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,"Austr. Vet. 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, A2b, 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 B, a and 20% 22, 23-dihydroavermectin B, b. Ivermectin has been tested on a variety of laboratory and domestic animals for control of nematodes, ticks, and heartworms.

Avermectin B2a is active against the root-knot nematode, Meloidogyne 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, I. 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 Bla (major component) and avermectin B, b. 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

December24, 1985 ; EPO applicationsO 094 779and 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) 0,-NH-CO-NRl-Alk-R2 (Formula I) wherein Ar is aryl or heteroaryl optionally substituted by one or more RI groups ; each Alk is a linear or cyclic alkylene radical of up to 8 C atoms ; R'is isHorCl6alkyl ; Alk = Aklayl R2 is heteroaryl or heterocycloalkyl optionally substituted by Ar, or forms such a group by cyclisation with R1; and R3 is OH, halogen, CF3, OCF, or a group selected from NH2, SO2-Cl 6 alkyl, C6 l0 aryl, C610 aryloaxy, C56 cycloalkyl, Cl 5 alkoxy, and Cl 6 alkyl, said group being optionally substituted by OH, C1-6 alkoxy, C1-6 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 RI 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, SO2 alkyl, C6 o aryl, Cl 6 alkoxy, and Cl 6 alkyl, said group being optionally substituted by OH, Cl 6 alkoxy, Cl 6 alkyl, phenyl, halogen, or CF3.

Particularly preferred anthelmintic compounds according to Formula II 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 Cl 6 alkyl, aryl, or heteroaryl optionally substituted by RI ; or R and Y together form a cycloalkyl or heterocycloalkyl ring ; and R3 is OH, halogen, CF3, OCF3, or a group selected from NH2, SO2 alkyl, C6 l0 aryl, Cl 6 alkoxy, and Cl 6 alkyl, said group being optionally substituted by OH, Cl 6 alkoxy, Cl 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) o.,-NHR (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 ; R is H or R3,

R3 is OH, halogen, CF3, OCF3, or a group selected from NH2, SO2 alkyl, C6 l0 aryl, Cl 6 alkoxy, and Cl 6 alkyl, said group being optionally substituted by OH, C1-6 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 Het2 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 l0 aryl, C1-6 alkoxy, and Cl 6 alkyl, said group being optionally substituted by OH, C 6 alkoxy, Cl 6 alkyl, phenyl, halogen, or CF3.

Particularly preferred anthelmintic compounds according to Formula V are exemplifiedby 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, 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 anthelmintic compound AKC 2154 of the class represented in Figure 47.

Figure 49 depicts anthelmintic compound AKC 2155 of the class represented in Figure 47.

Figure 50 depicts anthelmintic compound AKC 121 of the class represented in Figure 47.

Figure 51 depicts anthelmintic compound AKC 2157 of the class represented in Figure 47.

Figure 52 depicts anthelmintic compound AKC 2158 of the class represented in Figure 47.

Figure 52 depicts anthelmintic compound AKC 119 of the class represented in Figure 47.

Figure 54 depicts anthelmintic compound AKC 2159 of the class represented in Figure 47.

Figure 55 depicts anthelmintic compound AKC 2160 of the class represented in Figure 47.

Figure 56 depicts anthelmintic compound AKC 2162 of the class represented in Figure 47.

Figure 57 depicts anthelmintic compound AKC 2161 of the class represented in Figure 47.

Figure 58 depicts anthelmintic compound AKC 2163 of the class represented in Figure 47.

Figure 59 depicts anthelmintic compound AKC 2164 of the class represented in Figure 47.

Figure 60 depicts anthelmintic compound AKC 2166 of the class represented in Figure 47.

Figure 61 depicts anthelmintic compound AKC 2167 of the class represented in Figure 47.

Figure 62 depicts anthelmintic compound AKC 2168 of the class represented in Figure 47.

Figure 63 depicts anthelmintic compound AKC 2165 of the class represented in Figure 47.

Figure 64 depicts anthelmintic compound AKC 2171 of the class represented in Figure 47.

Figure 65 depicts anthelmintic compound AKC 2172 of the class represented in Figure 47.

Figure 66 depicts anthelmintic compound AKC 2169 of the class represented in Figure 47.

Figure 67 depicts anthelmintic compound AKC 2170 of the class represented in Figure 47.

Figure 68 depicts anthelmintic compound AKC 2173 of the class represented in Figure 47.

Figure 69 depicts anthelmintic compound AKC 2176 of the class represented in Figure 47.

Figure 70 depicts anthelmintic compound AKC 2174 of the class represented in Figure 47.

Figure 71 depicts anthelmintic compound AKC 2175 of the class represented in Figure 47.

Figure 72 depicts anthelmintic compound AKC 2180 of the class represented in Figure 47.

Figure 73 depicts anthelmintic compound AKC 2177 of the class represented in Figure 47.

Figure 74 depicts anthelmintic compound AKC 2178 of the class represented in Figure 47.

Figure 75 depicts anthelmintic compound AKC 2181 of the class represented in Figure 47.

Figure 76 depicts anthelmintic compound AKC 2179 of the class represented in Figure 47.

Figure 77 depicts anthelmintic compound AKC 110 of the class represented in Figure 47.

Figure 78 depicts anthelmintic compound AKC 2077 of the class represented in Figure 47.

Figure 79 depicts anthelmintic compound AKC 2078 of the class represented in Figure 47.

Figure 80 depicts anthelmintic compound AKC 2079 of the class represented in Figure 47.

Figure 81 depicts anthelmintic compound AKC 2080 of the class represented in Figure 47.

Figure 82 depicts anthelmintic compound AKC 2081 of the class represented in Figure 47.

Figure 83 depicts anthelmintic compound AKC 2182 of the class represented in Figure 47.

Figure 84 depicts anthelmintic compound AKC 2183 of the class represented in Figure 47.

Figure 85 depicts anthelmintic compound AKC 2184 of the class represented in Figure 47.

Figure 86 depicts anthelmintic compound AKC 2185 of the class represented in Figure 47.

Figure 87 depicts anthelmintic compound AKC 2187 of the class represented in Figure 47.

Figure 88 depicts anthelmintic compound AKC 2186 of the class represented in Figure 47.

Figure 89 depicts anthelmintic compound AKC 2188 of the class represented in Figure 47.

Figure 90 depicts anthelmintic compound AKC 2189 of the class represented in Figure 47.

Figure 91 depicts anthelmintic compound AKC 2190 of the class represented in Figure 47.

Figure 92 depicts anthelmintic compound AKC 2191 of the class represented in Figure 47.

Figure 93 depicts anthelmintic compound AKC 2192 of the class represented in Figure 47.

Figure 94 depicts anthelmintic compound AKC 2193 of the class represented in Figure 47.

Figure 95 depicts anthelmintic compound AKC 2194 of the class represented in Figure 47.

Figure 96 depicts anthelmintic compound AKC 2084 of the class represented in Figure 47.

Figure 97 depicts anthelmintic compound AKC 2085 of the class represented in Figure 47.

Figure 98 depicts anthelmintic compound AKC 122 of the class represented in Figure 47.

Figure 99 depicts anthelmintic compound AKC 2082 of the class represented in Figure 47.

Figure 100 depicts anthelmintic compound AKC 2087 of the class represented in Figure 47.

Figure 101 depicts anthelmintic compound AKC 2086 of the class represented in Figure 47.

Figure 102 depicts anthelmintic compound AKC 2083 of the class represented in Figure 47.

Figure 103 depicts anthelmintic compound AKC 2195 of the class represented in Figure 47.

Figure 104 depicts anthelmintic compound AKC 2196 of the class represented in Figure 47.

Figure 105 depicts anthelmintic compound AKC 2197 of the class represented in Figure 47.

Figure 106 depicts anthelmintic compound AKC 2200 of the class represented in Figure 47.

Figure 107 depicts anthelmintic compound AKC 2198 of the class represented in Figure 47.

Figure 108 depicts anthelmintic compound AKC 2199 of the class represented in Figure 47.

Figure 109 depicts anthelmintic compound AKC 2201 of the class represented in Figure 47.

Figure 110 depicts anthelmintic compound AKC 2202 of the class represented in Figure 47.

Figure 111 depicts anthelmintic compound AKC 2203 of the class represented in Figure 47.

Figure 112 depicts anthelmintic compound AKC 2204 of the class represented in Figure 47.

Figure 113 depicts anthelmintic compound AKC 120 of the class represented in Figure 47.

Figure 114 depicts anthelmintic compound AKC 2206 of the class represented in Figure 47.

Figure 115 depicts anthelmintic compound AKC 2091 of the class represented in Figure 47.

Figure 116 depicts anthelmintic compound AKC 2088 of the class represented in Figure 47.

Figure 117 depicts anthelmintic compound AKC 2096 of the class represented in Figure 47.

Figure 118 depicts anthelmintic compound AKC 2090 of the class represented in Figure 47.

Figure 119 depicts anthelmintic compound AKC 2092 of the class represented in Figure 47.

Figure 120 depicts anthelmintic compound AKC 2095 of the class represented in Figure 47.

Figure 121 depicts anthelmintic compound AKC 2097 of the class represented in Figure 47.

Figure 122 depicts anthelmintic compound AKC 2089 of the class represented in Figure 47.

Figure 123 depicts anthelmintic compound AKC 2098 of the class represented in Figure 47.

Figure 124 depicts anthelmintic compound AKC 2093 of the class represented in Figure 47.

Figure 125 depicts anthelmintic compound AKC 2094 of the class represented in Figure 47.

Figure 126 depicts anthelmintic compound AKC 2207 of the class represented in Figure 47.

Figure 127 depicts anthelmintic compound AKC 2208 of the class represented in Figure 47.

Figure 128 depicts anthelmintic compound AKC 2211 of the class represented in Figure 47.

Figure 129 depicts anthelmintic compound AKC 2209 of the class represented in Figure 47.

Figure 130 depicts anthelmintic compound AKC 2210 of the class represented in Figure 47.

Figure 131 depicts anthelmintic compound AKC 2212 of the class represented in Figure 47.

Figure 132 depicts anthelmintic compound AKC 2213 of the class represented in Figure 47.

Figure 133 depicts anthelmintic compound AKC 2214 of the class represented in Figure 47.

Figure 134 depicts anthelmintic compound AKC 2215 of the class represented in Figure 47.

Figure 135 depicts anthelmintic compound AKC 2216 of the class represented in Figure 47.

Figure 136 depicts anthelmintic compound AKC 2217 of the class represented in Figure 47.

Figure 137 depicts anthelmintic compound AKC 2218 of the class represented in Figure 47.

Figure 138 depicts anthelmintic compound AKC 2109 of the class represented in Figure 47.

Figure 139 depicts anthelmintic compound AKC 2099 of the class represented in Figure 47.

Figure 140 depicts anthelmintic compound AKC 2103 of the class represented in Figure 47.

Figure 141 depicts anthelmintic compound AKC 2104 of the class represented in Figure 47.

Figure 142 depicts anthelmintic compound AKC 2100 of the class represented in Figure 47.

Figure 143 depicts anthelmintic compound AKC 2105 of the class represented in Figure 47.

Figure 144 depicts anthelmintic compound AKC 2107 of the class represented in Figure 47.

Figure 145 depicts anthelmintic compound AKC 2101 of the class represented in Figure 47.

Figure 146 depicts anthelmintic compound AKC 2106 of the class represented in Figure 47.

Figure 147 depicts anthelmintic compound AKC 2108 of the class represented in Figure 47.

Figure 148 depicts anthelmintic compound AKC 2102 of the class represented in Figure 47.

Figure 149 depicts anthelmintic compound AKC 2220 of the class represented in Figure 47.

Figure 150 depicts anthelmintic compound AKC 2219 of the class represented in Figure 47.

Figure 151 depicts anthelmintic compound AKC 2115 of the class represented in Figure 47.

Figure 152 depicts anthelmintic compound AKC 2110 of the class represented in Figure 47.

Figure 153 depicts anthelmintic compound AKC 2119 of the class represented in Figure 47.

Figure 154 depicts anthelmintic compound AKC 2116 of the class represented in Figure 47.

Figure 155 depicts anthelmintic compound AKC 2111 of the class represented in Figure 47.

Figure 156 depicts anthelmintic compound AKC 2120 of the class represented in Figure 47.

Figure 157 depicts anthelmintic compound AKC 2117 of the class represented in Figure 47.

Figure 158 depicts anthelmintic compound AKC 2112 of the class represented in Figure 47.

Figure 159 depicts anthelmintic compound AKC 2121 of the class represented in Figure 47.

Figure 160 depicts anthelmintic compound AKC 2113 of the class represented in Figure 47.

Figure 161 depicts anthelmintic compound AKC 2114 of the class represented in Figure 47.

Figure 162 depicts anthelmintic compound AKC 2118 of the class represented in Figure 47.

Figure 163 depicts anthelmintic compound AKC 2122 of the class represented in Figure 47.

Figure 164 depicts anthelmintic compound AKC 2125 of the class represented in Figure 47.

Figure 165 depicts anthelmintic compound AKC 2130 of the class represented in Figure 47.

Figure 166 depicts anthelmintic compound AKC 2126 of the class represented in Figure 47.

Figure 167 depicts anthelmintic compound AKC 2127 of the class represented in Figure 47.

Figure 168 depicts anthelmintic compound AKC 2123 of the class represented in Figure 47.

Figure 169 depicts anthelmintic compound AKC 2124 of the class represented in Figure 47.

Figure 170 depicts anthelmintic compound AKC 2128 of the class represented in Figure 47.

Figure 171 depicts anthelmintic compound AKC 2131 of the class represented in Figure 47.

Figure 172 depicts anthelmintic compound AKC 2129 of the class represented in Figure 47.

Figure 173 depicts anthelmintic compound AKC 2132 of the class represented in Figure 47.

Figure 174 depicts anthelmintic compound AKC 2133 of the class represented in Figure 47.

Figure 175 depicts anthelmintic compound AKC 2134 of the class represented in Figure 47.

Figure 176 depicts anthelmintic compound AKC 2139 of the class represented in Figure 47.

Figure 177 depicts anthelmintic compound AKC 2135 of the class represented in Figure 47.

Figure 178 depicts anthelmintic compound AKC 2140 of the class represented in Figure 47.

Figure 179 depicts anthelmintic compound AKC 2136 of the class represented in Figure 47.

Figure 180 depicts anthelmintic compound AKC 2149 of the class represented in Figure 47.

Figure 181 depicts anthelmintic compound AKC 2151 of the class represented in Figure 47.

Figure 182 depicts anthelmintic compound AKC 2138 of the class represented in Figure 47.

Figure 183 depicts anthelmintic compound AKC 2141 of the class represented in Figure 47.

Figure 184 depicts anthelmintic compound AKC 2143 of the class represented in Figure 47.

Figure 185 depicts anthelmintic compound AKC 2145 of the class represented in Figure 47.

Figure 186 depicts anthelmintic compound AKC 2147 of the class represented in Figure 47.

Figure 187 depicts anthelmintic compound AKC 2137 of the class represented in Figure 47.

Figure 188 depicts anthelmintic compound AKC 2150 of the class represented in Figure 47.

Figure 189 depicts anthelmintic compound AKC 2152 of the class represented in Figure 47.

Figure 190 depicts anthelmintic compound AKC 2142 of the class represented in Figure 47.

Figure 191 depicts anthelmintic compound AKC 2144 of the class represented in Figure 47.

Figure 192 depicts anthelmintic compound AKC 2146 of the class represented in Figure 47.

Figure 193 depicts anthelmintic compound AKC 2148 of the class represented in Figure 47.

Figure 194 depicts anthelmintic compound AKC 2153 of the class represented in Figure 47.

Figure 195 depicts anthelmintic compound AKC 1501 of the class represented in Figure 47.

Figure 196 depicts anthelmintic compound AKC 1502 of the class represented in Figure 47.

Figure 197 depicts anthelmintic compound AKC 1503 of the class represented in Figure 47.

Figure 198 depicts anthelmintic compound AKC 1504 of the class represented in Figure 47.

Figure 199 depicts anthelmintic compound AKC 1505 of the class represented in Figure 47.

Figure 200 depicts anthelmintic compound AKC 1506 of the class represented in Figure 47.

Figure 201 depicts anthelmintic compound AKC 1507 of the class represented in Figure 47.

Figure 202 depicts anthelmintic compound AKC 1508 of the class represented in Figure 47.

Figure 203 depicts anthelmintic compound AKC 1509 of the class represented in Figure 47.

Figure 204 depicts anthelmintic compound AKC 123 of the class represented in Figure 47.

Figure 205 depicts anthelmintic compound AKC 1510 of the class represented in Figure 47.

Figure 206 depicts anthelmintic compound AKC 1511 of the class represented in Figure 47.

Figure 207 depicts anthelmintic compound AKC 1512 of the class represented in Figure 47.

Figure 208 depicts anthelmintic compound AKC 1513 of the class represented in Figure 47.

Figure 209 depicts anthelmintic compound AKC 1514 of the class represented in Figure 47.

Figure 210 depicts anthelmintic compound AKC 1520 of the class represented in Figure 47.

Figure 211 depicts anthelmintic compound AKC 1515 of the class represented in Figure 47.

Figure 212 depicts anthelmintic compound AKC 1521 of the class represented in Figure 47.

Figure 213 depicts anthelmintic compound AKC 1518 of the class represented in Figure 47.

Figure 214 depicts anthelmintic compound AKC 1516 of the class represented in Figure 47.

Figure 215 depicts anthelmintic compound AKC 1517 of the class represented in Figure 47.

Figure 216 depicts anthelmintic compound AKC 1519 of the class represented in Figure 47.

Figure 217 depicts anthelmintic compound AKC 1522 of the class represented in Figure 47.

Figure 218 depicts anthelmintic compound AKC 1524 of the class represented in Figure 47.

Figure 219 depicts anthelmintic compound AKC 1523 of the class represented in Figure 47.

Figure 220 depicts anthelmintic compound AKC 104 of the class represented in Figure 47.

Figure 221 depicts anthelmintic compound AKC 1525 of the class represented in Figure 47.

Figure 222 depicts anthelmintic compound AKC 1527 of the class represented in Figure 47.

Figure 223 depicts anthelmintic compound AKC 1526 of the class represented in Figure 47.

Figure 224 depicts anthelmintic compound AKC 1528 of the class represented in Figure 47.

Figure 225 depicts anthelmintic compound AKC 1530 of the class represented in Figure 47.

Figure 226 depicts anthelmintic compound AKC 1531 of the class represented in Figure 47.

Figure 227 depicts anthelmintic compound AKC 105 of the class represented in Figure 47.

Figure 228 depicts anthelmintic compound AKC 1529 of the class represented in Figure 47.

Figure 229 depicts anthelmintic compound AKC 125 of the class represented in Figure 47.

Figure 230 depicts anthelmintic compound AKC 1532 of the class represented in Figure 47.

Figure 231 depicts anthelmintic compound AKC 1533 of the class represented in Figure 47.

Figure 232 depicts anthelmintic compound AKC 1534 of the class represented in Figure 47.

Figure 233 depicts anthelmintic compound AKC 1535 of the class represented in Figure 47.

Figure 234 depicts anthelmintic compound AKC 124 of the class represented in Figure 47.

Figure 235 depicts anthelmintic compound AKC 1536 of the class represented in Figure 47.

Figure 236 depicts anthelmintic compound AKC 1537 of the class represented in Figure 47.

Figure 236 depicts anthelmintic compound AKC 1538 of the class represented in Figure 47.

Figure 238 depicts anthelmintic compound AKC 1539 of the class represented in Figure 47.

Figure 239 depicts anthelmintic compound AKC 1540 of the class represented in Figure 47.

Figure 240 depicts anthelmintic compound AKC 1543 of the class represented in Figure 47.

Figure 241 depicts anthelmintic compound AKC 1541 of the class represented in Figure 47.

Figure 242 depicts anthelmintic compound AKC 1542 of the class represented in Figure 47.

Figure 243 depicts anthelmintic compound AKC 1544 of the class represented in Figure 47.

Figure 244 depicts anthelmintic compound AKC 1545 of the class represented in Figure 47.

Figure 245 depicts anthelmintic compound AKC 1546 of the class represented in Figure 47.

Figure 246 depicts anthelmintic compound AKC 1547 of the class represented in Figure 47.

Figure 247 depicts anthelmintic compound AKC 1500 of the class represented in Figure 47.

Figure 248 depicts one library scheme by which the skilled artisan can create the compounds represented by the structure depicted in Figure 47.

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. Particularly preferred is the compound represented by Structures 11-21, and compounds related thereto as represented by Structure 47 depicted in Figure 47, and as further exemplified by Structures 48-247 depicted in Figures 48 through 247.

Preferred anthelmintic compounds useful in accord with the subject invention are represented by Structure 47, wherein : R, is aryl (optionally substituted with OC, 5, halogen, phenyl, or NO2) ; arylacetal (optionally substituted with NO2) ; C,, 0 straight or branched alkyl ; naphthyl ; furan ; or thiophene ; CI-5 phenyl ether ; R2 is C,, 0 alkyl (optionally substituted with aryl (optionally substituted with halogen) ; aryl (optionally substituted with halogen, OC, 5 alkyl, amine, or OC, 5 Ar) ; or C,, 0 branched or straight alkyl ; R3 is aryl (optionally substituted with OC,. s) ; CI-10 straight or branched alkylptionally substituted with morpholine ; pyrrole ; pyrrolidone ; C15 phenyl ether, or ArOCF3) ; aryl (optionally substituted with halogen, OC, 5 alkyl (optionally halogenated) or C, ; alkyl ; amine (optionally substituted with H, C, 5 alkyl, or aryl)) ; C2 s alcohol with

ether linkage ; heterocycle (optionally substituted with carbonyl) ; or C3 s cycloalkyl (optionally substituted with alcohol) ; R4 is H, or Cl 5 straight or branched alkyl ; or R3 and R4 form a heterocycle (optionally substituted with NO2, NO2Cl 5, or NOCl 5).

Generally, the anthelmintic compounds of the subject invention can be unsubstituted or substituted, saturated or unsaturated. The anthelmintic component of a 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 od 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 2154","Compound 48", or"Structure 48" each refer to the same compounds and each 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 contemplated by the subject invention.

Anthelmintic compounds specifically exemplified herein include Compounds 1- 46 represented by Structures 1-46 above, and Compounds 48-247 represented by Structures 48-247 depicted in Figures 48-247.

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 treatiing 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 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 phytotoxicityto 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 B. t. s, 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 lb/acre to about 0. 1 lb/acre and, preferably, from about 0. 5 lb/acre to about 2 lbs/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 lb/acre. Preferably, the rate of avermectin is between about 0. 005 lb/acre and 0. 01 lb/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. Ivermectinmay 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 Bacillus thuringiensis 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.

Anthelmintic compounds having Formulae I, II, III, IV, and V, Structure 47 and most preferably Structures 1-46, and particularly Structures 11-21 and Structures 48-247, 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 subj ect 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, and more preferably the Structure 47 compounds, particularly as exemplified by Structures 11-21 and 48-247. 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, or

48-247 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 1, 000 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 1, 000 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 11-21 and Structure 48-247 compounds 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, skin, 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 pharmacologicallyacceptable 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, Structure 47, particularly Structures 1-46, and particularly Structures 11-21 and 48-247 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 11-21 and 48-247, 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-parasiticnematodes 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, Structure 47 compounds, and preferably the compounds of Structures 1-46, more preferably the compounds of Structures 11-21 and 48-247, 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 Diabroticai. 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 orientalis (cockroach) Spodoptera littoralis (cotton leafworm) Hellothis virescens (tobacco budworm) Chortiocetes terminifera (locust) Diabrotica spp. (rootworms) Agrotis spp. (cutworms) Chilo partellus (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, preferably compounds of Structure 47, more preferably as exemplified by Structures 11- 21 and 48-247, 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 triisopropylnaphthalenesulphonates. Suitable agents of the non-ionic type include, for example, the condensationproducts 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 concentratesmay 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 dispersible 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 ; g) 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 : DIC : Diisopropylcarbodiimide DMAP : Dimethylaminopyridine DCM : Dichloromethane DMF : Dimethylformamide NCS : N-chlorosuccinimide NMM : N-methylmorpholine LAMPS : Large scale manual multiple peptide synthesis apparatus SLE : Supported liquid extraction dH2O 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 Caenorhabditis elegans 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 L 1 larvae collected. See The Nematode Caenorhabditis elegans (1988) Cold Spring Harbor Laboratory Press. Using a Matrix Programmable Pipette, the L Is were distributed into 96-well tissue culture plates, 20 LI

in 50, u1 NGM per well. Antibiotic/Antimyoticwas added to each well, and 1 % by weight E. coli strain HB 101. The subject anthelmintic compounds were stored at 5mM in 100% DMSO. 0. 7, u1 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. 8, aM, 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 1 OOx 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,"L1/L2"would mean a mixture of L1's and L2's are present in the well."L4/&num AD/B"would mean that a mixture of L4'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 V I 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.

@P US 03/02843<BR> Dose Response Tracking 5 Day Visual Score<BR> DR&num HTS Tracking Library &num Structure&num Source 1' Well Address V1 V2 V3 V4 V5 V6<BR> 1575 AKC 111 1 N2 &num 93 5081:D10 Dead Dead L1 Dead(L3/L &num AD/B OK<BR> 1647 AKC 112 2 N2 &num 98 5090:A10 L2/L3 L2/L3 L2/L3 L3/L4 &num AD/#B OK<BR> 1466 AKC 113 3 N2 &num 85 5061:A10 Dead Dead Dead Dead(L2/L L2/Dead(A L4/&num AD/E<BR> 1469 AKC 107 4 N2 &num 86 5061:D10 Dead Dead(L2/L3) L2/L3 Dead(LA) L4/Dead(A L2/Dead(I<BR> 1477 AKC 114 5 N2 &num 86 5061:D11 Dead L3/Dead(L4) L3 L3/Dead(L L2/L3 -L1<BR> 1476 AKC 108 6 N2 &num 86 5061:C11 Dead Dead L1 L1 L1/L2 L2/L3<BR> 1473 AKC 115 7 N2 &num 86 5061:H10 Dead Dead(L2/L3) L2/Dead(L2) Dead(L2) Dead(L2/L L2/L<BR> 035 AKC 119 11 N2 &num 126 5393:B4 &num AD/B &num AD/B &num AD/B &num AD/B &num AD/B <<BR> 2059 AKC 110 12 N2 &num 128 5399:C4 L1 L1 L1 L1 L1 L1<BR> 2083 AKC 120 13 N2 &num 130 5419:C4 L1 L2/L3 L1 L1/L2 &num AD/#B OK<BR> 2032 AKC 121 14 N2 &num 126 5389:C4 L1 L1 L1/L2 &num AD/#B &num AD/B <<BR> 2029 AKC 2153 15 N2 &num 126 5379:C4 L1 L1 L1/L2 &num AD/B &num AD/B &num AD/B<BR> 1962 AKC 122 16 N2 &num 121 5373:B8 Dead L1 &num AD/B &num AD/B &num AD/B OK<BR> 1388 AKC 104 17 N2 &num 80 5022:C4 L1/L2 L1/L2 L1/L2 &num AD/B L1/L2 L1/L2<BR> 1372 AKC 123 18 N2 &num 79 5016:B8 L1 L1/L2 &num AD/B < &num AD/B &num AD/-B<BR> 1402 AKC 124 19 N2 &num 81 5033:D8 &num AD/B! &num AD/B! L2/L3 L4/&num AD/B L4/&num AD/B L4/&num AD/B<BR> 1396 AKC 125 20 N2 &num 80 5031:G8 L2/Dead(L3) L2/Dead(L4) L2 L2 &num AD/B <<BR> 1393 AKC 105 21 N2 &num 80 5031:G2 L2/Dead(L3) L2/Dead(L3) L2/Dead(L3) L2/Dead(L 1.2/Dead(L L2/Dead(@<BR> 1164 AKC 126 22 N2 &num 64 4724:E10 L1/L2 L1/L2 L1/L2 &num AD/#B #B &num AD/B<BR> 1174 AKC 102 23 N2 &num 65 4727:ER L1/L2 L1/L2 L2/L3 L3 &num AD/B B<BR> 806 AKC 103 24 N2 &num 149 4470:D10 L1/L2 Dead(L3/L4) Dead(L4) B L4/&num AD/B @<BR> 18 AKC 171 25 N2 &num 2 2606:A1 Dead L2 Dead(L4)/&num A L2/L3 < L2/L3<BR> 433 AKC 128 26 N2 &num 31 3313:A10 Dead Dead Dead L1 &num AD/#B OK @<BR> 506 AKC 129 27 N2 &num 37 3315:A10 Dead Dead L1/L2 L1/L2 OK L1/L2<BR> 484 AKC 130 28 N2 &num 35 3314:D10 Dead &num AD/#B &num AD/B &num AD/#B OK &num AD/B<BR> 486 AKC 131 29 N2 &num 35 3314:F10 Dead L1 &num AD/#B &num AD/#B &num AD/#B < @<BR> 568 AKC 132 30 N2 &num 41 3323:G4 Dead(L2) Dead Dead &num AD/B &num AD/B Dead<BR> 569 AKC 133 31 N2 &num 41 3323:H4 Dead Dead Dead Dead Dead Dead<BR> 187 AKC 340 32 N2 &num 14 2665:B5 L1 L1 &num AD/B! &num AD/B! &num AD/B B @<BR> 133 AKC 34 33 N2 &num 10 2640:A11 Dead Dead Dead &num AD/B &num AD/B OK<BR> 149 AKC 135 34 N2 3!11 2641:A8 Dead L1 L1/L2 &num AD/B #B &num AD/B!<BR> CONTROL OK OK OK OK OK OK

Example 3-Nematicidal Activity of Anthelmintic Compositions 32-46 The C. elegans 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 140, uM, 70, uM, 35, uM, 17. 5, uM, 8. 8µM, 4. 4, uM, 2. 2µM, and 1. 09, uM. The visual evaluation of viability was conducted at Day 4, and the results are presented in Table 2.

Table 2. Compound, uM Concentration 140 70 35 17. 5 8. 8 4. 4 2. 2 1. 09 AKC-138 L1 L1 L1 L2 NB OK OK OK AKC-144 L3/L4 L4/AD/B B #B OK OK OK OK AKC-141 L1 L1 L1 < OK OK OK OK AKC-116 L1/L2 L2/L3 L3 B ! B OK OK OK AKC-117 L1/L2 L2/L3 L3 B ! B < OK OK AKC-118 L2 L2/L3 L3 L4/AD/B ! B NB OK OK Control OK OK OK OK OK OK OK OK Example 4-Activity Against Nematode (C. elegans ! 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 CaCl2, 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 1L 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 1 L S basal (components should first be autoclaved separately) : 1 M potassium citrate (pH 6) 10 ml Trace metals solution (see below) 10 ml 1M CaCl2 3 ml 1M MgSO4 3 ml Trace Metals solution Na2EDTA 1. 86 g (to 5mM) Fe2SO4 7H20 0. 69 g (to 2. 5mM) MnCl2#4H2O 0. 20 g (to lmM) ZnSO4 7H20 0. 29 g (to ImM) CUS04-5H20 0. 025 g (to 0.1mM) <BR> <BR> <BR> <BR> dH20<BR> <BR> <BR> <BR> <BR> 1L Procedure : 1. Make anthelmintic compound dilutions as indicated in Examples 2-3.

2. To 500 p1 of each dilution, added 10 p1 of eggs (estimated >200 eggs/10 psi).

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 ul S Medium.

7. Centrifuge at 2000 rpm for 5 minutes at room temperature 8. Pipette off supernatant.

9. Re-suspend in 300 ul S Medium.

10. Transfer 300 1 into 24-well tissue culture bioassay tray.

11. Add 2 ul 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. elegans 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 ml S Medium.

7. Spin as above for 2 minutes.

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

9. Repeat #7.

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

11. Add 280 1ll of S Medium to each well of 24-well tissue culture plate.

12. Add 20 1 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 by Compounds 48-194

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 48-194, as shown in Figures 47 and 48-194. The general library scheme resulting in Compounds 47 is depicted in Figure 248.

Synthesis of Oximes : A solution of 4-fluorobenzaldehyde (248g, 2. 0 moles) in 300 mL of hot denatured ethanol was treated with a solution of sodium acetate (197 g, 2. 4 moles) in 400 mL of water, followed by a solution of hydroxyl amine hydrochloride (153 g, 2. 2 moles) in 400 mL of water. Within a few minutes, a white solid began to separate. The mixture was stirred over night, diluted with 500 mL of water, and filtered and then the solid was washed 3 times with water and dried at 50 °C. Yield was 269 g (97%) of a white solid.

The foregoing procedure was applied to the other precursors listed in Table 10. The oximes obtained are listing in Table 7.

When this procedure was applied to 4-methoxybenzaldehyde, the product separated as an oil and was isolated by. extraction with ether, washed with bicarbonate solution, dried over sodium sulfate and then concentrated to give a solid.

Step A : Coupling a a, P-unsaturated acid to thiophenol resin An a, P-unsaturated acid (99 mmoles, 3 equiv.) (selected from those listed in Table 8) and DIC (99 mmoles, 15. 5 mL, 3 equiv.) were added to a 600 mL Pyrex glass bottle with 180 mL DCM (6 mL/g resin). The mixture was stirred for 10 minutes. 30 g of thiophenol resin (1. 1 mmoles/g, 33 mmoles, 1 equiv.) was then added followed by DMAP (1. 21g, 9. 9 mmoles, 0. 3 equiv.) and NMM (11 mL, 99 mmoles, 3 equiv.). Additional DCM was added to minimally swell the resin. The slurry was shaken at room-temperature for 24 hours, then transferred to a 600 mL LAMPS vessel, washed with DCM (4X) and MeOH (IX) twice, and dried in a vacuum oven for 3 days. The resin was qualitatively analyzed by a FeCl3/pyridine Table 7 Oximes 10 plates per oxime, 6 oximes, total 5280 compounds Entry MOLE MW ACD Name Structure MOLE N-OH plates 62. 6g 139. 13 N/A 4-fluorobenzaldehyde oxime eN-OH 1-10 plat s 11-68. Og 151. 16 N/A 4-methoxy benzaldehyde/N-OH 20 oxime 3-OH plates 21-70. 0g 155. 6 N/A 2-chloro- 30 dehyde oxime 4-oh -N-OH plates 31 62. 6g 139 13 N/A 2-fluorobenzaldehyde oxime F 5 NOS g NOH plates 41-45. 52g 101. 15 N/A 2, 2-dimethyl propionaldehyde 50 oxime 6 plates 51-60 115. 8g 257. 28 N/A 3-benz) coo beiizaldeliyde oxime-OH Table 10 Precursors, Reagents, Solvents Entry Amt. MW ACD Name Structure ~ ~ ~~ ~/O precursor 200g 124. 11 3378 4-fluorobenzaldehyde F -/O precursor 200g 136. 15 3385 4-methoxy benzaldehyde m//° precursor 200g 140. 57 3304 2-chlorobenzaldehyde > ce, precursor 200g 124. 11 3302 2-fluorobenzaldehyde F 0 precursor 150g 86. 13 6962 Trimethyl acetaldehyde et precursor 300g 242. 27 3386 3-benzyloxy-4-methoxy o benzaldehye \/ 600g 82 sodium acetate 2 L ethanol, denatured cnn < : n c hydroxy) amine ,, ,,,, , precursor 500g 69. 5 hydroxyl amine NH2OH-HCI hydrochloride 7 L methanol 5 L ether

test (Breitenbucher et. al., 1998). Approximately 10 mg of resin was suspended in CH2CL2 (0. 5 mL). Pyridine (5 drops) was added to the resin followed by FeCl3 (10 drops of a 0. 5 M soln. in CHCl3). The resin was washed 5X with CH2Cl2 A purple color on the resin indicated phenol.

Step B : 1, 3-Dipolar cycloaddition of nitrile oxides to a, (3-unsaturated esters A stirred solution of 0. 45 moles of oxime in dry DMF was treated portion wise with 59. 5 g (0. 445 moles) of NCS (total volume 900 mL, based on 10 plates and 20 extra wells) and shaken at room-temperature for 4 hours. Caution : this reaction may be exothermic. For the two fluorobenzaldehyde oximes and 2-chlorobenzaldehyde oxime, the mixture was heated at 50°C for 3 hours. The resin was partitioned into 2. 7 u gf fritted Polyfiltronics plates (-100 mg/well, 10 plates per oxime) according to the microtiter plate layout. The plate was placed into a clamp with a Teflon sheet to seal at the bottom. The chlorinated oxime solutions were added to the appropriate wells (1. 0 mL /well) by a Robbins 96 Hydra, followed by NMM (55 uL/well, 0. 5 mmol). The plates were clamped shut, tipped on their sides and shaken at room-temperature for 30 minutes.

The plates were then heated at 55 ° C for 48 hours. The resin was rinsed with DMF (2x) and DCM (2X) repeating three times.

Step C : Cleavage by Aminolysis The plate was placed into an open clamp with Teflon sheet to seal at the bottom.

A solution of the appropriate cleaving amine (0. 4 M, 1. 0 mL) in anhydrous pyridine was added to each well of the plate. Each amine was used for 264 wells. For 275 mL of 0. 4M solution, 0. 11 mole was weighed out and diluted to 275 mL with pyridine. For dimethylamine, 1. 0 mL of commercial solution (2. 0 M in THF) was added to the appropriate wells. The plate was clamped shut and shaken at room-temperature for 24 hours. The plate was frozen with dry ice for 20 minutes, unclamped and placed on top of a 2-mL Beckman deepwell microtiter plate. The plate thawed and drained into the collection plate. The plates containing amines 1-8 were rinsed with 0. 5 mL of MeOH. For the other plates containing amines 9-40, the resin was rinsed with 0. 5 mL of 10% MeOH

in CHCL). The second rinse was with MeOH (0. 5 mL) for all wells. The plate was concentrated on the Savant (high heat) until dryness.

Step D : SLE The cleaving amines were divided into two groups. Amines 1-8 on the list below are the"water soluble"set, and Amines 9-40 constitute the"hydrophobic"set. The SLE conditions for the two sets differed as described below.

For the water-soluble set, a Polyfiltronics plate with 10 micron polypropylene fritts was filled to within 5 mm of the top with Varian hydromatrix diatomaceous earth.

To each well was added 0. 4 mL of water and then they were allowed to equilibrate for 15 minutes. The residue was taken up in 0. 8 mL of 20% THF/chloroform and transferred to the top of the SLE plate. The residue plate was rinsed two more times with 0. 5 mL of the 20% THF/chloroform solution and transferred to the top of the SLE plate.

The SLE plate was rinsed with 0. 5 mL of chloroform once. The elute was collected into another Beckman plate. The plate was concentrated on the Savant (medium heat) until dryness.

For the hydrophobic set, the SLE material was activated with 0. 4 mL of 2 N Hcl Example 7-Preparation of Anthelmintic Compounds 47 as specifically exemplified by Compounds 195-247 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 195-247, as shown in Figures 195-247.

Synthesis of Oximes : A solution of 4-chlorobenzaldehyde (211g, 1. 5 moles) in 900 mL of hot denatured ethanol was treated with a solution of sodium acetate (148g, 1. 8 moles) in 400 mL of water followed by a solution of hydroxylamine hydrochloride (114g, 1. 65 moles) in 250 mL of water. Within a few minutes a white solid began to separate. The mixture was stirred over night, diluted with 600 mL of water, and filtered. The solid was then washed three times with water and dried at 75 °C. Yield was 209g (89%) of a white

solid. The foregoing procedure was applied to the other precursors listed in Table 12.

The oximes obtained are listed in Table 11.

When this procedure was applied to 1-heptaldehyde, the product separated as an oil and was isolated by extraction with ether, and then dried over sodium sulfate and concentrated to give a solid.

Step A : Coupling a a, (3-unsaturated Acid to Thiophenol Resin An a, p-unsaturated acid (99 mmoles, 3 equiv.) (selected from those listed in Table 8) and DIC (99 mmoles, 15. 5 mL, 3 equiv.) were added to a 600 mL Pyrex glass bottle with 180 mL DCM (6 mL/g resin). The mixture was stirred for 10 minutes. 30 g of thiophenol resin (1. 1 mmoles/g, 33 mmoles, 1 equiv.) was then added followed by DMAP (1. 21g, 9. 9 mmoles, 0. 3 equiv.) and NMM (11 mL, 99 mmoles, 3 equiv.).

Additional DCM was added to minimally swell the resin. The slurry was shaken at room temperature for 24 hours, then transferred to a 600 mL LAMPS vessel, washed with DCM (4X) and MeOH (1 X) twice, and dried in a vacuum oven for 3 days. The resin was qualitatively analyzed by a FeCl3/pyridine test (Breitenbucher et. al., 1998).

Approximately 10 mg of resin was suspended in CH2Cl2 (0. 5 mL). Pyridine (5 drops) was added to the resin followed by FeCl3 (10 drops of a 0. 5 M soln. in CHCl3). The resin was washed 5X with CH2Cl2. A purple color on the resin was indicative of phenol.

Step B : 1, 3-Dipolar Cycloaddition of Nitrile Oxides to a, p-unsaturated Esters A stirred solution of 0. 45 moles of oxime in dry DMF was treated portion wise with 58. 8 g (0. 445 moles) of NCS (total volume 900 mL, based on 10 plates and 20 extra wells) and shaken at room temperature for 4 hours. Caution : this reaction may be exothermic. For the two chlorobenzaldehydeoximes, the mixture was heated at 50 : ; C for 4 to 5 hours. The resin was partitioned into 2. 7 n gf fritted Polyfiltronics plates (-100 mg/well, 10 plates per oxime) according to the microtiterplate layout (Section 9). The plate was placed into a clamp with a Teflon sheet to seal at the bottom. The chlorinated oxime solutions were added to the appropriate wells (1. 0 mL/well) by a Robbins 96 Hydra, followed by NMM (55 nL Insert table 8 Table 8 α, -Unsaturated Acids-Resin Precursors Entry ACD MW resin Name Structure resin H HA 1 16837 227 22. 5g 2, 4-dimethoxycinnamic acid IN LJ/f.. 2 4369 148 14. 65g cinnamic acid O 0 O 3 16839 192 l 9. 0g 2-ethoxycinnamic acid HO Xt RIZ O HO 4 14010 224 22. 2g 4-phenylcinnamic acid HOS I O HO 5 143427 245 24. 26g 5-bromo-2-fluorocinnamic acid If 0 O HO OCH3 6 14376 238 23. 6g 2, 3, 4-trimethoxycinnamic acid VOCH3 -OCHg 0 Ho 7 t 6544 190 18. 8g 4-isopropylcinnamic acid If O HO 8 16848 192 19. Og 4-ethoxycinnamic acid zoot 0 O 9 63250 237 23. 5g 4, 5-methylenedioxy-2- nitrocinnamic acid ou Entry ACD MW For 30g Name Structure resin 10 17017 253 25. 05 4, 5-dimethoxy-2-nitrocinnamic H OCH3 acid acrid O 2-bromo-4, 5-dimethoxycinnamic HO acid HO. F B OCH3 O I H 12 51582 200 19. 8g 2-chloro-6-fluorocinnamic acid l I Cl 0 13 14317 198 19. 6g 3-(1-naphthyl) acrylic acid W 0 O 14 17596 257 25. 4Ag 5-bromo-2-methoxycinnamic acid HO. O 15 14535 154 15. 25g 3- (3-thienyl) acrylic acid O 0 0 JL JL 16 154. 2 15. 3g Crotonic anhydride I O 0 w 0 19. 8g HO I 17 236027 200 2-chloro-4-fluorocinnamic acid -''-O F O 18 75074 138 13. 66g 3-furanacrylic acid HO "CL 19 16843 192 19. 01 3-ethoxycinnamic acidv H°J% °Et riz O HO 20 4375 238 23. 56g 2-methoxycinnamic acid IN H3C0 Entry ACD MW For 30g Name Structure resin HA 21 6331 I 227 22. Sg 4-chloro-3-nitrocinnamic acid'\ N02 CI Cri HO F 22 143287 200 19. 8 g 4-chloro-2-fluorocinnamic acid C ! CI Table 11 Oximes 10 plates per oxime, 6 oximes, total 5280 compounds Entry Amt. MW ACD Name Structure 1 54. 5 g 121. 1 benzaldehyde NOH plates oxime < 1-10 2 70 g 155. 6 4-NOH plates chlorobenzaldehyd < 11-20 e oxime C 3 73. 9 g 164. 24- (N, N-NOH plates dimethylamino) 21-30 benzaldehyde J) J oxime N 4 74. 3 g 165. 2 2-NOH plates ethoxybenzaldehyd 31-40 e oxime \4OEt 5 70 g 155. 6 3-NOH plates chlorobenzaldehyd 41-50 e oxime Ci 6 58. 1 g 129. 2 1-heptaldehyde NOH plates oxime VJJ 51-60 Table 12 Precursors, Reagents, Solvents Entry Amt. MW ACD Name Structure precursor 200g 140. 6 3379 4-chlorobenz-O aldehyde c precursor 200g 3381 4-N, N- dimethylamino benzaidehyde precursor 200g 3316 2-ethoxybenz -. 0 aldehyde OEt precursor 200g 140. 6 3350 3-chlorobenz aldehyde cr precursor 00g 114 7028 l-heptaldehyde ~ w 600g 82 sodium acetate 2 L ethanol, denatured precursor 500g 69. 5 hydroxyl amine NH2OH HCI hydrochloride precursor 0. 5 mole 155. 6 16412 4-chlorobenzald N-OH oxime CI precursor 0. 5 mole 164. 2 16413 4- (N, N- NOH dimethylamino) benzaldehyde oxime N" I precursor 0. 5 mole 165. 2 none 2-ethoxybenz NOH aldehyde oxime < O eut precursor 0. 5 mole 155. 6 none 3-chlorobenz NOH aldehyde oxime ci CI precursor 0. 5 mole 129. 2 none l-heptaldehyde NOH oxime 7 L methanol 5L ether

/well, 0. 5 mmol). The plates were clamped shut, tipped on their sides and shaken at room temperature for 30 minutes. The plates were then heated at 55 °C for 48 hours. The resin was rinsed with DMF (3x) and DCM (3X) repeating three times.

Step C : Cleavage by Aminolysis The plate was placed into an open clamp with Teflon sheet to seal at the bottom.

A solution of the appropriate cleaving amine (0. 4 M, 1. 0 mL) (selected from those listed in Table 9) in anhydrous pyridine was added to each well of the plate. Each amine was used for 264 wells. For 275 mL of 0. 4 M solution, 0. 11 mole was weighed out and diluted to 275 mL with pyridine. For dimethylamine, 1. 0 mL of commercial solution (2. 0 M in THF) was added to the appropriate wells. The plate was clamped shut and shaken at room temperature for 48 hours. The plate was frozen with dry ice for 30 minutes, unclamped and placed on top of a 2-mL Beckman deepwell microtiter plate. The plate thawed and drained into the collection plate. The plates containing Amines 1-8 were rinsed with 0. 5 mL of MeOH. The plates containing Amines 9-40 were rinsed with 0. 5 mL of 10% MeOH in CHCl3 The second rinse was with MeOH (0. 5 mL) for all wells.

The plate was concentrated on the Savant (high heat) until dryness.

Step D : SLE The cleaving amines were divided into two groups. Amines 1-8 are the"water soluble"set, and Amines 9-40 are the"hydrophobic"set. The SLE conditions for the two sets differed as follows : For the water-soluble set, a Polyfiltronics plate with 10 micron polypropylene frits was filled to within 5 mm of the top with Varian Hydromatrix diatomaceous earth.

0. 4 mL of water was added to each well and allowed to equilibrate for 15 minutes. The residue was taken up in 0. 8 mL of 20% THF/chloroform and transferred to the top of the SLE plate. The residue plate was rinsed two more times with 0. 5 mL of the 20% THF/chloroform solution and transferred to the top of the SLE plate. The SLE plate was rinsed with 0. 5 mL Insert table 9 Table 9 Cleaving Amines Entry ACD MW Name Structure mole 14. 10 14100 128. 2 3-pyrrol idinopropyiqmine H2N, ~rND A2 17. 30 14616 157. 3 l-(3-aminopropyl)-4- methylpiperazine NH2 i A3 11. 24 14812 102. 2 1-dimeth lamino-2-propylamine NH21 ou A4 9. 80 59196 89. 1 1-amino-2-butanol H 2N 1-1 A5 14. 3 6182 130 1-aminoethylmorpholine Non A6 12. 76 8176 116 N, N-diethylethylenediamine N NH2 'NH A7 11. 55 8181 105 2-aminoethoxyethanol HO O~NH2 A8 6. 71 8183 61 aminoethanol HONH2 A9 8. 03 11690 73 butylamine H2N A F A10 15. 29 134208 139 4-fluorophenethylamine NHp All 7. 7 14820 70 3-aminopropionitrile H2NN A 12 9. 79 14831 89 methoxypropylamine HZN''O' NHz A13 12. 65 191368 115 2-aminocyclohexanol \/=\ A14 18. 04 191602 164 N-aminopropyl-N-methylanaline H2N A15 20. 46 25572 186 2-bromobenzylamine I NH2 bu A16 15. 62 3201 142 aminopropylpyrolidine-2-one H2N~N ;, 0 A 17 17. 93 40754 163 4-tert-butylbenzylamine NH2 (H3C3C A 18 15. 29 41323 139 4-fluoro-a-methylbenzylamine NH Fuzz A 19 3 L 4731 117 L-isoleucinol OH A20 16. 61 4732 151 S-2-amino-3-phenyl-l-propanol MOH Entry mole ACD MW Name Structure mole A21 15. 07 52975 137 2-phenoxyethylamine oNH2 0 H A22 14. 08 59019 128 3-acetamidopyrrolidine 'N--NH 0 H OH A23 20. 46 59040 186 3-(tert-butoxy °t N=GNH carbonylamino) pyrrolidine A24 9. 68 5972 88 morpholine HNuO A25 11. 33 59847 103 3-amino-2, 2-dimethyl-l-HO) < NH2 propanol cl A26 20. 9 60612 190 2, 6-dichlorophenethylamine NH2 OCH3 OCH3 A27 19. 91 60613 181 2, 3-dimethoxyphenethylamine OCH3 iNH2 OCH3 A28 19. 91 60614 181 2, 5-dimethoxyphenethylamine H3cO NH2 CH3 OCH3 A29 19. 91 60618 181 3, 5-dimethoxyphenethylamine ETC Et0 A30 1815 60620 165 4-ethoxyphenethylamine Et° NH2 NH2 A31 21. 01 61237 l 91 4-(trifluoromethoxy) F3COn] A32 21. 01 61267 191 3- (trifluoromethoxy) benzylamine F3C0 O H A33 14. 08 64475 128 2-aminocaprolactam H2N-\J A34 13. 42 75408 122 2-aminobenzylamine H2N Han F A35 15. 29 75502 139 2-fluorophenethylamine 61, NU2 A36 21. 45 79755 195 3-ethoxy-4-methoxy M eOPt phenethylamine EtOeNH2 Q A37 15. 07 8110 137 2-methoxybenzylamine I Entry ACD MW Name Structure mole A38 15. 07 8137 137 2-am ino-l-phenylethanol NH2 OH ou A39 19. 91 8188 181 3, 4-dimethoxyphenethylamine H 2N O A40 263 mL 8288 45 Dimethyl amine H (2. 0 M in THF)

of chloroform once. The elute was collected into another Beckman plate. The plate was concentrated on the Savant (medium heat) until dryness.

For the hydrophobic set, the SLE material was activated with 0. 4 mL of 2 N HCI.

Example 8 Nematicidal Activity of Anthelmintic Compositions 48-247 The nematicidal activity of anthelmintic Compositions 48-247 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 AKC # MP # Well mOD % Run SVisual Score mOD% Run Visual Score Well Addr V1 V2 V3 V4 V5 V6 2077 5372 A5 193 100% L1 161 95% L1/L2 5372:A5 L1 #AD/B OK OK OK OK 2078 5372 C5 213 110% L1 178 105% L1 5372:C5 L1 #AD/B OK OK OK OK 2079 5372 E5 210 109% L1 178 105% 1AD/B! 5372:E5 L1 L1 < OK OK OK 2080 5372 F5 213 110% L1 189 112% L1/L2 5372:F5 L1 L1 #B OK OK OK 2081 5372 G5 223 116% L1 184 109% 2AD/B! 5372:G5 L1 #AD/B < OK OK OK B! B! B! OK OK OK 2082 5373 E4 231 120% L1/L2 195 115% L1/L2 5373:E4 Dead L1 #AD/B < OK OK 2083 5373 H4 270 140% L1/L2 232 137% 14/AD/B! 5373:H4 L1 #B OK OK OK OK 2084 5373 A5 194 101% L1/L2 185 109% L1/L2 5373:A5 L1 #AD/B #AD/B #AD/B < OK 2085 5373 B5 206 107% L2/L3 185 109% L1/L2 5373:B5 L1 #AD/B B #B OK OK 2086 5373 G5 215 111% L1/L2 176 104% B 5373:G5 3!AD/B < < OK OK OK 122 5373 B8 191 99% L1 176 104% L1/L2 5373;B8 Dead L1 #AD/B #AD/B #AD/B OK 2087 5373 F8 264 137% L2/L3 244 144% L4 - Husks 5373:F8 L2/L3 #AD/B #AD/B OK OK OK L1 L1 L1 L1 #AD/B! OK 2088 5374 B1 201 104% L1/L2 181 107% L1/L2 5374:B1 L1 L1 #AD/B < OK OK 2089 5374 G1 220 114% L2/L3 197 117% 5AD/B! 5374:G1 L1 < < OK OK OK 2090 5374 B4 179 93% L1/L2 142 84% 3AD/B 5374:B4 Dead < OK OK OK OK 2091 5374 A5 195 101% L1 165 98% L1/L2 5374:A5 Dead < OK OK OK OK 2092 5374 B5 205 106% Dead 185 109% L1/L2 5374:B5 Dead OK OK OK OK OK 2093 5374 G5 225 117% L2 168 99% 5AD/B 5374:G5 Dead B #B OK OK OK 2094 5374 H5 223 116% dead 169 100% 4AD/B 5374:H5 Dead OK < OK OK OK L1 B! B! OK OK OK 2095 5374 B8 191 99% L2/L3 158 93% 5AD/B 5374:B8 B OK OK OK OK OK 2096 5374 B11 219 113% L2 173 102% 3AD/B 5374:B11 L1/L2 #AD/B B OK OK OK 2097 5374 E11 215 111% 1AD/B! 152 90% B 5374:E11 #AD/B < OK OK OK OK 2098 5374 G11 220 114% L1/L2 159 94% 1AD/B 5374:G11 #AD/B! #B #B OK OK OK 2099 5375 B4 187 97% L1/L2 163 96% L1/L2 5375:B4 L1 Dead #1AD/B < < OK 2100 5375 F4 241 125% L1 212 125% L1/L2 5375:F4 L1 #AD/B #AD/B #B OK OK 2101 5375 G4 272 141% L1 232 137% L1/L2 5375:G4 L1/L2 L1 L1 #B OK OK L1 L1 L1 L1 #B OK 2102 5375 H4 264 137% Dead 234 138% 2AD/B! 5375:H4 Dead #AD/B! #AD/#B < OK OK 2103 5375 B5 213 110% L2/L3 169 100% 4AD/B 5375:B5 L1/L2 #AD/B < OK OK OK 2104 5375 E5 220 114% L2 163 96% B 5375:E5 L1 #AD/B < OK < OK 2105 5375 F5 220 114% Dead 191 113% L1/L2 5375:F5 Dead #AD/B #B OK OK OK 2106 5375 G5 235 122% L2 200 118% L1/L2 5375:G5 L1 L1/L2 #AD/B < < OK 2107 5375 F8 230 119% L1 194 115% 4AD/B 5375:F8 #AD/B #B < OK OK OK 2108 5375 G8 257 133% L1/L2 212 125% 1AD/B 5375:G8 L1 < #AD/B OK OK OK B! B! B! OK OK OK Table 3 2109 5375 A9 198 103% L1/L2 150 89% L4/2AD/B! 5375:A9 L4/#AD/B OK OK OK OK OK 2110 5376 B1 191 99% L1/L2 178 105% L2 5376:B1 L1/L2 B < OK OK OK 2111 5376 F1 219 113% L1 209 124% L1/L2 5376:F1 L1 L1 < OK OK OK 2112 5376 G1 214 111% Dead 199 118% L1 5376:G1 L1 3!AD/B B OK OK OK 2113 5376 G2 217 112% 2AD/B! 186 110% 3AD/B 5376:G2 #B < OK OK OK OK 2114 5376 G3 210 109% L3 157 93% B 5376:G3 B OK < OK #B OK 2115 5376 A5 188 97% L1/L2 181 107% L3 5376:A5 L1 #B < OK OK OK L1 L1 L1 L1 #B OK 2116 5376 B5 207 107% L3/L4 174 103% B! 5376:B5 L1 < OK OK OK OK 2117 5376 F6 224 116% L2/L3 167 99% B 5376:F6 L1 < OK OK OK OK 2118 5376 G6 230 119% L2/L3 182 108% 1AD/B! 5376:G6 #AD/B #AD/B < OK OK OK 2119 5376 B11 207 107% L1/L2 179 106% 1AD/B! 5376:B11 L1 #AD/B #B OK OK OK 2120 5376 F11 210 109% L1 186 110% L1/L2 5376:F11 L1 #AD/B #B OK OK OK 2121 5376 G11 208 108% L1 188 111% L1/L2 5376:G11 L1 L1 #AD/B OK OK OK 2122 5377 B2 228 118% L2/L3 177 105% 4AD/B! 5377:B2 L1 #AD/B OK OK OK OK B! B! B! OK OK OK 2123 5377 G2 237 123% L1/L2 190 112% 2AD/B! 5377:G2 < OK OK OK OK 2124 5377 G3 218 113% L2/L3 161 95% 4AD/B 5377:G3 OK < OK OK OK OK 2125 5377 C4 228 118% L2/L3 196 116% 5AD/B 5377:C4 L1 L1 #AD/B < < #AD/#B 2126 5377 D4 222 115% Dead 146 86% 5AD/B 5377:D4 #AD/B #AD/B #B < OK OK 2127 5377 E4 238 123% L1/L2 196 116% L2 5377:E4 L1/L2 #1AD/B #AD/B #AD/B < #B 2128 5377 G4 286 148% L2/L3 221 131% 5AD/B 5377:G4 #AD/B #B OK #B O OK 2129 5377 H4 277 144% L1/L2 226 134% 2AD/B! 5377:H4 L1 #AD/B #B OK OK < L1 L1 L1 L1 #B OK 2130 5377 C5 233 121% L3 190 112% 5AD/B 5377:C5 B < OK OK OK OK 2131 5377 G5 236 122% L2/L3 165 98% 3AD/B 5377:G5 L1 #AD/B OK OK OK OK 2132 5377 H5 236 122% L2/L3 184 109% L3/L4 5377:H5 L1 #AD/B #B #AD/B < OK 2133 5377 H4 249 129% L2/L3 204 121% 5AD/B 5377:H8 L1 #AD/8 < #B OK OK 2134 5378 B1 208 108% L1/L2 186 110% 2L4/1AD/B! - No P 5378:B1 L2/L3 #AD/B B < OK OK 2135 5378 F1 219 113% L2 197 117% B 5378:F1 #AD/B! OK OK OK OK OK 2136 5378 G1 219 113% L1 208 123% L1/L2 5378:G1 L1 L1 #B #AD/B OK OK B! B! B! OK OK OK 2137 5378 H1 229 119% L2 173 102% 4AD/B 5378:H1 #AD/B! OK OK OK OK OK 2138 5378 G3 218 113% L2/L3 170 101% B 5378:G3 < OK < OK OK OK 2139 5378 D5 207 107% Dead 168 99% L1/L2 5378:D5 L1 OK OK OK OK OK 2140 5378 F5 217 112% Dead 175 104% L1 5378:F5 L1 < OK OK OK OK 2141 5378 G5 214 111% Dead 192 114% 2AD/B 5378:G5 L1 #AD/B L1 #AD/B OK #B 2142 5378 H5 228 118% L1/L2 184 109% L3/L4 5378:H5 OK OK OK OK OK 2143 5378 G6 222 115% L2 202 120% L1 5378:G6 L1 #AD/B #B OK OK OK L1 L1 L1 L1 #AD/B B 2144 5378 H6 228 118% L1/L2 198 117% L1/L2 5378:H6 L1/L2 B #B OK OK OK Table 3 2145 5378 G7 237 123% L2/L3 199 118% L2 5378:G7 L4/AD/B < < OK OK OK 2146 5378 H7 222 115% L3 174 103% B 5378:H7 #B OK OK OK OK OK 2147 5378 G9 253 131% L2/L3 184 109% L2/L3 5378:G9 #1AD/B! #AD/B < OK OK OK 2148 5378 H9 247 128% L2/L3 156 92% B 5378:H9 #AD/B OK OK OK OK OK 2149 5378 G10 230 119% 1AD/B! 157 93% B 5378:G10 #AD/B OK OK OK OK OK 2150 5378 H10 236 122% L2/L3 186 110% B 5378:H10 L2/L3 #B OK OK OK OK B! B! B! OK OK OK 2151 5378 G11 214 111% L2/L3 177 105% L4/2AD/B! 5378:G11 L1/L2 < OK OK OK OK 2152 5378 H11 227 118% L1/L2 189 112% L1/L2 5378:H11 L1/L2 #AD/B #B < OK OK 2153 5379 C4 264 173% L1/L2 181 107% L2/L3 5379:C4 L1 L1 L1/L2 #AD/B #AD/B #AD/B 2154 5383 B8 204 133% L1/L2 162 96% L1/L2 5383:B8 L1 L1 #AD/B #AD/#B OK OK 2155 5388 G6 226 148% 5AD/B 184 109% L4/1AD/B! 5388:G6 #B OK OK OK OK OK 121 5389 C4 213 139% Dead(L1/L2/AD) 181 107% L1/L2 5389:C4 L1 L1 L1/L2 #AD/#B #AD/B < 2157 5391 E4 197 129% L4/AD/B 161 95% 3AD/B! - Few Eggs 5391:E4 #1AD/B < < OK OK OK L1 L1 L1 L1 #AD/B OK 2158 5393 B2 219 143% L1/L2 190 112% 1AD/B 5393:B2 L1 L1/L2 #AD/B OK OK OK 119 5393 B4 187 122% 5AD/B 158 93% 1AD/B 5393:B4 #1AD/B #AD/B #AD/B #AD/B #AD/B < 2159 5393 B8 209 137% L1/L2 182 108% Dead 5393:B8 L1 L1 L1 L1/L2 #B OK 2160 5394 B1 197 129% L1/L2 184 109% L2/L3 5394:B1 L1 L1 #AD/#B < OK OK 2161 5394 G5 230 150% L1/L2 201 119% 1AD/B - Husks 5394:G5 #AD/B! #AD/#B OK OK OK OK 2162 5394 B11 232 152% 5AD/B 176 104% L2/L3 5394:B11 L1 #B #AD/B < OK OK 2163 5395 A1 195 172% 2AD/B! 169 100% 2AD/B! 5395:A1 L1 #B OK OK OK OK B! B! B! OK OK OK 2164 5395 A5 195 127% 3AD/B! 172 102% 3AD/B! 5395:A5 L1 OK OK OK OK OK 2165 5395 B5 203 133% L4/AD/B! - Clear 181 107% L4/AD/B! 5395:B5 #AD/B! < OK < OK OK 2166 5395 A6 207 135% L3/L4/1AD/B! 190 112% L2/L3 5395:A6 L1 OK OK OK OK OK 2167 5395 A8 177 116% L2/L3 162 96% 2AD/B! 5395:A8 L1 OK OK OK OK OK 2168 5395 A9 226 148% L2/L3 178 105% L2/L3 5395:A9 OK OK OK OK OK 2169 5396 A2 182 119% 4AD/B 176 104% 1AD/B! 5396:A2 #AD/B! < OK OK OK OK 2170 5396 A9 205 134% 4AD/B 182 108% 1AD/B! 5396:A9 L1 OK OK OK OK OK L1 L1 L1 L1 #B < 2171 5396 A10 186 122% L1/L2 177 105% L1 5396:A10 L1 #AD/B OK OK OK OK 2172 5396 A11 188 123% B! 176 104% 2AD/B! 5396:A11 L1 OK OK OK OK OK 2173 5396 B11 207 135% L2/L3 176 104% 2AD/B! 5396:B11 L1/L2 L3/#AD/B! B OK OK OK 2174 5397 G2 246 161% L2/L3/L4 192 114% L2/L3 5397:G2 L1/L2 L1/L2 #AD/b! B #B OK 2175 5397 G6 242 158% B (Dead AD) 209 124% L4/AD/B! 5397:G6 L2/L3 #AD/B! B #B < OK 2176 5397 F8 224 146% L4/AD/B 210 124% 2AD/B! 5397:F8 L1 #AD/B #AD/#B OK OK OK 2177 5398 G5 215 141% L1/L2 197 117% L1/L2 5398:G5 L1 #AD/B #AD/#B #AD/#B OK OK B! B! B! OK OK OK 2178 5398 G7 214 140% L4/AD/B! - Clear 204 121% L4/AD/B! - Clear 5398:G7 L2/L3 #B OK OK OK OK 2179 5398 H7 186 122% L4/AD/B - Clear 186 110% B 5398:H7 OK OK OK OK OK Table 3 2180 5398 G11 232 152% L1/L2 195 115% Dead(L3) 5398:G11 Dead(L2/L3) L1/L2 OK OK OK OK 2181 5398 H11 224 146% L1/L2 184 109% L3 5398:H11 L1 #AD/B < < < OK 110 5399 C4 225 147% L1/L2 198 117% L1 5399:C4 L1 L1 L1 L1 L1 2182 5400 C1 227 148% L2/L3 192 114% L2/L3/L4 5400:C1 L1/L2 #AD/B B OK OK OK 2183 5401 F4 216 141% L2/1AD/B! 137 81% B! - Healthy 5401:F4 L1 < < OK OK OK L1 L1 L1 L1 L1 L1 2184 5403 B8 190 124% L1/L2 168 99% L1/L2 5403;B8 L1 L1 < OK OK OK 2185 5406 A1 182 119% L4/AD/B! 167 99% 1AD/B! 5406:A1 L1 < OK OK OK OK 2186 5406 A9 203 133% L3/L4 176 104% L1/L2 5406:A9 L1 B OK OK OK OK 2187 5406 A10 182 119% 5AD/B! 177 105% L1/L2 5406:A10 L1 < OK OK OK OK 2188 5407 H5 253 165% L4/AD/B! - Clear 174 103% L2/L3 5407:H5 L1 #AD/B! #B OK OK OK 2189 5408 G1 225 147% B! 189 112% B 5408:G1 #AD/B! #AD/B < OK OK OK 2190 5408 G11 219 143% L4/AD/B! - Dying 175 104% 2AD/B! 5408:G11 Dead #AD/#B B OK OK OK B! B! B! OK OK OK 2191 5408 H11 227 148% L2/L3 191 113% L1/L2 5408:H11 L1 L1 < OK OK OK 2192 5409 A4 171 112% L4/AD/B! - Clear 45 86% L3/2AD/B! 5409:A4 L3/L4 B < OK OK OK 2193 5409 C4 204 133% L1/L2 193 114% L1/L2 5409:C4 L1 L1 L1 #AD/B! #B < 2194 5409 C8 219 143% L2/L3 - Dying 195 115% L2 5409:C8 L1 L1 B < OK B 2195 5410 C11 202 132% L3/1AD/B! 176 104% 2AD/B! 5410:C11 L1 #AD/B OK OK OK OK 2196 5413 B8 201 131% L2/Dead(L3) 161 95% L4/AD/B! - No Eggs 5413:B8 L1 L1 #AD/#B OK OK OK 2197 5414 B11 209 137% L1/L2 182 108% 1AD/B! 5414:B11 L1 L1 < OK OK OK L1 L1 L1 #AD/B < OK 2198 5416 A3 194 127% L1/L2 173 102% 1AD/B! 5416:A3 L1 #AD/B OK OK OK OK 2199 5416 A4 170 111% 4AD/B 160 95% L1 5416:A4 L1 B OK OK OK OK 2200 5416 A10 179 117% 4AD/B 163 96% L1/L2 5416:A10 L1 B OK OK OK OK 2201 5417 G2 234 153% L3/L4 - Dying 183 108% 3AD/B 5417:G2 L1 L1 < OK OK OK 2202 5417 G5 236 154% L1/L2 198 117% L1/L2 5417:G5 #AD/B! #AD/#B < OK OK 2203 5418 G11 225 147% L1/L2 185 109% L1/L2 - Dying 5418:G11 L1 L1/L2 < < OK OK 2204 5418 H11 223 146% L2/L3/L4 192 114% L3/L4/2AD/B! - No 5418:H11 L1 L4/#AD/B < OK OK OK B! B! B! OK OK OK 120 5419 C4 190 124% 4AD/B 215 127% 1AD/B! 5419:C4 L1 L2/L3 L1 L1/L2 #AD/#B OK 2206 5419 H4 241 158% 2AD/B 238 141% 3AD/B 5419:H4 Dead < OK OK OK OK 2207 5423 B8 175 114% 1AD/B 159 94% 1AD/B 5423:B8 Dead < #B OK OK OK 2208 5428 B11 207 135% L1/L2 175 104% L1/L2 5424:B11 Dead L1 #B < OK OK 2209 5425 A3 201 131% 5AD/B! 177 105% L1/L2 5425:A3 L1 OK OK OK OK OK 2210 5425 A7 187 122% L4/AD/B! 179 106% L1/L2 5425:A7 L1 OK OK OK OK OK 2211 5425 A10 189 124% B! - Dying 173 102% L1/L2 5425:A10 OK OK OK OK OK L1 L1 L1 L1 L1 #AD/B 2212 5426 A10 183 120% 2L3/2AD/B! 168 99% L1/L2 5426:A10 L1 #AD/B OK OK OK OK 2213 5426 E11 213 139% L2/L3 176 104% L2/L3 5426:E11 L1 L1 OK OK OK OK 2214 5427 G5 164 107% B - Unhatched Eggs 151 89% B! 5427:G5 L1/L2 #B OK OK OK OK Table 3 2215 5427 H5 212 139% L3/L4 174 103% L3/L4 5427:H5 L1 L1/L2 #B OK OK OK 2216 5428 G11 221 144% L2/L3 179 106% L2/L3 5428:G11 #B #B < #B OK OK 2217 5428 H11 230 150% L1/L2 178 105% L2/L3 5428:H11 L1 L1 #AD/#B OK < OK 2218 5429 H4 231 151% 4AD/B 199 118% 4AD/B 5429:H4 Dead < OK OK OK OK B! B! B! OK OK OK 2219 5430 E5 123 80% B! - Unhatched Eggs. He 143 85% B! - Unhatched Eg 5430:E5 OK OK OK OK OK 2220 5430 C11 217 142% L1/L2 189 112% Dead 5430:C11 L1 L1 #B < OK OK

Example 9Sheep 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 used 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 # 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.

GROUP10 : 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 will continue to be 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 will have intestinal material collected for nematode recovery and identification.

Nematodes are recovered, identified, and enumerated according to Zimmerman Research SOP#NEMRECOVID. 99. 01. All individualsperformingnematoderecoveries 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 will be slaughtered using humane methods recommended by the AVMA. The neck blood vessels are severed and after the animal is completely exsanguinated, the abdomen are 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.

Table 4 Akkadix Trial - 1 Sheep AKK 101 Strongyles Strongyles ! ! Strongyles ! Sheep Weight/lbs Total 17-Jan 22-Jan 24-Jan Number 1/12/2000 EPG-pre EPG-pre EPG-5day % change EPG-7day %Change Group 59 79 4210 4130 4640 14370 -247.94 AKC 104 72 42 1540 1510 1800 3580 -137.09 1. 4mg/kg 67 2001 31. 03 Total / Mean 179 2013.33 1976.67 2190.99 -10.79 6050.00 -206. 07 Group 57 67 3220 2980 510 2160 27.52 ~~ 2160 ! P I AKC 105 79 86 790 790 390 1230 -55.70 1.4mg/kg 73 47 140 140 80 490 -250.00 Total/Mean 200 2075.00 1955.00 490. 00 74. 94 1940.00 0. 77 Group 83 91 2310 2000 10 0 100.00 Ivermectin 92 113 570 570 0 0 100. 00 . 2mg/kg 53 77 90 70 0 0 100.00 Total/Mean 281 990.00 880.00 3.33 99.62 0.00 100.00 Group 1 81 74 2240 2240 780 770 65. 63 Negative 75 109 370 300 260 1360 -353.33 Control 56 80 40 40 30 30 25.00 Total/Mean 1 263 883.33 860.00 356.67 58.53 720.00 16.28

Example 10-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 compounds 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 individualsperformingnematoderecoveries 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 Result are depicted in Tables 5 and 6.

Table 5 Akkadix |AKK-102 Sheep 24-May Sheep Weight/tbs worm Counts Number 5/17/2000 Abomasum Abomasum Small Intestine Small Intest. Large Intest. Haemonchus Ostertagia Trichostrongylus Nematodirus Trichuris Group 530 47 60 580 40 8500 5 Negative 1341 57 60 20 20 2520 15 Control 524 54 20 80 0 0 10 Mean Ct. 47 227 20 3673 10 Group 1347 45 0 0 0 100 0 Ivermectin 1336 58 20 0 0 100 0 200mcg/kg 539 47 0 0 0 0 0 Mean Ct. 7 0 0 67 0 %Efficacy 86 100 100 98 Group 523 31 720 1740 AKC 104 1335 41 0 120 1 mg/kg 1340 53 60 200 Mean Ct. 260 687 %Efficacy -457 Table 6 Akkadix Trial-2 Sheep ! AKK 102 Strongyles Strongyles Strongyles Sheep Weight/lbs Total 15-May 22-May 24-Ma Number j 5/17/2000 EPG-pre EPG-pre JEPG-5dayf% Changer EPG-7day% Change Group 1337 45 1160 1070 900 890 16. 82 AKC 105 1343 47 390 180 130 310 -72. 22 1. 5mg/kg 537 36 160 140 160 430 -207. 14 Total/Mean 128 570.00 463.33 396.67 14. 39 543.33 -17. 27 Group Group 311 10301 8301 6101 1080-30. 12 Group 523 31 1030 830 610 1080 -30.12 AKC 104 1335 41 370 220 170 60 72.73 1mg/kg 1340 53 120 100 NS 120 -20.00 Total/Mean 125 506.67 383.33 390.00 -1.74 420.00 -9.57 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 15.85 Group 1334 41 560 530 690 290 45.28 AKC 110 536 40 280 220 850 1510 -586.36 -183. 33 3.5mg/kg 1339 46 70 40 0 30 25.00 Group 127 303.33 263.33 513.33 -94.94 610.00 -131.65 Total/Mean 127 303.33 263.33 513.33 -94.94 610.00 -1 Group 13471 451 560 450 0, 0 j 100. 00 Ivermectin 1336 58 220 170 0 0 100.00 200mc/kg 539 47 100 40 0 0 100.00 Total/Mean 150 293.33 220.00 0.00 100.00 0.00 100.00 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.