations are presented in the ?gures beloW. Results dem 12 285.0 8.1 onstrate that both formulations resulted in almost identical 13 289.0 8.3 plasma pro?les for both levamisole and eprinomectin. FIGS. 2 and 3 summarize the data. US 2011/0281920 A1 Nov. 17, 2011
TABLE 8
Excipients used in the Eprinomectin/Levamisole iniectable formulations
Formulation 1 Formulation 2 % W/v % W/v Ingredient CAS No. Composition Composition Function
BHT 128-37-0 0.1 0.1 Antioxidant Dimethylacetamide 127-19-5 5 .0 5 .0 Solvent Methyl Paraben 99-76-3 0.15 0.15 Preservative Polysorbate 80 9005-65-6 5.0 i Surfactant Cremophor EL 61796-12-6 i 5.0 Surfactant Water For Injection 7789-20-0 q.s. to 100% q.s. to 100% Vehicle
Formulation Development the stress study monitored the assay of the tWo actives. Den sity and pH values Were also monitored. Density Was in the [0151] A number of solvents Were evaluated including Glycerol Formal, 2-Pyrol, Propylene Glycol, Miglyol 840 range 1.05-1.10 and pH in the range 3.0-5.0. Visual assess and DMA from 2-20% (see Example 2). The preferred ment shoWs the product to be ?oWable With a Water-like batches contained a surfactant such as Cremophor EL, appearance. Polysorbate 80 and modi?ed Lecithin in the range 2-5%. [0154] The bulk of the WFI Was added to the main manu Vehicles trialed included Water, Glycerol Formal and modi facturing vessel and methyl paraben, the Water soluble pre ?ed vegetable oil. The combination of 5% DMA and 5% servative, Was added ?rst to maintain microbial resistance. CREMAPHOR EL or POLYSORBATE 80 Was demon Levamisole Phosphate Was added next as it Water soluble and strated to be quite useful for the levamisole/ivermectin for at a relatively high concentration. In a separate vessel, epri mulations according to the present invention. nomectin and BHT Were dissolved in DMA and the surfactant [0152] Formulations comprising eprinomectin instead of added next. This combination Was then added to the bulk ivermectin Were evaluated using CREMAPHOR EL and sev aqueous phase With mixing. Eprinomectin is not Water eral other surfactants. The stability data provided above indi soluble, so this carried the eprinomectin into and maintained cated that eprinomectin performed at least as Well as ivermec it in solution. Finally the batch Was made to Volume With WFI tin. A series of batches Were prepared comparing the addition and mixed. All mixing Was accomplished using simple of BHT as an antioxidant to batches Without antioxidant. mechanical agitation. Heat Was only required to dissolve the Stress studies shoWed a signi?cant improvement in stability Methyl Paraben, and once all soluble components Were dis of the actives When BHT Was added to the formulation. solved, the WFI Was cooled before further processing occurs. Finally, Methyl Paraben Was selected for a preservative as the marketed product may be provided as a multi-dose pack. Example 2 Physicochemical Properties [0155] Preliminary Work focused on stability of ivermec [0153] Assessment of developmental formulation batches tin/levamisole-containing formulations, though subsequent Was generally performed on the basis of appearance, includ studies (summarized in Example 1) shifted to the related ing color, of the solution and the assay content of both of the macrocyclic lactone eprinomectin. As mentioned, epri active ingredients. The description of appearance for the pro nomectin offers a signi?cant advantage in terms of short posed formulation is that of a clear light yelloW to yelloW Withdrawal time due its relatively loW milk to plasma parti solution. The physicochemical properties investigated during tion ratio. Table 9 summarizes batches 2-13.
TABLE 9
Formulation summary: IVM is ivermectin and LEV is levamisole phosphate
Source and batch number of Batch Batch Formulation APIs Purpose, method of
No siZe % W/v IVM LEV manufacture and comments
02 100 mL Ivermectin 0.70 Hisun Guilin Purpose: Preliminary Levamisole P04 22.3 080340 020903 screening trial to determine a Glycerol Formal 20.0 suitable base for Ivermectin BenZyl Alcohol 1.0 and Levamisole Phosphate Polysorbate 80 5.0 550 C. 5 days WFI to 100 Assay compared 03 100 mL Ivermectin 0.70 Hisun Guilin to initial
Levamisole P04 22.3 080340 020903 IVM LEV
DMA 2.0 02 78.0% 94.4% Cremophor EL 2 0 03 101.2% 94.1% WFI to 100 04 96.5% 86.3% US 2011/0281920 A1 Nov. 17, 2011 10
TABLE 9-continued
Formulation summary: IVM is ivermectin and LEV is levamisole phosphate
Source and batch number of Batch Batch Formulation APIs Purpose, method of
No size % W/v IVM LEV manufacture and comments
04 100 mL Ivermectin 0.70 Hisun Guilin 06 99.6% 99.7% Levamisole P04 22.3 080340 020903 07 98.0% 94.5% Propylene Glycol 20.0 08 125.3% 96.1% BenZyl Alcohol 1.5 Result: Batch 03 (07) batch 06 WFI to 100 most suitable formulations. 06 100 mL Ivermectin 0.70 Hisun Guilin Batch 03, a solution preferred Levamisole PO4 22.3 080340 090601 over batch 06 an emulsion. Miglyol 840 20.0 Phospholipon 1.8 Glycerol Formal 2.2 WFI to 100 07 100 mL Ivermectin 0.70 Hisun Guilin (Repeat Levamisole PO4 22.3 070403 090601 03) DMA 2.0 Cremophor EL 2.0 WFI to 100 08 100 mL Ivermectin 0.70 Hisun Guilin Levamisole P04 22.3 070403 090601 WFI 30.0 Montanide to 100 09 100 mL Ivermectin 0.70 Hisun Guilin Purpose: Repeat batch 03 Levamisole PO4 22.3 080340 090601 using tWo different batches of DMA 2.0 Ivermectin Cremophor EL 2.0 55° C. 5 days WFI to 100 Percent initial
IVM LEV
10 100 mL Ivermectin 0.70 Hisun Guilin 09 83.6% 97.9% Levamisole P04 22.3 070403 090601 10 58.4% 97.6% DMA 2.0 Result: Crystallization Cremophor EL 2.0 observed. WFI to 100 11 100 mL Ivermectin 0.70 Hisun Guilin Purpose: Repeat batch 03 but Levamisole PO4 22.3 080340 090601 increase surfactant and solvent DMA 5.0 levels. Use tWo different Cremophor EL 5.0 batches of Ivermectin. WFI to 100 55° C. 5 days 12 100 mL Ivermectin 0.70 Hisun Guilin Percent initial
Levamisole P04 22.3 070403 090601 IVM LEV
DMA 5.0 11 85.6% 97.2% Cremophor EL 5.0 12 93.1% 98.3% WFI to 100 Result: Stability improved. 13 300 mL Ivermectin 0.70 Hisun Guilin Purpose: Repeat batch 12 With Levamisole PO4 22.3 070403 090601 stress testing at 40° C. and DMA 5.0 55° C. Cremophor EL 5.0 5 days WFI to 100 Percent initial
IVM LEV
40° C. 98.3% 99.6% 55° C. 85.2% 97.3% Result: Stability improved.
Example 3 order to arrive at the proper blend of actives and excipients to _ _ produce an effective, pharmaceutically acceptable formula Altematlve Formulations t1on.- lnjectable- formulatlons- tend to be partlcularly- challeng [0156] The following example illustrates the signi?cant 111% to develop, as factors Such as eXCeSSlVe Vlscoslty and time and effort that Was invested in developing the stable, injection site irritation can preclude the use of otherwise inj ectable formulations according to the present invention. A effective excipients (cg. excipients Which provide good sta person of ordinary skill Will immediately appreciate the rig bility for the combination of active pharmaceutical ingredi orous experimentation that Was, and is very often required in ents). US 2011/0281920 A1 Nov. 17, 2011
[0157] Pre-formulation development efforts commenced [0158] Early indications Were that CAPMUL MCM (me using the hydrochloride salt of Levamisole. Preliminary solu dium chain mono and diglycerides) aqueous based formula bility data strongly contra-indicated the use of NMP and tions might provide a clinically acceptable base for injection. Glycerin as a single solvent system to solubiliZe Levamisole Batches lNJ0003-01 to lNJ0003-11 Were therefore formu and the use of vegetable oil to solubiliZe the lvermectin. A lated With Levamisole using either the hydrochloride or the large number of surfactants and stabilizers including CAP phosphate salts thereof (formulation compositions are sum MUL MCM, TWEEN 80, SPAN 20, CREMAPHOR RH 40, marized in Table 10). CAPMUL MCM and TWEEN 80 With PVP K-30 and SODIUM CMC Were trialed. Also trialed Were a citrate buffer Was the basis of this aqueous formulation. several buffers, including phosphate and acetate salts before Although considered stable at the planned storage of 2-8° C., citrate Was ?nally selected based upon the desirability/stabil unacceptable site reactions occurred during clinical investi ity of the resulting formulations. gation.
TABLE 10
Formulation summary — batches INJ003-01 through 1NJ003-11
Batch Batch Formulation
No size % W/v Purpose, method of manufacture and comments
INJ003 — 400 mL Ivermectin 0.48 Purpose: Preliminary screening trial to determine a 01 Levamisole HCl 20.0 suitable base for Ivermectin and Levamisole HCl Capmul MCM 25.0 25° C. Tween 80 15.0 Assay compared to initial
Tri Sodium Citrate 1.5 Ivermectin Levamisole HCl
Conc. HCl 0.25 mL 1 M 99% 98% WFI to 100 2 M 98% 99% 4 M 97% 97% 6 M 95% 98% 9 M 93% 97% 12 M 92% 97% 40° C. Assay compared to initial
Ivermectin Levamisole HCl
1 M 95% 95% 2 M 91% 96% 4 M 86% 92% 6 M 78% 90% 55° C. Assay compared to initial
Ivermectin Levamisole HCl
2 Wk 84% 95% 4 Wk 70% 93% INJ003- 400 mL Ivermectin 0.48 Purpose: Preliminary screening trial to determine a 02 Levamisole HCl 20.0 suitable base for Ivermectin and Levamisole HCl Capmul MCM 10.0 25° C. Tween 80 10.0 Assay compared to initial
Tri Sodium Citrate 2.0 Ivermectin Levamisole HCl
Conc. HCl 0.25 mL 1 M 97% 97% WFI to 100 2 M 95% 96% 4 M 93% 97% 6 M 91% 95% 9 M 90% 96% 12 M 87% 96% 40° C. Assay compared to initial
Ivermectin Levamisole HCl
1 M 90% 94% 2 M 83% 91 % 4 M 79% 91 % 6 M 75% 88% US 2011/0281920 A1 Nov. 17, 2011 12
TABLE 10-c0ntinued
Formulation summary — batches INJ003-01 through INJ003-11
Batch Batch Formulation
No siZe % W/v Purpose, method of manufacture and comments
55 ° C. Assay compared to initial
Ivermectin Levamisole HCl
2 Wk 74% 89% 4 Wk 69% 83% INJ003- 1000 mL Ivermectin 0.48 Purpose: Batch submit for site reaction study 03 Levamisole HCl 16.11 25° C. Capmul MCM 25.0 Assay compared to initial
TWeen 80 15.0 Ivermectin Levamisole HCl
Tri Sodium Citrate 1.5 1 M 98% 99% Conc. HCl 0.25 mL 2 M 94% 96% WFI to 100 4 M 94% 99% INJ003- 500 mL Ivermectin 0.48 Purpose: Same formula as batch 1NJ003-01 04 Levamisole HCl 20.0 25° C. Capmul MCM 25.0 Assay compared to initial
TWeen 80 15.0 Ivermectin Levamisole HCl
Tri Sodium Citrate 1.5 1 M 97% 100% Conc. HCl 0.25 mL 2 M 97% 99% WFI to 100 4 M 96% 98% INJ003- 500 mL Ivermectin 0.48 Purpose: Same formula as batch 1NJ003-01 05 Levamisole HCl 20.0 25° C. Capmul MCM 25.0 Assay compared to initial
TWeen 80 15.0 Ivermectin Levamisole HCl
Tri Sodium Citrate 1.5 1 M 89% 79% Conc. HCl 0.25 mL 2 M 92% 81% WFI to 100 4 M 87% 77% INJ003- 500 mL Ivermectin 0.48 Purpose: Same formula as batch 1NJ003-01 — With 06 Levamisole HCl 20.0 parabens Capmul MCM 25.0 25° C. TWeen 80 15.0 Assay compared to initial
Tri Sodium Citrate 1.5 Ivermectin Levamisole HCl
Methylparaben 0.18 1 M 99% 99% Propylparaben 0.20 2 M 100% 98% Conc. HCl 0.25 mL 4 M 96% 99% WFI to 100 INJ003- 1500 mL Ivermectin 0.57 Purpose: Batch With Levamisole PO4 & BHT 07 Levamisole PO4 20.0 Results: Sample kept at 55° C. Were separated into Capmul MCM 25.0 tWo layers in three Weeks TWeen 80 15.0 BHT 0.02 Methylparaben 0.18 Propylparaben 0.20 Tri Sodium Citrate 1.5 Propyl gallate 0.02 Conc. HCl 0.25 mL WFI to 100 INJ003- 1500 mL Ivermectin 0.57 Purpose: Batch With Levamisole PO4 & nil BHT 08 Levamisole PO4 20.0 Results: Sample kept at 55° C. Were separated into Capmul MCM 25.0 tWo layers in three Weeks TWeen 80 15.0 Methylparaben 0.18 Propylparaben 0.20 Tri Sodium Citrate 1.5 Propyl gallate 0.02 Conc. HCl 0.25 mL WFI to 100 US 2011/0281920 A1 Nov. 17, 2011 13
TABLE 10-c0ntinued
Formulation summary — batches INJ003-01 through INJ003-11
Batch Batch Formulation
No siZe % W/v Purpose, method of manufacture and comments
INJ003- 1500 mL Ivermectin 0.57 Purpose: Batch With Levamisole PO4 & BHT With 09 Levamisole PO4 20.0 pH 4.5 Capmul MCM 25.0 2-8° C. TWeen 80 15.0 Assay compared to initial
BHT 0.03 Ivermectin Levamisole PO4
Methylparaben 0.18 1 M 103 104 Propylparaben 0.20 2 M 105 100 Tri Sodium Citrate 1.5 3 M 100 99 Propyl gallate 0.02 6 M 103 101 Conc. HCl 0.25 mL 9 M 101 99 WFI to 100 25° C. Assay compared to initial
Ivermectin Levamisole P04
1 M 103 103 2 M 103 99 3 M 100 95 6 M 101 94 9 M 93 90 INJ003- 1500 mL Ivermectin 0.57 Purpose: Batch With Levamisole PO4 & BHT With 10 Levamisole PO4 20.0 pH 4.0 Capmul MCM 25.0 2-8° C. TWeen 80 15.0 Assay compared to initial
BHT 0.03 Ivermectin Levamisole PO4
Methylparaben 0.18 1 M 103 103 Propylparaben 0.20 2 M 103 99 Tri Sodium Citrate 1.5 3 M 96 102 Propyl gallate 0.02 6 M 104 102 Conc. HCl 0.25 mL 9 M 100 98 WFI to 100 25° C. Assay compared to initial
Ivermectin Levamisole P04
1 M 103 102 2 M 103 99 3 M 99 96 6 M 99 94 9 M 92 89 INJ003- 1500 mL Ivermectin 0.57 Purpose: Batch With Levamisole PO4 & BHT With 11 Levamisole PO4 20.0 pH 5.0 Capmul MCM 25.0 2-8° C. TWeen 80 15.0 Assay compared to initial
BHT 0.03 Ivermectin Levamisole PO4
Methylparaben 0.18 1 M 101 104 Propylparaben 0.20 2 M 102 102 Tri Sodium Citrate 1.5 3 M 101 10 Propyl gallate 0.02 6 M 102 100 Conc. HCl 0.25 mL 9 M 98 99 WFI to 100 25° C. Assay compared to initial
Ivermectin Levamisole P04
1 M 102 102 2 M 102 101 3 M 95 96 6 M 98 94 9 M 97 86 US 2011/0281920 A1 Nov. 17, 2011
INJ0003/11 Method of Preparation [0159] 1. Ivermectin Was dissolved in CAPMUL MCM TABLE 12-continued under stirring. Formulation ingredients [0160] 2. To this Tween 80 Was added under stirring. [0161] 3. BHT Was added to step 1 under stirring. % W/v [0162] 4. Methyl paraben and Propyl paraben dissolved Ingredient Composition Function in 600 ml of Water under heating. Methyl Paraben 0.15 Preservative [0163] 5. Levamisole phosphate Was added to step 3 Polysorbate 80 5.0 Surfactant under stirring at room temperature. Water For Injection q.s. to 100% Vehicle [0164] 6. Propyl gallate Was added to step 4 under stir *Corrected for potency ring. [0165] 7. Trisodium citrate and 50% HCl solution Were [0171] The results demonstrated that abamectin, moxidec added to step 5 under stirring. tin and doramectin are suitable macrocyclic lactones for for [0166] 8. Step 2 Was added to step 6 under stirring mulations according to the instant invention. When compared [0167] 9. Adjusted the pH to around 5.00 using 20% HCl to the eprinomectin/levamisole formulation, the general solution, if necessary. physical parameters are very similar. [0168] 10. The resulting solution Was made up to volume With Water. TABLE 13a
Formulation of various macrocyclic lactones levamisole TABLE 11 Eprinomectin/ Abamectin/ Site reaction test for ivermectin/levamisole Levamisole Inj. Levamisole Inj. injection for cattle — IN] 0003/11 B. No.
Species: Bovine SOW21-08 SOW21-23 Number: 2 animals Type: mixed Condition 2-8° C. 25° C. 40° C. 2-8° C. 25° C. 40° C. Age: ~6 months Sex: either Storage 5 Days 5 Days 5 Days 5 5 5 Weight: Greater than 125 kg Period Days Days Days Source: Commercial farm Description * * * * * * Health status: Normal animals in good body condition. Wt/mL 1.0843 1.0843 1.0843 1.0831 1.0831 1.0831 Special requirements: No anthelmintic treatment in the past 4 Weeks Viscosity 3.5 cP 3.5 cP 3.5 cP 3.3 cP 3.3 cP 3.3 cP Medication history: Herd-based treatments as part of normal pH 4.22 4.21 4.06 4.17 4.16 4.05 animal husbandry practice. Housing and The study animals Were maintained as a management: single mob and rotationally grazed on pasture With continuous access to Water throughout the experimental period. TABLE 13b Formulation of various macrocyclic lactones levamisole [0169] Results: TWo calves Were treated. No sting in either Moxidectin/ Doramectin/ animal Was noted upon injection. A site reaction occurred in Levamisole Inj. Levamisole Inj. both animals Within 1 hour of injection, lasting for the entire B. No. 28 days of the trial. The reaction Was a slightly raised area at the point of injection getting as large as 150x170 mm and SOW 21-24 SOW 21-25 110x140 mm. Such reactions Would be unacceptable in a Condition 2-8° C. 25° C. 40° C. 2-8° C. 25° C. 40° C. commercial farming situation. Storage 5 5 5 5 5 5 Period Days Days Days Days Days Days Example 4 Description * * * * * * Wt/mL 1.0825 1.0825 1.0825 1.0837 1.0837 1.0837 Formulations Including Other Macrocyclic Lactones Viscosity 4.2 cP 4.2 cP 4.2 cP 3.3 cP 3.3 cP 3.3 cP pH 4.15 4.14 4.02 4.13 4.12 4.01 [0170] Tests Were conducted to determine Whether the invention Would be able to be used as a vehicle for delivering * Clear very light yelloW solution a Wider range of macrocyclic lactone active ingredients. The test formulations Were preparedusing POLYSORBATE 80 as the surfactant of choice according to Table 12. Example 5 Other Formulations TABLE 12 [0172] Formulations including novel avermectins Would be Formulation ingredients prepared in accordance With Table 12, and could include the % W/v compounds and amounts recited in Table 14. The formula Ingredient Composition Function tions could also include any one of the avermectins or aver mectin monosaccharides, modi?ed in the 4' or 4" position, Macrocyclic lactone active 070* Active Levamisole Phosphate 223* Active describedinU.S. Pat. No. 7,704,961,U.S. Pat. No. 7,521,429, BHT 0.1 Antioxidant US 2006-0105970A1, US. Pat. No. 7,678,740, US. Pat. No. Dimethylacetamide 5 .0 S olvent 7,632,820, US. Pat. No. 7,678,773, US. Pat. No. 7,605,134, and US. Pat. No. 6,933,260 (each to Merial). US 2011/0281920 A1 Nov. 17, 2011 15
liver and peri-renal fat. Both primary and reserve samples TABLE 14 Were stored frozen until shipment of the “primary samples” to the lab. “Reserve” samples remained in froZen storage. No Amounts and identities of novel avermectin compounds detectable concentrations of eprinomectin nor levamisole % W/v Were found in any tissues from untreated cattle indicating no Avermectin Composition contamination at the ?eld or analytical phases. Statistical 4"-diallylaminosulfonyloxy-avermectin B1 0.5-0.9 analyses and Withholding periods Were calculated. No 4"-(2-Hydroxyethylaminosulfonyloxy)-avermectin B1 0.5-0.9 adverse effects Were noted in any of the treated cattle for the 4"-dipropargylaminosulfonyloxy-avermectin Bl 0.5-0.9 4"-bis(cyanomethyl)aminosulfonyloxy-avermectin B1 0.5-0.9 duration of the study. 4"-Dicyanomethylaminosulfonyloxy-avermectin B1 0.5-0.9 4"-pyrrolidinosulfonyloxy-avermectin B 1 05-09 TABLE 16 4"-benZoylaminosulfonyloxy-avermectin B1 0.5-0.9 4"-epi-sulfamoyloxy-avermectin B 1 05-09 Study design 4"-methylaminosulfonyloxy-avermectin B 1 05-09 4"-ethylaminosulfonyloxy-avermectin B 1 05-09 Parameter Description 4"-sulfamoyloxy-avermectin B1 0.5-0.9 4"-allylaminosulfonyloxy-avermectin B1 0.5-0.9 Number 24 Any dimeric avermectin 0.5-0.9 Species Bovine Any hetero-dimeric avermectin/milbemycin Breed Short Horn cross (Hereford x Friesian dams, Any 4" substituted avermectin 0.5-0.9 Short Horn sires) Any 4'-4" dually substituted avermectin 0.5-0.9 Sex Females and castrated males Any avermectin monosaccharide 0.5-0.9 Age Approximately 8 Months Body Weight Range 200-246 kg Condition Excellent condition, healthy Example 6 Determination of the Chemical Residues and Deple [0175] The ?eld study Was conducted according to the tion Pro?les for Eprinomectin and Levamisole Fol details outlined in Table 16. In this GLP tissue residue study, lowing Subcutaneous Injection of Cattle tWenty (20) treated cattle and tWo (2) untreated control cattle Were sacri?ced for tissue collection folloWing the subcutane [0173] These studies Were conducted to determine the resi due levels and depletion pro?le of eprinomectin and levami ous injection of a developmental parasiticide. TWo (2) further sole in the tissue of cattle, following the subcutaneous inj ec “contingency” animals Were treated and managed With the tion of an inj ectable formulation according to the instant treated mob. All animals Were chosen from a mob of thirty invention, containing 7 g/ L eprinomectin and 223 g/ L levami (30) cattle. Upon arrival at the research facility, all cattle Were sole phosphate. Thirty Short Horn cross cattle Were sourced already individually identi?ed With an ear tag With a unique from a commercial farm. On the day of treatment (Day 0) all number in their left car so it Was unnecessary to remove that cattle Were Weighed, and then, based on live Weight, 24 ani tag and/ or replace it With another. On the day of treatment mals Were allocated to treatment and tissue collection groups. (Day 0) all cattle Were Weighed and ranked from highest to The treatment groups and sampling times are presented in loWest based on live Weight. In order to keep the live Weight Table 15. range as narroW as possible, the heaviest four and lightest tWo
TABLE 15
Treatment groups, active ingredients and dose rates
Treatment Active ingredient Number of cattle treated group concentration Dose rate (tissue collection times)
1. Untreated i i 2 cattle (2 on Day 21) 2. Test Item 7 gL eprinomectin + 1.25 mL/35 kg 20 cattle (5 on each of Days 223 gL levamisole phosphate 21, 28, 35 and 42) 3. Test Item 7 gL eprinomectin + 1.25 mL/35 kg 2 cattle contingency 223 gL levamisole phosphate
[0174] Residues. The untreated cattle Were removed from animals from the 30 potential study animals Were removed the site of treatment before application of the test item. A from the study. Labelled discs (20>
TABLE 17 TABLE 19
Average live Weight of each tissue collection day for treated and untreated cattle Presents the maximum residue limits for eprinomectin (mgkg), and levamisole (mg/kg) limit (PQL) for eprinomectin (mg/kg) and levamisole Treatment Collection day (mg/kg) in animal meat, edible offal and fat tissue Group 0 21 28 35 42
Untreated 223 Levamisole Treated 231 228 227 227 I I Contingency 215 Eprinomectin (mgkg) (mg/kg)
[0176] Cattle that Were treated received a White uniquely Fat 0'25 0'01 numbered tag in their right car while the untreated control Llver 1-5 0-1 cattle received a red uniquely numbered tag in their right ear. Kidney 0.3 0.01 Contingency animals received a blue numbered tag in their Muscle 0_05 001 right ear. Treatment details are presented in Table 18.
TABLE 18 TABLE 20 Active ingredient, dose rates and cattle numbers in treatment groups in accordance With ACVAM guidelines for maximum alloWable residues in animal meat edible offal and fat tissue Limit of quantitation (LOQ), limit of detection (LOD) and practical quantitation limit (PQL) for eprinomectin (mgkg) and Treatment Active ingredient Number levamisole (mgkg) in animal meat, edible offal and fat tissue. group concentration Dose rate of cattle 1- Untmat?d i i 2 Eprinomectin (mgkg) Levamisole (mg/kg) 2. Test Item 7 gL eprinomectin and 1.25 mL/35 20 223 g/L levamisole kg LOD LOQ PQL LOD LOQ PQL phosphate 3- Contingency 7 g/L 6Prinomectin and 1-25 mL/35 2 Fat tissue 0.001 0.005 0.01 0.001 0.005 0.01 kg . 223 gL l?vamisol? Edible offal & 0.001 0.005 0.01 0.002 0.006 0.01 phosphate malt
TABLE 21
Eprinomectin B1a and levamisole concentrations in liver tissue
Eprinomectin B1 a (mg/kg) Levamisole (mgkg)
Ear Tag Interval Product (Corrected) (Uncorrected) (Corrected) (Uncorrected)
1 21 UTC