60/179, 005, filed January 28, 2000.

Background of the Invention Nematodes are important plant pests which cause millions of dollars of damage each year to turf grasses, ornamental plants, and food crops. Efforts to eliminate or minimize damage caused by nematodes in agricultural settings have typically involved the use of soil fumigation with materials such as chloropicrin, methyl bromide, and dazomet, which volatilize to spread the active ingredient throughout the soil. Such fumigation materials can be highly toxic and may create an environmental hazard. Various non- fumigant chemicals have also been used, but these too create serious environmental problems and can be highly toxic to humans.

The accepted methodology for control of nematodes afflicting animals has centered around the use of the drug benzimidazole and its congeners. The use of these drugs on a wide scale has led to many instances of resistance among nematode populations (Prichard, R. K. et al. [1980]"The problem of anthelmintic resistance in nematodes,"Allstr. L'et. J. 56 : 239-251 ; Coles, G. C. [1986]"Anthelmintic resistance in sheep,"In Veterinary Clinics of North America : Food Animal Practice, Vol 2 : 423-432 [Herd, R. P., Eds.] W. B. Saunders, New York).

The pesticidal activity of avermectins is well known. The avermectins are disaccharide derivatives of pentacyclic, 16-membered lactones. They can be divided into four major compounds : Ala, A2a, Bla, and B2a ; and four minor compounds : A, b, A, b, B, b, and B2b.

The organism which produces avermectins was isolated and identified as Streptomyces avermitilis MA-4680 (NRRL-8165). Characteristics of the avermectin producing culture and the fermentation process are well documented and known to those

skilled in the art (Burg, R. W. et al. [1979]"Avermectins, New Family of Potent Anthelmintic Agents : Producing Organism and Fermentation,"Antimicrob. Agents Chemother. 15 (3) : 361-367). The isolation and purification of these compounds is also described in U. S. Patent No. 4, 310, 519, issued January 12, 1982.

Another family of pesticides produced by fermentation are the milbemycins, which are closely related to the avermectins. The milbemycins can be produced by a variety of Streptomyces and originally differed from the avermectins only in the C-13 position. The milbemycins and their many derivatives are also well known to those skilled in the art and are the subject of U. S. patents. See, for example, U. S. Patent No. 4, 547, 520.

While the avermectins were initially investigated for their anthelmintic activities, they were later found to have other insecticidal properties, although the degree varies.

The activity of avermectins must generally be determined empirically.

22, 23-dihydroavermectin B, is a synthetic derivative of the avermectins and has been assigned the nonproprietary name of ivermectin. It is a mixture of 80% 22, 23- dihydroavermectin Bla and 20% 22, 23-dihydroavermectin B lb Ivermectin has been tested on a variety of laboratory and domestic animals for control of nematodes, ticks, and heartworms.

Avermectin B is active against the root-knot nematode, Meloidogytie incognita.

It is reported to be 10-30 times as potent as commercial contact nematicides when incorporated into soil at 0. 16-0. 25 kg/ha (Boyce Thompson Institute for Plant Research 58th Annual Report [1981] ; Putter, 1. et al. [1981]"Avermectins : Novel Insecticides, Acaracides, and Nematicides from a Soil Microorganism,"Experientia 37 : 963-964).

Avermectin B2a is not toxic to tomatoes or cucumbers at rates of up to 10 kg/ha.

Avermectin B, is a combination of avermectin B, a (major component) and avermectin Blb.

It has demonstrated a broad spectrum of insecticidal activities. The data indicate that avermectin B, is primarily a miticide, although it is also effective on the Colorado potato beetle, potato tuberworm, beet armyworm, diamondback moth, gypsy moth, and the European corn borer.

The use of avermectins in various agricultural applications has been described in publications and patents. The use of avermectin with spray oils (lightweight oil compositions) has been described. See, for example, U. S. Patent No. 4, 560, 677 issued

December 24, 1985 ; EPO applications 0 094 779 and 0 125 155 ; and Anderson, T. E., J. R.

Babu, R. A. Dybas, H. Mehta (1986) J. Econ. Entomol. 79 : 197-201.

There is a continuing need for new, alternative materials and methods useful for killing nematodes.

Brief Summary of the Invention The subject invention concerns substituted compositions and processes for controlling nematodes. In one embodiment, the subject invention comprises the use of certain substituted organic compounds to control nematodes which infest and afflict animals. Nematodes which infest plants or the situs of plants can also be controlled using the methods and compositions of the subject invention, as can other acarid and arthropod pests.

Preferred compounds useful according to the subject invention can be represented by the Formulae I, II, III, IV, and V as further described herein.

1. A urea derivative of the following Formula I : Ar-(Alk) 01-NH-CO-NR'-Alk-R2 (Formula I) wherein Ar is aryl or heteroaryl optionally substituted by one or more R3 groups ; each Alk is a linear or cyclic alkylene radical of up to 8 C atoms ; R'is H or C, 6 alkyl ; W is heteroaryl or heterocycloalkyl optionally substituted by Ar, or forms such a group by cyclisation with R' ; and R3 is OH, halogen, CF3, OCF, or a group selected from NH2, SO2-CI 6 alkyl, C610 aryl, C610 aryloaxy, C56 cycloalkyl, Cl s alkoxy, and C16 alkyl, said group being optionally substituted by OH, Cl 6 alkoxy, C16 alkyl, phenyl, halogen, or CF3.

Particularly preferred anthelmintic compounds according to Formula I are exemplified herein by compounds represented by structures 1-10 (depicted in Figures 1- 10, respectively), which have been assigned the respective reference numbers : AKC 111 (STRUCTURE 1), AKC 112 (STRUCTURE 2), AKC 113 (STRUCTURE 3), AKC 107 (STRUCTURE 4),

AKC 114 (STRUCTURE 5), AKC 108 (STRUCTURE 6), AKC 115 (STRUCTURE 7), AKC 116 (STRUCTURE 8), AKC 117 (STRUCTURE 9), and AKC 118 (STRUCTURE 10).

2. A heterocycle-substituted amide of the following Formula II : Ar-(Alk) 0 l-NH-CO-Het (Formula II) wherein Ar is aryl or heteroaryl optionally substituted by one or more R3 groups ; each Alk is an optionally cyclic alkylene radical of up to 8 C atoms ; Het is heteroaryl or heterocycloalkyl optionally substituted by Ar and/or R3 ; and R3 is OH, halogen, CF3, OCF3, or a group selected from NH2, SO alkyl, C6 aryl, Cl4 alkoxy, and C, alkyl said group being optionally substituted by OH, C, 6 alkoxy, C16 alkyl, phenyl, halogen, or CF3.

Particularly preferred anthelmintic compounds according to Formula 11 are exemplified herein by compounds represented by structures 11-25 (depicted in Figures 11- 25 respectively), which have been assigned the respective reference numbers : AKC 119 (STRUCTURE 11), AKC 110 (STRUCTURE 12), AKC 120 (STRUCTURE 13), AKC 121 (STRUCTURE 14), AKC 2153 (STRUCTURE 15), AKC 122 (STRUCTURE 16), AKC 104 (STRUCTURE 17), AKC 123 (STRUCTURE 18), AKC 124 (STRUCTURE 19), AKC 125 (STRUCTURE 20), AKC 105 (STRUCTURE 21), AKC 126 (STRUCTURE 22), AKC 102 (STRUCTURE 23), AKC 103 (STRUCTURE 24), and

AKC 171 (STRUCTURE 25).

3. A secondary arylamine of the following Formula III : Ar-NH-CHR-CH2-CO-Y (Formula III) wherein Ar is aryl or heteroaryl optionally substituted by one or more R3 groups ; R is aryl, heteroaryl, or heterocycloalkyl optionally substituted by R3 ; Y is C1-6 alkyl, aryl, or heteroaryl optionally substituted by R3 ; or R and Y together form a cycloalkyl or heterocycloalkyl ring ; and is is OH, halogen, CF3, OCF3, or a group selected from-NH2, SO, alkyl, C6~, o aryl, C, 4 alkoxy, and C, -6alkyl, said group being optionally substituted by OH, C, 6 alkoxy, C, 6 alkyl, phenyl, halogen, or CF3.

Particularly preferred anthelmintic compounds according to Formula III are exemplified herein by compounds represented by structures 26-31 (depicted in Figures 26- 31, respectively), which have been assigned the respective reference numbers : AKC 128 (STRUCTURE 26), AKC 129 (STRUCTURE 27), AKC 130 (STRUCTURE 28), AKC 131 (STRUCTURE 29), AKC 132 (STRUCTURE 30), and AKC 133 (STRUCTURE 31).

4. A diaryl amine of the following Formula IV : Ar- Z) o--Ar- (CH2) 0-uNHR (Formula IV) wherein Ar is aryl or heteroaryl optionally substituted by one or more R3 groups ; Z is NH, O, S, or Alk ; and Alk is a linear or cyclic alkylene radical of up to 8 C atoms wherein said radical optionally includes one or more heteroatoms ; RisHorR3, R3 is OH, halogen, CF3, OCF3, or a group selected from NH2, SO, alkyl, C,-,, aryl, C4 alkoxy, and Cl4 -6 alkyl, said group being optionally substituted by OH, C, 6 alkoxy, C, 6 alkyl, phenyl, halogen, or CF3.

Particularly preferred anthelmintic compounds according to Formula IV are exemplified by compounds represented by structures 32-37 (depicted in Figures 32-37, respectively), which have been assigned the respective reference numbers : AKC 109 (STRUCTURE 32), AKC 134 (STRUCTURE 33), AKC 135 (STRUCTURE 34), AKC 136 (STRUCTURE 35), AKC 137 (STRUCTURE 36), and AKC 138 (STRUCTURE 37).

5. A substituted heteropolycyclic compound of the following Formula V : Het2-Q (Formula V) wherein Hertz is two or three fused aromatic rings including one or more heteroatoms selected from N, O and S, and Q includes at least one substituent selected from OH, COOR3 and CONHR3, and optionally also another substituent selected from alkyl and alkenyl of up to 10 C atoms ; wherein R3 is OH, halogen, CF3, OCF3, or a group selected from NH2, SO2 alkyl, C6, 0 aryl, C,-6 alkoxy, and C 16alkyl, said group being optionally substituted by OH, C 1-6 alkoxy, C, 6 alkyl, phenyl, halogen, or CF3.

Particularly preferred anthelmintic compounds according to Formula V are exemplified by compounds represented by structures 38-43 (depicted in Figures 38-43, respectively), which have been assigned the respective reference numbers : AKC 139 (STRUCTURE 38), AKC 140 (STRUCTURE 39), AKC 141 (STRUCTURE 40), AKC 142 (STRUCTURE 41), AKC 143 (STRUCTURE 42), and AKC 144 (STRUCTURE 43).

For the foregoing Formulae I, II, III, IV, and V, as well as throughout this disclosure, the following definitions apply.

"Aryl"refers to an aromatic group, typically of 6-10 C atoms, such as phenyl or naphthyl.

"Alk"includes, for example, (CH2) n wherein n is an integer of up to 6, e. g. 1, 2, 3, or 4, or cyclohexylene.

"Heteroaryl"means an aromatic group including one or more heteroatoms selected from O, S and N. It will typically have 5 or 6 ring atoms. It may also be fused to one or more aryl groups. Examples are in the illustrated compounds.

"Heterocycloalkyl"means a cycloalkyl group in which one or more C atoms are replaced by one or more heteroatoms selected from O, S and N. It will typically have 5 or 6 ring atoms. Examples are in the illustrated compounds of structures 1-43.

Other preferred anthelmintic compounds useful according to the subject invention are represented by structures 44, 45, and 46 (depicted in Figures 44-46, respectively), and have been assigned the respective reference numbers : AKC 145 (STRUCTURE 44), AKC 146 (STRUCTURE 45), and AKC 147 (STRUCTURE 46).

The invention process is particularly valuable to control nematodes which are pests to animals, as well as nematodes attacking the roots of desired crop plants, ornamental plants, and turf grasses. The desired crop plants can be, for example, cotton, soybean, tomatoes, potatoes, grapes, strawberries, bananas or vegetables.

In one embodiment of the subject invention, the subject anthelmintic compounds are used in conjunction with one or more other nematicidal agents. The other nematicidal agents may be, for example, a biological agent, an avermectin, a milbemycin, or a fatty acid.

In another embodiment, the subject invention further provides methods for killing the eggs of nematodes. Thus, the subject invention further relates to the surprising discovery that certain compounds have ovicidal activity against nematode eggs.

Compositions comprising the anthelmintic compounds of the subject invention are particularly useful for replant applications in nematode-control schemes.

Description of the Drawings Figure 1 depicts Structure 1 which represents anthelmintic compound AKC 111.

Figure 2 depicts Structure 2 which represents anthelmintic compound AKC 112.

Figure 3 depicts Structure 3 which represents anthelmintic compound AKC 113.

Figure 4 depicts Structure 4 which represents anthelmintic compound AKC 107.

Figure 5 depicts Structure 5 which represents anthelmintic compound AKC 114.

Figure 6 depicts Structure 6 which represents anthelmintic compound AKC 108.

Figure 7 depicts Structure 7 which represents anthelmintic compound AKC 115.

Figure 8 depicts Structure 8 which represents anthelmintic compound AKC 116.

Figure 9 depicts Structure 9 which represents anthelmintic compound AKC 117.

Figure 10 depicts Structure 10 which represents anthelmintic compound AKC 118.

Figure 11 depicts Structure 11 which represents anthelmintic compound AKC 119.

Figure 12 depicts Structure 12 which represents anthelmintic compound AKC 110.

Figure 13 depicts Structure 13 which represents anthelmintic compound AKC 120.

Figure 14 depicts Structure 14 which represents anthelmintic compound AKC 121.

Figure 15 depicts Structure 15 which represents anthelmintic compound AKC 2153.

Figure 16 depicts Structure 16 which represents anthelmintic compound AKC 122.

Figure 17 depicts Structure 17 which represents anthelmintic compound AKC 104.

Figure 18 depicts Structure 18 which represents anthelmintic compound AKC 123.

Figure 19 depicts Structure 19 which represents anthelmintic compound AKC 124.

Figure 20 depicts Structure 20 which represents anthelmintic compound AKC 125.

Figure 21 depicts Structure 21 which represents anthelmintic compound AKC 105.

Figure 22 depicts Structure 22 which represents anthelmintic compound AKC 126.

Figure 23 depicts Structure 23 which represents anthelmintic compound AKC 102.

Figure 24 depicts Structure 24 which represents anthelmintic compound AKC 103.

Figure 25 depicts Structure 25 which represents anthelmintic compound AKC 171.

Figure 26 depicts Structure 26 which represents anthelmintic compound AKC 128.

Figure 27 depicts Structure 27 which represents anthelmintic compound AKC 129.

Figure 28 depicts Structure 28 which represents anthelmintic compound AKC 130.

Figure 29 depicts Structure 29 which represents anthelmintic compound AKC 121.

Figure 30 depicts Structure 30 which represents anthelmintic compound AKC 132.

Figure 31 depicts Structure 31 which represents anthelmintic compound AKC 133.

Figure 32 depicts Structure 32 which represents anthelmintic compound AKC 109.

Figure 33 depicts Structure 33 which represents anthelmintic compound AKC 134.

Figure 34 depicts Structure 34 which represents anthelmintic compound AKC 135.

Figure 35 depicts Structure 35 which represents anthelmintic compound AKC 136.

Figure 36 depicts Structure 36 which represents anthelmintic compound AKC 137.

Figure 37 depicts Structure 37 which represents anthelmintic compound AKC 138.

Figure 38 depicts Structure 38 which represents anthelmintic compound AKC 139.

Figure 39 depicts Structure 39 which represents anthelmintic compound AKC 140.

Figure 40 depicts Structure 40 which represents anthelmintic compound AKC 141.

Figure 41 depicts Structure 41 which represents anthelmintic compound AKC 142.

Figure 42 depicts Structure 42 which represents anthelmintic compound AKC 143.

Figure 43 depicts Structure 43 which represents anthelmintic compound AKC 144.

Figure 44 depicts Structure 44 which represents anthelmintic compound AKC 145.

Figure 45 depicts Structure 45 which represents anthelmintic compound AKC 146.

Figure 46 depicts Structure 46 which represents anthelmintic compound AKC 147.

Figure 47 depicts a basic structure, Structure 47, of a preferred class of anthelmintic compound.

Figure 48 depicts one library scheme by which the skilled artisan can create the compounds represented by the structure depicted in Figure 47.

Figure 49 depicts a hydroxyamidine synthesis.

Figure 50 depicts BOC-amino acid prepared by catalytic hydrogenation of a pyridine-containing acid and subsequent BOC-protection.

Detailed Disclosure of the Invention The process of the subject invention concerns the use of certain organic compounds to control the infestation of plants or animals by nematodes. These organic compounds comprise Formulae I, II, III, IV, and V, as well as Structures 44, 45, and 46.

In a particularly preferred embodiment of the subject invention, the anthelmintic compound is selected from the group consisting of Compounds 1-46 represented by Structures 1-46. Preferred anthelmintic compounds useful in accord with the subject invention are represented by Structure 47, wherein : R, is aryl (optionally substituted with OC15 or C15 straight or branched alkyl) ; R, is Cl, 0 branched, straight, or cyclic alkyl ; R3 is aryl (optionally substituted with halogen or OCF3).

Generally, the anthelmintic compounds of the subject invention can be unsubstituted or substituted, saturated or unsaturated. The anthelmintic component of an anthelmintic composition used according to the subject invention may be a single anthelmintic compound or a mixture of two or more anthelmintic compounds. The subject compounds may be used in conjunction with other anthelmintic compounds, including the free acids and salts of the anthelmintic compounds of the present invention.

The salts may be, for example, sodium or potassium salts, or ammonium salts. As would be apparent to the ordinary skilled artisan, physiologically acceptable acids and salts of the subject anthelmintic compounds can readily be made and used in accord with the teachings herein, and are hereby expressly included by reference to each compound or group of compounds. For example,"AKC 111","Compound 1", or"Structure 1"is intended to include the physiologically acceptable acids and salts thereof. In addition, the subject anthelmintic compounds may have an assymetrical carbon atom, i. e., optically active site.

These compounds exist in (R) and (S) enantiomeric forms. Both the (R) and (S) enantiomers of the subject compounds are contempated by the subject invention.

Anthelmintic compounds specifically exemplified herein include Compounds 1-46 represented by Structures 1-46 above.

The subject compounds used in the invention can be applied to animals, the living and feeding areas of animals, plants, or to the situs of plants needing nematode control.

The anthelmintic compositions may be applied by, for example, drip and drench techniques. With the drip application, the subject compositions can be applied directly to the base of plants or to the soil root zone. The composition may be applied through already existing drip irrigation systems. This procedure is particularly applicable for ornamental plants, strawberries, tomatoes, potatoes, grapes, and vegetables.

Alternatively, a drench application can be used. For treating plants, a sufficient quantity

of the anthelmintic composition is applied such that the composition drains to the root area of the plants. An important aspect of the subject invention is the surprising discovery that certain 4-phenoxy-6-amino-pyrimidine compounds have excellent nematicidal activity at concentrations which are not phytotoxic.

The drench technique can be used for a variety of crops and for turf grasses. The drench technique can also be used for animals. Preferably, for administration to animals the anthelmintic composition would be administered orally to facilitate activity against internal nematode parasites. The compositions of the subject invention can readily be applied using the teachings provided herein.

In a preferred embodiment of the subject invention, an anthelmintic compound will be applied as an aqueous microemulsion. As described herein, the concentration of the active ingredient should be sufficient to control the nematode infestation without causing phytotoxicity to the desired plants. The concentration of anthelmintic compound may be, for example, from about 0. 0001% to about 2%, preferably from about 0. 025% to about 1%, and, most preferably, from about 0. 05% to about 0. 5%.

The anthelmintic composition used according to the subject invention can be applied in conjunction with one or more other nematicidal agents. The other nematicidal agent may, for example, be applied simultaneously or sequentially with the anthelmintic.

Such other nematicidal agents include, for example, avermectins, the Bts, and fatty acids.

The avermectin compound used according to the subject invention may be any of the avermectins, milbemycins, or derivatives of either, having activity against nematodes. The avermectin's activity will be enhanced when combined with an anthelmintic compound as described herein. Thus, the specific combination of ingredients can be manipulated to provide the optimal composition for a particular application.

Standard concentrations of avermectins are well known to those skilled in the art.

For example, the avermectin compounds can be employed in the combination of the subject invention at concentrations of from about 0. 03 to about 110 parts per million (ppm). Preferably, from about 1 to about 5 ppm are employed.

As would be readily appreciated by a person skilled in the art, the delivery of the subject anthelmintic and/or avermectin compound can be calculated in terms of the active ingredient applied per unit area. For example, the subject anthelmintic may be applied at a rate of about 0. 02 Ib/acre to about 0. 1 lb/acre and, preferably, from about 0. 5 Ib/acre to

about 2 Ibs/acre. Similarly, the avermectin product can be applied at a rate of up to about 16 oz. of formulated product ("AVID,"available from Merck) per acre. Preferably, about 4 oz. to about 8 oz. formulated"AVID"per acre would be used. Thus, the avermectin compound can be applied up to about 0. 02 Ib/acre. Preferably, the rate of avermectin is between about 0. 005 Ib/acre and 0. 01 Ib/acre. A person of ordinary skill in the art would readily appreciate that the desired application rate of the active ingredients could be achieved using a great variety of different concentrations of active ingredients while varying the application rate of the solution. Thus, a large quantity of dilute solution could be applied or a smaller quantity of a more concentrated solution.

A variety of different avermectins or related compounds can be used according to the subject invention. Ivermectin may also be used according to the subject invention, as may the milbemycins. For brevity, the term"avermectin"is used herein to refer to all the avermectins and their derivatives as well as related compounds such as the milbemycins and the ivermectins."Derivatives"refer to chemical modifications of the avermectins or milbemycins which are well known and available to those skilled in this art. Such derivatives are described, for example, in U. S. Patent No. 4, 560, 677. Avermectin is readily available under a variety of tradenames including"AVID,""ZEPHYR," "VERTIMEC,"and"AGRI-MEK." The anthelmintic compositions of the subject invention may also be used in conjunction with nematicidal agents other than the avermectins. For example, the anthelmintic compounds may be used with biological agents such as Bacillits thitritigietisis or with nematicidal fungi. In this context, the anthelmintic composition could be applied at concentrations which would not antagonize the action of the biological agent. The biologically active agent may be in a live proliferative form or may be in a dead stabilized form as described, for example, in U. S. Patent Nos. 4, 695, 462 and 4, 695, 455.

Furthermore, the anthelmintic compositions of the subject invention may be used with plants which are specifically bred or engineered for nematode resistance. The plants may, for example, be transformed with B. t. genes which confer nematode resistance or may simply be hybrids or varieties selected for such resistance. The anthelmintic compositions of the subject invention are particularly effective against free-living ectoparasitic nematodes and, therefore, combined use with plants selected for endoparasitic nematode resistance is highly advantageous.

The subject invention further relates to the surprising discovery that the anthelmintics of the subject invention have ovicidal activity against nematode eggs. Thus, in another embodiment, provided are methods for killing the eggs of nematodes, including those within cysts or egg masses that are commonly formed by Heterodera, Globodera, and Meloidogyne (cyst and root-knot) species.

The ovicidal compositions according to the subject invention are particularly useful for preplant applications in nematode-control schemes. In addition, the ovicidal compositions of the subject invention can be advantageously used as postplant nematicides, especially because of their relatively low phytotoxicity. In the latter embodiments, ovicidal compositions of the subject invention can be delivered, after planting and at appropriate, essentially non-phytotoxic concentrations of anthelmintic compounds, along with irrigation water and/or plant nutrients to ensure a continuous zone of nematode protection to the enlarging plant root mass. Thus, when applied using these techniques, which include drench or drip systems as are known in the art, phytopathogenic nematodes in their vermiform (wormlike) and egg stages are controlled.

Antheimintic compounds having Formulae I, II, III, IV, and V, Structure 47 and most preferably Structures 1-46, particularly Structures 1-7 and 35, 36, and 37, are used in preferred embodiments for killing nematode eggs. In addition, microemulsions of the subject compounds are highly preferred for ovicidal applications. In preferred embodiments, the anthelmintic compound (s) will be present in a concentration of greater than about 150 ppm. More preferably, the concentration will be greater than about 200 ppm ; most preferably it will be about 250 ppm or more. For certain conditions, the anthelmintic compounds should be applied at high concentrations of about 1, 000 ppm to about 5, 000 ppm or more.

In light of the subject disclosure, one skilled in the art could readily use a variety of application techniques and formulations to prevent the hatching of nematode eggs in a variety of agricultural, farm-related, and garden-related settings.

Examples of animal parasitic nematodes against which the subject compounds can be used include the following : Amblyomma spp.

Babesia spp. (RBC) Bunostomum spp.

Calliphorid larvae

Capillaria spp.

Chabertia ovina Chorioptes Cooperia spp.

Cryptosporidium sp.

Damalinia ovis Damalinia caprae Demodex Dermacentor spp.

Dicrocoelium dentriticum Dictyocaulus filaria Echinococcus hydatid cyst Eimeria spp.

Elaeophora schneideri Fasciola hepatica Fasciola gigantica Fascioloides magna Giardia sp.

Gongylonema spp.

Haematobia irritans Haemonchus contortus contortus Ixodes Linguatula serrata larvae Linguatula serrata nymphs Linognathus spp.

M. domestica Marshallagia marshalli Melophagus ovinus Moniezia benedeni Moniezia expansa Muellerius capillaris Musca autumnalis Nematodirus spp.

Oesophagostomum spp.

Oestrus ovis Ornithodoros Ostertagia circumcincta Ostertagia trifurcata Otobius Paramphistomum sp.

Parelaphostrongylus tenuis Protostrongylus sp.

Psoroptes Rhipicephalus spp.

Sarcoptes scabiei Sarcocystis spp.

Sarcocystis spp. cysts Schistosoma spp.

Stomoxys calcitrans Strongyloides papillosus Taenia hydatigena cysticerci Taenia multiceps coenurus Taenia ovis cysticerci Thelazia Thysanosoma actinoides Theileria spp. C) Toxocara vitulorum Toxoplasma gondii Toxoplasma gondii cysts Trichostrongylus axei Trichostrongylus spp.

Trichuris ovis Trypanosoma spp. (plasma) It has been found that helminth, acarid and arthropod endo-and ectoparasitic infestations may be controlled, prevented or eliminated, by applying to, injecting or orally dosing said animals with an endo-or ectoparasiticidally effective amount of the subject anthelmintic compounds, preferably the above-described Structure 1-46 compounds. This may be achieved by applying the compound to the skin, hide and/or hair of the animals, or injecting or orally dosing said animals with a solid or liquid formulated composition.

For control of flea infestations, treatment of the infested animal to control adults in conjunction with treatment of the area occupied by the infested animal to control flea larvae is recommended. The compositions of the present invention may be admixed with suitable carriers for application to interior and/or exterior areas for control of flea larvae.

The compositions of the present invention may be employed as animal feeds, animal feed premixes or feed concentrates. Feed concentrates and feed premixes, useful in the practice of the invention, may be prepared by admixing about 0. 25% to 35% by weight of a subject anthelmintic compound, preferably a Structure 1-46 compound, with about 99. 75% to 65% by weight of a suitable agronomic carrier or diluent. Carriers suitable for use include 0. 75% to 35% by weight of a physiologically acceptable alcohol such as benzyl alcohol phenethyl alcohol or propylene glycol, 0 to about 10% by weight of a vegetable oil such as corn oil or soybean oil, or propylene glycol and about 30% to 95% by weight of a sorptive, edible organic carrier such as corn grits, wheat middlings, soybean meal expanded corn grits, extracted corn meal or the like or a sorptive silica or a silicate. These feed premixes or concentrates may be admixed with the appropriate amount of animal feed to provide the animals with about 0. 5 ppm to 1000 ppm and

preferably about 1 ppm to 500 ppm of the compound in the animal's diet. These premixes or concentrates may also be used as top dressings for the animal's daily ration and applied across the top of the daily ration in sufficient amount to provide the animal with about 0. 5 ppm to 1000 ppm and preferably about 1 ppm to 500 ppm of the active ingredient, based on the animal's total feed.

The subject anthelmintic compounds, and particularly the Structure 1-46 compounds, most particularly Structures 1-7, 35, 36, and 37, may be administered to the animals in or with their drinking water.

The compound may also be administered in the form of a pill, tablet, bolus, implant, capsule, or drench, containing sufficient anthelmintic compound to provide the treated animal with about 0. 01 mg/kg to 100 mg/kg of animal body weight per day of the compound. These dosage forms are prepared by intimately and uniformly mixing the active ingredient with suitable finely divided diluents, fillers, disintegrating agents and/or builders such as starch, lactose, talc, magnesium stearate, vegetable gums, or the like.

These unit dosage formulations may be varied with respect to the total weight and content of anthelmintic compound depending upon the kind and size of the animal to be treated, the severity or type of infection encountered and the weight of the host.

Alternatively, the anthelmintic compound may be administered to animals parenterally, for example, by intraruminal, intramuscular, or subcutaneous injection in which the active ingredient is dissolved or dispersed in a liquid carrier. For this type administration the compound may be dispersed in a physiologically acceptable solvent for subcutaneous injection, or it may be dispersed in a fat or wax or mixture thereof containing an oil, buffer, surfactant, stabilizer, preservative and salt. Components useful in these preparations include carbowax, aluminum monostearate gel, diethyl succinate, soya oil, glyceral dioleate, saline, and capric/caprylic triglycerides.

The subject anthelmintic compounds may also be applied topically to the larger animals such as swine, sheep, cattle, and horses and companion animals such as dogs and cats in the form of aqueous dips or sprays. For this type administration, the active compound is generally prepared as a wettable powder, emulsifiable concentrate, aqueous flowable, or the like, which is mixed with water at the site of treatment and applied topically to the hide, skm, or hair of the animal. Such sprays or dips usually contain about 0. 5 ppm to 5, 000 ppm and preferably about 1 ppm to 3000 ppm of the compound.

Advantageously, the subject anthelmintic compounds may also be prepared as pour-on formulations and poured on the backs of the animals such as swine, cattle, sheep, horses, poultry, and companion animals to protect them against infestation by nematodes, acarids, and arthropod endo-and ectoparasites. Such pour-on compositions are generally prepared by dissolving, dispersing, or emulsifying the anthelmintic compound in a suitable nontoxic pharmacologically acceptable diluent for pour-on and administration. The diluent must be compatible with the compound and should not be a source of irritation or damage to the animals hide, skin, or hair. Such diluents include vegetable oils, spreading oils, polyhydric alcohols, aliphatic or aromatic hydrocarbons, esters of fatty acids, and lower alkyl ketones.

A typical pour-on formulation includes about 0. 5% to 30% by weight of the anthelmintic compound, about 30% to 60% by weight of an aliphatic or aromatic hydrocarbon, mono or polyhydric alcohol, lower alkyl ketone or mixtures thereof, 0 to about 20% by weight of a vegetable or mineral oil and about 0. 5% to 30% by weight of a spreading oil. Another typical pour-on contains about 45% by weight of xylene, about 15% by weight of the anthelmintic compound, about 10% by weight of corn oil or mineral oil, about 25% by weight of cyclohexanone and about 5% by weight of other pharmacologically acceptable spreading agents, antifoam agents, surfactants, or the like.

The subject anthelmintic compounds may also be prepared as ear tags for animals, particularly quadrupeds such as cattle and sheep. The tags may be prepared by stirring together about 55% to 60% by weight of a vinyl dispersion resin, having an inherent viscosity of about 1. 20 and an average particle size of about 0. 75 microns, a curing temperature range of about 120°C to 180°C, with about 28% by weight of butylbenzylphthalate. Stirring is continued, and about 1. 5% by weight of ca/Zn stearate stabilizer is added along with about 7. 0% of the compound and 2. 8% of epoxidized soybean oil. The resulting mixture is deaerated for 15 to 20 minutes at 125 mm/Hg. This mixture can be coated on an ear tag blank by dipping and the resulting tag cured at about 145°C to 150°C for about five minutes.

The compounds of Formulae I-V and particularly Structures 1-46 are nematicidal and can be used to control nematodes in crop plants. Therefore, in a further preferred aspect of the invention, there is provided a method for killing or controlling nematodes which comprises applying to the locus of the pests or to a plant susceptible to attack by

the pest an effective amount of a compound having any of Structures 1-46, preferably Structure 47, and particularly Structures 1-7, 35, 36, and 37, as defined herein.

The term"controlling"extends to non-lethal effects which result in the reduction or prevention of damage to the host plant or animal and the limitation of nematode population increase. These effects may be the result of chemical induced disorientation, immobilisation, or hatch prevention or induction. The chemical treatment may also have deleterious effects on nematode development, reproduction, or viability.

The compounds of the invention can be used against both plant-parasitic nematodes and nematodes living freely in the soil. Examples of plant-parasitic nematodes are : ectoparasites, for example Xiphinema spp., Longidorus spp :, and Trichodorous spp. ; semi-endoparasites, for example, Tylenchulus spp. ; migratory endoparasites, for example, Pratylenchus spp., Radopholus spp., and Scutellonema spp. ; sedentary endoparasites, for example, Heterodera spp., Globodera spp., and Meloidogyne spp. ; and stem and leaf endoparasites, for example, Ditylenchus spp., Aphelenchoides spp., and Hirshmaniella spp..

The Formulae I-V compounds, and preferably the compounds of Structures 1-46, display nematicidal activity against different types of nematodes including the cyst nematode. The subject compounds may also be used to combat and control infestations of insect pests such as Lepidoptera, Diptera, Homoptera, and Coleoptera (including Diabrotica i. e. corn rootworms) and also other invertebrate pests, for example, acarine pests. The insect and acarine pests which may be combated and controlled by the use of the invention compounds include those pests associated with agriculture (which term includes the growing of crops for food and fiber products), horticulture and animal husbandry, forestry, the storage of products of vegetable origin, such as fruit, grain and timber, and also those pests associated with the transmission of diseases of man and animals. Examples of insect and acarine pest species which may be controlled by the subject compounds include : Myzus persicae (aphid) Aphis gossypii (aphid) Aphis fabae (aphid) Megoura viceae (aphid) Aedes aegypti (mosquito)

Anopheles spp. (mosquitos) Culex spp. (mosquitos) Dysdercus fasciatus (capsid) Musca domestica (housefly) Pieris brassicae (white butterfly) Plutella maculipennis (diamond back moth) Phaedon cochleariae (mustard beetle) Aonidiella spp. (scale insects) Trialeuroides spp. (white flies) Bemisia tabaci (white fly) Blattella germanica (cockroach) Periplaneta americana (cockroach) Blatta orientals (cockroach) Spodoptera littorals (cotton leafworm) Hellothis virescens (tobacco budworm) Chortiocetes terminifera (locust) Diabrotica spp. (rootworms) Agrotis spp. (cutworms) Chilo pa. rtellus (maize stem borer) Nilaparvata lugens (planthopper) Nephotettix cincticeps (leafhopper) Panonychus ulmi (European red mite) Panonychus citri (citrus red mite) Tetranychus urticae (two-spotted spider mite) Tetranychus cinnabarinus (carmine spider mite) Phyllcoptruta oleivora (citrus rust mite) Polyphagotarsonemus latus (broad mite) Brevipalpus spp. (mites) In order to apply the compound to the locus of the nematode, insect, or acarid pest, or to a plant susceptible to attack by the nematode, insect, or acarid pest, the compound is usually formulated into a composition which includes in addition to at least

one of the subject anthelmintic compounds suitable inert diluent or carrier materials, and/or surface active agents. Thus, in two further aspects of the invention there is provided a nematicidal, insecticidal, or acaricidal composition comprising an effective amount of a subject anthelmintic compound and preferably of any of Structures 1-46 as defined herein and an inert diluent or carrier material and optionally a surface active agent.

The amount of active ingredient generally applied for the control of nematode pests is from 0. 01 to 10 kg per hectare and preferably from 0. 1 to 6 kg per hectare.

The compositions containing the active ingredient can be applied to the soil, plant or seed, to the locus of the pests, or to the habitat of the pests, in the form of dusting powders, wettable powders, granules (slow or fast release), emulsion or suspension concentrates, liquid solutions, emulsions, seed dressings, fogging/smoke formulations or controlled release compositions, such as microencapsulated granules or suspensions.

Dusting powders are formulated by mixing the active ingredient with one or more finely divided solid carriers and/or diluents, for example natural clays, kaolin, pyrophyllite, bentonire, alumina, montmorillonite, kieselguhr, chalk, diatomaceous earths, calcium phosphates, calcium and magnesium carbonates, sulphur, lime, flours, talc, and other organic and inorganic solid carriers.

Granules are formed either by absorbing the active ingredient in a porous granular material for example pumice, attapulgite clays, fullers earth, kieselguhr, diatomaceous earths, ground corn cobs, and the like, or on to hard core materials such as sands, silicates, mineral carbonates, sulphates, phosphates, or the like. Agents which are commonly used to aid in impregnation, binding or coating the solid carriers include aliphatic and aromatic petroleum solvents, alcohols, polyvinyl acetates, polyvinyl alcohols, ethers, ketones, esters, dextrins, sugars, and vegetable oils with the active ingredient.

Other additives may also be included, such as emulsifying agents, wetting agents, or dispersing agents.

Microencapsulated formulations (microcapsule suspensions CS) or other controlled release formulations may also be used, particularly for slow release over a period of time, and for seed treatment.

Alternatively the compositions may be in the form of liquid preparations to be used as dips, irrigation additives or sprays, which are generally aqueous dispersions or emulsions of the active ingredient in the presence of one or more known wetting agents,

dispersing agents or emulsifying agents (surface active agents). The compositions which are to be used in the form of aqueous dispersions or emulsions are generally supplied in the form of an emulsifiable concentrate (EC) or a suspension concentrate (SC) containing a high proportion of the active ingredient or ingredients. An EC is a homogeneous liquid composition, usually containing the active ingredient dissolved in a substantially non-volatile organic solvent. An SC is a fine particle size dispersion of solid active ingredient in water. To apply the concentrates they are diluted in water and are usually applied by means of a spray to the area to be treated. For agricultural or horticultural purposes, an aqueous preparation containing between 0. 0001% and 0. 1% by weight of the active ingredient (approximately equivalent to from 5-2000 g/ha) is particularly useful.

Suitable liquid solvents for ECs include methyl ketone, methyl isobutyl ketone, cyclohexanone, xylenes, toluene, chlorobenzene, paraffins, kerosene, white oil, alcohols, (for example, butanol), methylnaphthalene, trimethylbenzene, trichloroethylene, N-methyl-2-pyrrolidone, and tetrahydrofurfuryl alcohol (THFA).

Wetting agents, dispersing agents, and emulsifying agents may be of the cationic, anionic, or non-ionic type. Suitable agents of the cationic type include, for example, quaternary ammonium compounds, for example cetyltrimethyl ammonium bromide.

Suitable agents of the anionic type include, for example, soaps ; salts of aliphatic monoesters of sulphuric acid, for example sodium lauryl sulphate ; salts of sulphonated aromatic compounds, for example sodium dodecylbenzenesulphonate ; sodium, calcium or ammonium lignosulphonate ; or butylnaphthalene sulphonate ; and a mixture of the sodium salts of diisopropyl-and triisopropylnaphthalene sulphonates. Suitable agents of the non-ionic type include, for example, the condensation products of ethylene oxide with fatty alcohols such as oleyl alcohol or cetyl alcohol ; or with alkyl phenols such as octyl phenol, nonyl phenol, and octyl cresol. Other non-ionic agents are the partial esters derived from long chain fatty acids and hexitol anhydrides, the condensation products of the said partial esters with ethylene oxide, and the lecithins.

These concentrates are often required to withstand storage for prolonged periods and after such storage, to be capable of dilution with water to form aqueous preparations which remain homogeneous for a sufficient time to enable them to be applied by conventional spray equipment. The concentrates may preferably contain 1-85% by weight of the active ingredient or ingredients. When diluted to form aqueous preparations such

preparations may contain varying amounts of the active ingredient depending upon the purpose for which they are to be used.

The subject anthelmintic compounds may also be formulated as powders (dry seed treatment DS or water disperible powder WS) or liquids (flowable concentrate FS, liquid seed treatment LS), or microcapsule suspensions CS for use in seed treatments. The formulations can be applied to the seed by standard techniques and through conventional seed treaters. In use the compositions are applied to the nematodes, to the locus of the nematodes, to the habitat of the nematodes, or to growing plants liable to infestation by the nematodes, by any of the known means of applying pesticidal compositions, for example, by dusting, spraying, or incorporation of granules.

The compounds of the invention may be the sole active ingredient of the composition or they may be admixed with one or more additional active ingredients such as nematicides, agents which modify the behavior of nematodes (such as hatching factors), insecticides, synergists, herbicides, fungicides or plant growth regulators where appropriate.

Suitable additional active ingredients for inclusion in admixture with the compounds of the invention may be compounds which will broaden the spectrum of activity of the compounds of the invention or increase their persistence in the location of the pest. They may synergise the activity of the compound of the invention or complement the activity for example by increasing the speed of effect or overcoming repellency.

Additionally multi-component mixtures of this type may help to overcome or prevent the development of resistance to individual components.

The particular additional active ingredient included will depend upon the intended utility of the mixture and the type of complementary action required. Examples of suitable insecticides include the following : a) Pyrethroids such as permethrin, esfenvalerate, deltamethrin, cyhalothrin in particular lambda-cyhalothrin, biphenthrin, fenpropathrin, cyfluthrin, tefluthrin, fish safe pyrethroids for example ethofenprox, natural pyrethrin, tetramethrin, s-bioallethrin, fenfluthrin, prallethrin, and 5-benzyl-3-furylmethyl-(E)-(1 R, 3 S)-2, 2-dimethyl-3-(2-oxothiolan-3-ylidenem ethyl) cyclopropane carboxylate ;

b) Organophosphates such as profenofos, sulprofos, methyl parathion, azinphos-methyl, demeton-s-methyl, heptenophos, thiometon, fenamiphos, monocrotophos, profenophos, triazophos, methamidophos, dimethoate, phosphamidon, malathion, chloropyrifos, phosalone, terbufos, fensulphothion, fonofos, phorate, phoxim, pyrimiphos-methyl, pyrimiphos-ethyl, fenitrothion, or diazinon ; c) Carbamates (including aryl carbamates) such as pirimicarb, cloethocarb, carbofuran, furathiocarb, ethiofencarb, aldicarb, thiofurox, carbosulphan, bendiocarb, fenobucarb, propoxur, or oxamyl ; d) Benzoyl ureas such as triflumuron or chlorofluazuron ; e) Organic tin compounds such as cyhexatin, fenbutatin oxide, or azocyclotin ; f) Macrolides such as avermectins or milbemycins, for example such as abamectin, avermectin, and milbemycin ; Hormones and pheromones ; h) Organochlorine compounds such as benzene hexachloride, DDT, endosulphan, chlordane, or dieldrin ; i) Amidines, such as chlordimeform or amitraz ; j) Fumigant agents ; k) nitromethylenes such as imidacloprid.

In addition to the major chemical classes of insecticide listed above, other insecticides having particular targets may be employed in the mixture if appropriate for the intended utility of the mixture. For instance, selective insecticides for particular crops, for example stemborer specific insecticides for use in rice such as cartap or buprofezin, can be employed. Alternatively, insecticides specific for particular insect species/stages, for example, ovo-larvicides such as chlofentezine, flubenzimine, hexythiazox, and tetradifon ; motilicides such as dicofol or propargite ; acaricides such as bromopropylate or chlorobenzilate ; or growth regulators such as hydramethylon, cyromazin, methoprene, chlorfluazuron, and diflubenzuron may also be included in the compositions.

Examples of suitable synergists for use in the compositions include piperonyl butoxide, sesamax, safroxan, and dodecyl imidazole.

Suitable herbicides, fungicides, and plant-growth regulators for inclusion in the compositions will depend upon the intended target and the effect required.

An example of a rice selective herbicides which can be included is propanil, an example of a plant growth regulator for use in cotton is"Pix", and examples of fungicides for use in rice include blasticides such as blasticidin-S. The ratio of the compound of the invention to the other active ingredient in the composition will depend upon a number of factors including type of target, effect required from the mixture, etc. However in general, the additional active ingredient of the composition will be applied at about the rate as it is usually employed, or at a slightly lower rate if synergism occurs.

The anthelmintic compounds according to the invention also show fungicidal activity and may be used to control one or more of a variety of plant pathogens. In a further aspect the invention therefore includes a method of combating fungi which comprises applying to a plant, to a seed of a plant, or to the locus of the plant or seed a fungicidally effective amount of a compound as herein defined or a composition containing the same. The invention further includes a fungicidal composition comprising a fungicidally effective amount of a compound as herein defined and a fungicidally acceptable carrier or diluent therefor.

Examples of plant pathogens which the compounds or fungicidal compositions of the invention may control, methods by which fungi may be combatted and the form of suitable compositions, including acceptable carriers and diluents ; adjuvants such as wetting, dispersing, emulsifying, and suspending agents ; and other ingredients, such as fertilisers and other biologically active materials, are described, for instance, in International application No. WO 93/08180, the content of which is incorporated herein by reference.

All of the U. S. patents cited herein are hereby incorporated by reference.

Following are examples which illustrate procedures for practicing the invention.

These examples should not be construed as limiting. All percentages are by weight and all solvent mixture proportions are by volume unless otherwise noted. For clarity the following abbreviations shall be used throughout the examples : ACD : Available Chemicals Directory ACN : Acetonitrile

AcOH : Acetic Acid AMPS : (Aminomethyl) polystyrene AUC : Area under curve BAM Benzamidoxime BOC : t-Butoxycarbonyl CDI : 1, 1'-Carbonyldiimidazole CI : Chemical Ionization 1, 2-DCE : 1, 2-Dichloroethane DCM : Dichloromethane DIPEA : N, N-Diisopropylethylamine DIC : 1, 3-Diisopropylcarbodiimide DMAP 4-(Dimethylamino) pyridine EDC : 1- (3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride ESI : Electrospray ionization HCI : Hydrochloric Acid HOBt : 1-Hydroxybenzotriazole HPLC : High Performance Liquid Chromatography LLE : Liquid Liquid Extraction LC/MS : Liquid Chromatography/Mass Spectroscopy O/N : Overnight RT : Room Temperature SLE : Solid-supported liquid-liquid extraction THF : Tetrahydrofuran TFA : Trifluoroacetic acid QC : Quality Control dH, O : Distilled water Example 1-Preparation of Anthelmintic Compounds 1-46 The anthelmintic compounds of the subject invention can readily be produced using procedures well known to those skilled in the art.

A variety of anthelmintic compounds useful according to the subject invention can be readily prepared by a person skilled in this art having the benefit of the subject disclosure.

Example 2-Nematicidal Activity of Anthelmintic Compositions 1-31 Caerzorhabditis elegant adults were grown on Nematode Growth Medium (NGM) until they produced eggs, then the adults were removed.

The eggs were allowed to hatch, and the LI larvae collected. See The Nematode Caenorhabditis elegans (1988) Cold Spring Harbor Laboratory Press. Using a Matrix Programmable Pipette, the LIs were distributed into 96-well tissue culture plates, 20 LI in 501 NGM per well. Antibiotic/Antimyotic was added to each well, and I % by weight E. coli strainHB101. The subject anthelmintic compounds were stored at 5mM in 100% DMSO. 0. 741 of compounds 1-31 were added to the left-most column of wells to yield a final concentration of 70, uM in 1. 4% DMSO, with 1. 4% DMSO only as the control.

The compounds were then subjected to 5 more 3-fold dilutions from left to right to yield 6 column concentrations of 70, uM, 23. 3µM, 7. 8µM, 2. 6µM, 0. 9µM, and 0. 3µM. Plates were stored in air-tight Rubbermaid plastic boxes at 20°C. The nematodes had cleared all control wells by day 4, and nematode viability was scored by visual examination under a 100x dissecting microscope on day 5. A visual viability scoring system was used as follows :

WORM VISUAL SCORING-GUIDE Lethalitv : Dead only stiff L 1 s (no movement) Dead (L4) worms are dead, but at a later larval stage LI majority of worms are LI (based on size) worms move when plate is tapped L2 majority of worms are L2 (based on size) L3 majority of worms are L3 (based on size) L4 majority of worms are L4 (based on size) Partial Penetrance : AD majority of worms are adult #AD 5 adult worms or less Broodsize Reductions : B ! sterile (0-25 progeny) B low broodsize (25-100 progeny) -B moderate broodsize (100-250 progeny) < reduced broodsize (250-500 progeny) OK no effect (-1000+ progeny) If several classes of worms exist in a well. then all classes are scored. If adults are present. then the brood score is also recorded. Thus,"L 1/L2"would mean a mixture of Ll's and L2's are present in the well."L4/&num AD/B would mean that a mixture of LA s and adults are resent in the well. The"&num AD" would mean that there are 6 or less adults. and the"B"would mean that there were 100 progeny or less.

The results are reported in Table 1. Column VI has a compound concentration of 70, uM with sequential 3-fold dilutions reported in columns V2, V3, V4, V5, and V6, respectively, such that the V6 concentration was 0. 3, uM.

Table 1. Dosc Itcsyonsc Inacl, Gvb S lky vmn : a Scnrc Siiiiciiitcfi Siitircc I'% %'ell A, I, Iicss v 1 V2 V3 V.) vs 1575 AYC 111 N ? II'JJ tfIX1 : 17111 ntal i>c : ul 1. ? f) cal (1.)/1. Il/vl)/Il () f : 1. AaC 112 2 N2 1/98 5 () 9 () : AII) 1 2/1 3 1. 2/1. 3 1 2/1.) 1. 3/1-1 llA n (IfL ; AKC 117 J N ? ! ! 85 5l161 : A 1 (1 () e : nl I) tml (caJ 1) caJ (I /L l. 2/I) caJ (n I. a/ ! 1/vl>ri 1 (, 9 AKC 107 I N2 DXi (} 5nCl Dln l) cad Dcal1 (1. ?/I. J) I.'~/I, J 1) cavl (1. 4) 1. 4/l) cad (A 1. 2/l) c. z d77 AKC 114 5 N2 1186 1 I) C. I (l 1. 3 1. 311) cail (l. 1. ?/1. 3-11 176 Ar C 108 (1 N 2/186 1 1 I) cltl I) citl 1. 1 1. 1 1. 1/1. 2 1. 2/1 3 17 ; hKC 115 7 N2 118 {, Sn (l Illn DcaL l) cad, l 2/1 3) 1. 2/Dcad (1. 2) Dcid (1. 2) Dcall (1. 2/1. 1. 2/1. 3 '76 AXCt08N ; t ;) ! 6.'ifj. Cnnca'att).) L) L)/). ). /i. 3 tlSS AKC 119 1I N2 1/120 $3) 3 lil IlAD/lx IIAI)/II DAI)/II I/AI)/II IsAI)/I1 < n : <. AKcH9"Ntnf. jn)/<ADn/'An/n ; ; AD/ !)/<A [)/n « An/n< ,) ss > AKC llO 12 N2 i/l 2S 53 9/t 1 1. 1 1. 1 1. l 1 1 1 1 1 1 AKC AKC 120 13 N2 1/13 (1 S. N4) C>l 1. 1 1. 2/1. 3 1. t 1. 1/i. 2 IlAlll-ll () a 132 AIC 121 14 t42 5181) C-1 1. 1 1. 1 1. 111. 2//A I)/-11 It A 1)/l I 1 AKC 2153 15 N2 n12 (537'C-l 1. 1 1. 1 1. 1/1. 2 IIAI)/I) IIAI)/II/IAI) ll '62 AKC122'N22) : 373 : f) SUcnd). j « A))/HAD/) i//A))/ [tOK ) S : AKC10'7N2/' ! !') : 118 I) cad 1 1 IIAI)/II IIA51tI IlAI) IIì (18 ; ; ? S AKC 104 17 N2"g"5f'2 : C-1 1. 1/1.'1. 1112 LIJL2 IIAI)/II 1. 1/1. 2 i. l/1. 2 A ; C 123 IS N ? u79 >cW. sts 1. 1 I. in. ? IInI) I1 < nnl) 11 uvt) i-Il 102 AYC 124 11) N2 ItH 1 5 () 33. 1) 8//A 1)/I I !//A 1) 11) ! 1. 2/1.) I., I///A 1)/Il I., I/IIAI)/Il 1) 111 <) 6 AKC 125 2"N2 118 S () J ( (iH 1. 2/l) cind (1. 31 1. 2/l) cadll. ll 1. 2 1. 2 I/AI)/II ; '3 AKC 105 2'N2 IX (s ((l I (; 1. 2/l) citd. l. l) 1. 2'1) ci,-1". 3) 1. 2/1),., 1, 1. 3) 1. 2/licad (|. 1 2X)) c ; |s1, 1. 1. 2/D 1 AYC 125 211 N2/181) f)) I c ; h 1. 2 1. 2 IIA I) III AKC10221N2) 727) xr !/). 2) ! -2). 2/t.) LI/<A !)/) tn At ; C 102'1 N ? IOS. I7=7. I : K I. I/I. ? I. I/I. ? I.''/I.) I, 1 fo\I>/II II AfiC 103''I N ? Ill-Ii. . 17n I>In I. I/I. ? 11 : 1 (1. 7/1 : 1) 1) c : vl ( !. l) IS f. IlIIAI) l1S . At : C 171'S N ? Il' !'U (II. IvI I) c : m ! i : ? 1) cnl (1.. 1) ! ll/ 1.'~'/1. 7 < I. ?/1. J kKC 128 2 (, N2 1131 3113. Ali) I) e ; i, l I) citl I) c ; i, l AKC 129 27 N2 1/17 111S Aiii i) e ; itl I) c ; ill 1. 1/1. 2 AYC 130 2x N2 1X) 5 111 1 1) 111 I) c ; sd l/AI)/11/IAI)/II IIAI) I-II Cl. ; ItAI)/lt AKC 131 21) N2 1135 33 I) c : ttt 1. 1 IIAI)/-, I) IIAD/-Il IIAI)/-]) AKC 132 Jn t12 1 23 il l) cal1 (1. 2) 1) I) call nAl) m llAl) m AKC 133) 1 N ? Ii. l l J 1). I I I I) cml f hal I) c : nl f) caJ nr : nl I) c ; l, l AliC 340 32 N2 111-1 2tS (. S IIS 1. 1 1. 1 IIAI)/II ! I/AI)/II ! DAI)/lt 11 A1 ; C 134 J7 N ? IIIO' ? l. lll : I I l) cal 4) c : vt I) call IlAD/ll/1t1)/11 () 1 ; AKC 032N2/N-) 2<n).)).) MAn/H ! « At)/) i ! « A))/tt AJ) C13. 33N2/NO2.)"AUf) c) nc.'tt)) caAD/HMAU/)) OK Cl} nu CON) HQ ! OKU ! QH !. HOK"K

Example 3-Nematicidal Activity of Anthelmintic Compositions 32-46 The C. elegarts nematode activity assay for anthelmintic compounds 32-46 was similar to that described in Example 2 above, except for the following noted differences.

The compound concentrations were adjusted to 140 µM and subjected to 2-fold dilutions to yield 140ktM, 704M, 35, uM, 17. 5µM, 8. 8µM, 4. 4, aM, 2. 2, uM, and 1. 09, uM. The visual evaluation of viability was conducted at Day 4, and the results are presented in Table 2.

Table 2. Compund µM Concentration 140 70 35 17. 5 8. 8 4. 4 2. 2 1. 09 APC-934 L1 L1 L1 L2 #B OK OK OK APC-6972 L3/L4 L4/AD/B B #B OK OK OK OK APC-6897 Ll Ll Ll < OK OK OK OK APC-7076 L1/L2 L2/L3 L3 B ! B OK OK OK APC-7077 L1/L2 L2/L3 L3 B ! B < OK OK APC-7078 L2 L2/L3 L3 L4/AD/B! B #B OK OK Control OK OK OK OK OK OK OK OK Example 4-Activity Against Nematode (C. elega71s Eggs Compositions of the subject invention are surprisingly found to be ovicidal. The following procedures are used to test for lethal effects against nematode eggs.

Materials As referred to herein,"S Medium"refers to"S basal"supplemented with Cal2, MgSO4, and a trace metals solution as follow : S basal

NaCI 5. 857 g 1M potassium phosphate (pH 6) 50. 0 ml Cholesterol (5mg/ml in EtOH) 1. 0 ml dH20<BR> 1 L The above preparation is then autoclaved. S basal can be stored until needed.

Just prior to use, S Medium is made from S basal by adding, asceptically, the following components to 1L S basal (components should first be autoclaved separately) : 1M potassium citrate (pH 6) 10 ml Trace metals solution (see below) 10 ml 1M CaCI2 3 ml 1 M MgSO4 3 ml Trace Metals solution Na2EDTA 1. 86 g (t9 5mM) Fe2SO4 7H20 0. 69 g (to 2. 5mM) MnCl#4H2O 0. 20 g (to lmM) ZnS04-7H20 0. 29 g (to 1mM) CuSO4#5H2O 0. 025 g (to 0. 1mM) dH20 1L Procedure : 1. Make anthelmintic compound dilutions as indicated in Examples 2-3.

2. To 500 ul of each dilution, added 10 u1 of eggs (estimated >200 eggs/10 µl).

3. Mixed well and allowed to incubate at room temperature for from 30 minutes to 3 hours.

4. Centrifuge at 2000 rpm for 5 minutes at room temperature.

5. Pipette off supernatant.

6. Re-suspend in 500 tl S Medium.

7. Centrifuge at 2000 rpm for 5 minutes at room temperature

8. Pipette off supernatant.

9. Re-suspend in 300 gl S Medium.

10. Transfer 300, u1 into 24-well tissue culture bioassay tray.

11. Add 2 il of stationary phase E. coli to each well.

12. Score after 3 days at room temperature in the dark.

Example 5-Additional Observations of Activity Against Nematode (C. elegans) Eggs Additional tests are conducted to confirm the ovicidal activity. The following procedures are used.

1. Make anthelmintic compound dilutions to 2X. concentrations shown in Example 4.

2. Distribute 0. 5 ml of each dilution into 1. 5-ml Eppendorf tubes.

3. Add 0. 5 ml of C elegai7s egg preparation to 0. 5 ml 2X dilution to yield final exposure concentration.

4. Mix well and allow to incubate at room temperature for from 30 minutes to 3 hours.

5. Centrifuge at 2000 rpm for 5 minutes at room temperature.

6. Pipette off supernatant and re-suspend in 1. 5 mi S Medium.

7. Spin as above for 2 minutes.

8. Pipette off supernatant and re-suspend in 1. 5 ml S Medium.

9. Repeat &num 7.

10. Pipette off supernatant and re-suspend in 1. 0 ml S Medium.

11. Add 280 p1 of S Medium to each well of 24-well tissue culture plate.

12. Add 20 Vi of each treated (and control) sample in triplicate into the respective wells.

13. Score after 3 days at room temperature in the dark.

Example 6-Preparation of Anthelmintic Compounds 47. as Specifically Exemplified bv Compounds 1-7 While the anthelmintic compounds of the subject invention can readily be produced using procedures well known to those skilled in the art. The following is a

preferred method of producing anthelmintic Compounds 47, as exemplified by Compounds 1-7, as shown in Figures 47 and 1-7.

The experiments to optimize the reaction conditions as well as to synthesize the standards were generally performed in 12 x 75 mm test tubes, using a preheated VWR Brand Dry Block Heater (VWR Scientific, catalog #13259-030) with a 16-hole, 12-13 mm test tube heating block (VWR Scientific, catalog &num 13259-120). Later experiments and precursor validation were performed in Beckman 2 mL square-well microtiter plates.

Most reagent additions during the validation phase of development were accomplished with either single-channel pipettors (e. g. Oxford Benchmate or Eppendorf repeator pipets) or 8-channel Matrix pipettors. All the isocyanates were commercially available at the time this document was drafted and were used as received. EDC was purchased from Advanced ChemTech. DMAP was purchased from Aldrich. DIPEA and Et3N were from Acros. Dowex-1 anion exchange resin (hydroxide form, Sigma catalog number I-9880) was washed according to the following sequence : methanol, chloroform, 50% aqueous methanol, using one portion per solvent. SLE's were performed using bulk packing material ("hydromatrix") supplied by Varian (catalog number 0019-8003) and used as received. Some starting materials were synthesized and their preparation is described below.

Preparation of Starting Materials.

The ten amidoximes employed in the library (see Table 7) are either commercially available or can be prepared by treatment of the corresponding commercially available benzonitriles with hydroxylamine hydrochloride and base in ethanol in moderate to high yields. Six of the sixteen BOC-amino acids employed in the library (see Table 8) were purchased from various vendors. The remaining 10 were prepared in high yield by treatment of the corresponding commercially available amino acids with di-t-butyl dicarbonate and NaOH in aqueous THF. Table 10 lists commercially available building blocks useful for synthesis of the subject compounds. All synthesized precursors were characterized by HPLC, MS, and NMR. In cases where BOC-amino acids could not be satisfactorily characterized by these methods, these intermediates were subsequently characterized by conversion to the corresponding 1, 2, 4-oxadiazole.

General Procedure Method A : 4-Methoxybenzamidoxime.

To a stirred solution of 12. 0 g (0. 172 mol) of hydroxylamine hydrochloride and 22. 2 g (0. 172 mol) of DIPEA in 350 mL of ethanol was added 19. 2 g (0. 144 mol) of 4-methoxybenzonitrile. The resulting mixture was stirred at room temperature for 18 h overnight, then concentrated in vacuo. The oily residue was triturated with 300 mL of water and the resulting precipitate was filtered, washed with H20, then dried in vacuo to afford 17. 5 g (73%) of the desired product. MS (ESI) m/z 167 (M+H, 100%).'H-NMR (DMSO-d6) 6 3. 75 (s, 3H), 5. 75 (s, 2H), 6. 90 (d, 2H), 7. 65 (d, 2H), 9. 45 (s, 1H).

4-Methylbenzamidoxime (Method A). A mixture of 19. 5 g (0. 167 mol) of 4-methylbenzonitrile, 13. 9 g (0. 20 mol) of hydroxylamine hydrochloride, and 25. 8 g (0. 20 mol) of DIPEA in 350 mL of EtOH afforded 21. 6 g (86%) of a white solid. MS (ESI) m/z 150 (100%).'H-NMR (DMSO-d6) 6 2. 28 (s, 3H), 5. 77 (s, 2H), 7. 2 (d, 2H), 7. 6 (d, 2H), 9. 57 (s, 1H).

Piperonylamidoxime (Method A). A mixture of 20. 0 g (0. 136 mol) of piperonylonitrile, 11. 3 g (0. 163 mol) of hydroxylamine hydrochloride, and 21. 0 g (0. 163 mol) of DIPEA in 350 mL of EtOH gave 16. 4 g (67%) of light yellow crystals. MS (ESI) m/z 180 (100%).'H-NMR (DMSO-d6) 6 5. 75 (s, 2H), 6. 03 (s, 2H), 6. 88 (d, 1H), 7. 23 (d, 2H), 9. 51 (s, 1H).

3, 4-Dimethylbenzamidoxime (Method A). A mixture of 20. 0 g (0. 152 mol) of 3, 4-dimethylbenzonitrile, 12. 7 g (0. 183 mol) of hydroxylamine hydrochloride, and 23. 6 g (0. 183 mol) of DIPEA in 350 mL of EtOH to afford 17. 0 g (68%) of light yellow crystals after recrystallization from ethyl acetate. MS (ESI) m/z 165 (M+H, 100%).

'H-NMR (DMSO-d6) 6 2. 23 (s, 6H), 5. 75 (s, 2H), 7. 2 (d, 1H), 7. 36-7. 48 (m, 2H), 9. 5 (s, 1H).

4-Methylsulfonylbenzamidoxime (Method A). A solution of 22. 5 g (0. 124 mol) of 4-methylsulfonylbenzonitrile, 10. 4 g (0. 149 mol) of hydroxylamine hydrochloride, and 19. 3 g (0. 149 mol) ofDIPEA in 350 mL of EtOH afforded 25. 3 g (95%) of a white solid.

MS (ESI) m/z 215 (M+H, 100%).'H-NMR (DMSO-d6) 8 3. 23 (s, 3H), 6. 0 (s, 2H), 7. 87 (s, 4H), 9. 98 (s, 1H).

3-Methoxybenzamidoxime (Method A). The mixture from 20. 0 g (0. 150 mol) of 3-methoxybenzonitrile, 12. 5 g (0. 180 mol) of hydroxylamine hydrochloride, and 23. 2 g

(0. 180 mol) of DIPEA in 350 mL of EtOH was concentrated in vacuo and the residue was filtered through a short plug of silica gel (eluant : ethyl acetate). The filtrate was concentrated in vacuo and triturated with water. The resulting solid was filtered and dried to afford 19. 5 g (78%) of the desired product. MS (ESI) m/z 167 (M+H, 100%).

'H-NMR (DMSO-d6) 6 3. 77 (s, 3H), 5. 85 (s, 2H), 6. 9-7. 35 (m, 4H), 9. 7 (s, 1H).

3-Methylbenzamidoxime (Method A). The mixture from 20. 0 g (0. 171 mol) of 3-methylbenzonitrile, 14. 2 g (0. 205 mol) of hydroxylamine hydrochloride, and 26. 4 g (0. 205 mol) of DIPEA in 350 mL of EtOH was concentrated in vacuo and the residue was partitioned between DCM and a minimum amount of water. The layers were separated and the DCM layer was stirred with a few grams of silica gel for 10 min. The suspension was filtered then concentrated in vacuo to afford 21. 3 g (83%) of the desired product as a solid. MS (ESI) m/z 151 (M+H, 100%).'H-NMR (DMSO-d6) 6 2. 33 (s, 3H), 5. 75 (s, 2H), 7. 2-7. 5 (m, 4H), 9. 6 (s, 1H).

4-Z-Butoxybenzamidoxime (Method A). The residue from a mixture of 20. 0 g (0. 114 mol) of 4-n-butoxybenzonitrile, 9. 52 g (0. 137 mol) of hydroxylamine hydrochloride, and 17. 7 g (0. 137 mol) of DIPEA in 350 mL of EtOH was triturated with water. The resulting solid was suspended in hexane, stirred for 1 h at room temperature, and filtered to afford 21. 8 g (92%) of a white solid. MS (ESI) m/z 209 (M+H, 100%).

'H-NMR (DMSO-d6) 6 0. 85 (s, 3H), 1. 35-1. 74 (m, 4H), 3. 95 (m, 2H), 5. 8 (s, 2H), 6. 90 (d, 2H), 7. 64 (d, 2H), 9. 50 (s, 1H).

Method B : 2-Methoxybenzamidoxime.

A suspension of 11. 5 g (0. 165 mol) of hydroxylamine hydrochloride, 17. 5 g (0. 165 mol) of Na2CO3, and 20. 0 g of 2-methoxybenzonitrile in 350 mL of EtOH and 30 mL of H2O was heated at 80 °C for 10 h. After cooling to room temperature, the mixture was filtered, and the filter-cake washed with EtOH. The filtrate was concentrated in vacuo to give a semi-solid product, which was triturated with a mixture of ether/hexane, and the white solid was filtered, washed with hexane then dried to afford 17. 4 g (70%) of the desired product. MS (ESI) m/z 167 (M+H, 100%).'H-NMR (300 MHz, DMSO-d6) 6 3. 81 (s, 3H), 5. 6 (s, 2H), 6. 90 (t, 1H), 7. 10 (d, 1H), 7. 35 (t, 2H), 9. 40 (s, 1H).

General Procedure for BOC-protection : 4- (BOC-aminomethyl) benzoic acid.

To a solution of 5. 80 g (0. 145 mol) of NaOH in 250 mL of H20 was added 20. 0 g (0. 132 mol) of 4- (aminomethyl) benzoic acid. After the acid had completely dissolved, a solution of 31. 8 g (0. 145 mol) of di-t-butyl-dicarbonate in 100 mL of THF was added.

The mixture was stirred at room temperature overnight then concentrated in vacuo to remove most of the THF. The resulting aqueous layer was acidified to pH 2-3 with solid KHSO4. The mixture was extracted with ether and the combined extracts dried (MgSO4) and concentrated in vacuo to afford 32. 7 g (99%) of a white solid.'H-NMR (300 MHz, DMSO-d6) 6 1. 39 (s, 9H), 4. 20 (d, 2H), 7. 36 (d, 2H), 7. 48 (t, 1H), 7. 88 (d, 2H).

BOC-trans-4- (Aminomethyl) cyclohexanecarboxylic acid. According to the general procedure, a mixture of 15. 7 g (0. 10 mol) trans-4- (aminomethyl) cyclohexanecarboxylic acid, 4. 40 g (0. 110 mol) of NaOH, and 24. 0 g (0. 110 mol) of di-t-butyl-dicarbonate in 100 mL of THF and 250 mL of water gave 23. 2 g (90%) of the desired product as a white solid.'H-NMR (300 MHz, DMSO-d6) 6 0. 75-0. 95 (m, 2H), 1. 35 (s, 9H), 1. 22-1. 3 (m, 3H), 1. 73 (d, 2H), 1. 85 (d, 2H), 2. 13 (m, 1H), 2. 80 (t, 2H), 6. 79 (t, 1H).

BOC-DL-3-Aminocyclohexanecarboxylic acid. According to the general procedure, a mixture of 20. 0 g (0. 14 mol) of the 3-aminocyclohexanecarboxylic acid (stereochemistry undefined), 6. 16 g (0. 154 mol) of NaOH, 33. 49 g (0. 154 mol) of (BOC) 2O in 120 mL THF and 250 mL water afforded 28. 7 g (84. 4%) of a white solid.

BOC-4-Aminocyclohexanecarboxylic acid. According to the general procedure, a mixture of 20. 0 g (0. 140 mol) of the 4-aminocyclohexanecarboxylic acid (a cis/trans mixture), 6. 16 g (0. 154 mol) of NaOH, and 33. 5 g (0. 154 mol) of (BOC) 20 in 120 mL THF and 250 mL water afforded 24. 2 g (71%) of the desired product as a colorless solid.

BOC-DL-3-Aminobutyric acid. To a solution of 6. 40 g (0. 160 mol) of NaOH in 250 mL of water was added 15. 0 g (0. 145 mol) of DL-3-aminobutyric acid. To this solution was added 160 mL (0. 160 mol) of 1. 0 M solution of (BOC) 20 in THF. The resulting mixture was stirred at room temperature overnight, then processed according to the general procedure to afford 22. 5 g (76%) of the desired product as a white solid.

BOC-DL-B-Aminoisobutyric acid. According to the general procedure, a mixture of 20. 0 g (0. 194 mol) 6fDL- (-aminobutyric acid, 8. 52 g (0. 213 mol) of NaOH, 213 mL (0. 213 mol) of 1. 0 M (BOC) 20 in THF and 250 mL water gave 38. 6 g (98%) of a white

solid.'H-NMR (300 MHz, DMSO-d6) 6 0. 17 (d, 3H),, 0. 54 (s, 9H), 1. 60-1. 70 (m, 1H), 2. 02-2. 12 (m, 1H), 2. 25-2. 35 (m, 1H), 6. 00 (t, 1H), 11. 35 (s, lH) BOC-DL-3-Amino-3-phenylpropionic acid. According to the general procedure, a mixture of 20. 0 g (0. 121 mol) of DL-3-amino-3-phenylpropionic acid, 5. 32 g (0. 133 mol) of NaOH, 133 mL (0. 133 mol) of 1. 0 M (BOC) 20 in THF and 250 mL of water gave 25. 7 g (80%) of a white solid.'H-NMR (300 MHz, DMSO-d6) 6 0. 50 (s, 9H), 1. 71-1. 80 (m, 2H), 4. 05 (t, 1H), 6. 35-6. 45 (m, 5H), 6. 70 (d, 1H), 11. 37 (s, 1H).

BOC-DL-Nipecotic acid. To a stirred solution of 4. 19 g (0. 105 mol) of NaOH in 100 mL of water was added 13. 0 g (0. 101 mol) of DL-nipecotic acid. This solution was cooled in an ice water bath then treated with 100 mL (0. 100 mol) of 1. 0 M (BOC) 2O in THF. The resulting mixture was stirred at room temperature for 15 h, then concentrated in vacuo to remove most of the THF. The resulting aqueous solution was washed with ether, then acidified with H3PO4 (10 mL). The white precipitate was filtered, washed with water, and dried under high vacuum to afford 21. 5 g (93%) of a white powder.'H-NMR (300 MHz, DMSO-d6) 8 1. 38 (s, 9H), 1. 42-1. 63 (m, 2H), 1. 86-1. 91 (m, 2H), 2. 24-2. 32 (m, 1H), 2. 81 (dt, 1H), 3. 66 (br d, 1H), 3. 89 (br s, 2H), 12. 3 (s, 1H).

BOC-4-Piperidinoacetic acid. A suspension of 24. 3 g (0. 140 mol) of 4-pyridylacetic acid hydrochloride and 2. 07 g of Pt02 in 150 mL AcOH was hydrogenated at 50 psi. After hydrogen uptake had ceased, the mixture was kept at 50 psi for 30 min, then purged with nitrogen for 15 min. The mixture was filtered and the catalyst was washed with water. CAUTION : THE CATALYST MUST BE KEPT WET WITH WATER AT ALL TIMES, OTHERWISE A FIRE WILL RESULT. DO NOT WASH THE CATALYST WITH FLAMMABLE ORGANIC SOLVENTS SUCH AS METHANOL OR ETHANOL. The filtrate and washings were concentrated in vacuo to give a colorless semisolid mixture which was triturated with 250 mL of diethyl ether and the resulting suspension was stirred for a few hours. The solid was filtered, washed with ether and hexane, then dried in vacuo to give 25. 4 g (100%) of 4-piperidineacetic acid hydrochloride as a white powder.

To a solution of 12. 0 g (0. 300 mol) of NaOH in 300 mL water was added the solid isolated above. The resulting solution was cooled in an ice water bath and treated with 100 mL of THF, followed by 140 mL (0. 140 mol) of 1. 0 M (BOC) O in THF. The

resulting solution was stirred at room temperature overnight. The THF was removed in vacuo and the resulting aqueous solution was washed with ether, then acidified to pH 1-2 with 85% H3PO4. The solution was extracted with ethyl acetate, then the combined organic extracts were washed with brine, dried (Na, SO,), and concentrated in vacuo to give 29. 1 g (86%) of a colorless, viscous oil which solidified upon drying in vacuo to give a white solid.'H-NMR (300MHz, DMSO-d) 1. 01 (dq, 2H), 1. 37 (s, 9H), 1. 60 (br d, 2H), 1. 73-1. 82 (m, 1H), 2. 12 (d, 2H), 2. 67 (br s, 2H), 3. 88 (br d, 2H), 12. 1 (s, 1H).

BOC-DL-3- (3-piperidino) propionic acid. A suspension of 24. 8 g (0. 166 mol) of 3- (3-pyridyl) acrylic acid in DCM was treated with 45 mL of 4N HCI in dioxane for 2h, then diluted with ether and filtered. The solid was washed with ether, and dried in vacuo to afford 31. 0 g of a colorless solid. The solid was suspended in 150 mL of acetic acid and 2. 71 g of PtO2 was added. The suspension was hydrogenated at 50-55 psi until hydrogen uptake had ceased. The mixture was diluted with 50 mL of water, filtered, and the catalyst washed with water, keeping the catalyst wet at all times. The combined filtrate and washings were concentrated in vacuo to give 31. 0 g (96%) of the piperidinopropionic acid as a white powder.

The above solid was added to a stirred solution of 13. 1 g (328 mmol) of sodium hydroxide in 250 mL of water, then the reaction mixture was cooled in an ice water bath.

After the solids had dissolved, 160 mL (160 mmol) of a 1. 0 M solution of (BOC) 2O in THF was added via an addition funnel. An additional 80 mL of THF was used to wash the addition funnel. The reaction mixture was stirred for 68 hours, allowing the ice bath to warm up to room temperature. The mixture was concentrated in vacuo to remove most of the TIF and the resulting aqueous solution washed with diethyl ether (300 mL). The aqueous phase was acidified to pH 2-3 with 15 mL of 85% phosphoric acid, the solution was then extracted with ethyl acetate (300 mL). The extract was washed with saturated aqueous NaCl (2x100 mL), dried (Na2SO4) and concentrated in vacuo to afford 39. 5 g (96%) of a white solid.'H-NMR (300 MHz, DMSO-d6) 6 1. 00-1. 44 (m, with s at 1. 37 ppm, 13H), 1. 52-1. 57 (m, 1H), 1. 70-1. 75 (m, 1H), 2. 23 (t, 2H), 2. 78 (br t, 2H), 3. 68 (br s, 2H), 12. 1 (s, 1H).

Library Synthesis.

Step A : O-Acylation and Cyclization.

Caution : Moisture Sensitive Reactions.

The following chemistry is moisture-sensitive. All solutions must be prepared from anhydrous solvents (e. g. Aldrich"Sure/Seal"), ideally just before they are to be added to the plates. Furthermore, reagent additions should be done as quickly as possible to minimize moisture accumulation from the atmosphere on standing.

To 2-mL square well Beckman plates was added 700 L (0. 140 mmol) of a 0. 2 M solution of the BOC-amino acids in 1, 4-dioxane using a Robbins HydraT""96-well dispenser (Robbins Scientific, catalog number 1029-80-1) to the assigned wells. Each BOC-amino acid solution was then treated with, in the following sequence, 80 L (0. 04 mmol) of a 0. 5 M solution of 4-DMAP in 1, 4-dioxane and 140 L (0. 140 mmol) of a 1. 0 M solution of EDC in CHC13, using the Robbins HydraTX'to add both reagents. The resulting mixtures were shaken on an IKL orbital shaker (VWR Scientific, catalog number 33994-220) for 5-10 min followed by 700 VL (0. 140 mmol) of a 0. 2 M solution of the appropriate hydroxyamidine in 1, 4-dioxane. Each plate was covered with a teflon sheet, clamped and shaken on a Lab line reciprocal shaker (VWR Scientific, catalog number 57008-195 ; setting 6) for a minimum of 18 hours.

The plates were removed from the shaker and unclamped. To each well was added 20 L (0. 140 mmol) of neat Et3N. The plates were then shaken, unclamped, on a reciprocal shaker (setting 5) for 4-5 minutes, then the plates were heated, uncovered, in a preheated (100°-105°C) nitrogen-purged oven (VWR Scientific, catalog number 52201-656) for 7 hours. The plates were removed from the oven and allowed to cool to room temperature. Generally the solvents will have evaporated when the plates are removed from the oven.

The contents of each well was dissolved in 1. 0 mL of CHCl3 then 300 L of 10% aqueous citric acid solution was added to each well. The plates were shaken on a reciprocal shaker for 2 h. The two-phase mixtures were transferred to Polyfiltronics plates (type PP, 10 lm) with wells previously half-filled with hydromatrix material and pre-activated with 500 L of 10% aq. citric acid and the plates were placed over 2-mL square-well Beckman plates. Each source well was rinsed once with 250 L of CHCI3 then transferred to the Polyfiltronics plate. Another 2x250 uL of CHCl3 were added to each well of the Polyfiltronics plate. After the contents of the wells were allowed to drain,

the collection plates were concentrated in a Genevac evaporator for 3-4 h (Atlas, catalog number HT-12-CDOP).

Step B : BOC removal.

Each well was treated with 1. 0 mL of a 1 : 1 mixture (v : v) of TFA in DCM. A teflon sheet was placed on top of each plate secured with a rubber band and was shaken on a reciprocal shaker for 2 hours. The plates were concentrated in the Genevac evaporator for 3-4 h using the ramping function. After evaporation, the resulting well contents of the plates were redissolved in 1. 0 mL of 50% aqueous ACN, and the plates were shaken on an IKL Works microtiter plate shaker (VWR Scientific, catalog number 33994-220) for 30 min or the well contents were agitated in parallel using a modified Chiron Mimetopes"PIN"holder with fitted with 96 pegs to dissolve the samples before being frozen in a-80°C freezer (Revco, catalog ULT-2586-7 A) for at least 5 h (preferably overnight). The plates were then lyophilized in a tray lyophilizer (Virtis Unitop, catalog number 800L ; tray temperature : 20°C) for 18 h.

Step C : Urea Formation.

Using the Robbins Hydra', the lyophilized products were treated with 450 gL (0. 182 mmol) per well of a 0. 4 M solution of DIPEA in CHCl3 and shaken for 5-10 min.

To each mixture was added 840 uL (0. 126 mmol) of the appropriate isocyanate (see Table 9) as a 0. 15 M solution in CHCl3, employing the Robbins Hydra for the reagent additions. Each plate was covered with a Teflon sheet, clamped, and shaken on a reciprocal shaker for 18 h.

The plates were removed from the shaker and 300 L of a 10% aqueous Na, C03 solution was added to each well using the Robbins Hydra". The plates were shaken on a reciprocal shaker for 2 h, then the mixtures were transferred using the Robbins HydraTM to Polyfiltronics plates (PP, 10 um) with wells previously half-filled with hydromatrix material and pre-activated with 500 (L of 10% aqueous Na2CO3. The plates were placed over 2-mL square-well Beckman plates. Each well was rinsed once with 250 uL of CHC13 that was collected in the Beckman plates. Another 2x250 PL of CHCi3 was added to each well of the Polyfiltronics plates and allowed to drain into the Beckman plates. The Beckman plates should be about 3/4 full (ca. 1. 5 mL) with solvent.

To each well was added 300 L of 2 N aqueous HC1 and the plates were shaken on a reciprocal shaker for 2 h. The mixtures were transferred to Polyfiltronics plates (PP, 10 um) with wells previously half-filled with hydromatrix material and pre-activated with 500 uL of 2 N HCl per well. The plates were placed over a 2-mL square-well Beckman plates with wells previously loaded with 100-120 mg of Dowex-1 anion exchange resin.

Each source well was rinsed with 2x250 L ofCHCIg then transferred to the Polyfiltronics plates. Another 250 uL of CHCl3 was added to each well of the Polyfiltronics plates.

After the plates were allowed to drain, the Beckman collection plates were put into a plastic container that was tightly-capped and shaken on a reciprocal shaker overnight.

The mixtures were transferred, using the Robbins Hydra fitted with small gauge needles to prevent clogging by the resin, to Polyfiltronics plates (PP, 10 m) with wells previously loaded with a thin layer of silica gel (ca 30-40 mg ; Baxter Scientific Products, 60A, 230-400 mesh ; catalog number C4582-85). The Polyfiltronics plates were placed on top of 2-mL Beckman plates. Each well of the reaction plates were rinsed with CHCl3 (2x250 pL) and transferred to the Polyfiltronics plates. The solvent was evaporated on the Genevac evaporator for 3-4 hours.

ACN (1. 25 mL/well) was added and the plates were shaken on an orbital shaker for 30 min then sonicated for another 15-20 min. The plates were centrifuged for 30 min in either the Savant or Genevac evaporators without applying heat or vacuum. The resulting solutions were transferred by the Robbins Hydra'to a set of second, TARED 2-mL square-well Beckman plates. The plates were placed in the-80°C freezer for at least 5 h (preferably overnight), then lyophilized in the tray lyophilizer (tray temperature : 20°C) for 18 h overnight.

Note on Solvents for QC. Samples submitted for direct injection QC analysis must be diluted with a mixture of 90% ACN and 10% of a 2. 0 molar solution of ammonia in methanol (Aldrich ; catalog number 34, 142-8). The use of ammonia in methanol as a buffer for QC analysis improved ionization during direct injection analysis of samples for this library.

Development General Comments.

Early development experiments were run in 12x75 mm test tubes and reactions were heated in a VWR Scientific Dry Block heater. All SLE and other purification experiments were done in Polyfiltronics plates.

Hydroxyamidine Synthesis.

In most cases these precursors were prepared using literature methods. Since only one hydroxamidine (ACD #31485) was commercially available, the remaining 9 were synthesized as shown in Figure 49 from a benzonitrile (1) and hydroxylamine hydrochloride. Heating was sometimes necessary to drive the reaction to completion.

Other conditions explored for hydroxyamidine preparation included several bases (K2CO3, Na2CO3, NaOCH3, Et3N and DIPEA), and solvents (methanol or ethanol). The most suitable conditions identified were NH20H-HCl/DIPEA/ethanol at room temperature as described above or NH20HvHCI/Na2CO3/aqueous ethanol at 80°C.

BOC-amino Acids.

While most commercially available BOC-a-amino acids failed to give product with sulfonyl chlorides, the use of acid chlorides did afford the desired products. Additional BOC-amino acids needed to expand the diversity for the library were synthesized from commercially available amino acids with the exception of two cases which were prepared by catalytic hydrogenation of a pyridine-containing acid and subsequent BOC-protection.

An example is shown in Figure 50.

Acid Coupling and Cyclization (Step A).

There was ample literature precedent for the coupling of the hydroxyamidines to the BOC-amino acid and subsequent cyclization to afford the 1, 2, 4-oxadiazole ring. For development of a suitable production method for parallel synthesis in microtiter plates, various coupling agents were explored including CDI, DIC and EDC. The latter reagent was preferred since the SLE conditions were able to remove byproducts derived from the EDC. Bases such as DBU and triethylamine were explored in addition to catalysts such as 4-DMAP and HOBt. Lastly, several solvents and solvent mixtures were also investigated, including toluene, 1, 4-dioxane, p-xylene, 1, 2-dichloroethane,

THF/1, 4-dioxane, DCM/1, 4-dioxane, and CHC13/1, 4-dioxane. Ultimately EDC and 4-DMAP in a mixture of CHC13 and 1, 4-dioxane were established as the best conditions for 0-acylation. Cyclization of the O-acylated hydroxyamidines was done in situ in this solvent mixture by heating for a minimum of seven hours. Incomplete cyclization was noted with shorter heating times ; however prolonged heating (>24 hours) resulted in the formation of additional byproducts that were not identified.

Purification at this stage was accomplished with 10% aqueous citric acid using standard SLE material. The extraction solvent employed was CHC13. Other solvents for the SLE step were not investigated.

BOC Removal and Acylation (Step B).

Use of both TFA in DCM or 4 N HC1 in 1, 4-dioxane cleanly gave the desired salts of the amines. While the latter conditions were expected to be easier to use in production since the solvent could be removed by lyophilization, this turned out to be more difficult in practice, primarily due to solubility problems of the resulting HC1 salts. The use of TFA/DCM for the deprotection step, while avoiding the salt solubility issue, had other problems. Removal of the residual TFA by simple evaporation on the Genevac or Savant gave erratic results due to the presence of excess TFA which was not being completely neutralized when the base and acylator were added. Lyophilization of the evaporated plates from aqueous ACN circumvented this problem. The TFA salts were generally more soluble in CHCI3 used in the acylation step.

Step C was optimized for solvent and base. Solvents explored included 1, 2-DCE, DCM, ethyl acetate, THF, and CHC13 ; bases included DIPEA and NMM The best combination was DIPEA in CHC13. Purification of the final products was accomplished first with a basic SLE with 1 N aqueous KOH or 10% aqueous Na2CO3. The latter was preferred due to possible destruction of the SLE material with KOH. This purification step was followed by an acidic SLE with 2 N HCI. Both acidic (Amberlite IR-120) and basic (Amberlite IRA-67) ion exchange resins were evaluated as alternatives to the SLE steps but results were inconsistent. Use of aminomethylpolystyrene resin to scavenge unreacted isocyanate was also evaluated but recovery of material was slightly lower with little difference in purity. Due to the number of SLE steps required for the library, the

scale of the library was established at 140 mol per we4l to compensate for losses in the purification steps.

Filtration of the final products through a thin layer of silica gel reduced the amounts of unacylated amines to less than 8% as determined by HPLC-LJV at 214 nm by AUC. In addition, this step eliminated many strongly charging minor byproducts as seen by LC-MS.

Example 7 Nematicidal Activity of Anthelmintic Compositions 48-118 The nematicidal activity of anthelmintic Compositions 48-118 were determined in accordance with the procedure outlined in Example 2. The results are reported in Table 3.

Table 3 ! HTS Data HTS Tracking Initial HTS Run Follow-up HTS Run 5 Day Visual Score i mOD 1% Run Standard Visual Score Well Address V1 V2 V3 V4 V5 V6 113 ! 5061 A10 221 136%Dead (L2/L3) ! 175 139%Dead (L3) 5061 : A10 Dead IDead Dead ; Dead (L2/L3) L2/Dead (AD) L4/#AD/B ! ! B ! B !'B ! IOK OK OK 107'5061 D10 235 144% Dead (L3) 188 1i9% Dead (i3) 506i : D10 Dead Dead (L2/ii2/L3 IDead (L4) LiiDead (AD) L2/Dead (L4) 115 5061 H10 254 156%Dead (L3) i 211 ! 167%Dead (L3), 5061 : H10Dead ! bead (L2/LL2/bead (L2) bead (L2)'bead (L2/L3) L2/L3 r"j''L1Tl L1"L4/#Ab/B OK''6K 10815061 C11 223 137% ! Dead (L2/L3) 215 171%Dead 061 : C11 Dead iDead L1 IL1. L1/L2 L2JL3 114J506lbl1'221 136%'Dead (L3) 204 162%'L1/L2'5061 : D11 Dead L3/Dead (LL3 IL3/Dead (L4) i2/i3-B 11115081 D10 145%'Dead (L3) (L3) 2001 159% Dead (L3) 5081 : D10 Dead, Dead L2 Dead (L3/L4) #AD/B OK 112'5090 195 120% ! pead (L3) 183 ! 145%Dead (L3) 5090 : A10L2/L3'L2/L3 L2/L3'L3/L4 #AD/-B ok

Example 8-Sheep Test I Experimental Procedure Sheep naturally infected with a variety of gastrointestinal nematodes are purchased from local sources and are transported to the test site. The animals are housed in a manner to preclude further infection by nematode larvae. The animals are evaluated for the presence of adequate nematode burdens by performing a standard fecal egg per gram (EPG) count. Eggs are differentiated into the following groups : trichostrongyle (strongyle), Strongyloides, Trichuris, or Nematodinis. Only sheep judged by the study parasitologist to have adequate nematode infections are retained as test subjects.

The sheep are fed good quality hay (no concentrated rations) and water ad libitum.

Following a five-day acclimation period, the sheep are randomly assigned by EPG count into treatment groups which include non-treated Negative control (placebo) ; Positive Control (commercially available ivermectin for sheep) : and various anthelmintic compounds of the present invention (test compound) dissolved in DMSO. The first replicate of 10 animals is randomly assigned to groups 1-10 ; the second replicate of 10 animals is randomly assigned to groups 1-10 ; and the third replicate of 10 animals is randomly assigned to groups 1-10. Thus 10 groups of 3 animals each is created.

The randomization is performed on fecal samples collected 24-48 hours prior to scheduled treatment. The EPG counts are performed according to Zimmerman Research SOP &num NMEPG. 99. 01 On treatment day, the animals are weighed and divided into groups with three animals per group as follows : GROUP 1 : Non-treated negative control (placebo) of 10 ml of DMSO.

GROUP 2 : Positive Control treatment of 200 mcg/kg commercially available ivermectin for sheep.

GROUP 3 : Compound @ dissolved in DMSO.

GROUP 4 : Compound @ dissolved in DMSO.

GROUP 5 : Compound @ dissolved in DMSO.

GROUP 6 : Compound @ dissolved in DMSO.

GROUP 7 : Compound @ dissolved in DMSO.

GROUP 8 : Compound @ dissolved in DMSO.

GROUP 9 : Compound @ dissolved in DMSO.

GROUP 10 : Compound @ dissolved in DMSO.

The placebo (DMSO), the commercially available drug, and the test anthelmintic compounds are administered in a 3ml volume by subcutaneous injection using a sterile syringe fitted with a proper needle. The animal is adequately immobilized for injection of the placebo, commercially available drug, or test anthelmintic compound.

Following treatment, the animals are observed at hourly intervals for the first 8 hours, then daily until necropsy. They are housed in a manner to prevent further nematode infections. Fecal samples are taken for EPG counts on the 5th day and 7th day after treatment.

Seven days following treatment the sheep are humanely slaughtered in accordance with the Guide for the Care and Use of Laboratory Animals (DHEW Publication No.

86-23). Necropsy procedures are according to Zimmerman Research SOP # NCRGIH. 99. 01, Necropsy for Helminth Recovery, specifically for gastrointestinal nematodes. Fecal samples are taken for EPG counts during the sample collection process on this day. All animals are necropsied, but only the animals from the experimental treatment groups that have a significant egg count reduction on day 5 or day 7 have intestinal material collected for nematode recovery and identification.

Nematodes are recovered, identified, and enumerated according to Zimmerman Research SOP # NEMRECOVID. 99. 01. All individuals performing nematode recoveries are blinded to treatment versus control animals. Preliminary estimates of total nematodes recovered from each gut sample are provided prior to identification and enumerations by the study parasitologist. At the discretion of the study parasitologist, seven days after the drug administration fecal egg counts are performed and all animals showing 90% or better trichostrongylid egg reduction are slaughtered using humane methods recommended by the AVMA. The neck blood vessels are severed and after the animal is completely exsanguinated, Table 7 Hydroxyamidines.

The quantities specified below are for a set of six plates according to the library layout.

380 mL of a 0. 2 M solution of each hydroxyamidine is required for each set of plates. Entry ACD MW Amt. () Name Structure I 19952 150 11. 4 4-methylbenzamidoxime form - 2 31485 136 10. 34 benzamidoxime MOH I \, NH2 / 3 NA 166 12. 62 4-methoxybenzamidoxime OH NH, 0 4 119015 180 13. 68 piperonyloamidoxime OH rNH2 n 5 NA 166 12. 62 2-methoxybenzamidoxime OH Nu2 / 6 NA 208 15. 81 4-n-butoxybenzamidoxime OH Fez O 7 NA 164 12. 46 3, 4-dimethylbenzamidoxime OH NH2 / 8 NA 214 16. 26 4-methylsulfonylbenzamidoxime oH NH2 02 02 9 NA 166 12. 62 3-methoxybenzamidoxime OH NH / 10 NA150 114 3-methytbenzamidoxime H I \, NHz Table 8 BOC-amino acids The quantities specified below are for a set of six plates according to the library layout.

30 mL of a 0. 2 M solution of each BOC-amino acid is required for each set of plates. Entry ACD MW Amt. (Name Structure 1 NA 257 1. 542 BOC-trans-4- (aminomethyl) cyclohexanecarboxylic acid,,. HOOT 2 63356 245 I. 47 BOC-7-aminoheptanoic acid HOOC ~<N-BOC H 3 76999 229 1. 374 BOC-isonipecotic acid C N, BOC HOOC 4 NA 243 1. 458 BOC-4-aminocyclohexane H carboxylic acid (cis/trans S BOC mixture) H 5 37291 189 1. 134 BOC-beta-Ala-OH HOOC---N'BOC 6 37313 203 1. 218 BOC-4-aminobutyric acid H H 7 27033 203 1. 218 BOC-DL-3-aminobutyric acid 8 HOOCH BOC 8 37798 231 1386 BOC-6-aminohexanoic acid HOOCN'BC H 9 22818 251 1. 506 BOC-4- (aminomethyi) benzoic N-BOC 2 acid H HOOT 10 NA 229 1. 374 BOC-nipecotic aicd HOOCK BOC 11 NA 243 1. 458 BOC-3-aminocyclohexane HOON BC carboxylic acid (stereochemistry c) BOC undefined) v 12 NA 203 1. 218 BOC-DL-beta-aminobutyric acid HOOCH BOC 13 76903 217 1. 302 BOC-5-aminopentanoic acid HOOCN BOC 14 NA 243 1. 458 BOC-4-piperidinoacetic acid BOC HO (X) 15NA 265r59Boc-DL-3-amino-3- phenylpropionic acid HOOCv BOC C 16 NA 257 1. 542 BOC-3- (3-piperidino) propionic HOOCHS, BOC acid Table 9 Isocyanates The quantities specified below are for a set of six plates according to the library layout.

17 mL of a 0. 15 M solution of each isocyanate is required for each set of plates. Entry ACD MW Amt. (g) Name Structure 1 8882 203 0. 518 2, 6-diisopropylphenyl isocyanate ! neo 2 37076 165 0. 421 2-(methylthio) phenyl X isocyanate ! neo 37072 0. 385 5-fluoro-2-methylphenyl isocyanate NCO F F 4 13863 165 0. 421 3-(methylthio) phenyl oSgNCO isocyanate t !) [ 5 13867 147 0. 375 3, 4-dimethylphenyl NCO isocyanate | l l w 6 13879 161 0. 411 4-acetylphenyl NCO isocyanate OX Zu 7 13887 155 0. 396 octyl isocyanate 8 1994 119 0. 304 phenyl isocyanate NCO 9 19909 161 0. 411 2, 4, 6-trimethylphenyl isocyanate NEO I 10 2029 133 0. 339 p-tolyl isocyanate NCO 11 14705 178 0. 454 4-methyl-2-nitrophenyl N02 isocyanate NCO 12 19912 175 0. 447 2, 6-diethylphenyl isocyanate NCO 13 144846 113 0. 289 n-amyl isocyanate 14 1996 137 0. 350 2-fluorophenyl < NCO isocyanate F 15 1999 153 0. 392 2-chlorophenyi NCO isocyanate au 1 CI 16 2004 149 0. 380 2-methoxyphenyl NCO isocyanate 17 2005 179 0. 457 2, 4-dimethoxyphenyl isocyanate ! NCO 0 18 2006 179 0. 457 2, 5-dimethoxyphenyl 0, NCO isocyanate // 19 2007 163 0. 416 2-ethoxyphenyl N isocyanate o0m 20 2008 187 0. 477 2-trifluoromethylphenyl NCO isocyanate caf3 CPg 21 35702 203 0. 518 4- (trifluoromethoxy) NCO phenyl isocyanate F 3Csow O 22 2022 198 0. 505 4-bromophenyl NCO isocyanate B ria 23 2042 71 0. 1 81 ethyl isocyanate NCO 24 2045 85 0. 217 n-propyl isocyanate 25 2046 99 0. 253 n-butyl isocyanate wNCO 26 209615 155 0. 396 tert-octyl isocyanate- < NCO 27 2019 149 0. 380 3-methoxyphenyl O NCO isocyanate W 28 41744 211 0. 539 dodecyl isocyanate CH3 (CH2) » NCO 29'37042 191 0. 488 4-n-butoxyphenyl NCO isocyanate O 30 7046 178 0. 454 2-methyl-3-nitrophenyl NCO isocyanate N02 NOZ 31 37047 127 0. 324 hexyl isocyanate NCO NLU 32 13858 167 0. 426 3-chloro-4-methylphenyl NCO isocyanate ! ! i w ') 7087 195 0. 498 2-biphenyl isocyanate I NCO Table 10 Commercially available building blocks for the preparation of the Hydroxyamidines and BOC-amino acids. Entry ACD MW Name Structure 1 1827 117 p-tolunitrile CN i 2 1818 133 4-methoxybenzonitrile ACN 3 5820 147 piperonylonitrile <0 CN 0 4 1783 133 2-methoxybenzonitrile . CN rl i 5 43482 208 4-n-butoxybenzonitrile CN 6 16380 164 3, 4-dimethybenzonitrile CN i 7 216489 214 4-methylsulfonylbenzonitrile CN xi 8 1801 166 3-methoxybenzonitrile i 9 1808 117 m-tolunitrile CN 10 1466 157 trans-4- (aminomethyl) NH2 cyclohexanecarboxylic acid ! J HOOT 11 6004 129 isonipecotic acid NH HOOT 12 59562 143 3-aminocyclohexanecarboxylic acid HOOCNH2 (stereochemistry undefined) X) 13 191601 143 4-aminocyclohexanecarboxylic acid NH (cis/trans mixture) HOOT 14 10203 151 4- (aminomethyl) benzoic acid HOOC NH2 15 8087 103 DL-3-Aminobutyric acid HOOCH NHz 16 8145 103 DL- -Ammolsobutyricacid HOOCsf NH 17 8064 165 DL-3-Phenylpropionic acid i HOOC NH NH2 18 5992 129 DL-Nipecotic acid HOOC 19 12827 174 4-Pyridylacetic acid hydrochloride HCI-N I COSH 20 6410 149 Trans-3- (3-Pyridyl) acrylic acid N i i C02H

the abdomen is opened. The abomasum, the small and large intestines are tied at the omasal and pyloric openings, the duodenum, the end of the small intestine and at the end of the large intestine. Each section is transferred in a separate bucket containing warm water and is slit open and thoroughly washed. The epithelium is inspected before it is removed. The thus prepared washings are saved in gallon jars. An appropriate preservative is added. If preservative is not available, all the intestinal washing should kept in a refrigerator. These washings are passed through a 100-mesh sieve (pore size 149 pm), and the residue is examined for the presence of worms under a dissecting microscope.

Lugol's solution may be used to stain the worms. All worms are picked up counted and identified as to the species. An effort should be made to recover any immature forms present. The efficacy should be calculated using the controlled anthelmintic test.

(Mean number of worms in controls minus Mean number of worms in treated animal) Percentage efficacy = X 100 Mean number of worms in controls Results are depicted in Tables 4 and 5.

For Example, Sheep Test I results for AKC-107 are listed in Table 4.

Table 4 Akkadix Trial-1 Sheep AKK 101 Strongyles Strongyles ; Strongyles Sheep WeighUlbs Total 17-Jan 22-Jan 24-Jan Number 1/12/2000 EPG-pre EPG-pre EPG-5day % Change EPG-7day % Change if Group 55 76 4200 4070 3750 oouo-62. 16 AKC 107 89'90 1000 800 320. z 360 55. 00 1. 4mg/kg 80 82 180 180 130 ! 380-111. 11 Total/Mean 248 1793. 33 1683. 33 2035. 00-20. 89 2446. 67-45. 35 ; ; , I i Group 54 92 2880 2820 920 2370 15. 96 AKC108 69 59 560 480 2090 81. 25 14mg/kg 61 85 110 80 10 50 37. 50 Total/Mean 236 1183. 33 1126. 67 470. 00 58. 28 836. 67 25. 74 .., I Group 83i 91 2310 2000 10 0 100. 00 Jvermectin 92 113 570 570 00 100. 00 . 2mglkg 53 77 90 70 0 0, 100. 00 Totaf/Mear) 281 990. 00 880. 00 3. 33 99. 62 0. 00 100. 00 ! Group 81 74 2240 2240 780 770 65. 63 Negative i 75j 109 370 300 260 1360'-353. 33 Control 56 ! 80'40 40 30 30 : 25. 00 Total/Mean 263 883. 33 860. 00 356. 67 58. 531 720. 00 16. 28 , ''

Example 9-Sheep Test II Experimental Procedure Sheep naturally infected with a variety of gastrointestinal nematodes are purchased from local sources and are transported to the test site. The animals are housed in a manner to preclude further infection by nematode larvae. The animals are evaluated for the presence of adequate nematode burdens by performing a standard fecal egg per gram (EPG) count. Eggs are differentiated into the following groups : trichostrongyle (strongyle), Strongyloides, Trichuris, or Nematodiris. Only sheep judged by the study parasitologist to have adequate nematode infections are retained as test subjects.

The sheep are fed good quality hay (no concentrated rations) and water ad libitum.

Following a five day acclimation period, the sheep are randomly assigned by EPG count into the following treatment groups : Groups 1-9, various anthelmintic compounds of the present invention (test compound) dissolved in DMSO : Group 10, Positive Control (commercially available ivermectin for sheep) ; Group 11, non-treated Negative control (DMSO only). The first replicate of 11 animals is randomly assigned to groups 1-11 ; the second replicate of 11 animals is randomly assigned to groups 1-11 ; and the third replicate of 11 animals is randomly assigned to groups 1-11. Thus 11 groups of 3 animals each are created.

The randomization is performed on fecal samples collected 24-48 hours prior to scheduled treatment. The EPG counts are performed according to Zimmerman Research SOP &num NMEPG. 99. 01.

GROUP 1 : AKKADIX compound dissolved in DMSO.

GROUP 2 : AKKADIX compound dissolved in DMSO.

GROUP 3 : AKKADIX compound dissolved in DMSO.

GROUP 4 : AKKADIX compound dissolved in DMSO.

GROUP 5 : AKKADIX compound dissolved in DMSO.

GROUP 6 : AKKADIX compound dissolved in DMSO.

GROUP 7 : AKKADIX compound dissolved in DMSO.

GROUP 8 : AKKADIX compound dissolved in DMSO.

GROUP 9 : AKKADIX compound dissolved in DMSO.

GROUP 10 : Positive Control treatment of 200 mcg/kg commercially available ivermectin for sheep.

GROUP 11 : Non-treated negative control (placebo) of 3 ml of DMSO.

On treatment day, the animals are weighed, tagged, and divided into groups of three animals per group as follows : The placebo (DMSO), the commercially available drug, and the test anthelmintic compounds are administered in a 3ml volume of DMSO by subcutaneous injection using a sterile syringe fitted with a sterile needle. The site of injection is clipped and swabbed with alcohol prior to injection. The animal is adequately immobilized for injection of the placebo, commercially available drug, or experimental compound.

Following treatment, the animals are observed at hourly intervals for the first 8 hours, then daily until necropsy. They are housed in a manner to prevent further nematode infections.

On the fifth day following treatment, fecal samples are obtained from each animal, properly labeled and used for EPG counts.

Seven days following treatment, all the sheep are weighed and humanely slaughtered in accordance with the Guide for the Care and Use of Laboratory Animals (DHEW Publication No. 86-23). Necropsy procedures are according to Zimmerman Research SOP # NCRGIH. 00. 01, Necropsy for Helminth Recovery, specifically for gastrointestinal nematodes. Fecal samples are taken for EPG counts during the sample collection process on this day.

Nematodes are recovered, identified, and enumerated according to Zimmerman Research SOP # NEMRECOVID. 00. 01. All individuals performing nematode recoveries are blinded to treatment versus control animals.

Anthelmintic efficacy is calculated using the controlled test procedure : Mean number of worms in controls minus mean number of worms in treated % Efficacy=----------x 100 Mean number of worms in controls Results are depicted in Tables 6 and 7.

For Example, Sheep Test II results for AKC 107 are listed in Tables 5 and 6.

Insert Table 5 and 6.

Table 5 Akkadix AKK-102 1 Sheep 24-May Sheep IWeighUlbs Worm Coünts Number 5/17/2000 Abomasum Abomasum Small Intestine Small Intest. Large Intest. . Haemonchus Ostertag ! a Thchostrongytus Nematodirus, Trichuris Group 5301 471 60 5801 40 8500'5 Negative 1341 ; i 571 601 201 15 Control ! 20 801 0 0 10 Mean Ct. I, 471 227 20 3673 i0 Group 1347 45 0 0 0 100 0 Ivermectin 1336 5820'0 0 100 0 200mcglkg 5391 47'0 i 0 0 0 0 Mean Ct. 0 I 0 7 0 ; 0 67 0 % Efficacy'86 i00 ; 100 98 j t,, I ~, i Group 540 29 i 20 3660 360 7220 i 0 AKC107 ! 531 4960 ! 340 80 8900 10 . 8mg/kg 533 43 20 480 : 20 7220 15 Mean Ct. 67 1493 153 7780 12 zu Efficacy 43 : I.

Table 6 Akkadix Trial-2 Sheep AKK 102 Strongyles Strongyles Strongyles Sheep Weight/lbs Total 15-May 22-May t 24-May ; Number 5/17/2000 EPG-pre EPG-pre EPG-5day % Change EPG-7day%Change Group 540 29 3740 3260 2300 970 70. 25 AKC 107 ! 531 49 550 340 300 130 61. 76 . 8mg/kg ! 53343 200100 130190-90. 00 Tota !/Mean 1211496671233. 33 910. 0026. 22430. 00 65. 14 ~ ~~. ~~ ~.. ~ ; ~ ; ~.. ~~. ~~. ~,, ; ;. ~~ ~ i Group i 53450 ; 28202770r 1640 ! ! 590 78. 70 AKC108 1349 ! 33' 520 ! 390''50 ! 670-71. 79 8mg/kg 1 1344 36 i70 i70 1470 590 !-24i. 06 Total Mean 119 1170. 001 1110. 00 1053. 335. 11 ; 616. 67 44. 44 Group 530 47 710 640 690 300 53. 13 Negative 1341 57 160 120 160'340'-183. 33 Control 524 54 70 60 : 140-50 16. 67 Total/Mean 158 313. 33 273. 33 330. 00-20. 73 230. 00 i5. 85 i i Group 1347 560 450' 0i 01 100. 00 Ivermectin ! 133658 220170 0 ! 0' 100. 00 200mcg/kg 539 47 100 40 0 01 i00. 00 Tota)/Mean ! ! 150 293. 33220. 00 0. 00 100. 0 It should be understood that the examples and embodiments described herein are for illustrative purposes only and that various modifications or changes in light thereof will be suggested to persons skilled in the art and are to be included within the spirit and purview of this application and the scope of the appended claims.