Abstract:
The present invention provides non-peptide organic compounds that have a structure analogous to or reminiscent of the TMOF structure and have pesticidal activity. Thus the present invention concerns pesticidal compounds that inhibit digestion in pests by terminating or otherwise blocking synthesis of digestive enzymes by activating a TMOF receptor (collectively referred to herein as “pesticidal compounds”). The pesticidal compounds and other compounds of the present invention are usefully employed in the control of pests, particularly insect pests Such as mosquitoes, which ingest blood.

Description:
RELATED APPLICATIONS  
       [0001]    This application is a continuation of U.S. patent application Ser. No. 09/876,502 filed Jun. 7, 2001, which is a continuation-in-part of commonly owned, copending application Ser. No. 09/457,509, filed Dec. 8, 1999, the disclosures of which are incorporated by reference herein in their entirety. 
     
    
     
       FIELD OF THE INVENTION  
         [0002]    The present invention concerns functionalized alkenes, along with alkane and alkyne analogs thereof, that have pesticidal activity, along with methods of use thereof.  
         BACKGROUND OF THE INVENTION  
         [0003]    Many blood-ingesting pests are known to feed on humans and animals, and many pests are vectors for pathogenic microorganisms that threaten human and animal health, including commercially important livestock, pets and other animals. Various species of mosquitoes, for example, transmit diseases caused by viruses, and many are vectors for disease-causing nematodes and protozoa. Mosquitoes of the genus Anopheles transmit Plasmodium, the protozoan which causes malaria, a devastating disease which results in approximately I million deaths annually. The mosquito species  Aedes aegypti  transmits an arbovirus that causes yellow fever in humans. Other arboviruses transmitted by Aedes species include the causative agents of dengue fever, eastern and western encephalitis, Venezuelan equine encephalitis, St. Louis encephalitis, chikungunya, oroponehe and bunyarnidera. The genus Culex, which includes the common house mosquito  C. pipiens,  is implicated in the transmission of various forms of encephalitis and filarial worms. The common house mosquito also transmits  Wuchereria banuffi  and  Brugia malayi,  which cause various forms of lymphatic filariasis, including elephantiasis.  Trypanasomas cruzi,  the causative agent of Chagas&#39; disease, is transmitted by various species of blood-ingesting Triatominae bugs. The tsetse fly (Glossina spp.) transmits African trypanosomal diseases of humans and cattle. Many other diseases are transmitted by various blood-ingesting pest species. The order Diptera contains a large number of blood-ingesting and disease-bearing insects, including, for example, mosquitoes, black flies, no-see-ums (punkies), horse flies, deer flies and tsetse flies.  
           [0004]    Various pesticides have been employed in efforts to control or eradicate populations of disease-bearing pests, such as disease-bearing blood-ingesting pests. For example, DDT, a chlorinated hydrocarbon, has been used in attempts to eradicate malaria-bearing mosquitoes throughout the world. Other examples of chlorinated hydrocarbons are BHC, lindane, chlorobenzilate, methoxychlor, and the cyclodienes (e.g., aldrin, dieldrin, chlordane, heptachlor, and endrin). The long-term stability of many of these pesticides and their tendency to bioaccumulate render them particularly dangerous to many non-pest organisms.  
           [0005]    Another common class of pesticides is the organophosphates, which is perhaps the largest and most versatile class of pesticides. Organophosphates include, for example, parathion, Malathion, diazinon, naled, methyl parathion, and dichlorvos. Organophosphates are generally much more toxic than the chlorinated hydrocarbons. Their pesticidal effect results from their ability to inhibit the enzyme cholinesterase, an essential enzyme in the functioning of the insect nervous system. However, they also have toxic effects on many animals, including humans.  
           [0006]    The carbamates, a relatively new group of pesticides, include such compounds as carbamyl, methomyl, and carbofuran. These compounds are rapidly detoxified and eliminated from animal tissues. Their toxicity is thought to involve a mechanism similar to the mechanism of the organophosphates; consequently, they exhibit similar shortcomings, including animal toxicity.  
           [0007]    A major problem in pest control results from the capability of many species to develop pesticide resistance. Resistance results from the selection of naturally-occurring mutants possessing biochemical, physiological or behavioristic factors that enable the pests to tolerate the pesticide. Species of Anopheles mosquitoes, for example, have been known to develop resistance to DDT and dieldrin. DDT substitutes, such as Malathion™, propoxur and fenitrothion are available; however, the cost of these substitutes is much greater than the cost of DDT.  
           [0008]    There is clearly a longstanding need in the art for pesticidal compounds that are pest-specific, that reduce or eliminate direct and/or indirect threats to human health posed by presently available pesticides, that are environmentally compatible in the sense that they are biodegradable, are not toxic to non-pest organisms, and have reduced or no tendency to bioaccummulate.  
           [0009]    Many pests, including for example blood-imbibing pests, must consume and digest a proteinaceous meal to acquire sufficient essential amino acids for growth, development and the production of mature eggs. Adult pests, such as adult mosquitoes, need these essential amino acids for the production of vitellogenins by the fat body. These vitellogenins are precursors to yolk proteins which are critical components of oogenesis. Many pests, such as house flies and mosquitoes, produce oostatic hormones that inhibit egg development by inhibiting digestion of the protein meal and thereby limiting the availability of the essential amino acids necessary for egg development.  
           [0010]    Serine esterases such as trypsin and trypsin-like enzymes (collectively referred to herein as “TTLE”) are important components of the digestion of proteins by insects. In the mosquito,  Aedes aegypti,  an early trypsin that is found in the midgut of newly emerged females is replaced, following the blood meal, by a late trypsin. A female mosquito typically weighs about 2 mg and produces 4 to 6 ug of trypsin within several hours after ingesting a blood meal. Continuous boisynthesis at this rate would exhaust the available metabolic energy of a female mosquito; as a result, the mosquito would be unable to produce mature eggs, or even to find an oviposition site. To conserve metabolic energy, the mosquito regulates TTLE biosynthesis with a peptide hormone named Trypsin Modulating Oostatic Factor (TMOF). Mosquitoes produce TMOF in the follicular epithelium of the ovary 12-35 hours after a blood meal; TMOF is then released into the hemolymph where it binds to a specific receptor on the midgut epithelial cells, signaling the termination of TTLE biosynthesis. This regulatory mechanism is not unique for mosquitoes; flesh flies, fleas, sand flies, house flies, dog flies and other insect pests which need protein as part of their diet have similar regulatory mechanisms.  
           [0011]    In 1985, Borovsky purified an oostatic hormone 7,000-fold and disclosed that injection of a hormone preparation into the body cavity of blood imbibed mosquitoes caused inhibition of egg development and sterility (Borovsky, D. [1985 ] Arch. Insect Biochem. Physiol.  2:333-349). Following these observations, Borovsky (Borovsky, D. [1988 ] Arch. Ins. Biochem. Physiol.  7:187-210) reported that injection or passage of a peptide hormone preparation into mosquitoes inhibited the TTLE biosynthesis in the epithelial cells of the gut. This inhibition caused inefficient digestion of the blood meal and a reduction in the availability of essential amino acids translocated by the hemolymph, resulting in arrested egg development in the treated insect. Borovsky observed that this inhibition of egg development does not occur when the oostatic hormone peptides are inside the lumen of the gut or other parts of the digestive system (Borovsky, D. [1988], supra).  
           [0012]    Following the 1985 report, the isolated hormone, (a ten amino acid peptide) and two TMOF analogues were disclosed in U.S. Pat. Nos. 5,011,909 and 5,130,253, and in a 1990 publication (Borovsky et al. [1990]  FASEB J.  4:3015-3020). Additionally, U.S. Pat. No. 5,358,934 discloses truncated forms of the full length TMOF which have prolines removed from the carboxy terminus, including the peptides YDPAP, YDPAPP, YDPAPPP, and YDPAPPPP.  
           [0013]    D. Borovsky and R. Linderman, U.S. patent application Ser. No. 09/295,996, filed Apr. 21, 1999, discloses additional novel peptides and the use thereof to control insect pests.  
           [0014]    TMOF analogs that have been identified to date are primarily peptide analogs. In order to provide a greater diversity of new pesticidal compounds, it would be desirable to possess compounds that are TMOF analogues, yet are not peptides.  
         SUMMARY OF THE INVENTION  
         [0015]    The present invention is based on the discovery of non-peptide organic compounds that have a structure analogous to or reminiscent of the TMOF structure and have pesticidal activity. Thus the present invention concerns pesticidal compounds that inhibit digestion in pests by terminating or otherwise blocking synthesis of digestive enzymes by activating a TMOF receptor (collectively referred to herein as “pesticidal compounds”). The pesticidal compounds and other compounds of the present invention are usefully employed in the control of pests, particularly insect pests such as mosquitoes, which ingest blood.  
           [0016]    Thus, a first aspect of the present invention is a method of controlling a pest such as an insect pest, comprising administering to said pest a pesticidally effective amount of a non-peptide TMOF analog (that is, an organic compound that has TMOF activity). This definition is specifically intended to exclude the peptide TMOF agonists or analogs disclosed in inter alia U.S. Pat. Nos. 5,011,909; 5,130,253; and 5,358,934, the disclosures of which are incorporated by reference herein in their entirety.  
           [0017]    Particular pesticidal compounds/non-peptide TMOF analogs of the present invention have the formula I below:  
                         
 
           [0018]    wherein:  
           [0019]    X is selected from the group consisting of —CHCH—, —CH 2 CH 2 —, and —CC—;  
           [0020]    Z is selected from the group consisting of —OH, —NH 2  and —OR 6  wherein R 6  is loweralkyl;  
           [0021]    m≧0 and n≧1 and together total an integer from 1, 2, 3 or 4 to 6, 8, 10 or 12;  
           [0022]    the ring bearing substituents R 1 , R 2 , R 3 , R 4  and R 5  is a phenyl ring or a cyclohexyl ring;  
           [0023]    R 1 , R 2 , R 3 , R 4 , and R 5  are each independently selected from the group consisting of —H, —OH, halo, loweralkyl, loweralkoxy and phenyl; subject to the proviso that:  
           [0024]    when the ring is a phenyl ring, a pair of R 1  and R 2 , R 2  and R 3 , or R 3  and R 4  on the phenyl ring may together represent —CR 7 ═CR 8 —CR 9 ═CR 10 —, to form with the phenyl ring illustrated above a naphthyl ring system, wherein R 7 , R 8 , R 9 , and R 10  are each independently selected from the group consisting of —H, —OH, halo, loweralkyl, and loweralkoxy.  
           [0025]    A second aspect of the present invention is a method of initiating a TMOF receptor-mediated biological response. The method comprises contacting to a TMOF receptor in vivo or in vitro for a time and in an amount sufficient to initiate a TMOF receptor-mediated biological response a compound of Formula I as described herein. The biological response may be any suitable biological response mediated by the TMOF receptor, including but not limited to inhibition of biosynthesis of a digestive enzyme such as trypsin.  
           [0026]    As noted above, the pesticidal compounds of the present invention have advantageous biological activity against pests. The novel compounds of the invention are particularly active against blood-sucking insects, particularly against species of mosquitoes such as  Aedes aegypti  that are common vectors of arthropod-borne viral diseases, such as arboviruses. Other biting pests such as flies, fleas, ticks, and lice can also be controlled using compounds and methods of the subject invention. These pests utilize TTLE as their primary blood-digesting enzymes.  
           [0027]    The subject compounds can also be used to control pests of agricultural crops, for example by applying the compounds to the agricultural crops. These pests include, for example, coleopterans (beetles), lepidopterans (caterpillars), and mites. The compounds of the subject invention can also be used to control household pests including, but not limited to, ants and cockroaches.  
           [0028]    Another aspect of the subject invention pertains to a method for controlling pests, particularly insect pests, comprising administering to said pest a pesticidally effective amount of a pesticidal compound of the subject invention.  
           [0029]    The subject invention provides pest control compositions comprising pesticidal compounds and a suitable pesticidal carrier. The pest control compositions are formulated for application to the target pests or their situs.  
           [0030]    The methods and materials of the subject invention provide a novel approach to controlling insects and insect-transmitted diseases. The compounds of the subject invention have advantageous activity and increased resistance to proteolysis over previously disclosed compounds.  
           [0031]    While the present invention is in part explained with reference to TMOF activity, it will be appreciated that this statement of underlying activity is provided for explanation, and that applicants do not intend to be bound by any particular theory of the invention. For example, the compounds of the invention may also be used as pest or insect repellants, and the compounds of the invention may control pests or insects by mechanisms in addition to, or different from, TMOF activity.  
           [0032]    The present invention is explained in greater detail below.  
         DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS  
         [0033]    As used herein, the term “pesticidally effective” is used to indicate an amount or concentration of a pesticidal compound which is sufficient to reduce the number of pests in a geographical locus as compared to a corresponding geographical locus in the absence of the amount or concentration of the pesticidal compound.  
           [0034]    The term “pesticidal” is not intended to refer only to the ability to kill pests, Such as insect pests, but also includes the ability to interfere with a pest&#39;s life cycle in any way that results in an overall reduction in the pest population. For example, the term “pesticidal” includes inhibition of a pest from progressing from one form to a more mature form, e.g., transition between various larval instars or transition from larva to pupa or pupa to adult. Further, the term “pesticidal” is intended to encompass anti-pest activity during all phases of a pest&#39;s life cycle; thus, for example, the term includes larvacidal, ovicidal, and adulticidal activity.  
           [0035]    The term “loweralkyl” as used herein means C 1  to C 4  alkyl, preferably methyl, ethyl or propyl.  
           [0036]    The term “loweralkoxy” as used herein means C 1  to C 4  alkoxy, preferably methoxy, ethoxy, or propoxy.  
           [0037]    The term “halo” as used herein means halogen, preferably fluoro, chloro, bromo or iodo, most preferably fluoro.  
           [0038]    1. Pesticidal Compounds.  
           [0039]    Compounds useful in the present invention have the general formula I below:  
                         
 
           [0040]    wherein:  
           [0041]    X is selected from the group consisting of —CHCH—, —CH 2 CH 2 —, and —CC—;  
           [0042]    Z is selected from the group consisting of —OH, —NH 2  and —OR 6  wherein R 6  is loweralkyl;  
           [0043]    m≧0 and n≧1 and together total an integer from 1 to 12; and  
           [0044]    R 1 , R 2 , R 3 , R 4 , and R 5  are each independently selected from the group consisting of —H, —OH, halo, loweralkyl, loweralkoxy and phenyl; subject to the proviso that:  
           [0045]    a pair of R 1  and R 2 , R 2  and R 3 , R3 and R 4 , or R 4  and R 5  on the phenyl ring may together represent —CR 7 ═CR 8 —CR 9 ═CR 10 —, to form with the phenyl ring illustrated above a naphthyl ring system, wherein R 7 , R 8 , R 9 , and R 10  are each independently selected from the group consisting of —H, —OH, halo, loweralkyl, and loweralkoxy.  
           [0046]    In one preferred group of compounds of formula I above, n and m are each at least 1 and together total an integer from 2, 3 or 4 to 6, 8, 10 or 12.  
           [0047]    In another preferred group of compounds of formula I above, the ring is a phenyl ring.  
           [0048]    In another preferred group of compounds of formula I above, the ring is a cyclohexyl ring.  
           [0049]    In another preferred group of compounds of formula I above, m is 0.  
           [0050]    In another preferred group of compounds of formula I above, at least one of R 1 , R 2 , R 3 , R 4  or R 5  is phenyl.  
           [0051]    Another preferred group of compounds of formula I above have the general formula II as given below:  
                         
 
           [0052]    wherein Z, n, m, R 1 , R 2 , R 3 , R 4 , and R 5  are as described above. In compounds of formula II, the phenyl ring and the carbonyl carbon may be either cis (Z) or trans (E) with respect to one another. The trans configuration is preferred.  
           [0053]    Specific examples of compounds as described above include:  
           [0054]    E-7-phenylhept-4-enoic acid, which has the structure:  
                         
 
           [0055]    E-7-(4-methoxyphenyl)hept-4-enoic acid, which has the structure:  
                         
 
           [0056]    Methyl E-7-phenylhept-4-enoate, which has the structure:  
                         
 
           [0057]    E-7-phenylhept-4-enoic acid amide, which has the structure:  
                         
 
           [0058]    Z-7-phenylhept-4-enoic acid, which has the structure:  
                         
 
           [0059]    E-7-(2,4-difluorophenyl)hept-4-enoic acid, which has the structure:  
                         
 
           [0060]    E-10-phenyldec-6-enoic acid, which has the structure:  
                         
 
           [0061]    and E-10-(4-methoxyphenyl)dec-4-enoic acid, which has the structure:  
                         
 
           [0062]    Additional examples of acids of the present invention include:  
           [0063]    E-7-(4-hydroxyphenyl)hept-4-enoic acid;  
           [0064]    E-7-(2,4-dibromophenyl)hept-4-enoic acid;  
           [0065]    E-7-(4-methylphenyl)hept-4-enoic acid;  
           [0066]    E-7-(2,4-diethylphenyl)hept-4-enoic acid;  
           [0067]    E-7-(2-ethoxyphenyl)hept-4-enoic acid;  
           [0068]    E-7-(2,4,-dipropoxyphenyl)hept-4-enoic acid: and  
           [0069]    E-10-(2,4-difluorophenyl)dec-4-enoic acid.  
           [0070]    Additional examples of amides of the present invention include:  
           [0071]    E-7-(2,4-difluorophenyl)hept-4-enoic acid amide;  
           [0072]    E-7-(4-methoxyphenyl)hept-4-enoic acid amide;  
           [0073]    E-10-phenyldec-6-enoic acid amide;  
           [0074]    E-7-(2,4-difluorophenyl)dec-6-enoic acid amide;  
           [0075]    E-7-(4-methoxyphenyl)dec-6-enoic acid amide; and  
           [0076]    Z-7-phenylhept-4-enoic acid amide.  
           [0077]    Additional examples of esters of the present invention include:  
           [0078]    Methyl E-7-(2,4-difluorophenyl)hept-4-enoate;  
           [0079]    Methyl E-7-(4-methoxyphenyl)hept-4-enoate;  
           [0080]    Methyl E-10-phenyldec-6-enoate;  
           [0081]    Methyl E-7-(2,4-difluorophenyl)dec-6-enoate;  
           [0082]    Methyl E-7-(4-methoxyphenyl)dec-6-enoate;  
           [0083]    Methyl Z-7-phenylhept-4-enoate;  
           [0084]    Ethyl E-7-(2,4-difluorophenyl)hept-4-enoate;  
           [0085]    Ethyl E-7-(4-methoxyphenyl)hept-4-enoate;  
           [0086]    Ethyl E-10-phenyldec-6-enoate;  
           [0087]    Propyl E-7-(2,4-difluorophenyl)dec-6-enoate;  
           [0088]    Propyl E-7-(4-methoxyphenyl)dec-6-enoate;  
           [0089]    Propyl Z-7-phenylhept-4-enoate;  
           [0090]    Ethyl E-7-phenylhept-4-enoate;  
           [0091]    Propyl E-7-phenylhept-4-enoate;  
           [0092]    Butyl E-7-phenylhept 4-enoate;  
           [0093]    Ethyl E-10-phenyldec-6-enoate;  
           [0094]    Propyl E-10-phenyldec-6-enoate; and  
           [0095]    Butyl E-10-phenyldec-6-enoate.  
           [0096]    Additional examples of compounds of the invention include the following:  
                         
 
           [0097]    Still additional examples of compounds of the invention are as follows:  
                         
 
           [0098]    In all of the specific alkene active compounds named or illustrated above, the double bond can be replaced with a single bond to produce an analogous series of active compounds that are alkanes.  
           [0099]    In all of the specific alkene active compounds named or illustrated above, the double bond can be replaced with a triple bond to produce an analogous series of active compounds that are alkynes.  
           [0100]    In all of the specific alkene, alkane and alkyne compounds named or illustrated above, the phenyl ring can be replaced with a naphthyl ring to produce an analogous series of active compounds based upon a naphthyl ring system.  
           [0101]    Compounds of the present invention can be made by the techniques described in the Examples below, or variations thereof that will be apparent to those skilled in the art.  
           [0102]    Compounds of the present invention can be made by the technique described in M. Ansell and J. Ducker, Reduced cyclic Compounds. Part XI. The Cyclisation of ω-Arylalkanoic Acids.  J. Chem. Soc.  206-212 (1961) or variations thereof that will be apparent to those skilled in the art.  
           [0103]    A further aspect of the subject invention are addition salts, complexes, or prodrugs such as esters of the compounds described herein, especially the nontoxic pharmaceutically or agriculturally acceptable acid addition salts. The acid addition salts can be prepared using standard procedures in a suitable solvent from the parent compound and an excess of an acid, such as hydrochloric, hydrobromic, sulfuric, phosphoric, acetic, maleic, succinic, ethanedisulfonic or methanesulfonic acids. Esterification to form derivatives such as the methyl or ethyl esters, can also be performed using standard procedures. Tartarate salts can be prepared in accordance with standard procedures.  
           [0104]    Also, derivation of the pesticidal compounds with long chain hydrocarbons will facilitate passage through the cuticle into the pest body cavity. Therefore, in a further embodiment, the subject invention provides compositions comprising the pesticidal compounds bound to lipids or other carriers.  
           [0105]    2. Methods and Formulations for Control of Pests.  
           [0106]    The subject invention concerns novel pest control compounds and methods for using such compounds. Specifically exemplified are novel pesticidal compounds, compositions comprising said pesticidal compounds and the use of such pesticidal compounds and compositions in controlling pests, particularly insect pests such as mosquitoes  
           [0107]    Preferably, the subject compounds have an LD 50  against mosquito larvae of less than 3.0 μmole/ml. More preferably, the compounds have an LD 50  of less than 2.0 μmole/ml, and, most preferably, the compounds have an LD 50  of less than 1.0 μmole/ml. As used herein, “LD 50 ” refers to a lethal dose of a peptide able to cause 50% mortality of larvae maintained on a diet of 1 mg/ml autoclaved yeast supplemented with the pesticidal polypeptide.  
           [0108]    Control of pests using the pest control compounds of the subject invention can be accomplished by a variety of methods known to those skilled in the art. The plant pests that can be controlled by the compounds of the subject invention include pests belonging to the orders Coleoptera, Lepidopterans, Hemiptera and Thysanoptera. These pests all belong to the phylum Arthropod. Other pests that can be controlled according to the subject invention include members of the orders Diptera, Siphonaptera, Hymenoptera and Phthiraptera. Other pests that can be controlled by the compounds of the subject invention include those in the family Arachnida, such as ticks, mites and spiders.  
           [0109]    The use of the compounds of the subject invention to control pests can be accomplished readily by those skilled in the art having the benefit of the instant disclosure. For example, the compounds may be encapsulated, incorporated in a granular form, solubilized in water or other appropriate solvent, powdered, and included into any appropriate formulation for direct application to the pest or to a pest inhabited locus.  
           [0110]    Formulated bait granules containing an attractant and the pesticidal compounds of the present invention can be applied to a pest-inhabited locus, such as to the soil. Formulated product can also be applied as a seed-coating or root treatment or total plant treatment at later stages of the crop cycle. Plant and soil treatments may be employed as wettable powders, granules or dusts, by mixing with various inert materials, such as inorganic minerals (phyllosilicates, carbonates, sulfates, phosphates, and the like) or botanical materials (powdered corncobs, rice hulls, walnut shells, and the like). The formulations may include spreader-sticker adjuvants, stabilizing agents, other pesticidal additives, or surfactants.  
           [0111]    Liquid formulations may be aqueous-based or non-aqueous (i.e., organic solvents), or combinations thereof, and may be employed as foams, gels, suspensions, emulsions, microemulsions or emulsifiable concentrates or the like. The ingredients may include rheological agents, surfactants, emulsifiers, dispersants or polymers.  
           [0112]    As would be appreciated by a person skilled in the art, the pesticidal concentration will vary widely depending upon the nature of the particular formulation, particularly whether it is a concentrate or to be used directly. The pesticidal compound will be present in the composition by at least about 0.0001% by weight and may be 99 or 100% by weight of the total composition. The pesticidal carrier may be from 0.1% to 99.9999% by weight of the total composition. The dry formulations will have from about 0.0001-95% by weight of the pesticide while the liquid formulations will generally be from about 0.0001-60% by weight of the solids in the liquid phase. These formulations will be administered at about 50 mg (liquid or dry) to 1 kg or more per hectare.  
           [0113]    The formulations can be applied to the pest or the environment of the pest, e.g., soil and foliage, by spraying, dusting, sprinkling or the like.  
           [0114]    The pest control compounds may also be provided in tablets, pellets, briquettes, bricks, blocks and the like which are formulated to float, maintain a specified depth or sink as desired. In one embodiment the formulations, according to the present invention, are formulated to float on the surface of an aqueous medium; in another embodiment they are formulated to maintain a depth of 0 to 2 feet in an aqueous medium; in yet another embodiment the formulations are formulated to sink in an aqueous environment.  
           [0115]    The term “control” is intended to include control by repelling insects, as discussed in greater detail below.  
           [0116]    3. Repellant Methods and Compositions.  
           [0117]    In insect repellant methods, the insect is administered the active agent by applying the active agent to a subject or substrate in an amount sufficient to repel insects.  
           [0118]    Subjects to be treated with compounds of the present invention include both human and animal subjects (e.g., dogs, cats, horses, cattle). Subjects may be directly or indirectly treated, such as by applying the active compound to the skin of the subject, or by applying the active compound to an article worn by or otherwise protecting the subject.  
           [0119]    Substrates to be treated with compounds of the present invention include, but are not limited to, floors, plants, containers, walls, pools or open bodies of water, etc.  
           [0120]    Insects that may be repelled by the methods of the present invention include ticks, fleas, cockroaches, and biting flies, typically of the order diptera, and further including mosquitoes, horse flies, deer flies, black flies, gnats, no-see ums, chiggers, etc.  
           [0121]    The term “mosquito” as used herein concerns any type of mosquito (e.g., Anopheles, Aedes, and Culex), including but not limited to Tiger mosquitoes,  Aedes aboriginis, Aedes Aegypti, Aedes, albopictus, Aedes cantator, Aedes sierrensis, Aedes sollicitans, Aedes squamiger, Aedes sticticus, Aedes vexans, Anopheles quadrimaculatus, Culex pipiens,  and  Culex quinquefaxciatus.    
           [0122]    The term “tick” as used herein includes any type of tick, including but not limited to, deer ticks, the American dog tick ( Dermacentor variabilis ),  Ornithodoros parkeri, O. moubata,  and  Dermacentor andersoni.    
           [0123]    The term “cockroach” as used herein refers to any type of cockroach, including but not limited to the American cockroach ( Periplaneta americana ), German cockroach ( Blattella germanica ), oriental cockroach ( Blatta orientalis ), wood cockroach ( Parcoblatta pennsylvanica ), brownbanded cockroach ( Supella lonigipalpa ), and smokybrown cockroach ( Periplaneta fuliginosa ).  
           [0124]    Other insect that can be treated by the repellant methods of the present invention include, but are not limited to: lice (Order Phthiraptera), such as head and body lice of humans,  Pediculus humanus capitis  and  P. H. humanus;  Fleas (Order Siphonaptera), such as cat and dog fleas, Ctenocephalides sp. human fleas, Echidnophaga, Pulex sp. Bees, wasps and ants (Order Hymenoptera) mites such as  Sarcoptes scabei  (human itch mite) the North American chigger or red bug, Trombicula sp. nematodes such as human parasitic nematodes, Silverfish (Order Thysanura), such as  Lepisma saccharina,  firebrat,  Thermobia domestica;  Termites (Order Isoptera) such as  Reticulitermes flavipes, Incisitermes minor, Marginitermes hubbardi,  and  Cryptotermes brevis;  Earwigs (Order Dermaptera); Psocids (Order Psocoptera) such as booklice; Beetles (Order Coleoptera), particularly wood eating beetles; Centipedes such as Lithobius, Geophilus, Scutigera and millipides such as  Julus terrestris;  Scorpions such as  Centruroides sculpturatus  and  Mastigoproctus gianteus;  etc.  
           [0125]    Liquid repellant formulations may be aqueous-based or non-aqueous (i.e., organic solvents), or combinations thereof, and may be employed as foams, gels, suspensions, emulsions, microemulsions or emulsifiable concentrates or the like. The ingredients may include rheological agents, surfactants, emulsifiers, dispersants or polymers.  
           [0126]    In one embodiment, a floor wax composition may include repellant compounds as described herein, in an amount effective to repel cockroaches that might otherwise feed upon the composition once applied to floors, or to simply repel cockroaches from floor surfaces to which they are applied.  
           [0127]    As will be appreciated by a person skilled in the art, the repellant concentration will vary widely depending upon the nature of the particular formulation, particularly whether it is a concentrate or to be used directly. The repellant compound will be present in the composition in a concentration of at least about 0.0001% by weight and may be 10, 50, 99 or 100% by weight of the total composition. The repellant carrier may be from 0.1% to 99.9999% by weight of the total composition. The dry formulations will have from about 0.0001-95% by weight of the pesticide while the liquid formulations will generally have from about 0.0001-60% by weight of the solids in the liquid phase.  
           [0128]    The formulations may be applied to the subject&#39;s skin, or may be applied to garments, belts, collars, or other articles worn by the subject from whom insects are to be repelled. The formulation may be applied to netting or screening that protects a subject, particularly a sleeping subject. The formulations may be applied to non-animal substrates from which insects are to be repelled, such as plants. Application to subjects or substrates may be carried out by spraying, dusting, sprinkling or the like.  
           [0129]    The compounds according to the present invention may be employed alone or in mixtures with one another and/or with such solid and/or liquid dispersible carrier vehicles as described herein or as otherwise known in the art, and/or with other known compatible active agents, including, for example, insecticides, acaricides, rodenticides, fungicides, bactericides, nematocides, herbicides, fertilizers, growth-regulating agents, etc., if desired, in the form of particular dosage preparations for specific application made therefrom, such as solutions, emulsions, suspensions, powders, pastes, and granules as described herein or as otherwise known in the art which are thus ready for use.  
           [0130]    The repellant compounds may be administered with other insect control chemicals, for example, the compositions of the invention may employ various chemicals that affect insect behavior, such as insecticides, attractants and/or repellents, or as otherwise known in the art. The repellant compounds may also be administered with chemosterilants.  
           [0131]    The repellant compounds are suitably applied by any method known in the art including, for example, spraying, pouring, dipping, in the form of concentrated liquids, solutions, suspensions, sprays, powders, pellets, briquettes, bricks and the like, formulated to deliver a repellant effective concentration of the repellant compound. The repellant formulations may be applied in a repellant effective amount to an area of pest infestation or an area susceptible to infestation, a body of water or container, a barn, a carpet, pet bedding, an animal, clothing, skin, and the like.  
           [0132]    4. Computational Chemistry Methods.  
           [0133]    Methods of identifying a non-peptide TMOF agonist from a peptide TMOF agonist are a further aspect of the present invention. Such methods typically comprise the steps of:  
           [0134]    modeling in a computer (preferably with simulated annealing) a model peptide TMOF agonist selected from the group consisting of TMOF and peptide TMOF analogs;  
           [0135]    determining in said computer spatial orientations for at least one key feature of said model peptide compound;  
           [0136]    generating in said computer a putative non-peptide TMOF agonist structure, said structure including (i) said at least one key feature and (ii) spatial orientations for said at least one key feature corresponding to said spatial orientations for said at least one key feature of said model peptide compound; then  
           [0137]    synthesizing said putative non-peptide TMOF agonist; and then  
           [0138]    screening said putative non-peptide TMOF agonist to determine the presence of TMOF activity therein.  
           [0139]    Peptide TMOF agonists or analogs that may be used as a basis for modeling of the instant invention include but are not limited to those disclosed in, inter alia, U.S. Pat. Nos. 5,011,909; 5,130,253; and 5,358,934, the disclosures of which are incorporated by reference herein in their entirety.  
           [0140]    TMOF and other TMOF peptide agonists (preferably tripeptide fragments of TMOF) may be modeled by the INSIGHT™ software (available through the North Carolina Super Computing Center for academic use) using a simulated annealing protocol as described by Lovas and Murphy (S. Lovas and R. Murphy, Molecular Modeling of Neuropeptides, in  Neuropeptide Protocols,  pp. 209-217 (Irvine, G. B.; Williams, C. H., Eds. Humana Press, N.J. 1997)) and Damewood (J. Damewood,  Rev. Computational Chem.  9, 1-79 (1996). Peptide Mimetic Design with the Aid of Computational Chemistry). Composite structural data was then used to determine approximate spatial orientations for what were deemed to be key structural features, particularly as a phenyl ring and a carboxylate functional group. These data indicated a greater degree of flexibility in the N-terminus than in the C-terminus of the peptide structures. Non-peptide analogs were then subjected to energy minimization by molecular mechanics and distances between functional groups compared to the data obtained in the data set of the TMOF fragments. These data indicated a starting point for analog synthesis as exemplified by E-7-phenylhept-4-enoic acid. Actual compounds having structures corresponding to structures generated by the computational techniques are then synthesized in accordance with known techniques, and screened for activity in a bioassay such as described below. Thus, compounds produced by the method that have TMOF agonist activity and can be used in the methods described herein generally have the structure P—R—C, where P is a phenyl group, R is an alkane, alkene, or alkyne, and C is a carboxylate group, all of which may be substituted or unsubstituted.  
           [0141]    The following examples are illustrative of the practice of the present invention and should not be construed as limiting. All percentages are by weight and all solvent mixture proportions are by volume unless otherwise noted. 
       
    
    
     EXAMPLE 1  
     Preparation of E-7-Phenylhept-4-enoic Acid  
       [0142]    E-7-Phenylhept-4-enoic acid was prepared from commercially available dihydrocinnamaldehyde in five steps. Dihydrocinnamaldehyde was combined with the lithium salt of ethyl diethoxyphosphonioacetate in tetrahydrofuran at room temperature. The resulting unsaturated ester, ethyl 5-phenylpent-2-enoate, was purified by chromatography. The unsaturated ester was then reduced to the allylic alcohol, E-5-phenylpent-2-en-1-ol, by reaction with excess diisobutylaluminum hydride in tetrahydrofuran at −78° C. The purified alcohol was then converted to the bromide, E-1-bromo-5-phenylpent-2-ene, by reaction with triphenylphosphine and carbon tetrabromide in methylene chloride at 0° C. The purified bromide was then used to alkylate the sodium salt of diethyl malonate to provide the diester, methyl E-1-carbomethoxy-5-phenylhept-4-enoate. The diester was purified by chromatography and then subjected to saponification using methanolic sodium hydroxide. The diacid was purified by acid/base extraction and then used directly in the next step of the reaction sequence. Decarboxylation was effected by heating the neat diacid at 170° C. (under Ar) for 30 minutes. The product, E-7-phenylhept-4-enoic acid, was obtained in greater than 95% purity. Any residual diacid was removed by dissolving the acid in hexane and filtering. All compounds in the reaction sequence were fully characterized by spectral analysis (infrared and nuclear magnetic spectroscopy) and new compounds were analyzed by combustion analysis.  
       EXAMPLE 2  
     7-Phenylheptanoic Acid  
       [0143]    7-Phenylheptanoic acid was prepared from E-7-phenylhept-4-enoic acid by hydrogenation at 40 psi using 5% palladium on carbon as the catalyst. Quantitative conversion of the alkene to the alkane was observed by thin layer chromatographic analysis. The product acid was purified by chromatography on silica gel and fully characterized by spectroscopic methods (infrared and nuclear magnetic resonance) and combustion analysis.  
       EXAMPLE 3  
     E-7-(4-Methoxyphenyl)hept-4-enoic Acid  
       [0144]    E-7-(4-Methoxyphenyl)hept-4-enoic acid was prepared in the same fashion as E-7-phenylhept-4-enoic acid by substituting 3-(4-methoxyphenyl) propional for dihydrocinnamaldehyde as the starting material in the synthetic sequence. 3-(4-methoxyphenyl)propional was combined with the lithium salt of ethyl diethoxyphosphonioacetate in tetrahydrofuran at room temperature. The resulting unsaturated ester, ethyl 5-(4-methoxyphenyl)pent-2-enoate, was purified by chromatography. The unsaturated ester was then reduced with excess diisobutylaluminum hydride in tetrahydrofuran at −78° C. The purified alcohol was then converted to the bromide, E-1-bromo-5-(4-methoxyphenyl)pent-2-ene, by reaction with triphenylphosphine and carbon tetrabromide in methylene chloride at 0° C. The purified bromide was then used to alkylate the sodium salt of diethyl malonate to provide the diester, methyl E-1-carbomethoxy-5-(4-methoxyphenyl)hept-4-enoate. The diester was purified by chromatography and then subjected to saponification using methanolic sodium hydroxide. The diacid was purified by acid/base extraction and then used directly in the next step of the reaction sequence. Heating the neat diacid at 170° C. (under Ar) for 30 minutes effected decarboxylation. The product, E-7-(4-methoxyphenyl)hept-4-enoic acid, was obtained in greater than 95% purity. Any residual diacid was removed by dissolving the acid in hexane and filtering. All compounds in the reaction sequence were fully characterized by spectral analysis and new compounds were analyzed by combustion analysis.  
       EXAMPLE 4  
     E-7-(4-hydroxyphenyl)hept-4-enoic Acid  
       [0145]    E-7-(4-hydroxyphenyl)hept-4-enoic acid was prepared from E-7-(4-methoxyphenyl)hept-4-enoic acid by removal of the methyl ether with boron tribromide. Boron tribromide was added to a solution of E-7-(4-methoxyphenyl)hept-4-enoic acid in methylene chloride at −78° C. and stirred for four hours. The product, E-7-(4-hydroxyphenyl)hept-4-enoic acid, was obtained by aqueous work-up of the reaction mixture followed by chromatography on silica gel, and was fully characterized by spectroscopic methods (infrared and nuclear magnetic resonance) and combustion analysis.  
       EXAMPLE 5  
     Bioassay of Compounds  
       [0146]    Mosquito larval mortality was followed for three days in microtiter plates containing 160 μL sterile water, 1-4 μL of the test compound dissolved in dimethylsulfoxide, and 10 μL of 2% Brewer&#39;s yeast. Controls were run under the same conditions without the test compounds. Larval mortality in the controls was 3% for 1-3 μL dimethylsulfoxide, and up to 20% for 4 μL dimethylsulfoxide. Data for various compounds of the invention as an LD 50  are given in Table 1 below.  
                                               TABLE 1                           Bioassay of Active compounds.                Compound   Activity (LD 50 )                            7-Phenylheptanoic acid   0.19 ± 0.02   mM           E-7-(4-Hydroxyphenyl)hept-4-enoic acid   0.59 ± 0.03   mM           E-7-Phenylhept-4-enoic acid   &lt;0.08   mM           E-7-(4-Methoxyphenyl)hept-4-enoic acid   1.28   mM                      
 
       EXAMPLE 6  
     Ethyl E-7-(Cyclohexyl)hept-4-enoate (IBI218)  
       [0147]    Ethyl E-7-(cyclohexyl)hept-4-enoate was prepared in two steps from the known compound 3-cyclohexylpropanal. A tetrahydrofuran solution of 3-cyclohexylpropanal was treated with vinyl magnesium bromide (as a solution in tetrahydrofuran) at 0° C. The resulting allyl alcohol derivative, 5-cyclohexyl-3-hydroxypent-1-ene, was purified by chromatography. The allylic alcohol was then dissolved in triethyl orthoacetate and heated to 140° C. to effect a Claisen rearrangement. The product, ethyl E-7-(cyclohexyl)hept-4-enoate, was obtained in &gt;95% purity after chromatography. The product was fully characterized by spectral analysis(infrared and nuclear magnetic spectroscopy) and combustion analysis.  
       EXAMPLE 7  
     Ethyl E-7-((4-phenyl)phenyl)hept-4-enoate (IBI 221)  
       [0148]    Ethyl E-7-((4-phenyl)phenyl)hept-4-enoate was prepared in two steps from the known compound 3-((4-phenyl)phenyl)-propanal. A tetrahydrofuran solution of 3-((4-phenyl)phenyl)propanal was treated with vinyl magnesium bromide (as a solution in tetrahydrofuran) at 0° C. The resulting allyl alcohol derivative, 5-((4-phenyl)phenyl)-3-hydroxypent-1-ene, was purified by chromatography. The allylic alcohol was then dissolved in triethyl orthoacetate and heated to 140 C. to effect a Claisen rearrangement. The product, ethyl E-7-((4-phenyl)phenyl)hept-4-enoate, was obtained in &gt;95% purity after chromatography. The product was fully characterized by spectral analysis(infrared and nuclear magnetic spectroscopy) and combustion analysis.  
       EXAMPLE 8  
     E-7-(Cyclohexyl)hept-4-enoic Acid (IBI172)  
       [0149]    Ethyl E-7-(cyclohexyl)hept-4-enoate was saponified in methanolic sodium hydroxide at room temperature. The resulting acid, E-7-(cyclohexyl)hept-4-enoic acid, was obtained in &gt;95% purity after chromatography. The product was fully characterized by spectral analysis (infrared and nuclear magnetic spectroscopy) and combustion analysis.  
       EXAMPLE 9  
     E-7-((4-phenyl)phenyl)hept-4-enoic Acid (IBI165)  
       [0150]    Ethyl E-7-((4-phenyl)phenyl)hept-4-enoate was saponified in methanolic sodium hydroxide at room temperature. The resulting acid, E-7-((4-phenyl)phenyl)hept-4-enoic acid, was obtained in &gt;95% purity after chromatography. The product was fully characterized by spectral analysis infrared and nuclear magnetic spectroscopy) and combustion analysis.  
       EXAMPLE 10  
     Repellent Assay for Ticks  
       [0151]    A repellent assay for ticks is carried out as follows:  
         [0152]    (1) Prepare 15 mg/ml test solution of test compound in 100% acetone in individual 500 uL bullet tubes. Prepare control solution (1 uL olive oil plus 100 uL 100 acetone).  
         [0153]    (2) prepare half circle test papers (radious=2.25 cm) from Whatman filter paper (#2 qualitative).  
         [0154]    (3) Saturate test papers with 60 uL of 100% acetone, control, or test solution. Test papers are in individual sterile petri dishes during this process.  
         [0155]    (4) Allow test papers to air dry at room temperature for one hour.  
         [0156]    (5) Select and place ten ticks into separate sterile glass petri dishes.  
         [0157]    Allow ticks to acclimate to covered dishes for one hour.  
         [0158]    (6) Transfer one test paper treated with a test solution and one test paper treated with the control solution to each of separate sterile petri dish bottoms. On control included two test papers treated with acetone. A second control included two test papers treated with the control solution.  
         [0159]    (7) Place ten ticks in between the two test papers. Place the test arenas in a dark room at room temperature.  
         [0160]    (8) Record the location of ticks 15 minutes, 1 hour and 2 hours after being placed in the test arenas.  
         [0161]    Results with a variety of different active compounds are given in the Table 2 below.  
                                                                                             TABLE 2                           Ornithodoros parkeri repellent assay.                15 minutes   1 hour   2 hours            Time   # ticks on   # ticks on   # ticks on   # ticks on   # ticks on   # ticks on       Compound   compound   acetone   compound   acetone   compound   acetone                    152       8   1   9   1   9       156   1   7   1   8   1   9       172   2   8       10       10       218       10       10       10       219               10   2   8       220   2   7   6   4   2   8       221   5   5   4   4   2   8       olive oil   8   2   6   4   5   5       acetone   3   2   5   5   4   6                  
 
         [0162]    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.