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. Met. J 56 : 239-251 ; Coles, G. C. [1986]"Anthelmintic resistance in sheep,"In Veterinary Clinics ofNorth, 4merica : 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, Bia, and B2,, ; and four minor compounds : Alb, A2b, Blb, 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 Chemotlçer. 15 (3) : 361-367). The isolation and purification of these compounds is also described in U. S. Patent No. 4, 310, 519, issued January 12, 1982.

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

4, 547, 520.

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

The activity of avermectins must generally be determined empirically.

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

AvermectinB2a 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 [1981J ; 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 Bl is a combination of avermectin Bla (major component) and avermectin Blb. It has demonstrated a broad spectrum of insecticidal activities. The data indicate that avermectin B, is primarily a miticide, although it is also effective on the Colorado potato beetle, potato tuberworm, beet armyworm, diamondback moth, gypsy moth, and the European corn borer.

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

December24, 1985 ; EPO applicationsO 094 779 andO 125 155 ; and Anderson, T. E., J. R.

Babu, R. A. Dybas, H. Mehta (1986) J. Ecopn. 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 hydroxyproline 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 d 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) l-NH-CO-NR1-Alk-R2 (Formula I) wherein Ar is aryl or heteroaryl optionally substituted by one or more R3 groups ; each Alk is a linear or cyclic alkylene radical of up to 8 C atoms ; Rl is H or C16 alkyl ; R2 is heteroaryl or heterocycloalkyl optionally substituted by Ar, or forms such a group by cyclisation with R' ; and R3 is OH, halogen, CF3, OCF, or a group selected from NH2, SO2-C,~6 alkyl, C6~lo aryl, C6. l0 aryloaxy, Cs 6 cycloalkyl, Cl 5 alkoxy, and Cl 6 alkyl, said group being optionally substituted by OH, C16 alkoxy, Cl 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) o-,-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 heterocycloalkyloptionally substitutedby Ar and/or R3 ; and R3 is OH, halogen, CF3, OCF3, or a group selectedfromNH2, SO2 alkyl, C610 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 C1 6 alkyl, aryl, or heteroaryl optionally substituted by R3 ; or R and Y together form a cycloalkyl or heterocycloalkyl ring ; and R3 is OH, halogen, CF3, OCF3, or a group selectedfromNH2, SO2 alkyl, C6 l0 aryl, Cl 6 alkoxy, and Cl 6 alkyl, said group being optionally substituted by OH, CI~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)0-1-Ar-(CH2)-01-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 $3,

R3 is OH, halogen, CF3, OCF3, or a group selected from NH2, SO2 alkyl, C6-10 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 IV are exemplifiedby 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, CI 6 alkoxy, Cl 6 alkyl, phenyl, halogen, or CF3.

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

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

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

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

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

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

Other preferred anthelmintic compounds useful according to the subj ect 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 subj ect anthelmintic compounds are used in conjunctionwith 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 subj ect 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 preplant 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 1297 of the class represented in Figure 47.

Figure 49 depicts anthelmintic compound AKC 1299 of the class represented in Figure 47.

Figure 50 depicts anthelmintic compound AKC 13 00 of the class represented in Figure 47.

Figure 51 depicts anthelmintic compound AKC 1298 of the class represented in Figure 47.

Figure 52 depicts anthelmintic compound AKC 1301 of the class represented in Figure 47.

Figure 53 depicts anthelmintic compound AKC 1302 of the class represented in Figure 47.

Figure 54 depicts anthelmintic compound AKC 126 of the class represented in Figure 47.

Figure 55 depicts anthelmintic compound AKC 1303 of the class represented in Figure 47.

Figure 56 depicts anthelmintic compound AKC 1314 of the class represented in Figure 47.

Figure 57 depicts anthelmintic compound AKC 1305 of the class represented in Figure 47.

Figure 58 depicts anthelmintic compound AKC 1315 of the class represented in Figure 47.

Figure 59 depicts anthelmintic compound AKC 13 06 of the class represented in Figure 47.

Figure 60 depicts anthelmintic compound AKC 1316 of the class represented in Figure 47.

Figure 61 depicts anthelmintic compound AKC 1307 of the class represented in Figure 47.

Figure 62 depicts anthelminticcompoundAKC 1317 of the class represented in Figure 47.

Figure 63 depicts anthelmintic compound AKC 1308 of the class represented in Figure 47.

Figure 64 depicts anthelmintic compound AKC 1318 of the class represented in Figure 47.

Figure 65 depicts anthelmintic compound AKC 1304 of the class represented in Figure 47.

Figure 66 depicts anthelmintic compound AKC 1309 of the class represented in Figure 47.

Figure 67 depicts anthelmintic compound AKC 102 of the class represented in Figure 47.

Figure 68 depicts anthelmintic compound AKC 1313 of the class represented in Figure 47.

Figure 69 depicts anthelmintic compound AKC 1319 of the class represented in Figure 47.

Figure 70 depicts anthelmintic compound AKC 1310 of the class represented in Figure 47.

Figure 71 depicts anthelmintic compound AKC 1320 of the class represented in Figure 47.

Figure 72 depicts anthelmintic compound AKC 1311 of the class represented in Figure 47.

Figure 73 depicts anthelmintic compound AKC 1321 of the class represented in Figure 47.

Figure 74 depicts anthelmintic compound AKC 1312 of the class represented in Figure 47.

Figure 75 depicts anthelmintic compound AKC 1322 of the class represented in Figure 47.

Figure 76 depicts anthelmintic compound AKC 1323 of the class represented in Figure 47.

Figure 77 depicts anthelmintic compound AKC 1326 of the class represented in Figure 47.

Figure 78 depicts anthelmintic compound AKC 1324 of the class represented in Figure 47.

Figure 79 depicts anthelmintic compound AKC 1327 of the class represented in Figure 47.

Figure 80 depicts anthelmintic compound AKC 1325 of the class represented in Figure 47.

Figure 81 depicts anthelmintic compound AKC 1328 of the class represented in Figure 47.

Figure 82 depicts anthelmintic compound AKC 1331 of the class represented in Figure 47.

Figure 83 depicts anthelmintic compound AKC 1329 of the class represented in Figure 47.

Figure 84 depicts anthelmintic compound AKC 1330 of the class represented in Figure 47.

Figure 85 depicts anthelmintic compound AKC 1332 of the class represented in Figure 47.

Figure 86 depicts anthelmintic compound AKC 1333 of the class represented in Figure 47.

Figure 87 depicts anthelmintic compound AKC 1334 of the class represented in Figure 47.

Figure 88 depicts anthelmintic compound AKC 1335 of the class represented in Figure 47.

Figure 89 depicts anthelmintic compound AKC 1337 of the class represented in Figure 47.

Figure 90 depicts anthelmintic compound AKC 1336 of the class represented in Figure 47.

Figure 91 depicts anthelmintic compound AKC 1339 of the class represented in Figure 47.

Figure 92 depicts anthelmintic compound AKC 1345 of the class represented in Figure 47.

Figure 93 depicts anthelmintic compound AKC 1340 of the class represented in Figure 47.

Figure 94 depicts anthelmintic compound AKC 1346 of the class represented in Figure 47.

Figure 95 depicts anthelmintic compound AKC 1338 of the class represented in Figure 47.

Figure 96 depicts anthelmintic compound AKC 1341 of the class represented in Figure 47.

Figure 97 depicts anthelmintic compound AKC 1342 of the class represented in Figure 47.

Figure 98 depicts anthelmintic compound AKC 1347 of the class represented in Figure 47.

Figure 99 depicts anthelmintic compound AKC 1343 of the class represented in Figure 47.

Figure 100 depicts anthelmintic compound AKC 1348 of the class represented in Figure 47.

Figure 101 depicts anthelmintic compound AKC 1344 of the class represented in Figure 47.

Figure 102 depicts anthelmintic compound AKC 1349 of the class represented in Figure 47.

Figure 103 depicts anthelmintic compound AKC 1353 of the class represented in Figure 47.

Figure 104 depicts anthelmintic compound AKC 1350 of the class represented in Figure 47.

Figure 105 depicts anthelmintic compound AKC 1351 of the class represented in Figure 47.

Figure 106 depicts anthelmintic compound AKC 1352 of the class represented in Figure 47.

Figure 107 depicts anthelmintic compound AKC 1354 of the class represented in Figure 47.

Figure 108 depicts anthelmintic compound AKC 1355 of the class represented in Figure 47.

Figure 109 depicts anthelmintic compound AKC 1356 of the class represented in Figure 47.

Figure 110 depicts anthelmintic compound AKC 1357 of the class represented in Figure 47.

Figure 111 depicts anthelmintic compound AKC 1358 of the class represented in Figure 47.

Figure 112 depicts anthelmintic compound AKC 1359 of the class represented in Figure 47.

Figure 113 depicts anthelmintic compound AKC 1360 of the class represented in Figure 47.

Figure 114 depicts anthelmintic compound AKC 1368 of the class represented in Figure 47.

Figure 115 depicts anthelmintic compound AKC 1361 of the class represented in Figure 47.

Figure 116 depicts anthelmintic compound AKC 1369 of the class represented in Figure 47.

Figure 117 depicts anthelmintic compound AKC 1362 of the class represented in Figure 47.

Figure 118 depicts anthelmintic compound AKC 1363 of the class represented in Figure 47.

Figure 119 depicts anthelmintic compound AKC 1364 of the class represented in Figure 47.

Figure 120 depicts anthelmintic compound AKC 1365 of the class represented in Figure 47.

Figure 121 depicts anthelmintic compound AKC 1366 of the class represented in Figure 47.

Figure 122 depicts anthelmintic compound AKC 1367 of the class represented in Figure 47.

Figure 123 depicts anthelmintic compound AKC 1377 of the class represented in Figure 47.

Figure 124 depicts anthelmintic compound AKC 1370 of the class represented in Figure 47.

Figure 125 depicts anthelmintic compound AKC 1378 of the class represented in Figure 47.

Figure 126 depicts anthelmintic compound AKC 1371 of the class represented in Figure 47.

Figure 127 depicts anthelmintic compound AKC 1372 of the class represented in Figure 47.

Figure 128 depicts anthelmintic compound AKC 1379 of the class represented in Figure 47.

Figure 129 depicts anthelmintic compound AKC 1373 of the class represented in Figure 47.

Figure 130 depicts anthelmintic compound AKC 1380 of the class represented in Figure 47.

Figure 131 depicts anthelmintic compound AKC 1374 of the class represented in Figure 47.

Figure 132 depicts anthelmintic compound AKC 1375 of the class represented in Figure 47.

Figure 133 depicts anthelmintic compound AKC 1381 of the class represented in Figure 47.

Figure 134 depicts anthelmintic compound AKC 1376 of the class represented in Figure 47.

Figure 135 depicts anthelmintic compound AKC 1382 of the class represented in Figure 47.

Figure 136 depicts anthelmintic compound AKC 1390 of the class represented in Figure 47.

Figure 137 depicts anthelmintic compound AKC 1383 of the class represented in Figure 47.

Figure 138 depicts anthelmintic compound AKC 1384 of the class represented in Figure 47.

Figure 139 depicts anthelmintic compound AKC 1391 of the class represented in Figure 47.

Figure 140 depicts anthelmintic compound AKC 1385 of the class represented in Figure 47.

Figure 141 depicts anthelmintic compound AKC 1392 of the class represented in Figure 47.

Figure 142 depicts anthelmintic compound AKC 1386 of the class represented in Figure 47.

Figure 143 depicts anthelmintic compound AKC 1393 of the class represented in Figure 47.

Figure 144 depicts anthelmintic compound AKC 1387 of the class represented in Figure 47.

Figure 145 depicts anthelmintic compound AKC 1388 of the class represented in Figure 47.

Figure 146 depicts anthelmintic compound AKC 1394 of the class represented in Figure 47.

Figure 147 depicts anthelmintic compound AKC 1389 of the class represented in Figure 47.

Figure 148 depicts anthelmintic compound AKC 1395 of the class represented in Figure 47.

Figure 149 depicts anthelmintic compound AKC 1396 of the class represented in Figure 47.

Figure 150 depicts anthelmintic compound AKC 1397 of the class represented in Figure 47.

Figure 151 depicts anthelmintic compound AKC 1259 of the class represented in Figure 47.

Figure 152 depicts anthelmintic compound AKC 1260 of the class represented in Figure 47.

Figure 153 depicts anthelmintic compound AKC 1261 of the class represented in Figure 47.

Figure 154 depicts anthelmintic compound AKC 1262 of the class represented in Figure 47.

Figure 155 depicts anthelmintic compound AKC 1268 of the class represented in Figure 47.

Figure 156 depicts anthelmintic compound AKC 1263 of the class represented in Figure 47.

Figure 157 depicts anthelmintic compound AKC 1267 of the class represented in Figure 47.

Figure 158 depicts anthelmintic compound AKC 1269 of the class represented in Figure 47.

Figure 159 depicts anthelmintic compound AKC 1264 of the class represented in Figure 47.

Figure 160 depicts anthelmintic compound AKC 1270 of the class represented in Figure 47.

Figure 161 depicts anthelmintic compound AKC 1265 of the class represented in Figure 47.

Figure 162 depicts anthelmintic compound AKC 1271 of the class represented in Figure 47.

Figure 163 depicts anthelmintic compound AKC 1266 of the class represented in Figure 47.

Figure 164 depicts anthelmintic compound AKC 1406 of the class represented in Figure 47.

Figure 165 depicts anthelmintic compound AKC 1398 of the class represented in Figure 47.

Figure 166 depicts anthelmintic compound AKC 1407 of the class represented in Figure 47.

Figure 167 depicts anthelmintic compound AKC 1399 of the class represented in Figure 47.

Figure 168 depicts anthelmintic compound AKC 1408 of the class represented in Figure 47.

Figure 169 depicts anthelmintic compound AKC 1400 of the class represented in Figure 47.

Figure 170 depicts anthelmintic compound AKC 1409 of the class represented in Figure 47.

Figure 171 depicts anthelmintic compound AKC 1401 of the class represented in Figure 47.

Figure 172 depicts anthelmintic compound AKC 1410 of the class represented in Figure 47.

Figure 173 depicts anthelmintic compound AKC 1402 of the class represented in Figure 47.

Figure 174 depicts anthelmintic compound AKC 1411 of the class represented in Figure 47.

Figure 175 depicts anthelmintic compound AKC 1403 of the class represented in Figure 47.

Figure 176 depicts anthelmintic compound AKC 1412 of the class represented in Figure 47.

Figure 177 depicts anthelmintic compound AKC 1404 of the class represented in Figure 47.

Figure 178 depicts anthelmintic compound AKC 1413 of the class represented in Figure 47.

Figure 179 depicts anthelmintic compound AKC 1405 of the class represented in Figure 47.

Figure 180 depicts anthelmintic compound AKC 1422 of the class represented in Figure 47.

Figure 181 depicts anthelmintic compound AKC 1414 of the class represented in Figure 47.

Figure 182 depicts anthelmintic compound AKC 1423 of the class represented in Figure 47.

Figure 183 depicts anthelmintic compound AKC 1415 of the class represented in Figure 47.

Figure 184 depicts anthelmintic compound AKC 1424 of the class represented in Figure 47.

Figure 185 depicts anthelmintic compound AKC 1416 of the class represented in Figure 47.

Figure 186 depicts anthelmintic compound AKC 1425 of the class represented in Figure 47.

Figure 187 depicts anthelmintic compound AKC 1417 of the class represented in Figure 47.

Figure 188 depicts anthelmintic compound AKC 1418 of the class represented in Figure 47.

Figure 189 depicts anthelmintic compound AKC 1426 of the class represented in Figure 47.

Figure 190 depicts anthelmintic compound AKC 1419 of the class represented in Figure 47.

Figure 191 depicts anthelmintic compound AKC 1427 of the class represented in Figure 47.

Figure 192 depicts anthelmintic compound AKC 1420 of the class represented in Figure 47.

Figure 193 depicts anthelmintic compound AKC 1428 of the class represented in Figure 47.

Figure 194 depicts anthelmintic compound AKC 1421 of the class represented in Figure 47.

Figure 195 depicts anthelmintic compound AKC 1437 of the class represented in Figure 47.

Figure 196 depicts anthelmintic compound AKC 1429 of the class represented in Figure 47.

Figure 197 depicts anthelmintic compound AKC 1438 of the class represented in Figure 47.

Figure 198 depicts anthelmintic compound AKC 1430 of the class represented in Figure 47.

Figure 199 depicts anthelmintic compound AKC 1439 of the class represented in Figure 47.

Figure 200 depicts anthelmintic compound AKC 1431 of the class represented in Figure 47.

Figure 201 depicts anthelmintic compound AKC 1440 of the class represented in Figure 47.

Figure 202 depicts anthelmintic compound AKC 1432 of the class represented in Figure 47.

Figure 203 depicts anthelmintic compound AKC 1441 of the class represented in Figure 47.

Figure 204 depicts anthelmintic compound AKC 1433 of the class represented in Figure 47.

Figure 205 depicts anthelmintic compound AKC 1442 of the class represented in Figure 47.

Figure 206 depicts anthelmintic compound AKC 1434 of the class represented in Figure 47.

Figure 207 depicts anthelmintic compound AKC 1443 of the class represented in Figure 47.

Figure 208 depicts anthelmintic compound AKC 1435 of the class represented in Figure 47.

Figure 209 depicts anthelmintic compound AKC 1444 of the class represented in Figure 47.

Figure 210 depicts anthelmintic compound AKC 1436 of the class represented in Figure 47.

Figure 211 depicts anthelmintic compound AKC 1445 of the class represented in Figure 47.

Figure 212 depicts anthelmintic compound AKC 1446 of the class represented in Figure 47.

Figure 213 depicts anthelmintic compound AKC 1272 of the class represented in Figure 47.

Figure 214 depicts anthelmintic compound AKC 1273 of the class represented in Figure 47.

Figure 215 depicts anthelmintic compound AKC 1274 of the class represented in Figure 47.

Figure 216 depicts anthelmintic compound AKC 1275 of the class represented in Figure 47.

Figure 217 depicts anthelmintic compound AKC 1276 of the class represented in Figure 47.

Figure 218 depicts anthelmintic compound AKC 1280 of the class represented in Figure 47.

Figure 219 depicts anthelmintic compound AKC 1277 of the class represented in Figure 47.

Figure 220 depicts anthelmintic compound AKC 1278 of the class represented in Figure 47.

Figure 221 depicts anthelmintic compound AKC 1279 of the class represented in Figure 47.

Figure 222 depicts anthelmintic compound AKC 1455 of the class represented in Figure 47.

Figure 223 depicts anthelmintic compound AKC 1447 of the class represented in Figure 47.

Figure 224 depicts anthelmintic compound AKC 1456 of the class represented in Figure 47.

Figure 225 depicts anthelmintic compound AKC 1457 of the class represented in Figure 47.

Figure 226 depicts anthelmintic compound AKC 1448 of the class represented in Figure 47.

Figure 227 depicts anthelmintic compound AKC 1458 of the class represented in Figure 47.

Figure 228 depicts anthelmintic compound AKC 1449 of the class represented in Figure 47.

Figure 229 depicts anthelmintic compound AKC 1459 of the class represented in Figure 47.

Figure 230 depicts anthelmintic compound AKC 1450 of the class represented in Figure 47.

Figure 231 depicts anthelmintic compound AKC 1454 of the class represented in Figure 47.

Figure 232 depicts anthelmintic compound AKC 1460 of the class represented in Figure 47.

Figure 233 depicts anthelmintic compound AKC 1451 of the class represented in Figure 47.

Figure 234 depicts anthelmintic compound AKC 1461 of the class represented in Figure 47.

Figure 235 depicts anthelmintic compound AKC 1452 of the class represented in Figure 47.

Figure 236 depicts anthelmintic compound AKC 1453 of the class represented in Figure 47.

Figure 237 depicts anthelmintic compound AKC 1469 of the class represented in Figure 47.

Figure 238 depicts anthelmintic compound AKC 1462 of the class represented in Figure 47.

Figure 239 depicts anthelmintic compound AKC 1470 of the class represented in Figure 47.

Figure 240 depicts anthelmintic compound AKC 1463 of the class represented in Figure 47.

Figure 241 depicts anthelmintic compound AKC 1471 of the class represented in Figure 47.

Figure 242 depicts anthelmintic compound AKC 1464 of the class represented in Figure 47.

Figure 243 depicts anthelmintic compound AKC 1472 of the class represented in Figure 47.

Figure 244 depicts anthelmintic compound AKC 1465 of the class represented in Figure 47.

Figure 245 depicts anthelmintic compound AKC 1466 of the class represented in Figure 47.

Figure 246 depicts anthelmintic compound AKC 1473 of the class represented in Figure 47.

Figure 247 depicts anthelmintic compound AKC 1467 of the class represented in Figure 47.

Figure 248 depicts anthelmintic compound AKC 1474 of the class represented in Figure 47.

Figure 249 depicts anthelmintic compound AKC 1468 of the class represented in Figure 47.

Figure 250 depicts anthelmintic compound AKC 1475 of the class represented in Figure 47.

Figure 251 depicts anthelmintic compound AKC 1486 of the class represented in Figure 47.

Figure 252 depicts anthelmintic compound AKC 1478 of the class represented in Figure 47.

Figure 253 depicts anthelmintic compound AKC 1487 of the class represented in Figure 47.

Figure 254 depicts anthelmintic compound AKC 1479 of the class represented in Figure 47.

Figure 255 depicts anthelmintic compound AKC 1488 of the class represented in Figure 47.

Figure 256 depicts anthelmintic compound AKC 1480 of the class represented in Figure 47.

Figure 257 depicts anthelmintic compound AKC 1489 of the class represented in Figure 47.

Figure 258 depicts anthelmintic compound AKC 1476 of the class represented in Figure 47.

Figure 259 depicts anthelmintic compound AKC 1477 of the class represented in Figure 47.

Figure 260 depicts anthelmintic compound AKC 1481 of the class represented in Figure 47.

Figure 261 depicts anthelmintic compound AKC 1490 of the class represented in Figure 47.

Figure 262 depicts anthelmintic compound AKC 1482 of the class represented in Figure 47.

Figure 263 depicts anthelmintic compound AKC 1491 of the class represented in Figure 47.

Figure 264 depicts anthelmintic compound AKC 1483 of the class represented in Figure 47.

Figure 265 depicts anthelmintic compound AKC 1492 of the class represented in Figure 47.

Figure 266 depicts anthelmintic compound AKC 1484 of the class represented in Figure 47.

Figure 267 depicts anthelmintic compound AKC 1493 of the class represented in Figure 47.

Figure 268 depicts anthelmintic compound AKC 1485 of the class represented in Figure 47.

Figure 269 depicts anthelmintic compound AKC 1494 of the class represented in Figure 47.

Figure 270 depicts anthelmintic compound AKC 1495 of the class represented in Figure 47.

Figure 271 depicts anthelmintic compound AKC 1496 of the class represented in Figure 47.

Figure 272 depicts anthelmintic compound AKC 1497 of the class represented in Figure 47.

Figure 273 depicts anthelmintic compound AKC 1281 of the class represented in Figure 47.

Figure 274 depicts anthelmintic compound AKC 1282 of the class represented in Figure 47.

Figure 275 depicts anthelmintic compound AKC 1283 of the class represented in Figure 47.

Figure 276 depicts anthelmintic compound AKC 1289 of the class represented in Figure 47. °

Figure 277 depicts anthelmintic compound AKC 1284 of the class represented in Figure 47.

Figure 278 depicts anthelmintic compound AKC 1290 of the class represented in Figure 47.

Figure 279 depicts anthelmintic compound AKC 1285 of the class represented in Figure 47.

Figure 280 depicts anthelmintic compound AKC 1286 of the class represented in Figure 47.

Figure 281 depicts anthelmintic compound AKC 1287 of the class represented in Figure 47.

Figure 282 depicts anthelmintic compound AKC 1291 of the class represented in Figure 47.

Figure 283 depicts anthelmintic compound AKC 1288 of the class represented in Figure 47.

Figure 284 depicts anthelmintic compound AKC 1498 of the class represented in Figure 47.

Figure 285 depicts anthelmintic compound AKC 1499 of the class represented in Figure 47.

Figure 286 depicts anthelmintic compound AKC 1292 of the class represented in Figure 47.

Figure 287 depicts anthelmintic compound AKC 1296 of the class represented in Figure 47.

Figure 288 depicts anthelmintic compound AKC 1293 of the class represented in Figure 47.

Figure 289 depicts anthelmintic compound AKC 1294 of the class represented in Figure 47.

Figure 290 depicts anthelmintic compound AKC 1295 of the class represented in Figure 47.

Figure 291 depicts one library scheme by which the skilled artisan can create the compounds represented by the structure depicted in Figure 47.

Figure 292 depicts Scaffold 1.

Figure 293 depicts Scaffold 2.

Figure 294 depicts Scaffold 3.

Figure 295 depicts Scaffold 4.

Figure 296 depicts Scaffold 5.

Figure 297 depicts Scaffold 6.

Figure 298 depicts a preferred pathway for synthesis of hydroxyproline derivatives.

Figure 299 depicts a preferred alternate pathway for synthesis of hydroxyproline derivatives.

Figure 300 depicts a synthetic pathway using diamines on nitrophenol carbonate linker.

Figure 301 compares the original base structure of intended compounds with the preferred base structure of the subject compounds.

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 are the compounds represented by Structures 22 and 23, and compounds related thereto as represented by Structure 47 depicted in Figure 47, and as further exemplifiedby Structures 48-290 depicted in Figures 48 through 290.

Preferred anthelmintic compounds useful in accord with the subject invention are represented by Structure 47, wherein : Rl is C1~5 branched or straight alkyl (optionally substituted with phenyl) ; C3 S cyclic alkyl ; aryl (optionally substituted with Cl l0 branched or straight alkyl, or Oui 5) ; R2 is C1-5 branched or straight alkyl which is optionally substituted with aryl (optionally substituted with Oc 5 or OAr) ; NCH2R5wherein R5 is Ar (optionally substituted with OCF3) ; OCl 5 ; CH2OR6wherein R6 is Cl 5 alkyl or C38 cyclic alkyl (optionally substituted with Cl 5 straight or branched alkyl, or OC1~5) ; aryl (optionally substituted with Cl 5 straight or branched alkyl ; OC1~5 ; halogen ; naphthyl (optionally substituted with OCI 5 or an amine) ; or a 3 ring fused polycyclic group ;

R3 is H ; Ci. s branched or straight alkyl which is optionally substituted with aryl (optionally substituted with halogen, OC, 5, Cl 5 branched or straight alkyl) ; OC, ~5 ; C28 ether ; cyclic alkyl (optionally substituted with Ci. branched, straight, or cylcic alkyl) ; aryl (optionally substituted with halogen, Cl-5 straight or branched alkyl, OCF3) ; R4 is H ; Cl 5 branched or straight alkyl which is optionally substituted with aryl (optionally substituted with halogen, OC, C ; ; branched or straight alkyl) ; OC ; C28 ether ; cyclic alkyl (optionally substituted with Ci. jo branched, straight, or cylcic alkyl) ; aryl (optionally substituted with halogen, Cl 5 straight or branched alkyl, OCF3) ; and X is CO or S02- Generally, the anthelmintic compounds of the subject invention can be unsubstituted or substituted, saturated or unsaturated. The anthelmintic component of an anthelmintic compounds used according to the subject invention may be a single anthelmintic compound or a mixture of two or more anthelmintic compounds. The subject compounds may be used in conjunction with other anthelmintic compounds, including the free acids and salts of the anthelmintic compounds of the present invention.

The salts may be, for example, sodium or potassium salts, or ammonium salts. As would be apparent to the ordinary skilled artisan, physiologically acceptable acids and salts of the subject anthelmintic compounds can readily be made and used in accord with the teachings herein, and are hereby expressly included by reference to each compound or group of compounds. For example,"AKC 1297","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 48-290 represented by Structures 48-290 depicted in Figures 48-290.

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

The anthelmintic compositions may be applied by, for example, drip and drench techniques. With the drip application, the subject compositions can be applied directly

to the base of plants or to the soil root zone. The composition may be applied through already existing drip irrigation systems. This procedure is particularly applicable for ornamental plants, strawberries, tomatoes, potatoes, grapes, and vegetables.

Alternatively, a drench application can be used. For treating plants, a sufficient quantity of the anthelmintic composition is applied such that the composition drains to the root area of the plants. An important aspect of the subject invention is the surprising discovery that certain 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 conjunctionwith 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 subj ect invention. Ivermectin may also be used according to the subject invention, as may the milbemycins. For brevity, the term"avermectin"is used herein to refer to all the avermectins and their derivatives as well as related compounds such as the milbemycins and the ivermectins."Derivatives"refer to chemical modifications of the avermectins or milbemycins which are well known and available to those skilled in this art. Such derivatives are described, for example, in U. S. Patent No. 4, 560, 677.

Avermectin is readily available under a variety of tradenames including"AVID," "ZEPHYR,""VERTIMEC,"and"AGRI-MEK." The anthelmintic compositions of the subject invention may also be used in conjunction with nematicidal agents other than the avermectins. For example, the anthelmintic compounds may be used with biological agents such as 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 replant 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 22, 23 and Structures 48-290, are used in preferred embodiments for killing nematode eggs. In addition, microemulsions of the subj ect 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. This may be achieved by applying the compound to the skin, hide and/or hair of the animals, or injecting or orally dosing said animals with a solid or liquid formulated composition.

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

The compositions of the present invention may be employed as animal feeds, animal feed premixes or feed concentrates. Feed concentrates and feed premixes, useful in the practice of the invention, may be prepared by admixing about 0. 25% to 35% by weight of a subject anthelmintic compound, preferably a Structure 1-46 compound, with about 99. 75% to 65% by weight of a suitable agronomic carrier or diluent. Carriers suitable for use include 0. 75% to 35% by weight of a physiologically acceptable alcohol such as benzyl alcohol, phenethyl alcohol or propylene glycol, 0 to about 10% by weight of a vegetable oil such as corn oil or soybean oil, or propylene glycol and about 30% to 95% by weight of a sorptive, edible organic carrier such as corn grits, wheat middlings, soybean meal, expanded corn grits, extracted corn meal or the like or a sorptive silica or a silicate. These feed premixes or concentrates may be admixed with the appropriate amount of animal feed to provide the animals with about 0. 5 ppm to 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 Structure 22, 23, and Structure 48-290 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, disintegratingagents 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 3, 000 ppm of the compound.

Advantageously, the subject anthelmintic compounds may also be prepared as pour-on formulations and poured on the backs of the animals such as swine, cattle, sheep, horses, poultry, and companion animals to protect them against infestation by nematodes, acarids, and arthropod endo-and ectoparasites. Such pour-on compositions are generally prepared by dissolving, dispersing, or emulsifying the anthelmintic compound in a suitable nontoxic pharmacologically acceptable diluent for pour-on and administration.

The diluent must be compatible with the compound and should not be a source of irritation or damage to the animals hide, skin, or hair. Such diluents include vegetable oils, spreading oils, polyhydric alcohols, aliphatic or aromatic hydrocarbons, esters of fatty acids, and lower alkyl ketones.

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

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

The compounds of Formulae I-V, Structure 47, particularly Structures 1-46, and particularly Structures 22, 23, and 48-290 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 22, 23, and 48-290, 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 22, 23, and 48-290, 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 22, 23, and 48-290, 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 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 ingredientwith 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 (S C) 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-ionictype 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 disperible powder WS) or liquids (flowable concentrate FS, liquid seed treatment LS), or microcapsule suspensions CS for use in seed treatments.

The formulations can be applied to the seed by standard techniques and through conventional seed treaters. In use the compositions are applied to the nematodes, to the locus of the nematodes, to the habitat of the nematodes, or to growing plants liable to infestation by the nematodes, by any of the known means of applying pesticidal compositions, for example, by dusting, spraying, or incorporation of granules.

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

Suitable additional active ingredients for inclusion in admixture with the compounds of the invention may be compounds which will broaden the spectrum of activity of the compounds of the invention or increase their persistence in the location of

the pest. They may synergise the activity of the compound of the invention or complement the activity for example by increasing the speed of effect or overcoming repellency. Additionally multi-component mixtures of this type may help to overcome or prevent the development of resistance to individual components.

The particular additional active ingredient included will depend upon the intended utility of the mixture and the type of complementary action required. Examples of suitable insecticides include the following : a) Pyrethroids such as permethrin, esfenvalerate, deltamethrin, cyhalothrin in particular lambda-cyhalothrin, biphenthrin, fenpropathrin, cyfluthrin, tefluthrin, fish safe pyrethroids for example ethofenprox, natural pyrethrin, tetramethrin, s-bioallethrin, fenfluthrin, prallethrin, and 5-benzyl-3-furylmethyl-(E)-(l 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) Carbamate (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) Amides, 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 fertilizers and other biologically active materials, are described, for instance, in International applicationNo. 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 : AC : Acylator ACD : Available Chemicals Directory ACN : Acetonitrile AcOH : Acetic Acid AM : Amine AUC : Area under curve BOC : t-Butoxycarbonyl Hyp-OH : Hydroxyproline DCM : Dichloromethane DIEA : N, N-Diisopropylethylamine DIC : 1, 3-Diisopropylcarbodiimide DMAP : Dimethylaminopyridine DMF : N, N-Dimethylformamide DMSO : Dimethylsulfoxide ESI : Electrospray ionization ESMS : ElectroSprayMassSpectrometry Fmoc : 9-Fluorenylmethoxycarbonyl HOBT : 1-Hydroxybenzotriazole KOH : Potassium hydroxide LC/MS : Liquid Chromatography/Mass Spectroscopy MS : Mass Spectroscopy

NMM : N-Methylmorpholine NMP : N-Methylpyrrolidinone Pyr : Pyridine S : Scaffold SLE : Solid liquid-liquid extraction THF : Tetrahydrofuran TFA : Trifluoroacetic acid TLC : Thin layer chromatography 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 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 L 1 in 50, u1NGMperwell. Antibiotic/Antimyoticwas addedto eachwell, and 1 % by weight E. coli strain HB 10 1. 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. 3, uM, 7. 8, uM, 2. 6, uM, 0. 9, uM, and 0. 3, uM. 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 Lethalit : Dead only stiff L Is (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 alsorecorded. Thus,"Ll/L2"would mean amixture of Ll's andL2's arepresentin 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 V1 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., uM.

Table 1. Dose Response Track))) ; ; X Day Visnat Score V4 V5 V6 DR//Wr'tTrack'oet. it'faryjfS) n (ctnre// Source)'Wet) AddressVtV2V3V4V5V6 1575 AKC 111 I N2 093 5081 : 1) 11) Dcd Dcacl I ? Deal (1. 3/l. IIAI) 113 OK 1G17 AKC 112 3 N2 I198 5090 : AI (1 1. 2/1, 3 1, 2/13 1, 2/1, 3 1, 3/1, 1 IIAI)/-[3 UK ).) 66AKC 1133N2//85506) : AIn 0DeadDeadDeadDead (L2/LL2/Dead (AL4///AD/) ; 14fi9 AKC 107 4 N2 186 5061 : 1) 10 Oeal !) eacl (I ?/1. 3) I'/i. 3 Oead (I. 4) 1. 4/Dead (A 1. 2/Deal (I 1477 477AKC 1145N2//865 () 6hDt) DeadL3/Dead (L')) L3L3/Dcad (LL2/). 3- !) 1^176 AKC 108 6 N2 IIS6 51) 61 : C I ! Dca ! head I. I I. I I. I/I ? 1. 1/I. J 7 73AKC 1157N2//86506) : m0DeadDead (t. 2/L3) L2/Dead (L2) Dead (L2) Dead ()-2/L !. 2/L3 035 AKC 119 II N2lll3 (5) 3 : 11-1 1lAD/13 IIAI) l (i IIAI)/13 IlAf)/Il llAOl (1 < 20S9AKC 110'2N2M) 28539') : C4U'.)) JL) t. t) J 2083 AKC 120 13 N2 #130 5419 2032, AKC 121'''N2//) 26S389 : Cd i. l !. ! i. l/1 ? IIAD/-. l3 IIAI)/13 < 2029 AKC 2153 15 N2 til26 5379 : C4Lft.)) J/L2MD/H//AD/) !//A))/n 1962 AKC 122 16 N ? N2131 5373 : H8DeadU//AD/n//AD/R//AR/HOK 1388 AKC 104 17 N2 #81) 502 : Cd 1, 1/1.'1. 1/1,'Ll/L2 #AI)/n 1, 111, 2 1, 1/1, 2 1372 AKC 123 18 N2 979 5016 : 119 I. I 1. 1/1. 3 IIAI) II3 < 1lAU/f3 IfAI)/-13 1402 AKC 124 1') N'fl81 51133 : 1) 8 ItA D/I 11 IIAI) tlll 1, 2/1. 3 I., IIIJAI) II) 1, 4///Ai)/Il f396AKC 12520N2//805<) 3) : (i8 1. 2/Deiul (1, 3) 1./I) cat1 (1, 4) 1. 2 1. 2 IIAD/13 1393 AKC 105 ? I N'NNII 51l. 1l : (7' 1, ?/1) c ; ul (1. 3) 1. 2/I) oal (1, 3) I ?/i) eut (1. 3) I ? ll) cd (I. I ?/Icacl (f. i. 2/ () cad (i 1393 AKC 105 1, 1/1. 2//A I)/-11 1 G1 AKC 126- 17, 1 AKc 102 21 N2 11 (ii. 1727 : 1 : 8 1. 1/1. 2 1"1/1. 2 1. 2/1. 3 i3 I : 1/lIAI)/I) S06 AKC I03'1 N31111') 11711 : 11111 1. 1/I ? Ieal/1. 3/1 ; 11 1) ctul (I. 1)' 18 AKC 171 25 N2 IJ2 201) 6 : A 1 1. 2 1, 2/1. 3 < 433 AKC 128 26 N2 931 3313 : A It l) e ; De : ul DeilKl l, l IIAI)/--13 () K 4-0 5 () G AKC 129 ? 7 N ? 1117 l1 I S : A I11 t) cal Ih ; u1 !. I/ !. 3 !. I/I ? (7K I. III. 18I AKC 130 28 N'835 331-11) 111 ) eilil IIAI)/-II/IAI)/II RAI)/-II () K IIAD# ! -'S-) AKC 13028N2//333t-) : D) fDead//AD/-) !//AD/t)//AD/-nOK//A)) ! 18G AKC 131"). N31135 33II : F) 0Dead !.)//AD/-H//AD/-B//AD/-H< 568AKC 13230N2//4) 3323 : (i : 1 f ? eatcf (1. 2) 1) cad 1) cacl IIAD/f3 IIADIIi (7en 69AKC 1333'N2//4) 3323 :)).) DeadDeadDeadDeadDeadDea a 187 AKC 340 32 N2 #14 2665 : 115 1. 1 IIAD/ii ! flAD/B IIAD/B OK ) 33AKC 13433N2//t02640 : A I I 1) eai ! 1) cci Dcal IIAD/I3 IIAf)/f3 OK 149 AKC 135 34 N2 If 11 264 I : Ag I) cad I'l 1, 111, 2 #AD/B (1'

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, uM and subj ected to 2-fold dilutions to yield 140, aM, 70, uM, 35, uM, 17. 5, uM, 8. 8µ, 4. 4uM, 2. 2µM, and 1. 09µM. 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 ~B OK OK OK AKC-144L3/L4L4/AD/BB-BOKOKOKOK 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 #B OK OK Control OK OK OK OK OK OK OK OK Example 4-Activity Against Nematode 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 :

NaCl 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 1L S basal (components should first be autoclaved separately) : 1M potassium citrate (pH 6) 10 ml Trace metals solution (see below) 10 ml 1M CaCl2 3 ml 1M MgS04 3 ml Trace Metals solution Na2EDTA 1. 86 g (to 5mM) Fe2SO4#7H20 0. 69 g (to 2. 5mM) MnCl2 4H20 0. 20 g (to 1mM) ZnSO4 7H20 0. 29 g (to ImM) CuSO4#5H20 0. 025 g (to 0. 1mM) dH20 1 L Procedure : 1. Make anthelmintic compound dilutions as indicated in Examples 2-3.

2. To 500 ttl of each dilution, added 10 ul of eggs (estimated >200 eggs/I 0 ici).

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 1ll S Medium.

10. Transfer 300 fil 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 3 0 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 &num 7.

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

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

12. Add 20 ul 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 48-290

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 producinganthelminticCompounds47, and exemplifiedCompounds 48-290, as shown in Figures 48-290. The general library scheme resulting in Compounds 47 is depicted in Figure 291. Scaffolds 1-6, depicted in Figures 292-297, are convenient intermediates for generating Compounds 47, and can be made from boc-hydroxyproline.

Scaffold Synthesis.

In a 2 L Morton three-neck flask equipped with mechanical stirrer, finely powdered 85% KOH (93. 50 g, 1. 666 mol, EM Biosciences) was dissolved in DMSO (400 mL, Aldrich, HPLC grade) under an atmosphere of nitrogen. After stirring at ambient temperature for 15 min, the apparatus was stirred and cooled with an ice-water bath for 10 min. After this 10 min period, Boc-Hyp-OH (48. 59 g, 0. 210 mol, Novabiochem) was added in one portion. DMSO (ca. 10 mL) was used to rinse residual Boc-Hyp-OH off the neck of the flask. After stirring for 5 min, the completely homogeneous solution was treated with one portion of 3-methoxybenzyl bromide (187 g, 0. 928 mol) to initiate symthesis of Scaffold 6 (Figure 297). DMSO (10 mL) was used to rinse residual 3-methoxybenzyl bromide off the neck of the flask. Other precursors useful in scaffold synthesis are listed in Table 8, where the"Entry"column corresponds to the Scaffold number desired. Caution : Alkylating agents are lacrymators and highly corrosive. Solutions of DMSO can potentially be highly toxic. After stirring at 0 °C for 15 min, the reaction mixture was warmed up to ambient temperature for 4 h. The reaction was monitored by the following procedure : the reaction mixture (0. 5 mL) was aliquoted to a 4 mL glass vial and diluted with water (1 mL). The aliquot was acidified with 1 M aqueous KHS04 (0. 5 mL) to pH 2-3 (tested by pH paper). Diethyl ether (2 mL) was added to the acidic mixture. The sample in the organic phase was analyzed by TLC (9 : 1 CH2Cl2/MeOH) by W light and Ninhydrin spray. After the reaction was judged complete after 4 h by TLC, the reaction mixture was poured into water (1. 2 L) in a 2 L Erlenmeyerflask. The Morton flask was rinsed with an additional portion of water (400 mL), and the aqueous wash was transferred into the Erlenmeyer flask. After stirring at ambient temperature for 5 min, the suspension was washed with diethyl ether (2x 1. 2 L).

The aqueous phase was acidified with 87% concentrated H3PO4 (150 mL) to pH 2-3 (tested by pH paper). This solution was then extracted diethyl ether (2x 1. 2L). The combined ether layers were washed with water (2x 650 mL), and saturated aqueous NaCl (2x 700 mL). The organic layer was dried over anhydrous MgSO4 (VWR) for 30 min, filtered, and concentrated in vacuo. The crude material was then triturated with hexane twice and concentrated in vacuo overnight.

The following procedure was used for scaffold 1, 2, and 6, Figures 292, 293, and 297, but was not used for scaffold 3, 4, and 5, Figure 294, 295, and 296. A solution of crude product in diethyl ether (1L) was treated with cyclohexylamine (18 mL, 0. 210 mol). Upon precipitation, the mixture was cooled with an ice bath, stirred for 20 min, and filtered through a Buchner funnel under vacuum. The complex was washed with diethyl ether (2x 250 mL) and hexane (2x 250 mL), and air-dried overnight.

A solution of the complex in water (500 mL) and diethyl ether (350 mL) was stirred at ambient temperature until all solid material dissolved. The solution was then acidified with 87% concentratedH3PO4 (40 mL) to pH 2-3. In a 1 L separatory funnel, the organic layer was separated from the aqueous layer. The aqueous layer was extracted with diethyl ether (350 mL). The combined organic layers were washed with 0. 5 M KHSO4 (350 mL), water (250 mL) and saturated aqueous NaCl (2x 300 mL). The combined organic layers were dried over anhydrous MgS04 for 15 min, and filtered.

Concentration in vacuo provided 72. 0 g (98%) of scaffold 6 as a yellow oil.

Step A. Carboxylic Acid Coupling.

DIC (0. 824 mL, 5. 265 mmol) was added to a CH2C12 solution (6 mL) of carboxylic acid from Step A (5. 265 mmol) and the solution was allowed to sit for 15 min with occasional swirling. The mixture was then added to the thiophenol resin (1. 5 g, 1. 755 mmol). The vial was rinsed with CH2CI2 (5 mL), and this solution was added to the resin. DMAP (0. 214 g, 1. 755 mmol) was then added to the slurry, and the mixture was shaken for at least 18 hr. The resin was then washed using the following solvents ; (CH2C12, THF) 4x ; CH2C12 4x, MeOH 3x. The resin was then dried on high vacuum overnight (ca. 16 h). The resin loading was determined by mass analysis as a percentage of theoretical. The resin was also qualitatively analyzed by IR, and an FeC13/pyr test.

Step B. Resin Plating.

The resin from Step A (ca. 100 mg per well using flat well Falcon Plates) was placed in the wells of a clampedpolyfiltronicsmicrotiterplate (2. 7 RM, GF/D) following the T. O. P #APD-602"Resin transfer with Falcon (Rev. Code : 20 Apr 1998 DGP)." Step C. Deprotection.

Both Steps C and D must be done sequentially on the same day. Using a Moduline, the resins were washed with CH2Cl2 2x. After placing plates in Hydra clamps, a solution (1 mL/well) of TFA/CH2Cl2/anisole (50 : 48 : 2) was added by a Robbins Hydra.

The plate was covered with a teflon sheet and clamped. Wells should point upward. The plates are shaken on a reciprocal shaker for 1 h. After freezing the bottom of the plates in dry ice, the plates are unclamped and TFA allowed to drain. Using a Moduline, the resin was then washed using the following solvents, CH2CI2 3x, 20% Et3N/CH2Cl2 3x, MeOH 3x, CH2C12 3x. The plates are placed back into a Hydra clamp with a Teflon sheet on the bottom.

Step D. Acylation.

Both Steps C and D must be done sequentially on the same day. Four different classes of reagents are used to acylate the unreactive 4-aminoalkyl substituent : sulfonyl chlorides (see Table 9), acid chlorides (see Table 9), isocyanates and isothiocyanates.

Each acylator is dissolved in 0. 702 M DIEA in CH2Cl2 and diluted to 0. 351 M with CH2Ct A solution of each acylator (1 mL/well) was added to the appropriate well. The plate was clamped, and shaken overnight (ca. 16 h) on a reciprocal shaker. Wells should point upward. Using a Moduline, the resins were washed using the following solvents, (CR, C12, then MeOH) repeat 4x, CH2Cl2 4x. The plate is placed back into a Hydra clamp with a Teflon sheet on the bottom.

Table 8 Alkyl bromides and iodides.

These precursors were used in the scaffold synthesis. Entry ACD MW Amount Name Structure 1 172 171. 04 342 g Benzyl bromide Br i 2 257 199. 1 255 g 1-Bromo-3-phenylpropane (i 3 1098 184. 02 259 g 1-Iodobutane/ 4 1509 177. 09 345 g (Bromomethyl) XBr cyclohexane 5 180 227. 15 396 g 4-tert-Butylbenzyl bromide ~Br 6 216590 901 G1 301 g 3-Methoxy benzylbromide Br it") Dr Table 9 Acid chlorides and sulfonyl chlorides.

The amount listed here is for one scaffold, which is a set of 12 plates, in the production library.

The final concentration is 0. 351 M and the total volume is 50 mL. Entry ACD MW Amount (Name Structure g) 1 653 140. 568 2. 467 Benzoyl chloride R d=1. 211 I w CI 0 2 665 200. 62 3. 521 2, 6-Dimethoxybenzoyl chloride °4f s W i 3 668 154. 595 2. 713 o-Toluoyl chloride 0 d1. 185 fi t 4 675 230. 646 4. 048 3, 4, 5-Trimethoxybenzoyl chloride 'CI -0 5 689 212. 675 3. 732 4-n-butoxybenzoyl 0 d= 1. 122 chloride HCI A~H 6 696 154. 595 2. 713 p-Toluoyl chloride d=1. 169 1 7 719 78. 4977 1. 378 Acetyl chloride 0 d=1. 104 8 728 108. 524 1. 905 Methoxyacetyl chloride 0 d=1. 197 9 745 92. 53 1. 624 Propionyl chloride d=1. 065 ~ vCI 'cl 10 1456 146. 616 2. 573 Cyclohexanecarbonyl d=1. 096 chloride Entry ACD MW Amount (Name Structure ) O CECI 11 3985 269. 751 4. 734 Dansyl chloride 0 N'cul .. 12 7450 190. 649 3. 346 p-Toluenesulfonyl R chloride XCI r 13 9674 214. 647 3. 767 3, 4- Dimethoxyphenylacetyl sO chloride XCI CI 14 9929 176. 549 3. 098 2, 5-Difluorobenzoyl d=1. 425 chloride F 'cl CRI 15 12046 214. 647 3. 767 2, 5- Dimethoxyphenylacetyl 1 chloride 0 CI 16 13655 230. 693 4. 049 Diphenylacetyl chloride d 0 cul o c 17 17527 200. 62 3. 521 2, 4-Dimethoxybenzoyl so o chloride XCI O Entry ACD MW Amount (Name Structure g) 18 24872 236. 674 4. 154 2, 5-o o Dimethoxybenzenesulfony 1 chloride '1-10 19 41509 246. 757 4. 331 4-test- Amylbenzenesulfonyl If chloride 0 20 44947 232. 749 4. 085 (-)-Menthoxyacetyl d=1. 04'chloride PO cri 21 46426 234. 681 4. 119 2-Ethoxy-1-naphthoyl 0 chloride [) j !) ! CI O 22 51569 198. 648 3. 486 2-Phenoxybutyryl chloride O Ci O 23 51769 236. 674 4. 154 3, 4-R ç Dimethoxybenzenesulfony o°+ I chloride 10, so v 24 74724 198. 691 3. 487 1-Adamantanecarbonyl 0 CI chloride Table 10 Amine Diversity Elements. Entry ACD MW Amount Name Structure (g) 1 3799 133. 19 1. 172 1-Aminoindane NH2 d=1. 038 'lez 2 4014 171. 242 1. 507 d=1. 063 Naphthyl) ethylamin H2N n2N 3 4731 117. 191 1. 031 L-Isolecinol HO d=1. 2 NH2 4 4732 151. 208 1. 331 L-Phenylalaninol / NH2 5 4734 117. 191 1. 031 D-Leucinol HO d=0. 917 NH2 6 8079 151. 208 1. 331 (IR, 2S)- (-)-Norephedrine OH 7 8085 75. 1101 0. 661 DL-2-Amino-l-propanol H2N H d=0. 943 8 8090 149. 236 1. 313 1-Methyl-3- d=0. 922 phenylpropylamine NH2 /NN2 8107 125. 15 1. 101 2-Fluorobenzylalnine F d=1. 095 euh2 NH2 10 8110 137. 18 1. 207 2-Methoxybenzylamine y d=1. 051 w, NH2 Entry ACD MW Amount Name Structure (g) 11 8137 137. 181 1. 207 2-Amino-1-phenylethanol OH NH2 12 8139 75. 1101 0. 661 1-Amino-2-propanol of NH2 d=0. 973 13 8183 61. 0833 0. 538 Ethanolamine d=1. 012 14 8188 181. 234 1. 595 3, 4-Dimethoxyphenethyl d=1. 074 amine J ! . 15 11690 73. 1379 0. 644 n-Butylamine , NH2 d=0. 740 16 14820 70. 0944 0. 617 3-Aminopropionitrile H 2N CN d=0. 914 17 17150 197. 232 1. 736 3, 4, 5-Trimethoxybenzyl NH2 d=1. 155 amine 0 . 1-0 18 21279 181. 321 1. 596 o-Aminobicyclohexyl d=0. 93 NH2 19 25572 186. 051 1. 637 2-Bromobenzylamine Br d=1. 480 20 25622 131. 22 1. 155 3-Butoxypropylamine d=0. 853 21 40754 163. 262 1. 437 4-tert-Butylbenzylamine w NH2 d=0. 881 Y' 22 41323 139. 172 1. 225 4-Fluoro-a- d=1. 059 methylbenzylamine NH2 23 41898 105. 204 0. 926 3- (Methylthio) propylamine gNH2 d=0. 953 Entry ACD MW Amount Name Structure 24 52392 167. 207 1. 471 2, 3-Dimethoxybenzyl amine d=1. 130 TT' 'k 25 52393 167. 207 1. 471 2, 4-Dimethoxybenzyl amine 0 d=1. 113 NH NH2 26 52975 137. 181 1. 207 2-Phenoxyethylamine H2N----10 d=1. 048 [) J 27 59019 128. 174 1. 128 3-Acetamidopyrrolidine H X XCNH 0 28 59040 186. 253 1. 639 3- (tert-butoxycarbonyl H amino) pyrrolidine N''CNH O ZNH v 29 60612 190. 072 1. 673 2, 6-Dichlorophenethyl amine Cl d=1. 28 H2N 30 60613 181. 234 1. 595 2, 3-Dimethoxy phenethyl sO d=1. 09 amine r+K0s ~ NH2 31 60614 181. 234 595 2, 5-Dimethoxy phenethyl d=1. 09 amine sowNH2 32 60618 181. 234 1. 454 3, 5-Dimethoxy phenethyl d=1. 07 amine NH2 33 60620 165. 234 1. 682 4-Ethoxyphenethylamine d=0. 99 NUI 34 61237 191. 151 1. 682 4- (Trifluoromethoxy) F NH2 d=1. 252 benzylamine F Entry ACD MW Amount Name Structure (g) 35 61267 191. 151 1. 349 3- (Trifluoromethoxy) d=1. 252 benzylamine F NH2 36 66934 153. 267 1. 075 1, 3, 3-Trimethyl-6- d=0. 902 azabicyclo (3. 2. 1) octane NH 37 75408 122. 17 1. 225 2-Aminobenzylamine Nl H2 NH2 38 75502 139. 172 2-Fluorophenethylamine d=1. 066 [J t NH2 39 75513 122. 17 1. 075 4-Aminobenzylamine I \ NH2 d=1. 078, 0 XJ H2N w 40 79755 195. 26 1. 718 3-Ethoxy-4-methoxy d=1. 05 phenethylamine "0 1 NH2 41 134208 139. 172 1. 225 4-Fluorophenethylamine d=1. 076 JJ j NH2 42 191368 115. 175 1. 014 2-Aminocyclohexanol HO H2N 43 191602 164. 25 1. 445 n- (3-Aminopropyl)-n- d=1. 00 methylaniline . 44 192238 153. 267 1. 349 (1R, 2R, 3R, 5S)-(-)- d=0. 909 Isopinocampheylamine., vNH2

Step E. Product Cleavage.

After washing with dioxane 2x, the resin from Step D was treated with the appropriate amine (0. 8 mL of a 0. 4 M solution in l, 4-dioxane, 0. 32 mmol), selected from those listed in Table 10. The plates were shaken on a reciprocal shaker in a clamped plate at ambient temperature for 48 h. Wells should point upward. After the bottoms of the plates are frozen, polyfiltronics plate is placed on a Beckman square well plate and the crude products collected. The resins were washed with 1, 4-dioxane (0. 35 mL, 2x).

After freezing the dioxane solutions in-80 °C freezer for 1 h, the products are lyophilized for at least 4 h.

Step F. SLE.

Removal of the excess amine starting material was accomplished by solid phase liquid-liquid extraction using Varian Chem Elut material packed into a polyfiltronics plate (10 micron PP/P). The Chem Elut (~2. 0 g) is treated with 2. 0 N HCl (0. 6 mL per well) followed by addition of the above product from Step E in 4 : 1 CH2Cl2/THF (1 mL).

After allowing the product solutionto elute for 15 min into the Beckmanplate, the source plate was washed with 4 : 1 CH2Cl2/THF (0. 350 mL), and each wash was transferred immediately to the SLE plate. Each 4 : 1 CH2C12/THF wash was allowed to drain for 15 min. The products were then concentrated in vacuo with Genevac following S. O. P.

&num APC-203-000"Genevac Evaporator (Rev. Code : 23 July 1998 BW)"and analyzed by LC and MS.

Development.

Early Development : Several synthetic pathways were explored for the library synthesis of hydroxyproline derivatives. After amine addition to bromomethyl Wang resin, Fmoc-Hyp-OH was coupled to solid-supported amine with DIC (5 eq.) and HOBt (2. 5 eq.) in NMP. PyBrop (2 eq.), and DIEA (4 eq.) in DCM also gave the desired product, as shown in Figure 298. However, the Mitsunobu reaction in polyfiltronics plates failed. Heat generated from the high reaction concentration and poor mixing contributed to the failure of this reaction. In some cases, intramolecular cyclization or elimination occurred during the Mitsunobu reaction. Cleavage of linker was also observed.

As shown in Figure 299, an alternate route required coupling Fmoc-Hyp-OH on bromomethyl Wang resin, and introducing the final diversity element in solution as the last step.

Figure 300 depicts the result of using diamines on nitrophenol carbonate linker, which gave low yield of the desired product.

Solution phase syntheses of scaffolds functionalized with the first diversity element were then considered as a strategy for library generation. After introduction of the phenols on the hydroxyproline scaffold via Mitsunobu reaction, extensive chromatographic purification and low yields of these scaffolds made this route less desirable for library generation. Accordingly, the original protocol was modified to allow for alkylation on the alcohol moiety of the hydroxyproline, and a preferred library synthesis was the result. (Figure 301).

Scaffold Synthesis.

A practical synthesis of alkylated hydroxyproline scaffolds was developed. The alkylation was driven to completion using 400 mol% instead of 200 mol% excess alkylating reagent. The reaction was executed under an atmosphere of nitrogen because KOH is hydroscopic. DMSO also needs to be anhydrous. In the work up procedure, H3PO4 is the preferred acidifying agent since the use of KHSO4 resulted in the precipitation of solids. Cyclohexylamine was also used to remove an unknown byproduct. No chromatography was involved in the syntheses.

Scaffold Coupling.

After scaffolds were coupled using DIC/DMAP, coupling was confirmed by FeCl3/pyridine test. The test was performed by dissolving about 50 mg of resin in pyridine (0. 5 mL) and adding a 0. 5 M chloroform solution of FeCl3 (0. 5 mL). The resin was washed with CH2Cl2and examined. Any color change to a dark color indicates free thiophenol. With free thiophenol resin, the beads will turn dark green-black. The test will indicate no free phenol if no color change from original resin color is observed. This is not a quantitative test, but a qualitative measure of resin loading.

Acylation.

Acid chlorides, sulfonyl chlorides, isocyanates, and isothiocyanates were generally soluble in DCM. Carboxylic acids were generally soluble in DMF.

Amine cleavage. a. Although pyridine is reported to be a suitable solvent for cleavage, 1, 4-dioxane also worked well. b. Secondary amine cleavage requires much more time (36-48 h) than the primary amine cleavage (24 h).

SLE. a. 4 : 1 CH2C12/THF used as elution solvent for SLE. In CH2C12, some products were insoluble. b. Priming volume increased from 0. 4 mL to 0. 6 mL of 2 N HC1 per well.

Plating.

After SLE, 4 : 1 ACN/MeOH used to dissolve products for QC samples.

Example 7 Nematicidal Activity of Anthelmintic Compositions 48-290 The nematicidal activity of anthelmintic Compositions 48-290 were determined in accordance with the procedure outlined in Example 2. The results are reported in 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 Add@ V1 V2 V3 V4 V5 V6 1259 4715 A7 169 83% Dead 181 118% 4AD/B 4715:A7 L1 < OK OK OK OK 1260 4715 B7 187 92% Dead 203 133% L1/L2 4715:B7 L1 < OK OK OK OK 1261 4715 C7 196 96% Dead 197 129% L1/L2 4715:C7 L1 #B OK OK OK OK 1262 4715 D7 182 89% Dead 191 125% L1/L2 4715:37 L1 #B OK OK OK OK 1263 4715 E7 200 98% Dead 207 135% 1AD/B! 4715:E7 L1 #AD/#B OK OK OK OK L1 L1 L1/l2 OK OK OK 1264 4715 F7 206 101% Dead 195 127% 1ADB! 4715:F7 L1 #AD/B OK OK OK OK 1265 4715 G7 203 100% Dead 188 123% L1/L2 4715:G7 L1 #AD/#B OK OK OK OK 1266 4715 H7 197 97% Dead 180 118% 2AD/B! 4715:H7 L1 < OK OK OK OK 1267 4715 E9 208 102% L1/L2/L3 201 131% B - AD Husks 4715:E9 L2/L3 -B OK OK OK OK 1268 4715 E10 198 97% Dead 198 129% 4AD/D 4715:E10 L1 OK OK OK OK OK 1269 4715 F10 193 95% Dead 202 132% 3AD/B 4715:F10 L1 < OK OK OK OK 1270 4715 G10 195 96% Dead/1AD 192 125% 4AD/B 4715:G10 L1 OK OK OK OK OK B! B! OK OK OK OK 1271 4715 H10 193 95% 1L3/1L4 171 112% B 4715:H10 #AD/B! OK OK OK OK OK 1272 4716 A7 166 81% Dead 191 125% Dead 4716:A7 L1/L2 < OK < OK OK 1273 4716 B7 189 93% Dead 212 139% L1/L2 4716:B7 L1 #AD/B OK OK OK OK 1274 4716 C7 183 90% Dead 200 131% 3AD/B! 4716;C7 L1 < OK OK OK OK 1275 4716 D7 180 88% Dead 195 127% 1AD/B! 4716:D7 L1 < OK OK OK OK 1276 4716 E7 184 90% Dead 198 129% L1/L2 4716:E7 L1 < OK OK OK OK 1277 4716 F7 197 97% Dead 207 135% 1AD/B! 4716:F7 L1 < OK OK OK OK L1 L1 L1 OK OK OK 1278 4716 G7 201 99% Dead 199 130% L1/L2 4716:G7 L1 #AD/B OK OK OK OK 1279 4716 H7 201 99% Dead 170 111% B 4716:H7 L1 OK OK OK OK OK 1280 4716 F10 201 99% Dead/1AD 182 119% B 4716:F10 L1/L2 OK OK OK OK OK 1281 4717 A7 169 83% Dead 190 124% 3AD/B! 4717:A7 L1 < OK < OK OK 1282 4717 B7 186 91% Dead 220 144% L1/L2 4717:B7 L1 #B OK OK OK OK 1283 4717 C7 195 96% Dead 204 133% 3AD/B! 4717:C7 L1 #B OK OK OK OK 1284 4717 D7 182 89% Dead 201 131% 1AD/B! 4717:D7 L1 #AD/B OK OK OK OK B! B! B! OK OK OK 1285 4717E7 197 97% Dead 218 142% 2AD/B! 4717:E7 L1 #B OK OK OK OK 1286 4717 F7 192 94% Dead 218 142% L1/L2 4717:47 L1 #AD/B OK OK OK OK 1287 4717 G7 206 101% Dead 198 129% 2AD/B 4717:G7 L1 < OK OK OK OK 1288 4717 H7 212 104% Dead 184 120% 5AD/B 4717:H7 L1 < OK OK OK OK 1289 4717 D10 192 94% L1/1L4 191 125% 0AD/B! - AD Husks 4717:D10 2/L3 L4/#AD/B! #B OK OK OK 1290 4717 E10 196 96% Dead 195 127% 2L4/3AD/B 4717:E10 #AD/B OK OK OK OK OK 1291 4717 H10 208 102% L1/1L4 227 148% B 4717:H10 L2/L3 OK OK OK OK OK L1 L1 L1 #AD/#B OK OK 1292 4718 E11 212 104% L3/L4 180 118% B 4718:E11 OK #AD/B OK OK OK OK 1293 4720 D2 205 100% AD/B! - No, P, No EggS, D 185 121% B 4720:D2 L3/L4 L3/L4 < OK OK OK 1294 4720 D3 209 102% L1/L2 206 136% Dead(L4/AD) 4720:D3 L3/L4 L3/L4 L3/L4 OK OK OK 1295 4720 D8 212 104% L1/L2Dead(L3/L4) 204 133% Dead(L4/AD) 4720DeadD8 L3/L4 L3/L4 L3/L4 #AD/B! OK OK 1296 4720 D10 202 99% Dead(L4) 212 139% Dead(AD) 4720:D10 L3/L4 L3/L4 L3/L4 #AD/#B OK OK 1297 4721 E1 220 108% L2 190 124% B - AD Husks 4721:E1 L3/L4 #AD/B #AD/B OK OK OK 1298 4721 E8 218 107% Dead/2L4 227 148% L2/L3 4721:E8 L2/L3 L3/L4 < OK OK OK B! B! B! OK OK OK 1299 4721 E10 210 103% 5AD/B 206 135% B - Dying AD 4721:E10 L1/L2 #AD/B #B OK OK OK 1300 4721 E11 209 102% L2/L3/1AD/B! 192 125% L4/AD/B 4721:E11 B #AD/B OK OK OK OK 1301 4722 F1 216 106% Dead 248 162% L3/L4Dead(L4) 4722:F1 L1 L1/L4 #AD/B! #AD/B OK OK 1302 4723 D7 200 98% L1/L2/L3 203 133% B! - Dead (L4/AD) 4723:D7 Dead(L3) L2/L3 #AD/B < OK OK 1303 4724 E9 207 101% L3/L4/2AD/B! - No P@ 1 B 212 139% Dead(L4/AD) 4724:E9 L1 L2/L3 #AD/B OK OK OK 126 4724 E10 210 103% L2/L3/L4 220 144% L2/L3 4724:E10 L1/L2 L1/L2 L1/L2 #AD/#B #B #AD/#B 1304 4724 E4 224 110% L1/L2 218 142% B! - Dying 4727:E4 L1 #AD/B OK OK OK OK L1 L1 L1 OK OK OK 1305 4727 A7 169 83% Dead 163 107% B 4727:A7 L1 #AD/#B OK OK OK OK 1306 4727 B7 187 92% Dead 195 127% L4/AD/B 4727:B7 L1 #B < OK OK OK 1307 4727 C7 186 91% Dead 207 135% 1AD/B! 4727:C7 L1 L1 < OK OK OK 1308 4727 D7 182 89% Dead 191 125% 5AD/B 4727:D7 L1 #AD/B < OK OK OK 1309 4727 E7 192 94% Dead 202 132% 5AD/B 4727:E7 L1 #AD/B < OK OK OK 1310 4727 F7 200 98% Dead 164 107% B 4727:47 L1 < < OK OK OK 1311 4727 G7 201 99% Dead 169 110% L4/AD/B 4727:G7 L1 #AD/#B OK OK OK OK B! B! B! OK OK OK 1312 4727 H7 215 105% Dead 157 103% B 4727:H7 Dead #AD/B OK OK OK OK 102 4727 E8 224 110% L2/L3/1AD 211 138% B - AD Dying 4727:E8 L1/L2 L1/L2 L2/L3 L3 #AD/B B 1313 4727 E9 215 105% L2/L3/1AD 204 133% B - AD Dying 4727:E9 L1 L2/Dead(L #B OK OK OK 1314 4727 A10 176 86% L1 177 116% B 4727:A10 L1 < OK OK OK OK 1315 4727 B10 184 90% L1 202 132% L3/L4/AD/B! 4727:B10 L1 #B OK OK OK OK 1316 4727 C10 189 93% L1 157 103% B 4727:C10 L1 OK OK OK OK OK 1317 4727 D10 175 86% L1 196 128% 2L4/3AD/B! 4727:D10 L1 OK OK OK OK OK L1 L1 L1 #AD/B! OK OK 1318 4727 E10 192 94% L1 172 112% B 4727:E10 L1 < OK OK OK OK 1319 4727 F10 190 93% L1 208 135% L1/L2 4727:F10 L1 #AD/B OK OK OK OK 1320 4727 G10 194 95% L1 204 133% 2L4/1AD/B! 4727:G10 L1 OK OK OK OK OK 1321 4727 H10 189 93% L1 205 134% B 4727:H10 L1 OK OK OK OK OK 1322 4728 C10 191 94% L1/L2 171 112% B 4728:C10 L1/L2 OK OK OK OK OK 1323 4728 D10 184 90% 1AD/B! 190 124% B 4728:D10 L1/L2 OK OK OK OK 1324 4729 D8 199 98% L1 189 124% 2AD/B 4729:D6 L2/Dead(L3) L4#AD/B < OK OK OK B! B! B! OK OK OK 1325 4729 E7 202 99% Dead 191 125% B 4729:E7 Dead < OK OK OK OK 1326 4729 D10 186 91% L1 185 121% B 4729:D10 L1 OK OK OK OK OK 1327 4729 E10 192 94% L1 161 105% B 4729:E10 #AD/B OK OK OK OK OK 1328 4739 D7 176 86% Dead 179 117% B 4739:37 L1 < OK OK OK OK 1329 4739 E7 194 95% Dead 208 136% 4AD/B 4739;E7 L1 < OK OK OK OK 1330 4739 E9 200 98% Dead 156 102% 3AD/#B 4739:E9 L1 OK OK OK OK OK 1331 4739 E10 210 103% L2/L3 207 135% 5AD/B - Husks 4739:E10 L1/L2 OK #1AD/#B OK OK OK L1 L1 L1 #B OK OK 1332 4740 B7 192 94% Dead 189 124% 4AD/B 4740:B7 L1 #AD/#B OK OK OK OK 1333 4740 C7 192 94% Dead 160 105% B 4740:C7 L1 < OK OK OK OK 1334 4740 D7 183 90% Dead 173 113% B 4740:D7 L1 < < OK OK OK 1335 4740 F7 210 103% Dead 172 112% B 4740:F7 L1 < OK OK OK OK 1336 4740 G7 219 107% Dead 147 96% B 4740:G7 #AD/B! B OK OK OK OK 1337 4740 G10 208 102% L1 202 132% B 4740:G10 L1 OK OK OK OK OK 1338 4741 D6 194 95% L1/L2 200 131% L2/L3 474:D6 L1 OK OK OK OK OK B! B! B! OK OK OK 1339 4741 A7 165 81% Dead 181 118% 4AD/B 4741:A7 L1 < OK OK OK OK 1340 4741 B7 188 92% Dead 165 108% B 4741:B7 #AD/B OK OK OK OK OK 1341 4741 D7 193 95% Dead 176 115% B 4741:D7 L1 OK OK OK OK OK 1342 4741 E7 195 96% Dead 209 137% 3AD/B! 4741:E7 L1 < OK OK OK OK 1343 4741 F7 216 106% Dead 211 138% 1AD/B! 4741:F7 L1 < OK OK OK OK 1344 4741 H7 208 102% Dead 176 115% B 4741:H7 L1 OK OK OK OK OK 1345 4741 B10 192 94% 1AD/B! 159 104% B 4741:B10 #AD/B! OK OK OK OK OK L1 L1 L1 OK OK OK 1346 4741 D10 198 97% L1/1L4 190 124% B - Dying AD 4741:D10 OK L3/L4 OK OK OK OK 1347 4741 F10 205 100% Dead 182 119% B 4741:F10 #AD/B! OK OK OK OK OK 1348 4741 G10 208 102% Dead/1AD/B! 190 124% B 4741:G10 #AD/B! OK OK OK OK OK 1349 4742 E11 218 107% L2/L3 221 144% L1/L2 4742:E11 L2/Dead(L3) L2/Dead(L B #AD/B #AD/B! OK 1350 4744 D2 196 96% Dead 200 131% L4/Dead(L4/AD) 4744:D2 Dead(L2/L3) OK OK OK OK OK 1351 4744 D3 208 102% L2/L3 192 125% B - AD Dying 4744:D3 L2/Dead(L3) Dead(L3/L8 B OK OK OK 1352 4744 D8 208 102% Dead 210 137% Dead(L3/L4) 4744:D8 Dead(L2/L3) #AD/B OK OK OK OK B! B! B! OK OK OK 1353 4744 D10 197 97% L2 203 133% Dead(L4/AD) 4744:D10 Dead(L2/L3) Dead(L3) OK OK OK OK 1354 4745 E1 220 108% L2/L3/L4 192 125% 4AD/B 4745:E1 L1/L2 #AD/#B OK OK OK OK 1355 4745 E10 210 103% L2/L3/L4 186 122% B 4745:E10 L2/L3 < OK OK OK OK 1356 4746 F1 200, 98% Dead 256 167% 4AD/B! - No P, AD 4746:F1 L1 #B OK OK OK OK 1357 4747 D1 203 100% Dead/2AD - Both Bag 174 114% 4AD/#B 4747:D1 L3/Dead(AD < < OK OK OK 1358 4747 D7 209 102% Dead 216 141% Dead(AD) 4747:D7 Dead(L2/L3) < OK OK OK OK 1359 4747 D8 206 101% 1AD/B 164 107% 5AD/#B 4747:D8 L2/L3 OK OK OK OK OK L1 L1 L1 L4#AD/E OK OK 1360 4751 A7 159 78% Dead 174 138% 1AD/B! 4751:A7 OK #AD/#B OK OK OK OK 1361 4751 B7 188 92% 1AD/B 190 151% 1AD/B! 4751:B7 L1 L1 OK OK OK OK 1362 4751 C7 192 94% Dead 199 158% L1/L2/L4 4751:C7 L1 #AD/#B OK OK OK OK 1363 4751 D7 185 91% Dead 194 154% L1/L2 4751:D7 L1 #AD/#B OK OK OK OK 1364 4751 E7 192 94% Dead 200 159% L1/L2 4751:E7 L1 #AD/B! OK OK OK OK 1365 4751 F7 192 94% Dead 29 166% 1AD/B! 4751:F7 B OK OK OK OK 1366 4751 G7 198 97% Dead 191 151% L1/L2 4751:G7 L1 < OK OK OK OK B! B! OK OK OK 1367 4751 H7 202 99% Dead 184 146% 3AD/B 4751:H7 L1 OK OK OK OK OK 1368 4751 B10 190 93% Dead 175 139% 2AD/B 4751:B10 L4/#AD/B OK OK OK OK OK 1369 4751 C10 188 92% 2AD/B 174 138% B 4751:C10 #AD/B! OK OK OK OK OK 1370 4752 B7 182 89% Dead 193 153% L1/L2 4752:B7 L1 #AD/B #B OK OK OK 1371 4752 C7 191 94% Dead 198 157% L1/L2 4752:C7 L1 #AD/B OK OK OK OK 1372 4752 D7 187 92% Dead 185 147% L2/L3 4752:D7 L1 #B OK OK OK OK 1373 4752 E2 197 97% Dead 198 157% L1/L2 4752:E7 L1 OK OK OK OK OK L1 L1 L1 L1 #AD/#B OK 1374 4752 F7 213 104% Dead 202 160% L1/L2 4752:F7 L1 #AD/B < OK OK OK 1375 4752 G7 200 98% Dead 200 159% 1AD/B! 4751:G7 L1 #AD/B < < OK OK 1376 4752 H7 199 98% Dead 208 165% L1/L2 4752:H7 L1 B B OK OK OK 1377 4752 B10 197 97% L2/L3 163 129% B 4752:B10 #AD/B OK OK OK OK OK 1378 4752 C10 195 96% L1/L2 172 137% 4AD/B 4752:C10 L1 OK OK OK OK OK 1379 4752 E10 195 96% 1AD/B! 167 133% B 4752:E10 L1/L2 OK OK OK OK OK 1380 4752 F10 205 100% Dead 179 142% B 4752:F10 L1 OK OK OK OK OK B! B! B! OK OK OK 1381 4752 H10 199 98% 2AD/B 209 166% L1/L2 4752:H10 L1 OK OK OK OK OK 1382 4753 A7 168 82% Dead 182 144% 1AD/B! 4753:A7 L1 OK OK OK OK OK 1383 4753 B7 191 94% Dead 191 152% 1AD/B! 4753:B7 L1 #B OK OK OK OK 1384 4753 C7 187 92% Dead 194 154% 1AD/B! 4753:C7 #AD/B OK OK OK OK OK 1385 4753 D7 196 96% Dead 195 155% L1/L2 4753:D7 L1 #AD/B OK OK OK OK 1386 4753 E7 194 95% Dead 197 156% L1/L2 4753:E7 L1 L4/#AD/B!#AD/#b OK OK OK 1387 4753 F7 182 89% Dead 207 164% L1/L2 4753:F7 L1/L2 #AD/B OK OK OK OK L1 L1 #AD/#B OK OK 1388 4753 G7 177 87% Dead 218 173% L1/L2 4753:G7 L1 #AD/B OK OK OK OK 1389 4753 H7 189 93% Dead 203 161% L1/L2 4753:H7 L1/L2 #AD/B < OK OK OK 1390 4753 B10 188 92% L1/L2 193 153% 5AD/B! 4753:B10 L1 OK OK OK OK OK 1391 4753 D10 193 95% Dead 177 140% 5AD/B 4753:D10 L4/#AD/B! OK OK OK OK OK 1392 4753 E10 192 94% Dead 202 160% L1/L2 4753:E10 L1/L2 OK OK OK OK OK 1393 4753 F10 203 100% Dead 212 168% L1/L2 4753:F10 L1/L2 OK OK OK OK OK 1394 4753 H10 207 101% L2/L3/L4 175 139% 5AD/B 4753:H10 L4/#AD/B! OK OK OK OK OK B! B! B! OK OK OK 1395 4756 D8 260 127% Dead 200 159% Dead(L4/AD) 4756:D8 Dead(L4) Dead(L4) OK OK OK OK 1396 4758 F1 197 97% Dead 161 128% B! - No P, Healthy, 4758;F1 #AD/B! OK OK OK OK OK 1397 4758 F2 224 110% 2AD/B! 157 125% B - HEALTHY, UNHATC 4758:F2 L4/#AD/B OK OK OK OK OK 1398 4763 A7 170 83% Dead 193 153% L1/L2 4763:A7 #AD/B OK OK OK OK OK 1399 4763 B7 191 94% Dead 205 163% 1AD/B! 4763:B7 L1 < OK OK OK OK 1400 4763 C7 192 94% Dead 192 152% 1AD/B! 4763:C7 L1/L2 OK OK OK OK OK 1401 4763 D7 183 90% Dead 187 148% L1/L2 4763:D7 #AD/B! < OK OK OK OK L1 L1 L1 #AD/#B OK OK 1402 4763 E7 189 93% Dead 199 158% L1/L2 4763:E7 L1 #AD/B OK OK OK OK 1403 4763 F7 190 93% Dead 202 160% L1/L2 4763:F7 #AD/B! OK OK OK OK 1404 4763 G7 201 99% Dead 193 153% 1AD/B! 4763:G7 L1 OK OK OK OK OK 1405 4763 H7 184 90% Dead 203 161% L1/L2 4763:H7 L1 < OK OK OK OK 1406 4763 A10 162 79% Dead 172 137% 2AD/B! 4763:A10 L1 OK OK OK OK OK 1407 4763 B10 173 85% Dead 190 151% L1/L2 4763:B10 L1 < OK OK OK OK 1408 4763 C10 191 94% Dead/2AD/B! - No P 195 155% L1/L2 4763:C10 L1 OK OK OK OK OK B! B! B! OK OK OK 1409 4763 D10 180 88% Dead 190 151% L1/L2 4763:D10 L1 OK OK OK OK OK 1410 4763 E10 183 90% Dead 199 158% L1/L2 4763:E10 L1 B OK OK OK OK 1411 4763 F10 193 95% Dead 199 158% L1/L2 4763:F10 L1 < OK OK OK < 1412 4763 G10 190 93% Dead 193 153% 3AD/B! 4763: G10 #AD/B! < OK OK OK OK 1413 4763 H10 206 101% 1AD/B! 200 159% 1AD/B! 4763:H10 L1/L2 OK OK OK OK OK 1414 4764 A7 216 106% Dead 182 144% 1AD/B! 4764:A7 L1 < OK OK OK OK 1415 4764 B7 228 112% Dead 194 154% 2AD/B 4764:B7 L4/#AD/B! < OK OK OK OK L1 L1 L1 L1 OK OK 1416 4764 C7 234 115% Dead 195 155% 1AD/B 4764;C7 L4/#AD/B! OK OK OK OK OK 1417 4764 D7 227 111% Dead 187 148% 1AD/B! 4764:D7 #AD/B! < OK OK OK OK 1418 4764 E7 240 118% Dead 195 155% 2AD/B! 4764:e7 #AD/B! < OK OK OK OK 1419 4764 F7 243 119% 1AD/B! 203 161% L1/L2 4764:F7 L1 B OK OK OK OK 1420 4764 G7 236 116% Dead 178 141% L1/L2 4764:G7 L1/L2 < OK OK OK OK 1421 4764 H7 230 113% Dead 203 161% L1/L2 4764:H7 L1 OK OK OK OK OK 1422 4764 A10 200 98% Dead 182 144% L1/L2 4764:A10 L1 OK OK OK OK OK B! B! B! OK OK OK 1423 4764 B10 196 96% Dead 198 157% 1AD/B! 4764:B10 L1/L2 < OK OK OK OK 1424 4764 C10 190 93% Dead 197 156% L1/L2 4764:C10 L1/L2 < < OK OK OK 1425 4764 D10 196 96% Dead 193 153% L1/L2 4764:D10 L1/L2 OK OK OK OK OK 1426 4764 F10 213 104% Dead 211 167% L1/L2 4764:F10 L1 < OK OK OK OK 1427 4764 G10 214 105% Dead 208 165% L1/L2 4764:G10 L1/L2 OK OK OK OK OK 1428 4764 H10 229 112% Dead 218, 173% L2/L3 4764:H10 L1/L2 < OK < OK OK 1429 4765 A7 175 86% Dead 181 144% 1AD/B! 4765:A7 #AD/B! OK OK OK OK OK L1 L1 L1 L1 OK OK 1430 4765 B7 185 91% Dead 197 156% 1AD/B! 4765:B7 L1 L4/#/AD/B OK OK OK OK 1431 4765 C7 195 96% Dead 196 157% L1/L4 4765:C7 L1 #AD/B OK OK OK OK 1432 4765 D7 187 92% Dead 192 152% L1/L2 4765:D7 L1 < OK OK OK OK 1433 4765 E7 190 93% Dead 201 160% 1AD/B! 4765:E7 L1 OK OK OK OK < 1434 4765 F7 190 93% Dead 193 153% 2AD/B! 4765:F7 #AD/B! OK OK OK OK OK 1435 4765 G7 192 94% Dead 195 155% L1/L2 4765:G7 L1 OK OK OK OK 1436 4765 H7 203 100% Dead 199 158% L1/L2 4765:H7 L1/L2 B < OK OK OK B! B B! OK OK OK 1437 4765 A10 161 79% Dead 176 140% 4AD/B! 4765:A10 L1 < OK OK OK OK 1438 4765 B10 165 81% Dead 196 156% 1AD/B! 4765:B10 L1 < OK OK OK OK 1439 4765 C10 180 88% Dead 197 156% 2L4/2AD/B! 4765:C10 L1/L2 OK OK OK OK OK 1440 4765 D10 193 95% Dead 194 154% L1/L2 4765:D10 L1 #AD/B! OK OK OK OK 1441 4765 E10 171 84% Dead 200 159% L1/L2 4765:E10 L1 #B OK OK OK OK 1442 4765 F10 192 94% Dead 205 163% 1AD/B! 4765:F10 L1 OK OK OK OK OK 1443 4765 G10 187 92% Dead 207 164% L1/L2 4765:G10 L1 OK OK OK OK OK L1/L2 L1 L1 L1 OK OK 1444 4765 H10 180 88% Dead 209 166% L1/L2 4765:H10 L1 OK OK OK OK OK 1445 4770 F1 203 100% Dead 201 160% B! - No P, Healthy, 4770:F1 L1/L2 #B OK OK OK OK 1446 4770 F8 223 109% L1/L2 172 137% B 4770:F8 L4/#AD/B! OK OK OK OK OK 1447 4775 A7 176 86% 1AD/B! 176 140% 1AD/B! 4775:A7 L1 < < < < OK 1448 4775 C7 192 94% Dead 196 156% 1AD/B! 4775:C7 L1 #B < OK OK OK 1449 4775 D7 184 90% Dead 188 149% 1AD/B! 4775:D7 L1 #B #B OK OK OK 1450 4775 E7 196 96% Dead 203 161% L1/L2 4775:E7 L1 #AD/B OK OK OK OK B! B! B! OK OK OK 1451 4775 F7 205 100% Dead 208 165% L1/L2 4775:F L1 OK OK OK OK OK 1452 4775 G7 200 98% Dead 184 146% 2AD/B! 4775:G7 L1 < OK OK OK OK 1453 4775 H7 190 93% Dead 185 147% 1AD/B 4775:H7 L1 OK OK OK OK OK 1454 4775 E9 205 100% L1/L2 181 144% L4/AD/B 4775:E9 #AD/B OK OK OK OK OK 1455 4775 A10 150 74% Dead 167 133% L1/L2 4775:A10 L1/L2 OK OK OK OK OK 1456 4775 B10 173 85% Dead 183 145% L1/L2 4775:B10 L1 OK OK OK OK OK 1457 4775 C10 178 87% 1AD/B! 183 145% L1/L2 4775:C10 L1 #B OK OK OK OK L1 L1 L1 L1 OK OK 1458 4775 D10 172 84% Dead 180 143% L1/L2 4775:D10 L1 #B OK OK OK OK 1459 4775 E10 171 84% Dead 190 151% L1/L2 4775:E10 L1 B #B OK OK OK 1460 4775 F10 188 92% Dead 196 156% L1/L2 4775:F10 L1 OK OK OK OK OK 1461 4775 G10 201 99% 1AD/B 181 144% 5D/B 4775:G10 L1 OK OK OK OK OK 1462 4776 A7 168 82% Dead 151 120% 3AD/#B 4776:A7 L1 < OK OK OK OK 1463 4776 B7 187 92% Dead 175 139% 4AD/B 4776:B7 L1 OK OK OK OK OK 1464 4776 C7 195 96% L1 179 142% 3AD/B 4776:C7 L1 < OK OK OK OK B! B! B! OK OK OK 1465 4776 D7 186 91% Dead 182 144% 2AD/B 4776:D7 L1 #B OK OK OK OK 1466 4776 E7 190 93% 1AD/B 198 157% 1AD/B! 4776:E7 L1 B OK OK OK OK 1467 4776 F7 200 98% L1 182 1445 4AD/B 4776:F7 #AD/B OK OK OK OK OK 1468 4776 G7 202 99% Dead 182 144% 1AD/B! 4776:G7 L1 < OK OK OK OK 1469 4776 A10 166 81% Dead 171 136% L1/L2 4776:A10 L1 < OK OK OK OK 1470 4776 B10 178 87% Dead 191 152% 1AD/B! 4776:B10 OK OK OK OK OK OK 1471 4776 C10 179 88% Dead 192 152% 3AD/B! 4776:C10 L1/L2 OK OK OK OK OK L1 L1 L1 < OK OK 1472 4776 D10 180 88% 1AD/B! 190 151% L1/L2 4776:D10 B OK OK OK OK OK 1473 4776 F10 190 93% 1AD/B 194 154% 4AD/B! 4776:F10 L1 #B OK OK OK OK 1474 4776 G10 196 96% L1/L2 200 159% 1AD/B! 4776:G10 L1/L2 OK OK OK OK OK 1475 4776 H10 193 95% 3AD/B 191 152% 5AD/B 4776:H10 L1/L2 OK OK OK OK OK 1476 4777 D3 200 98% L2/L3 185 147% Dead(AD) 4777:D3 Dead(L2/L3) < < OK OK L1 1477 4777 D6 199 98% L2/Dead(L3/L4) 172 137% B - AD Dying 4777:D6 Dead(L2/L3) #AD/B OK OK OK OK 1478 4777 A7 176 86% 1AD/B 175 139% 3AD/B 4777:A7 L1 OK OK < OK OK B! B! B! OK OK OK 1479 4777 B7 196 96% Dead 189 150% 3AD/B 4777:B7 B OK OK OK OK OK 1480 4777 C7 196 96% Dead 199 158% 2AD/B 4777:C7 #AD/B! < OK OK OK OK 1481 4777 D7 192 94% 1AD/B 189 150% 1AD/B! 4777:D7 L1 OK #B OK OK OK 1482 4777 E7 195 96% 1AD/B 194 154% L1/L2 4777:E7 L1 < OK OK OK OK 1483 4777 F7 203 100% Dead 203 161% 1AD/B! 4777:F7 L1 OK OK OK OK OK 1484 4777 G7 203 100% Dead 198 157% 1AD/B! 4777:G7 L1 OK OK OK OK OK 1485 4777 H7 207 101% Dead 199 158% 2AD/B! 4777:H7 L1/L2 OK OK OK OK OK L1 L1 L1 #AD/B B OK 1486 4777 A10 168 82% L1/L2 145 115% B 4777:A10 L1/L2 OK OK OK OK OK 1487 4777 B10 174 85% Dead 193 153% L1/L2 4777:B10 L1 OK OK OK OK OK 1488 4777 C10 182 89% Dead 190 151% L1/L2 4777:C10 L1 < OK OK OK OK 1489 4777 D10 192 94% Dead 193 153% L1/L2 4777:D10 L1 #B OK OK OK OK 1490 4777 E10 186 91% Dead 198 157% L1/L2 4777:E10 L1 OK < OK OK OK 1491 4777 F10 199 98% Dead 204 162% L1/L2 4777:F10 L1 OK OK OK OK OK 1492 4777 G10 209 102% L1/L2 219 174% 4AD/B! 4777:G10 L1 OK OK OK OK OK B! B! B! OK OK OK 1493 4777 H10 196 96% L1/L2 209 166% 1AD/B! 4777:H10 L1 < OK OK OK OK 1494 4778 E11 210 103% L1/L2 212 168% L2/L3 4778:E11 L2/L3 #AD/B OK OK OK OK 1495 4780 D2 200 98% L2 183 145% 3AD/B - Husks 4780:D2 Dead #AD/B OK OK OK OK 1496 4780 D3 202 99% L1/L2 193 153% Dead(AD) 4780:D3 L3/L4 L4/#AD/B OK OK OK OK 1497 4780 D8 213 104% Dead(AD) 197 156% L4/AD/B -Husks 4780:D8 #AD/B #AD/B OK OK OK OK 1498 4781 F1 211 103% L1/L2 214 170% L1/L2/L3 4781:F1 Dead B OK OK OK OK 1499 4782 D7 203 100% AD/B! - Dying, Bags 177 140% B-Dying, Husks 4782:D7 Dead(L4) Dead(AD) OK OK OK OK

Example 8-Sheep Test I Experimental Procedure Sheep naturally infected with a variety of gastrointestinal nematodes are purchased from local sources and are transportedto 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 &num NMEPG. 99. 01 On treatment day, the animals are weighed and divided into groups with three animals per group as follows : GROUP 1 : Non-treated negative control (placebo) of 10 ml of DMSO.

GROUP 2 : Positive Controltreatmentof200 mcg/kg commerciallyavailable 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 slaughteredin accordance with the Guide for the Care and Use of Laboratory Animals (DHEW Publication No.

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

Nematodes are recovered, identified, and enumerated according to Zimmerman Research SOP &num 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 is opened. The abomasum, the small and large intestines are tied at the omasal and pyloric openings, the duodenum, the end of the small intestine and at the end of the large intestine. Each section is transferred in a separate bucket containing warm water and is slit open and thoroughly washed. The epithelium

is inspected before it is removed. The thus prepared washings are saved in gallon jars. An appropriate preservative is added. If preservative is not available, all the intestinal washing should kept in a refrigerator. These washings are passed through a 100-mesh sieve (pore size 149 pm), and the residue is examined for the presence of worms under a dissecting microscope. Lugol's solution may be used to stain the worms. All worms are picked up counted and identified as to the species. An effort should be made to recover any immature forms present. The efficacy should be calculated using the controlled anthelmintic test.

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

Table 4 Akkadix AKK 101 Sheep Trial 17-Jan Rumen injection Sheep Weight/lbs Worm Counts Numbes 1/17/2000 Abomasum Abomasum Small Intestine Small Intest. Haemonchus Ostertagia Trichostrongylus Nematodirus Group 81 84 420 60 0 140 Negative 75 116 500 860 320 2300 Control 56 87 0 0 80 0 Mean Ct. 307 3071 133 813 Group 83 98 0 0 0 0 Ivermectin 92 129 0 0 0 0 200mcg/kg 53 82 0 0 0 0 Mean Ct. t Oj 0 0 %Efficacy 100 100 100 100 Group 58 74 0 80 380 1660 AKC 102 71 51 20 2440 3720 180 1. 4mg/kg60 10700480280 Mean Ct. 7 840 1527 707 %Efficacy 98 Table 5 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 1 58 73 4690 4640 300 260 94. 40 AKC 102 71 48 1670 1630 310 NS 80. 98 1. 4mg/kg 60 100 310 40 10 0 100. 00 Total/Mean 2223. 33 2103. 33 206 67 90. 17 86. 67 95. 88 l l Group 2 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 3 81 74 2240 2240 780 770 65.63 Negative 751 109 370 300 260 1360-353. 33 Control 56 80 40 40 30 30 25. 00 Total/Mean 263 883. 33 860.00 356. 67 58. 53 720. 001 16. 28 (

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

The sheep are fed good quality hay (no concentrated rations) and water ad libitum. Following a five day acclimation period, the sheep are randomly assigned by EPG count into the following treatment groups : Groups 1-9, various anthelmintic compounds of the present invention (test compound) dissolved in DMSO : Group 10, Positive Control (commercially available ivermectin for sheep) ; Group 11, non-treated Negative control (DMSO only). The first replicate of 11 animals is randomly assigned to groups 1-11 ; the second replicate of 11 animals is randomly assigned to groups 1-11 ; and the third replicate of 11 animals is randomly assigned to groups 1-11. Thus 1 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.

GROUP8 : 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 l l : 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 : I The placebo (DMSO), the commercially available drug, and the test anthelmintic compounds are administeredin 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 &num NEMRECOVID. 00. 01. All individualsperformingnematode recoveries are blinded to treatment versus control animals.

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

Table 6 Akkadix AKK-102 ! Sheep 24-May !) j Sheep Weight/lbs 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 524 541 20800010 Mean Ct. 47 227 20 3673 10 Group 1347 45 0 0 0 100 0 fvermectin133658 M001000 200mcg/kg 539 47 0 0 0 0 0 Mean Ct. 1 1 71 0 0 671 0 %Efficacy 86 100 100 98 Group 1348 45 20 340 AKC 102 1332 42 160 1380 tmg/kg 1338 33 0 100 Mean Ct. 1 607J %Efficacy Table 7 Akkadix Trial-2 Sheep AKK 102 Strongyles Strongyles Strongyles Sheep tWeight/lbs Total 15-May 22-May 24-May Number 5/17/2000 EPG-pre EPG-pre EPG-5dayj% Change EPG-7day%Change' ~ Group 1348 45 760 110 120 420 -281.82 AKC 102 1332 42 350 270 2332 570 -111.11 1mg/kg 1338 33 100 70 30 70 0.00 Total/Mean 120 403.33 150.00 827.33 -451.56 353.33 -135.56 Group 530 47 710 640 690 300 53.13 47 710 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 10 1334 41 560 530 690 290 45.28 AKC 110 536 40 280 220 850 1510 -586.36 3. 5mg/kg 1339 46 70 40 0 30 25. 00 Total/Mean 127 303.33 263.33 513.33 -94.94 610.00 -131.65 Group 1347 45 560 450 0 0 100.00 Ivermectin 1336 58 220 170 0 0 100. 00 200mcg/kg 539 47JOO 4oT 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.