ZA200402103B - Pesticidal formulations. - Google Patents

Description

: PCT/AU01/01169 §. DEC. 2003 13:31 SPRUSON & FERGUSON : Received 08 Deczmber 2003 (J 1

PESTICIDAL FORMULATIONS - ee . Technical Fleld ) The present invention relates (o combinations of pesticidally active compounds suitable for use as active agents in pesticidal formulations, the formuistions themselves «and to the various applications of those formulations as pesticides, specifically 1n controlling Phthiraptcra, Siphonapiera and Acarina pests. Such applications include the control of such external Phthiraptera, Siphonsptera and Acarina pests in domestic animels including but not limited to sheep, cattle, poultry, pigs, goals, camelids, hoyses, dogs and - cats, as well as thc household and rural applications of such formulations in control of” ww such pests. :

Background of the Invention

Historically, the greatest damage to domestic animals and crops has been caused and continues to be caused by pests such zs insects, fungi, nematodes and microbes.

Insects panicularly represent a cause for concern as hey are the most numerous of all 1s living organisms and constitute approximately 72% of all animal species. Approximately 1% of insects are considercd pests in thet they attack humans and/or domestic animals, ransmit human, animal and plant diseases, desuoy crops, objects and structures and compete for food and other necessities It is estimated that enonmous agricultural Josses result worldwide from insect presence. iu Domestic animals which include animals of agricultural worth such as sheep, cattie, © horses, goats, pigs and other ruminants and monogastrics are almost invariably subject to the activity of pests including inscets, acarides, ecanna, siphonaptcra, phthirapters, znoplura and mallophaga. External parasites such as licks, lice and fleas imitate the animals and can cause economic loss in the form of poor quality hide, wool or sheep skin, 2s poor quality meat/tissug, reduced weighl gain and even death as a result of the ammal carrying hannful parasites.

The losses resulting from inscct caused human and animal discases are also enormous. In fact, insects are considered to be the carriers of morc than 250 viruses which are pathogens of humans and higher animals. The numbers of human deaths jo caused by mosquito transmitted diseases such as malaria and lymphatic filanasis are . huge. Flies also transmit buman and animal reloted discases such as trachoma, trypanosomiasis and river blindness. o

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However, out of the nearly onc million species of arthropods which includes lice, ticks, flies and rites, only a small percentage require the application of control measures.

To date, the primary method for controlling insects and other pests, particularly in respect

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Cy le of domestic animals (such as sheep, cattle, goats, horses and hogs) has been by the application of synthetic chemical pesticide compositions. It is estimated that there are at least 35,000 formulated pesticide products worldwide with chemicals as the active . ingredients. Such pesticide products include antimicrobials, larvicides, insecticides, s animal dips, avicides and disinfectants. .

The extensive use of chemical insecticides since the 1940s has resulted in a large number of problems including widespread insect resistance, emergence of secondary pests, hazards to human and animal health as well as detrimental effects on fish and birds, environmental pollution and the increasing economic costs of new insecticides.

Many insect species have developed resistance to the action of specific insecticides so as to necessitate changes in control practices. There is an ever-widening pool of insect pests which are developing multiple resistance. The resistance genes having lengthy persistence in insect genomes which preclude successful reuse of an insecticide to control an insect population with resistant genes. is Pesticide/insecticide residues and their consequential many potential human, animal and environmental risks are also seen as one of the major problems resulting from chemical usage, particularly those formulations containing active agents which include organophosphates or synthetic pyrethroids. With the exception of microbial insecticides, : nearly all pesticides result in residues of various chemicals and their degradation products © 20 or metabolites which may be present in detectable amounts (ppb to ppm) in food despite food processing. Tissue/meat residues are also a major concern when considering use of insecticides on farm animals.

Potential human risks from the use of such insecticides include acute toxic reactions to the insecticide such as poisoning, skin and eye irritations, as well as possible 2s long term effects such as cancer, birth defects, and reproductive disorders. Acute inhalation toxicity as well as dermal penetration are also potential risks. Health hazards in humans may also arise from repeated exposure to a chemical over a limited period of time. :

In particular, the currently used actives of synthetic pyrethroids and : organophosphates which are commonly used in insecticidal formulations to control lice and flies, particularly on sheep, are not only toxic to animals but also to the human operator who applies them. Exposure in farmers or operators who handle both pesticide concentrates and the larger volumes of pesticide diluted for use, is a cause for concern.

Further, it is possible for the operator to ingest pesticides not only by mouth, but also by 3s breathing (eg spray drift) and by absorption through the skin (accidental spillage). Of hole @ particular concem has been the use of organophosphates where accidental exposure causes acute and chronic poisoning affecting the nervous system.

Accordingly, insect and other pest control has been sought to be directed away

B from exclusive reliance on insecticides and towards the optimisation of environmental , s and economic insect and pest control (integrated pest management). The application of microbial control in which insects are attacked by pathogens such as viruses, bacteria, fungi and protozoa are favoured as such microbial insecticides are highly selective for insect pests and do not leave toxic residues. However, such microbial insecticides are not without their problems such as the difficulty in applying as well as confining the natural enemy/parasite/disease to a large area. Further, they also have the disadvantage of short residual action and extreme specificity which limits general applicability.

Biological control has been recently applied in the area of insecticides/pesticides through the release of sterilised male insects. Genetic engineering has also recently been applied by way of mass introduction of deleterious mutations such as chromosomal 1s translocations. However, such procedures are very expensive and stringent criteria are required before release of sterile males is contemplated. Chemosterilants which sterilise large segments of insect pest populations are also known but are strong carcinogens which precludes their use.

The use of chemical insecticides and pesticides and their environmental and economic viability, the dangerous nature and magnitude of the persisting residues as well as increasing insect and pest resistance, together with high toxicity levels of many chemical insecticides, has resulted in the search for new substances or approaches to insect and other pest control.

There is therefore a need for compounds and combinations thereof which can be 2s used as active agents in pesticides, particularly against insects which afflict domestic animals or their environs, and which are effective at low application rates, selective in biologic action and have low toxicity and a high margin of safety to humans, crops, economic animals, aquatic organisms and birds. Such compounds and combinations must be both environmentally friendly in that there must be demonstrably low impacts on the ’ 3 environment, as well as economically viable to use on a large scale. Further, there must be none or little insect or other pest resistance to such compounds or combinations.

Fermentation product A83543, also known as spinosyn, includes a family of related compounds (spinosyns) produced by Saccharopolyspora spinosa. These are naturally derived fermentation products with a positive safety profile in contrast to currently used synthetic organically derived compounds (such as synthetic pyrethroids,

~ @ . 4 organophosphates, organochlorines and carbamates), and have previously been shown to exhibit excellent insecticidal activity. Accordingly by the term "A83343 compounds" which has the same scope as the phrase "spinosyn and derivatives and analogues thereof” . is meant components consisting of a 5,6,5-tricyclic ring system, fused to a 12-membered s macrocyclic lactone, a neutral sugar (2N,3N,4N-tri-O-methylrhamnose) and an amino sugar (forosamine). The family of natural components of A83543 include a genus taught in EPO patent application No. 0375316 and having the following general formula:

R! ORS? CH, ol OCH;

R?

H H 0) OR¢ 0)

RICH,” ~O H

H H

R3 wherein R! is H or a group selected from

CH; CH;

TITCAL RE NN

(a) (®)

CH;

TIAL, (©) or) (CH;);N } . rN

CH; (d) and R2, R4, R3, RS and R6 are hydrogen or methyl; or an acid addition salt thereof when

R! is other than hydrogen. = is The family of compounds from A83543 fermentation product has been shown to comprise individual compounds A83543A, A83453B, A83543C, A83453D, AB3543E, >

A83453F, A83543G, A83453H, AS83543], A83453L, A83543M, AB3453N, A83543Q,

AB83453R, A83543S, A83453T, A83453U, A83543V, A83453W, AB3453X. Boeck er al. described spinosyns A-H and J and salts thereof in US patent Nos 5,362,634, 5,496,932 re WO 03/024223 PCT/AU01/01169 ® 5 and 5,571,901 which are incorporated herein by reference. Mynderse et al. described spinosyns L-N, their N-demethyl derivatives and salts thereof in US patent No, 5,202,242 incorporated herein by reference. Tumer er al. described spinosyns Q-T, their N- *- demethyl derivatives and salts thereof in US patent Nos 5,591,606, 5,631,155 and , s 5,767,253 which are also incorporated herein by reference. Spinosyns K,0,P,U,V,W, and h Y are described in the article by DeAmicis, C.V. ef al. in American Chemical Society's

Symposium Series: Phytochemicals for Pest Control (1997), Chapter 11 "Physical and

Biological Properties of Spinosyns: Novel Macrolide Pest-Control Agents from

Fermentation” pp146-154.

Spinosyn A (A83543A) was the first spinosyn isolated and identified from the fermentation broth of Saccharapolyspora spinosa. Subsequent examination of the fermentation broth revealed that the parent strain of S.spinosa produced a number of spinosyns (A83543A to J). Compared to spinosyn A, spinosyns B to J are characterised by differences in the substitution patterns on the amino group of the forosamine, at 1s selected sites on the ring system and on the neutral sugar. The strains of S.spinosa produce a mixture of spinosyns which primary components are spinosyn A (~85%) and : spinosyn D (~15%). These are the two spinosyns that are currently known as the most active as insecticides.

Similar to the spinosyns, macrocyclic lactones have also previously been shown to exhibit excellent insecticidal activity. Macrocyclic lactones have a complex ring structure and include such well known anthelmintic compounds as avenmectins and milbemycins. The avermectins are isolated from fermentation products of Streptomyces avermitilis and ivermectin is a compound which is a semisynthetic chemical formed by modification of avermectin. The basic structure of the avermectins is a 16-membered 2s lactone ring to which are appended three main substituent groups: a hexahydrobenzofuran group, a disaccharide group (at C-13) and a spiroketal ring (C-17 to C-28). Doramectin is a novel avermectin. Milbemycins are other compounds which are not avermectins but which can be considered to come within the class of compounds which are macrocyclic lactones. The milbemycins differ structurally from the avermectin group, mainly in the ‘ 0 absence of a disaccharide group on C-13. Milbemycin D and milbemycin 5-oxime are two such macrocyclic lactones. Moxidectin is derived from the fermentation product ) nemadectin and possesses a methoxime substituent on C-23.

The present invention resides in the discovery of a synergistic combination of pesticidal compounds, the formulation and application of specific pesticidally active

Ca agents based on the synergistic combination and their use in pesticidal formulations against Phthiraptera, Siphonaptera and Acarina pests, particularly in domestic animals.

Objects of the invention .

Accordingly, it is an object of this invention to provide a pesticidal composition oo s which is systemically active against Phthiraptera, Siphonaptera and Acanna pests, i" containing a synergistic combination of at least one A83543 compound and at least one macrocyclic lactone.

It is also an object of this invention to provide a pesticidal composition which is systemically active against Phthiraptera, Siphonaptera and Acarina pests in domestic to animals including cattle, camellids, pigs, dogs, horses, cats, sheep, goats and poultry, containing a synergistic combination of at least one A83543 compound and at least one macrocyclic lactone.

It is another object of this invention to provide one or more pesticidal formulations systemically active against Phthiraptera, Siphonaptera and Acarina pests, containing a synergistic combination of at least one A83543 compound and at least one macrocyclic lactone as the active principles together with at least one acceptable carrier or diluent.

It is another object of this invention to provide one or more pesticidal formulations systemically active against Phthiraptera, Siphonaptera and Acarina pests in domestic animals including cattle, camellids, pigs, horses, dogs, cats, sheep, goats and poultry, containing a synergistic combination of at least one A83543 compound and at least one macrocyclic lactone as the active principles together with at least one acceptable camer or diluent.

It is also an object of the present invention to provide a method of eliminating and/or controlling Phthiraptera, Siphonaptera and Acarina pests in domestic animals including cattle, camellids, pigs, horses, dogs, cats, sheep, goats and poultry by applying or administering to said animals a pesticidally active combination of compounds alone or together with an acceptable carrier or diluent, such that said pesticidally active combination is systemically delivered to said Phthiraptera, Siphonaptera and Acarina i pests.

It is also an object of the present invention to provide a method of eliminating ; and/or controlling Phthiraptera, Siphonaptera and Acarina_pests by administering to said pests a pesticidally active combination of compounds alone or together with an acceptable carrier or diluent, such that said combination is present systemically in said pests.

: PCT/AU01/01169 © 6.DEC.2003 13:32 SPRUSON & FERGUSON Received 08 Decerz:ber 2003 ' 1@® 7

The term ‘Phthiraptera’ or ‘Phthirapiera pests’ 8s used herein defines members of the insect order Phthirapterz, which are parasitic during one or more stages of their life cycle, including the immature stage (Which is defined to include larval (nymph) foims), } : the adult stage or both stages 2nd further includes Phthiraptera insect ¢ggs. 3 The term *Siphonapiera’ or ‘Siphonzplera pests’ as used herein defines members of the insect order Siphonapiera which are parasitic during one or more stages of their life cycle, including the immature stage {which is defined to include larval forms), the adult stage or both stages and further includes Siphoneptcra insect eggs

The term ‘Acarina’ o1 ‘Acarina pests’ as used herein defines members of the H «Arachnida order Acarina which are parasitic during one or more stages of their life cycle. including the immature stage (which is defined to include larval (nymph) forms), the adult stage or both stages and further includes Acarina eggs.

Tt is further noted that for the purposes of the present application, the term spinosyn : : or analogue or derivative thereof is defined to include an individual spinosyn factor 1s (A83543A-H, J-W or Y) an N-demcthyl or other derivative of an individual spinosyn factor, or salt thereof, or a combination thereof, consistent with the disclosure of the abovementioned references which have been incorporated herein. As stated above, the term "AS83543 compound” is used herein to mean an individual spinosyn factor, or an analogue, a derivative or salt thereof, or a combination thereof.

The term ‘controlling or eradicating’ is used 10 refer to a decrease in the number of living insects or arachnids {adult or immaturc forms) or 10 a decrease in the number of visble insect or erachnid epps. The cxtemi of reduction somewhat depends on the application rate, actives used, prevailing weather conditions and other conditions known to those skilled in the ant as The term ‘effective amount’ also used hercin means the amount which is sufficient to cause 8 measurable reduction in the teated insect or arachnid population.

The word ‘carrier’ is used throughout the present specification to include carner blends, that is mixtures of more than one substance.

The term ‘synergistic’ as uscd herein is defined to mean a combination of su components wherein the activity of the combination is greater than the additive of the individual activities of cach component of the combination.

The term ‘macrocyclic lactone’ as used herein is defined to be compounds of the classes of milbemycins znd avermectins, including both naturally occurring compounds and synthetic derivatives thereof, especially those mentioned herein 2nd in the art cited 1s herein.

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: PUI/ALN1/01 169 © 8. DEC. 2003 13:32 SPRUSON & FERGUSON Received 08 Decerber 2003 §

The term ‘domestic animal’ as uscd herein is defined 10 include animals of agriculural worth and companion animale, including but not limited to catilc, camcllids, . pigs, dogs, cats, sheep, poultry, horses and goats as well as other ruminants and monogastrics. s The term ‘systemically active’ as used herein means having an effect or efficacy only when intracorporeally present within the target pest, such as after ingestion or other administzztion which results in the presence of both active agents in the target pest. The term does not mean having a deleterious effect when present within the system of a host domestic znimal, it is limited to activity or efficacy when intracorporcally present within wa pest. } v . Eg Summary of the Invention

A first aspect of the present invention provides an zttive composition for controlling or cradiczting Phthiraptera, Siphonaptera snd Acanna pests, said composition ) being systemically ective in said pests and comprising a synergistic combinaticn of at i+ least one A83543 compound and at least one compound which is 2 macrocyclic lactone.

A second sspect of the present invention provides 2 formulation for controlling or eradicating Phthirapiera, Siphonzptera and Acerina pests in demestic animals, said formulation including zn effective amount of a systemically active composition of the first aspect of the invention and a domestic animal physiologically acceptable carrier, s¢ diluent or excipient. ‘ : A third aspect of the present invention provides an exiemally applied formulation for contio) or erzdication of Phihirapiera, Siphonzplera &nd Acarina pesis in domestic animals, said formulation including an effective amount of a systemically active composition of the first aspect of the invention and & domestic animal physiologically 2s acceptable carrier.

A lourth aspect of the present invention provides a pesticidal bai formulation for i contro} or eradication of Phihirapiera, Siphonapiera and Acarina pests, said formulation including an effcctive amount of a systemically active compotition of the first aspect of the invention and an acceplable carrier. 3c A filth aspect of the present invention provides an orally administered formulation for control or eradication of Phthiraptera, Siphonaptera and Aczrine pests in domestic animals, said formulation including an cffeciive amount of a systemically active composition of the fust aspect of the invention and a domestic animal physiologically acceptable carrier, diluent or excipient.

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A sixth aspect of the present invention provides a parenterally administered {formulation for control or eradication of Phthitapiera, Siphonaptera and Acarina pests in © domestic animals, said formulation including an effective amount of a systemically active composition of the first aspect of the invention 2nd a domestic animal physiologically s acceptable carner, diluent or excipient. : A seventh aspect of the present invention provides a method of controlling or eradicaing Phthiraptera, Siphonspterz and Acarinz pests in domestic animals, said method including the external application of an effective amount of a systemically active composition zccording to the first aspect of the invention, or of a formulation according to ww the sccond or third aspects of the invention to a localised arca of the external surface of said animal.

An eiphth aspect of the present invention provides a method of controlling or cradicating Phthiraptera, Siphonaptera and Acarina pests, said method including the administration of an effective amount of a systemically active composition according to 1s the first aspect of the present invention, or of a formulation according to the second or fourth aspects of the invention directly to said pests.

A ninth aspect of the present invention provides a method of controlling or : eradicating Phthiraptera, Siphonaptera and Acerina pests in domestic animals, said method including the oral administration of an effcctive amount of a systemically active composition according to the first aspect of the invention, or of a formulation according to the second or fifth aspects of the invention to said animals,

A tenth aspect of the present invention provides a mcthod of controlling or . cradicating Phthiraptera, Siphonaptera znd Acanna pests in domestic animals, said method including the parenteral administrztion of an effective amount of a systemically 2s active composition according to the {ust aspect of the invention, or of a formulation according to the second or sixth aspects of the invention to said animals,

An eleventh aspect of the present invention provides the use of a systemically active composition of the first zspect of the present invention in the manufacture of a medicament Jor controlling or eradicating Phithiraptera, Siphonaptera and Acarina pests in an domestic animals.

A twelfth aspect of the present invention provides the use of a eystemically active composition of the first zspect of the present invention in the manufacture of 2 bait formulation for controlling or eradicating Phthiraptera, Siphonaptera and Acarina pests.

Another aspect of the present invention provides an sctive composition of the first 5s aspect of the present invention or a formulation of any one of the sccond to sixth aspects

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FULIA"UI/VL LOY © 8 DEC. 2003 13:33 SPRUSON & FERGUSON Received 08 December 2003 4 10 of the present invenion when used for controlling or climunating Phthiraptera,

Siphonzptera and Acarina pests in domestic animals.

A further aspect of the present invention provides an active compositfon of the first : aspect of the present invention or a formulation of the fourth aspect of the present s invention when used for conuolling or eliminzung Phthiraptera, Siphonaptera and

Acarine pests.

This invention is predicated upon the surprising discovery of a synergistic interaction between spinosyns and macrocyclic lactones. Whilst not wishing to be bound by theary, it is noted that macrocyclic lactones have a primary cfTect on the inscel - ww nervous system by activating inhibiiory glutamate receptors, wkile spinosyns primarily sctivale the nicotinic acetylcholine receptors in inscct neurones causing hyperactivity of neurones. However, both spinosyns and macrocyclic lactones have a secondary cffect on gamma 2minobutyric acid (GABA) gated chloride channels in arthropod neurones,

GABA being an inhibitory nevro-transmitter. It is therefore possible that when combined i+ together the spinosyns and macrocyclic lactones have a synergistic effect on the GABA ~ receptor leading to effects in &n insect’s nervous system, this being unrelated 10 the primary cffect of cither spinosyns or macrocyclic Jactones. in particular, in the present invention it is surprisingly demonstrated that synergism occurs when a spinosyn and a macrocyclic lactone are delivered together to Phthiraplera, Siphonaptera or Acarina animal pests in such a way that those chemicals are ingested or arc otherwisc present systemically in the pests. Surprisingly, this effect is not observed in in vitro 2ssays which rcly on contact such as treated paper or transient immersion in treated solutions. While not wishing to be bound by theory, it is speculated that this difference is related to the mechenisms of action and the nature of both chemicals which 2< have low vapour pressure and do not readily cross the cuticle of arthropods.

The two active agents are administered 10 achieve an intracorporeal presence in the pests This cffect is not observed in assays which rely on mere contact.

Typically, the first aspect of the present invention provides en active composition for control or eradication of Phthiraptera, Siphonaptcra and Acarina pests, typically in yo domestic animals, said composition being systemically active in ssid pests and being 2 synergistic combination of a spinosyn and & macrocyclic lactone compound, wherein the spinosyn : macrocyclic lactone compounds are present in the range of 1000:1 to 1:1000 w/w. In particular, the ratio of ivermectin:spinosad could be 1:1000 in 2 composition systemically active in heanworm (Dirofilaria immilis) ot fleas. :

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It is however observed that the synergism also operates at™ lower ratios

Accordingly, typically, in the systemically active composition of the invention, the spinosyn compound : macrocyclic lactone compound are present in the range of 100:1 to 1:100 w/w,

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ST a WO 03/024223 PCT/AU01/01169 ] PY 11

Also typically, in the systemically active composition of the invention, the spinosyn compound : macrocyclic lactone compound are present in the range of 10:1 to 1:10 w/w, a Further typically, in the systemically active composition of the invention, the

A s spinosyn compound : macrocyclic lactone compound are present in the range of 9:1 to 1:9 w/w.

More typically, in the systemically active composition of the invention, the spinosyn compound : macrocyclic lactone compound are present in the range of 8:1 to 1:8 w/w. :

Also typically, in the systemically active composition of the invention, the spinosyn compound : macrocyclic lactone compound are present in the range of 7:1 to 1:7 w/w.

Also typically, in the systemically active composition of the invention, the : spinosyn compound : macrocyclic lactone compound are present in the range of 6:1 to 1:6 1s wiw.

More typically, in the systemically active composition of the invention, the spinosyn compound : macrocyclic lactone compound are present in the range of 5:1 to 1:5 w/w,

Even more typically, in the systemically active composition of the invention, the spinosyn compound : macrocyclic lactone compound are present in the range of 4:1 to 1:4 w/w.

Most typically, in the systemically active composition of the invention, the spinosyn compound : macrocyclic lactone compound are present in the range of 3:1 to 1:3 w/w.

Also most typically, in the systemically active composition of the invention, the spinosyn compound : macrocyclic lactone compound are present in the range of 2:1 to 1:2 w/w.

Also most typically, in the systemically active composition of the invention, the spinosyn compound : macrocyclic lactone compound are present in the proportion of 1:1 ¢ 30 w/w. . One embodiment of the first aspect of the present invention provides a systemically active composition being a synergistic combination of spinosad and an avermectin.

Typically, the macrocyclic lactone of the first aspect of the invention is selected 3s from the group consisting of ivermectin (22,23-dihydroavermectin B, described in EP

© g.DEC, 2003 13:33 SPRUSON & FERGUSON Received 08 Dece:sber 2003 { @® 12 295117), abumectin, avermectin Ay, avermectin An, gvermecun Ag; avermectin Aap, avennectin B,,, avermectin Byy, avermectin By, and aveimnectin Ba. Also typically, the ) macrocyclic lactone of the first aspect of the invention can be selected from the group of compounds related to the naturally occurring avermectins above but which have a group sal the 25-substituent other than the isopropyl or (S)-sec-butyl groups, as set out in

European patent applications 0214731, 0284176, 0308145, 0317148, 0335541 and 0340832. Also typically, the macrocyclic Jacione of the first aspect of the invention can include moxidectin (and derivatives disclosed in EP 259779A), doramectin and its analogues (described in EP0214731B), selamectin, cprinomectin, milbemyecin including <* milbemyein oxime, milbemycin D (Antiobiotic B41D) and its analogues (described in

US3,050,360) and nemadectins (described in EP 170006A).

More typically, thc macrocyclic lactone of the fust aspect of the invention is selected from the gioup consisting of iveimecnn, moxidectin, doramectin, selamectin, milbemyein oxime, Milbemycin D, eprinomcctin and abamectin. 1s Even more typically, the macrocyclic Jactone of the first aspect of the invention ic ivermectin.

More 1ypically, the active composition is therefore a synergistic combination of spincsad and ivermectin which is systemically active in Phthirapiera, Siphonaptera and

Acaring pests. 2 Typically in the formulations of the present invention, the carrier can be non- aqueous Or aqueous and the active composition is suspended, dissolved or dispersed in the carrier. Also typically, the active composition can be admixed with a pharmaceutically or veterinarily accepiable diluent or carrier which is sclected with regard to the intended route of adminstration and in accordance with stendard practice In one embodiment, the is carriers of excipients used in the formulations of the third and fourth aspects of the picsent invention include dust carriers, solvents, emulsifiers, wetting and dispersing agents and water. In particular, in the bait formulations of the fourth aspect of the present invention, the carTiers or excipients include 2 food source, attractant or both such as blood or other tissue cxiracts and typically further include one or more of yeast, sugar, x pheromones, flavours, scents, solvents, wetting and dispersing agents.

In another embodiment, the carriers or excipients used in the orally delivered formulations of the fifth aspect of the present invention include tabletting excipients such as starch or lactose, capsule excipients or carriers and excipients commonly used in solutions or suspensions including water, flavouring and colouring agents. Sclection of 3s the carrier is of course meade on the basis of compatibility with the active composition.

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5. DEC. 2003 13:34 SPRUSON & FEREUSON : Received 08 December 2003 /® including such considerations as pH, moisture content and stability. “Selection of the carrier is slso made depending on the mode of application of the formulation-such es’ whether it is to be applied topically to 2 ‘domestic animal, or crally or parenterally administered to a domestic anima, or is instcad to be administered directly, such as via a s bait to the Phibiraptera, Siphonaptera or Acanna pests .

One embodiment of any one of the sccond to sixth aspects of the invention provides a formulation for controlling or eradicating Phthirapters, Siphonaptera and Acarins pests, said formulation including: . (2) from 0.1 10 40% by weight of an zctive composition of the first aspect of the N w present invention, and . (b) from 60-99.9% by weight of a suitable carrier.

Typically each dose of a fonnulztion of the invention would contain 30pg-2g of cach of thc spinosyn compound and macrocyclic lactone compound, more typically a formulation would contain 1mg-1g of cach of the spinosyn compound and macrocyclic 1s lactone compound.

Formulations of the third aspect of the present invention are typically liquids and can be made up as concentrates and then diluted prior lo use.

Formulations of the fourth aspect of the present invention being bait formulations arc typically solids including powders, granular or dessicated forms. It is also typical that 50 such bait formulations can be in the form of liquids or pastes. 1t has long been common practice to control extemal parasites on shecp, cattle and other domestic animals including but not limited 10 goats, pigs and horses by the localised topical application of a formulation containing an active insecticide/parasiticidc and 2 canier/vebicle. Typically therefore, a formulation of the third aspect of the invention is a 2s pour-on formulation including an effective zmount of an active composition of the first aspect of the invention and a topically acceplable camer.

It is also typical that a topically applied formulation can be a spray or dip or a solution such as a jetting fluid.

A pour-on formulation of the third aspect of the present invention is 1ypically liqmd w and is usually applied to the exterior of a domestic animal as a linc or 2 spot, which when ingested by such Phthiraptera, Siphonapterz and Acarina pests, for example afier feeding on a treated domestic animal cpidermis, acts systemically in those pests thereby protecting the animal apainst both immature and tdult forms of Phthiraplera,

Siphoneptera and Acarina pests such as ticks, flees and lice and can also act to decrease ys the number of viable Phthirapterz, Siphonaptera and Acarina cggs.

AMENDED SHEET

IPEA/AU

Co 2 WO 03/024223 ) PCT/AU01/0116Y ® 14

While the formulation is applied topically, to a localised area, the active agent migrates over the surface of the animal to cover its whole external surface area.

The carrier (also referred to herein as ‘vehicle’) present in such pour-on formulations of the third aspect of the present invention is formulated to achieve good a 5s spread around the skin and/or penetration of the epidermis of the animal. To date, B commercial pour-on formulations are suspensions, emulsifiable concentrates or solutions and are often comprised of at least one organic solvent. Solvents commonly used as carriers in such pour-on formulations include propylene glycol, paraffins, isoparaffins, aromatics, isopropylmyristate (IPM), glycol ethers, alcohols and n-propyl alcohol.

It is important to note that the topical formulations of the present invention can be used to control the arthropods of the orders Phthiraptera, Siphonaptera and Acarina in several ways. In respect of topical formulations of the present invention which just purely remain on the external surface of a domestic animal, these will be effective against pests such as biting lice and other pests which feed off the epidermis of the animal and will 1s thereby ingest the systemically active formulations. In respect of topical formulations of the present invention which after being topically applied at least partially penetrate : through the epidermis of a domestic animal, penetrating into the extracellular fluid and then draining via the lymphatic system of the animal into its blood stream, these formulations will be effective against blood sucking pests such as fleas, ticks and some lice. These pests will be specifically killed if they ingest blood containing the systemically active composition of the present invention. Accordingly, topical formulations of the present invention can also become (via penetration through the .epidermis of the host animal) systemically available in the host, as well as externally present on the host.

Another embodiment of the third aspect of the invention provides a pour-on formulation for control of Phthiraptera, Siphonaptera and Acarina pests in domestic animals, said formulation including: (a) from 0.1 to 40% by weight of at least one active agent of the first aspect of the present invention, and (b) from 60-99.9% by weight of a suitable carrier selected from the group consisting of TPM/alcohol, OP/IPM/OSU and GTCC/PMP/CAP where

TPM is Tripropylene glycol methyl ether;

P is octyl palmitate or 2-ethylhexyl palmitate which is an excellent lubricant, and can also be used as an emollient and a solvent; 3s P can be replaced by OS (octyl stearate or 2-ethylhexyl stearate);

. 5DEC.2003 13:34 SPRUSON & FERGUSON oo Received 08 December 2003 ¢ @® 1s

PM is isopropyl myristate which has excellent spreading and emollient properties - this can be used interchangeably with IPP or IPL; -

PP is isopropyl palmitate; -

PL is isopropyl} Jaurate, s MP is PPG 2 myristy| ether propionate which spreads repidly and promotes wetting of other matenals;

SU is di-2-ethylhexyl succinate and promotes wetting and spreading of lipophilic substances onto the skin;

CS is isocetyl siearate which can be used as an emollient, lubricant and spreading - 0 agent;

TCC is glyceryl ti czprylatc/caprate which is an excellent carrier or vehicle for : actives;

AP is a selected biend of branched chain esters which again acts as an emollient and spreading agent; 0s alcohol could be benzyl alcohol, propyl alcobol, diacctone alcobol or other suitable alcohol. other possible carriers which can be used in the formulations of the present invention include organic and inorganic alcohols, including propylene glycol which is a common ivermectin carrier, ethanol, propyl alcohol, benzyl alcohol, glycols and also 6 detergents.

Typically, the formulations of the third aspect of the present invention, can be in the form of a powder, cream, suspension, spray, emulsion, foam, pastc, aerosol, ointment, salve or ge] More typically, the formulation is 2 solution, and typically water soluble.

Typically, formulations of the second, third, Sfth or sixth aspects of the present ;« invention can be effectively epplicd to domestic animals such as sheep, cattle, goats, camelids, pigs, dogs, cats, poultry nd horses, other ruminants and monogastrics.

Typically, the bait formulations of the fourth aspect of the present invention will be administered by means other that on or onto domestic animals to afford a direct contact that will result in intracorporeal presence of the active agents in the three identified pest so orders, as is known by those skilled in the art. -

Typically a pour-on formulation of the third aspect of the present invention is applied by pouring in one or several lines or in a spot on the dorsal midline (back) or shoulder of 8 domestic animal. More typically, the pour-on formulation is applied by pouring along the back of thc enimal, following the spine. A pour-on fonnulation can 3» also be applied to the animal by other conventional methods including wiping an

AMENDED SHEET

CO PRARD mmm ment

. FU IAVULIVE LOY © g. DEC, 2003 13:35 SPRUSON & FERGUSON Received 08 December 2003 / 16 impregnated material over st least a small wea of the animal, by using commercially 2vailable epplicators, by means of a syringe. by spraying or by using a spray race.

Typically, approximately about 0.1:2000mg active composition/kg of anima) bodyweight is an effective emount for topical application to domestic znimals. Typically, s about 2-100mg of active composition of the first zspect of the present invention will be applied 10 8 cow or sheep (per kg bodyweight).

Typically, a formulstion of the present invention, such as a pour-on formulation, is formulated such that the aciive composition 1s present in & concentration of sbout 0.1- 40% weight / volume, more typically 0.1-20% weight / volume, preferably about 0.5 to « 5% depending on the poiency of the acuve.

Typically, an active composition or a fonnulation of the present invention is formulated for use such that each of the A83543 compound and the macrocyclic lactone arc present in a conccniration range of about 1-500ppm. More typically, cach of the

AB3543 compound and the macrocyclic lactone are present in a concentration range of 1s about 1-300ppm. Even more typically, cach of the A83543 compound and the macrocyclic lactone arc present in a concentration range of about 1-100ppm. The I- 500ppm concentration is most typical in respect of rcady to use formulations such as diluted dips and sprays. Pour-on formulations of the present invention will typically have 2 concentration of the actives in the range of from 1-100g/L, more typically S to 50 g/L, ws even more typically 10-25g/L. ln respect of oral formulations, such as lablets and paremieral formulations of the present invention, typically the administered dosc wil] be from 0.01 to 50mg/kg animal body weight, more typically 0.1 10 20 mg/kg. For bait formulations of the present invention, it is typical that the concentration is from 0.05 to 1000mg/kg, morc typically 1 to 100mg/kg. : 28 Typically only a small volume of a pour-on formulation is required in order to be effective against the Phthirapiera, Siphonapiera and Acarina pests, such as in the order of 0.5-80ml per application, with 10-60 ml pes applicztion being preferred for larger animals such as cattle and 1-20! per application for smaller animals such as sheep, dogs and cats.

W Typically a formulation of the fifth aspect of the present invention is in the form of 2 1ablet, capsule, bolus, solution, suspension or other elixiz. The formulations can also be sustained release formulations. Such formulations are prepared in a conventional manner in accordance with standard pharmaceutical and veterinary practice. Typically, capsules, boluses or tablet may be prepared by admixture of the active combination with a suitable

AMENDED SHEET ere AREAJAL oo oe -

oF PCT/AUN1/01169 8. DEC. 2003 13:35 SPRUSON & FERGUSON Received 08 December 2003 @ 16a fincly divided diluent or carrier, additionally containing 8 disintegrant gnd/or 2 binder <uch as talc, starch or lactose. }

AMENDED SHEET

IPEA/AU

Co “WO 03/024223 PCT/AU01/01169

Also typically, a formulation of the sixth aspect of the present invention is in the: form of a sterile aqueous solution or suspension of the active combination, with a parenterally acceptable carrier being water and other excipents such as salt or glucose a being optionally present. i 5 In the formulations of the present invention having pesticidal activity against

Phthiraptera, Siphonaptera and Acarina pests, the active agent is a combination of at least one compound selected from the class of spinosyn compounds (including spinosad) and at least one active agent selected from the macrocyclic lactones including ivermectin, abamectin, moxidectin, doramectin, eprinomectin and milbemycin,

The formulations of the present invention suitably can variously include one or more additional ingredients such as preservatives, spreading agents, adhesion promoters, active solubilisers such as oleic acid or lactic acid, viscosity modifiers, UV blockers or absorbers, colourants and stabilisers such as antioxidants. Suitably, surface active agents including anionic, cationic, non-ionic and ampholytic surface active agents can also be 1s included in the pour-on formulations of the present invention.

Isopropyl myristate (IPM), isopropyl! palmitate (IPP), caprylic/capric acid esters of saturated C,,-C,g fatty alcohols, oleic acid, oleyl ester, ethyl oleate, triglycendes, silicone oils and dipropylene glycol mono methyl ether (DPM) are common spreading agents used in pour-on formulations.

Typically, the methods of the seventh to tenth aspects of the present invention prevent ticks, lice, fleas, and other Phthiraptera, Siphonaptera and Acarina pest infestations of domestic animals, including but not limited to cattle, sheep, goats, pigs, horses, camelids, dogs, cats and poultry and other ruminants and monogastrics.

Typically, the active compositions, formulations and methods of the present 2s invention are effective against immature (including nymph) and adult forms of

Phthiraptera, Siphonaptera and Acarina pests. The pests do not have to be, although typically are present on a host domestic animal.

More typically, the active compositions, formulations and methods of the present invention are effective against immature and adult forms of Phthiraptera, Siphonaptera d 30 and Acanna pests in domestic animals. Also typically, the active compositions, i formulations and methods of the present invention are also effective in decreasing the number of viable Phthiraptera, Siphonaptera and Acarina eggs which may be present in domestic animals.

Mare typically, a topically, orally or parenterally administered formulation of the 3s present invention acts to control sheep body lice (Bovicola Ovis) in sheep, acts to control

® 18 similar lice in cattle, goats and camelids, acts to control ticks (eg Boophilus bovis), on cattle and acts to control Siphonaptera (Ctenocephalides felis and other fleas) in dogs, cats and other domestic animals. Also more typically, a topically, orally or parenterally administered formulation of the present invention acts to control lice and ticks in cattle oT s and fleas in both dogs and cats.

The formulations of the present invention are prepared according to known techniques. Where the formulation is a solution the parasiticide/insecticide is mixed with the carrier or vehicle, using heat and stirring where required. Auxiliary or additional ingredients can be added to the mixture of active and carrier or can be mixed with the active prior to the addition of the camer.

The formulations of the present invention can contain as little as 1ppm of each macrocyclic lactone compound and spinosyn compound per application and a synergistic effect is still observed.

The active compositions and formulations of the invention are non toxic to humans and animals as well as crops and plants, and residues in the wool, hides and tissue of animals treated or administered with the formulations are at environmentally acceptable levels. Further no skin irritation or other toxicity to end users results from the method and formulations of this invention. Environmental contamination is also minimised.

Also advantageously, as such spinosyn factors and macrocyclic lactones are very : efficacious at low levels due to their synergistic effect when combined together, the present invention is of utility against Phthiraptera, Siphonaptera and Acarina pest populations in domestic animals that have existing levels of resistance to both spinosyn compounds and macrocyclic lactones when these compounds are used separately.

Generally the administration of the formulations of the third aspect of the present invention, and active compositions of the present invention is by way of externally/topically to the domestic animals. Such topical application can take the form of dipping, showering, jetting, spraying, manually applying such as dusting, or otherwise placing or laying the formulation containing the active substance/s. Accordingly, typically the active compositions of the invention are formulated into a number of ) topically applied pesticidal formulations. :

Preferably such topical pesticidal formulations include spot-ons, pour ons, sprays, dips, dusts, lotions, gels, ointments, salves, dressings, towels, cremes, sticks, soaps, shampoos, collars, medallions, eartags and tail bands. Pour-on formulations including both aqueous and organic solvent based ones as well as emulsions and

. , PUL/AURLI/OT16Y . 6. DEC. 2003 13:35 SPRUSON & FERGUSON * Received 08 December 2003 / . suspensions are prefered. As stated above, the formulations can be im concentrated fonm which are diluted just prior (o application. -

More preferred are dip formulations, pour on formulations, jetting fluid formulations and jetting/spray race formulations. s Wenable powders are another formulation of the invention which are prepared by blending the active with 8 dust carrier which wets and suspends in water. A surface active agent cen be added. Sprays of wcttable powders can be applied to the environs of domestic animals including poultry houses, stables, dairy sheds and pig pens because of Cs their relative safety. : 16 Emulsions arc znother formulation of the invention which are solutions of the active in water-immiscible orgamc solvents, ccmmonly at 1-40%, with an optional surface active agent to promote emulsification, wetting and spreading. The choice of solvent is based on safety to plants, humans and animals, volatility, flammability and cost. Water cinulsion sprays from such cmulsion concentrates can be used in household Phthiraptera,

Siphonaptera and Acanna pest control. )

As stated above, the administrarion of the formulations of the fifth aspect of the . present invention is by way of oral administration while the administration of the formulations of the sixth aspect of the present invention is parenterally, for examplic intramuscularly, subcutaneously or intravenously.

The formulations of the fifth and sixth aspects of the present invention will vary with regard wo the weight of the active combinztion depending of the species of host domestic amma] to be treated as well as the body weight. The formulations may be administered as a dose of from 0.001mg 10 about SOmg per kg of animal body weight, more typically from 0.01 10 zbout 30mg per kg of animal body weight, and even more »s typically from 0.1 to about 20mg per kg of 2nimal body weight. - This can be given as a single dose or as several doses. Higher as well as lower dosage ranges are also conternplated within the present invention : : Ag the administration of the bait formulations of the fourth aspect of the present invention is by way of administration by meens other than on or onto domestic animals to w ufford a direct contact (such as by ingestion of the bait by the pest) that will result in iniracorporeal presence of the active agents in the three identified pest orders, as is known by those skilled in the art.

The spinosyn component of the active composition of the first aspect of the present invention may be present as a single compound, a mixture of two or more compounds, a 5s mixture including at least onc of A83543A and A83543D, or a mixture of at least one

AMENDED SHEET

IPFAIBYS

© , PCT/A1101/01169 6. DEC. 2003 13:36 SPRUSON & FEREUSON Received 08 Decerriver 2003 : 6 19a

A83543 compound together with the dried portion of the fermentation medium in which it is produced. .

AMENDED SHEET

IPEA/AU

© WO 03/024223 PCT/AUNI/01169

PY 20

The macrocyclic lactone compounds used in the present invention include such well known anthelmintic compounds as avermectins and milbemycins and derivatives and analogues thereof. As stated above, the avermectins are isolated from fermentation products of Streptomyces avermitilis and the isolation and chemical structure of the eight ’ s individual acomponents which make up the avermectins (otherwise known as C-076 - compounds) is described in detail in British patent specification 1573995. Ivermectin is a compound which is a semisynthetic chemical formed by modification of avermectin. : Commercially available ivermectin can include for example, the 25-isopropyl analogue of ivermectin. Avermectins being lipophilic can be prepared for the purposes of the formulations and methods of the present invention by dissolving an avermectin in an organic solvent such as chloroform, methylene chloride, acetone and alcohols.

Milbemycins as discussed above in detail, are other compounds which are not avermectins but which can be considered to come within the class of compounds which are macrocyclic lactones. The milbemycins differ structurally from the avermectin group, ts mainly in the absence of a disaccharide group on C-13. Examples of such compounds are described in UK patent 1390336 and EP patent applications 170006, 254583, 334484 and 410615 which are incorporated herein by cross reference.

The spinosyn compound may also be present as a salt in the active agent, formulations and methods of this invention. The salts would be prepared using standard procedures for salt preparation. For example, spinosyn A can be neutralised with an appropnate acid to form an acid additional salt. The acid addition salts of spinosyns which can be used in the present invention are useful and include salts formed by reaction with either an organic or inorganic acid such as, for example, sulfuric, hydrochlonc, phosphoric, acetic, succinic, citric, lactic, maleic, fumaric, cholic, pamoic, mucic, 2s glutamic, camphoric, glutaric, glycolic, phthalic, tartaric, formic, lauric, stearic, salicyclic, methanesulfonic, benzenesulfonic, sorbic, picric, benzoic, cinnamic and other like acids.

Generally, emulsifiable concentrates of the A83543 compounds comprise a convenient concentration of an A83543 compound dissolved in ani inert carrier which is either a water-miscible solvent or a mixture of a water immiscible organic solvent and emulsifiers. A preferred concentration range is 1-500g/L of said spinosyn compound, more preferably the concentration range is selected from the group consisting of 1- 400g/L, 1-350g/L, 1-300g/L, 1-250g/L, 1-200g/L, 1-150g/L, 1-100g/L, 1-90g/L, 1-80g/L, 1-70g/L, 1-60g/L, 1-50g/L, 1-40g/L, 1-30g/L, 1-20g/L, even more preferably 25g/L.

Useful organic solvents include aromatics including xylenes and petroleum fractions. 3s Other organic solvents may also be used, such as the terpenic solvents, including rosin

® derivatives, aliphatic ketones such as cyclohexanone and complex alcohols such as 2- ethoxyethanol.

Suitable emulsifiers for emulsifiable concentrates can be chosen from = conventional nonionic surfactants, including ethylene oxide adducts of alkylphenols and . s anionic surfactants, including sulphonate alkyl/aryl salts.

Aqueous suspensions (AS) comprise suspensions of an active water-insoluble spinosyn compound dispersed in an aqueous vehicle at a concentration in the range of from about 1-500g/L, preferably the concentration range is selected from the group consisting of about 1-400g/L, about 1-300g/L, about 1-250g/L, about 1-200g/L about 1- 150g/L, about 1-100g/L, about 1-50g/L, about 1-45g/L, about 1-40g/L, about 1-30g/L, more preferably about 25g/L. Generally the suspensions are prepared by finely grinding the spinosyn compound and mixing it into a vehicle comprised of water and surfactants chosen from such types as nonionic, sulfonated lignins and alkylsulfates. Inert ingredients may also be added. 1s The aqueous suspensions and emulsions are preferably diluted with water to obtain the desired spinosyn concentration in the final formulations of the invention.

In another preferred formulation, the one or more active substances take/s the form of a solution of the active/s in water. Sprays are the most common means of pesticide application on surfaces of structures such as stables, dairy sheds and pig pens. Sprays or dips are the most common means of pesticide application on small ruminant animal species with water generally as the principal carrier.

In the formulations of the present invention, particularly dip wash formulations it is preferred that the spinosyn compound and the macrocyclic lactone compound are each present in a concentration of about 500ppm or less. More typically, each are present in a 2s concentration of about 400ppm or less. Also typically, each are present in a concentration of about 300ppm or less, more typically 200ppm or less, even more typically 100ppm or less, most typically SOppm or less.

Best Mode and Other Modes of Performing the Invention

Preparation of the preferred formulations of the present invention can be made d 30 by conventional processes, several examples of which are found below. The preferred . process for preparing a spinosad and ivermectin combination of the present invention is to either co-formulate the combination or formulate each of the compounds separately then combine them together. The compounds could even exist in the combination as separate phases. For example, one active compound of the synergistic composition could be in

Claims (44)

4 PCT/AU01/01169 CLAIMS

1. A systemically active substance or composition for use in a method for controlling or eradicating Phthiraptera, Siphonaptera and Acarina pests, said substance or composition comprising a synergistic combination of at least one A83543 compound and at least one macrocyclic lactone, and said method comprising administering said substance or composition.

2. A systemically active substance or composition for use in a method of treatment as claimed in claim 1 wherein the ratio of A83543 compound: macrocyclic lactone is in the range of 1000:1 to 1:1000 w/w.

3. A systemically active substance or composition for use in a method of treatment as claimed in claim 1 wherein the ratio of A83543 compound: macrocyclic lactone is in the range of 10:1 to 1:10 w/w.

4. A systemically active substance or composition for use in a method of treatment as claimed in claim 1 wherein said A83543 compound is selected from the group consisting of any spinosyn compound and salts thereof, and a mixture of any two or more spinosyn compounds including spinosad.

5. A systemically active substance or composition for use in a method of treatment as claimed in claim 1 wherein said macrocyclic lactone is selected from the group consisting of ivermectin, abamectin, avermectin A,, avermectin A, avermectin A,,, : avermectin A,,, avermectin B,,, avermectin Bj, avermectin B,,, avermectin By, moxidectin, doramectin, selamectin, eprinomectin and milbemycin.

6. A systemically active substance or composition for use in a method of treatment as claimed in claim 1 wherein said A83543 compound is spinosad and said macrocyclic lactone is ivermectin.

7. A systemically active substance or composition for use in a method of treatment as claimed in claim 1 wherein said pests are present in domestic animals.

8. A formulation for use in a method of controlling or eradicating Phthiraptera, Siphonaptera and Acarina pests in domestic animals, said formulation including a systemically active substance or composition as claimed in claim 1 and a domestic animal physiologically acceptable carrier, diluent or excipient, and said method comprising administering an effective amount of said formulation. AMENDED SHEET y PCT/AU01/01169

9. The formulation as claimed in claim 8 wherein the active substance or composition is present in the formulation in a concentration of about 0.1-40% weight/volume.

10. The formulation as claimed in claim 8 wherein the formulation is orally administered to one or more domestic animals.

11. The formulation as claimed in claim 8 wherein the formulation is parentally administered to one or more domestic animals.

12. The formulation as claimed in claim 8 wherein said formulation is externally applied to one or more domestic animals.

13. The formulation as claimed in claim 8 wherein said formulation is a topical formulation for applying to domestic animals, said topical formulation being selected from the group consisting of spot-ons, pour ons, sprays, dips, dusts, lotions, gels, ointments, salves, dressings, towels, cremes, sticks, soaps, shampoos, collars, medallions, eartags, dip formulations, jetting fluid formulations, jetting/spray race formulations and tail bands.

14. The formulation as claimed in claim 8 wherein said formulation is a pour on formulation applied to a localised area of the external surface of a domestic animal.

15. The formulation as claimed in claim 8 wherein about 0.1-2000mg active agent/kg animal bodyweight is effective for topical application to domestic animals.

16. A bait formulation for use in a method for controlling or eradicating Phthiraptera, Siphonaptera and Acarina pests, said formulation including a systemically active substance or composition as claimed in claim 1 and an acceptable carrier, diluent or excipient, and said method comprising administering an effective amount of said bait formulation.

17. A method of preventing Phthiraptera, Siphonaptera and Acarina pests in domestic animals, said method including the external application of an effective amount of an active substance or composition as claimed in claim 1 or of a formulation as claimed in claim 8 to at least a localised area of the external surface of said animal.

18. A method of preventing Phthiraptera, Siphonaptera and Acarina pests, said method including the administration of an effective amount of an active substance or composition as claimed in claim 1 or of a formulation as claimed in claim 8 or claim 16 to said pests. AMENDED SHEET

» PCT/AU01/01169

19. A method of preventing Phthiraptera, Siphonaptera and Acarina pests in domestic animals, said method including the oral administration of an effective amount of an active substance or composition as claimed in claim 1 or of a formulation as claimed in claim 8 to said animals.

20. A method of preventing Phthiraptera, Siphonaptera and Acarina pests in domestic animals, said method including the parenteral administration of an effective amount of an active substance or composition as claimed in claim 1 or of a formulation as claimed in claim 8 to said animals.

21. Use of an active substance or composition as claimed in claim 1 for the manufacture of a medicament for preventing or controlling Phthiraptera, Siphonaptera and Acarina pests in domestic animals.

22. Use of an active substance or composition as claimed in claim 1 for the manufacture of a bait formulation for preventing or controlling Phthiraptera, Siphonaptera and Acarina pests.

23. An active substance or composition as claimed in claim 1 or a formulation as claimed in claim 8 when used for preventing or controlling Phthiraptera, Siphonaptera and Acarina pests in domestic animals.

24. A formulation for controlling or eradicating Phthiraptera, Siphonaptera and Acarina pests in domestic animals, said formulation including a systemically active substance or composition according to claim 1 and a domestically animal physiologically acceptable carrier, diluent or excipient for external application as a bait, and said method comprising oral or parenteral administration of an effective amount of said substance or composition to a domestic animal.

25. Use of a synergistic combination of at least one A83543 compound and at least one macrocyclic lactone in the manufacture of a systemically active medicament for controlling or eradicating Phthiraptera, Siphonaptera and Acarina pests.

26. Use as claimed in claim 25 wherein the ratio of A83543 compound: macrocyclic lactone is in the range of 1000:1 to 1:1000 w/w.

27. Use as claimed in claim 25 wherein the ratio of A83543 compound: macrocyclic lactone is in the range 10:1 to 1:10 w/w. AMENDED SHEET v PCT/AU01/01169

28. Use as claimed in claim 25 wherein said A83543 compound is selected from the group consisting of any spinosyn compound and salts thereof, and a mixture of any two or more spinosyn compounds including spinosad.

29. Use as claimed in claim 25 wherein said macrocyclic lactone is selected from the group consisting of ivermectin, abamectin, avermectin A, avermectin Ay, avermectin A,,, avermectin A,,, avermectin By,, avermectin B,,, avermectin B,, avermectin B,,, moxidectin, doramectin, selamectin, eprinomectin and milbemycin.

30. Use as claimed in claim 25 wherein said A83543 compound is spinosad and said macrocyclic lactone is ivermectin.

31. Use as claimed in claim 25 wherein said pests are present in domestic animals.

32. Use of a systemically active substance or composition as claimed in claim 1 in the manufacture of a systemically active medicament for controlling or eradicating Phthiraptera, Siphonaptera and Acarina pests in domestic animals.

33. Use as claimed in claim 32 wherein the active substance or composition is present in the formulation in a concentration of about 0.1-40% weight/volume.

34. Use as claimed in claim 32 wherein the medicament is effective when orally administered to one or more domestic animals.

35. Use as claimed in claim 32 wherein the medicament is effective when parentally administered to one or more domestic animals.

36. Use as claimed in claim 32 wherein said medicament is effective when externally applied to one or more domestic animals.

37. Use as claimed in claim 32 wherein said medicament is a topical formulation for applying to domestic animals, said topical formulation being selected from the group consisting of spot-ons, pour ons, sprays, dips, dusts, lotions, gels, ointments, salves, dressings, towels, cremes, sticks, soaps, shampoos, collars, medallions, eartags, dip formulations, jetting fluid formulations, jetting/spray race formulations and tail bands.

38. Use as claimed in claim 32 wherein said medicament is a pour on formulation that is effective when applied to a localised area of the external surface of a domestic animal.

39. Use as claimed in claim 32 wherein about 0.1-2000mg active agent/kg animal bodyweight is effective for topical application to domestic animals. AMENDED SHEET

PN PCT/AU01/01169

40. A systemically active substance or composition for use in a method of treatment according to any one of claims 1 to 7 or 23, substantially as herein described and illustrated.

41. A formulation for use in a method of treatment according to any one of claims 8 to 16 or 24, substantially as herein described and illustrated.

42. A method according to any one of claims 17 to 20, substantially as herein described and illustrated.

43. Use according to any one of claims 21, 22 or 25 to 39, substantially as herein described and illustrated.

44. A systemically active substance or composition for a new use in a method of treatment, a formulation for a new use in a method of treatment, a new non-therapeutic method of treatment, a new use of at least one A83543 compound and at least one : macrocyclic lactone as defined in claim 25 or of a substance or composition as claimed in claim 1, substantially as herein described. AMENDED SHEET