insecticides

Abstract

The present invention provides γ-pyrone compounds for use as insecticides. γ-pyrones have the general formula (1): JPEG2023537966000036.jpg3243 [wherein, R 1 , R 2 , R 3 and R 4 are each independently optionally substituted C-C 12 Alkyl, preferably C1-C6 alkyl, more preferably C1-C2 alkyl; H; -COOH; -OH; -OCH3 or -R 5 (CH2) n R 6 R 7 CH3; R 5 , R 6 and R 7 are each independently optionally substituted C-C 12 and n is 1 to 18.], salts, solvates, dimers or isomers thereof. Other embodiments of the present invention provide for the use of γ-pyrone compounds, pesticidal compositions comprising the compounds, and commercial embodiments include kits for sale on store shelves.

Description

[Technical Field]

[0001] Related Applications This application claims priority to Australian Provisional Patent Application No. 2020902941, filed August 18, 2020, the contents of which are to be construed as incorporated herein by reference.

[0002] FIELD OF THE INVENTION The present invention relates to pyrone derivative compounds, which may be of natural (eg, plant extracts) or synthetic origin, for use as insecticides. [Background technology]

[0003] Background of the Invention Any statement throughout this specification regarding prior art should in no way be taken as an admission that such prior art is commonly known in the art or forms part of the common general knowledge.

[0004] Pests have been a problem for crops and stored foods since the beginning of agriculture. Since the 1950s, synthetic pesticides have been the method of choice for controlling pest infestations in agricultural industries, including fields, stored grains, warehouses, and food processing facilities.

[0005] The Australian cotton industry also relies on repeated applications of synthetic pesticides to control crop pests. Concomitant problems related to insecticide resistance, beneficial insect species decline, high production costs, and environmental impacts currently require the investigation of alternative strategies for pest control. These include (but are not limited to) genetically modified cotton crops containing Bacillus thuringiensis ("Bt") and other host-plant-resistant insecticidal proteins, biopesticides, better management of beneficial insect species, trap crops, intercropping and interplanting, and behavioral manipulation of pests and beneficial insects. Genetically modified (transgenic) crops are currently being developed in Australia and many other countries to control lepidopteran pests, and their introduction has reduced the use of synthetic insecticides against these pests.

[0006] Although plant-derived insecticides are well known and in fact already being applied in the cotton industry, there is an ongoing need to discover new insecticidal compounds. Furthermore, the Australian cotton industry is challenged by unique environments, climates and pest species, such as aphids.

[0007] Approximately 5,000 species of aphids have been described, of which approximately 450 species colonize food and fiber crops. Aphids can cause significant economic damage in the agricultural industry. Not only can aphids damage crops, including cotton, and reduce yields, but they also have a greater impact by acting as vectors for plant viruses. The transmission of these viruses depends on the movement of aphids to different parts of the plant, to neighboring plants, and further into the field. In this respect, the exploratory behavior of aphids, which taste the host, is more damaging than the long-term aphid feeding and reproduction by resident individuals.

[0008] Control of aphids with insecticides is difficult because they reproduce quickly and even small areas of failure can allow rapid recovery of the population. Aphids can occupy the undersides of leaves that are missed by spraying, while systemic insecticides do not move satisfactorily to petals. Finally, some aphid species are resistant to common insecticide classes, including carbamates, organophosphates, and pyrethroids. Summary of the Invention [Problem to be solved by the invention]

[0009] It is an object of the present invention to overcome or mitigate one or more of the disadvantages of the prior art, or at least to provide a useful alternative.

[0010] It is an object of at least one preferred form of the present invention to provide compounds, pesticidal compositions or extracts that are capable of controlling pests in crop or ornamental plants.

[0011] Although the present invention has been described with reference to specific examples, it will be apparent to those skilled in the art that the present invention may be embodied in many other forms. [Means for solving the problem]

[0012] Summary of the Invention According to a first aspect of the present invention, there is provided a γ-pyrone compound for use as an insecticide. In a preferred embodiment, the γ-pyrone compound is podopyrone.

[0013] In some embodiments, the γ-pyrone has the general formula (1): [ka]

[0014] [During the ceremony,

[0015] R 1 , R 2 , R 3 and R 4 are each independently optionally substituted C-C 12 Alkyl, preferably C1-C6 alkyl, more preferably C1-C2 alkyl; H; -COOH; -OH; -OCH3 or -R 5 (CH2) n R 6 R 7 Selected from CH3;

[0016] R 5 , R 6 and R 7 are each independently optionally substituted C-C 12 alkyl, preferably C1-C6 alkyl, more preferably C1-C2 alkyl; -C=O; -COO-, N, S or O; and

[0017] n is 1 to 18.

[0018] or a salt, solvate, dimer or isomer thereof.

[0019] In some embodiments, the γ-pyrone has the general formula (2): [ka]

[0020] [During the ceremony,

[0021] R 1 , R 2 , R 3 are each independently C1-C 12 alkyl, preferably C1-C6 alkyl, more preferably C1-C2 alkyl, wherein the C1-C 12 Alkyl, C1-C6 alkyl and C1-C2 alkyl may be optionally substituted with epoxide, glycoside, acetoxy, halogen, cyano, amino, phenyl, heteroaryl; H; -COOH; -OH; or -OCH3;

[0022] R 5 , R 6 and R 7 are each independently C1-C 12 alkyl, preferably C1-C6 alkyl, more preferably C1-C2 alkyl, wherein the C1-C 12 Alkyl, C1-C6 alkyl and C1-C2 alkyl are optionally C1-C 12 alkyl, preferably C1-C6 alkyl, more preferably C1-C2 alkyl, epoxide, glycoside, acetoxy, halogen, cyano, amino, alcohol, phenyl, heteroaryl; optionally substituted with -C=O; -COO-, N, S, or O; and

[0023] n is 1 to 18.

[0024] or a salt, solvate, dimer or isomer thereof.

[0025] In some embodiments, the γ-pyrone has the general formula (3): [ka]

[0026] [During the ceremony,

[0027] n is 6, 7, or 8;

[0028] R 1 is C1-C 12 alkyl, preferably C1-C6 alkyl, more preferably C1-C2 alkyl; and

[0029] R 5 , R 6 and R 7 are each independently selected from —C═O and —CH—;

[0030] However, R 5 , R 6 and R 7 If one of the groups is -C=O, the remaining group is -CH2-. It is of the type.

[0031] In certain embodiments, the γ-pyrone is a compound selected from the group consisting of compounds (1a)-(1q) described herein, or a salt, solvate, dimer, or isomer thereof.

[0032] A second aspect of the present invention provides pesticidal compositions comprising the γ-pyrone compounds described herein.

[0033] Surprisingly, the present inventors have discovered that γ-pyrone compounds or insecticidal compositions containing these compounds can be selective for certain pests and have minimal impact on non-target organisms such as beneficial insects.

[0034] In a preferred embodiment, the present inventors have surprisingly discovered that γ-pyrone compounds or insecticidal compositions containing these compounds cause high mortality rates of aphids and mites without significantly affecting the genus Apis. The present inventors have discovered that γ-pyrone compounds have minimal impact on the European honeybee (Apis mellifera), which is considered the most important pollinator of agricultural crops worldwide due to its abundance and ease of handling by humans.

[0035] According to a third aspect of the present invention there is provided a formulation for controlling pests comprising:

[0036] one or more agriculturally acceptable diluents and / or carriers and / or other additives such as emulsifiers, wetting agents, surfactants, stabilizers, spreading agents, etc.; and

[0037] Compounds or insecticidal compositions comprising an insecticidally effective amount of one or more γ-pyrone compounds described herein A formulation comprising: The formulations herein, when used, exert insecticidal activity and / or repel pests and / or prevent pests from laying eggs and / or influence the egg-laying sites and / or prevent pests from feeding on plants.

[0038] According to a fourth aspect of the present invention, there is provided a compound or pesticidal composition comprising an insecticidally effective amount of one or more γ-pyrone compounds as described herein for controlling pests by exerting pesticidal activity and / or repelling pests and / or inhibiting pest oviposition and / or influencing oviposition sites and / or inhibiting pest feeding on plants.

[0039] According to a fifth aspect of the present invention, there is provided a method for controlling one or more pests by exerting insecticidal activity and / or repelling the pests and / or inhibiting pest oviposition and / or affecting oviposition sites and / or inhibiting pest feeding on plants, the method comprising treating a locus with a compound or insecticidal composition comprising an insecticidally effective amount of one or more γ-pyrone compounds described herein.

[0040] The present inventors have surprisingly discovered that a γ-pyrone compound or an insecticidal composition comprising the compound can affect at least two ion channels (e.g., neuronal ion channels) in pest insects. In some embodiments, the γ-pyrone compound or an insecticidal composition comprising the compound can affect at least two ion channels selected from the group consisting of sodium, potassium, and chloride ion channels.

[0041] In a preferred embodiment, the γ-pyrone compound or an insecticidal composition containing the compound can induce the outflow of sodium and potassium ions from target cells of insect pests. In a preferred embodiment, the γ-pyrone compound or an insecticidal composition containing the compound can induce the inflow of chloride ions from target cells of insect pests.

[0042] Typically, insecticidal compounds only affect a single ion channel. Surprisingly, the γ-pyrone compounds or insecticidal compositions of the present invention can have a dual mechanism of action, i.e., can affect at least two ion channels in insect pests, which can induce a neurotoxic mechanism of action.

[0043] According to a sixth aspect of the present invention there is provided a kit for sale on the shelf comprising:

[0044] a compound or pesticidal composition comprising one or more γ-pyrone compounds described herein;

[0045] one or more agriculturally acceptable diluents and / or carriers and / or other additives such as emulsifiers, wetting agents, surfactants, stabilizers, spreading agents, etc.;

[0046] Instructions for preparing a formulation comprising a pesticidally effective amount of one or more gamma-pyrone compounds or pesticidal compositions described herein per unit amount of one or more agriculturally acceptable diluents and / or carriers and / or other additives such as emulsifiers, wetting agents, surfactants, stabilizers, spreading agents, etc. A kit is provided comprising:

[0047] In some embodiments, the kit further comprises an application means, such as in the form of a sprayer, in which the formulation is optionally prepared.

[0048] According to a seventh aspect of the present invention there is provided an insecticidal composition for controlling pests, comprising one or more extracts from the genera Podorepis, Gonistylus and combinations thereof containing one or more secondary plant compounds (SPCs) which have insecticidal activity and / or repel pests and / or deter pest oviposition and / or influence oviposition sites and / or deter pests from feeding on the plant.

[0049] According to an eighth aspect of the present invention there is provided a formulation for controlling pests comprising:

[0050] one or more agriculturally acceptable diluents and / or carriers and / or other additives such as emulsifiers, wetting agents, surfactants, stabilizers, spreading agents, etc.; and

[0051] Insecticidal compositions comprising one or more extracts from the genera Podolepis, Gonistilus, and combinations thereof, containing an insecticidally effective amount of one or more secondary plant compounds (SPCs) described herein A formulation comprising:

[0052] The formulations herein, when used, exert insecticidal activity and / or repel pests and / or prevent pests from laying eggs and / or influence the egg-laying sites and / or prevent pests from feeding on plants.

[0053] According to a ninth aspect of the present invention there is provided the use of an insecticidal composition comprising one or more extracts from the genera Podorepis, Gonistylus and combinations thereof, comprising an insecticidally effective amount of one or more secondary plant compounds (SPCs) as described herein, for controlling pests by exerting insecticidal activity and / or repelling pests and / or inhibiting pest oviposition and / or influencing oviposition sites and / or inhibiting pest feeding on plants.

[0054] According to a tenth aspect of the present invention, there is provided a method of controlling one or more pests by exerting insecticidal activity and / or repelling the pests and / or inhibiting pest oviposition and / or affecting oviposition sites and / or inhibiting pest feeding on plants, the method comprising treating a locus with an insecticidal composition comprising one or more extracts from the genera Podorepis, Gonistylus, and combinations thereof, comprising an insecticidally effective amount of one or more secondary plant compounds (SPCs) as described herein.

[0055] As described herein, the inventors have surprisingly discovered that compositions comprising one or more extracts from the genera Podorepis, Gonistilus, and combinations thereof, including one or more secondary plant compounds (SPCs) described herein, can affect at least two ion channels (e.g., neuronal ion channels) in pest insects. In certain embodiments, the SPCs can affect at least two ion channels selected from the group consisting of sodium, potassium, and chloride ion channels.

[0056] In a preferred embodiment, the SPC is capable of causing sodium and potassium ions to flow out of the target cells of the pest. In a preferred embodiment, the SPC is capable of causing chloride ions to flow in of the target cells of the pest.

[0057] According to an eleventh aspect of the present invention there is provided a kit for sale on store shelves comprising:

[0058] an insecticidal composition comprising one or more extracts from the genera Podorepis, Gonistilus, and combinations thereof, containing one or more secondary plant compounds (SPCs) as described herein;

[0059] one or more agriculturally acceptable diluents and / or carriers and / or other additives such as emulsifiers, wetting agents, surfactants, stabilizers, spreading agents, etc.;

[0060] Instructions for preparing a formulation containing a pesticidally effective amount of one or more SPCs per unit amount of one or more agriculturally acceptable diluents and / or carriers and / or other additives such as emulsifiers, wetting agents, surfactants, stabilizers, spreading agents, etc. A kit is provided comprising:

[0061] In some embodiments, the kit further comprises an application means, such as in the form of a sprayer, in which the formulation is optionally prepared.

[0062] The present inventors have surprisingly discovered that extracts containing SPCs (in certain embodiments, γ-pyrones) have significant insecticidal activity against pests, particularly aphids and mites.

[0063] In particular, the present inventors have discovered that extracts from Podopilone-containing Podolepis jaceoides have significant insecticidal activity against pests, particularly aphids and mites.

[0064] In some embodiments, the SPC is γ-pyrone. In some embodiments, the γ-pyrone is podopyrone. In some embodiments, the podopyrone is selected from the group consisting of 10'-oxopodopyrone, 10'-oxo-8-methylpodopyrone, 9'-oxopodopyrone, 9'-oxo-8-methylpodopyrone, 1'-oxo-nor-podopyrone, 1'-oxopodopyrone, 1'-deoxo-8-methyl-1'-oxonorpodopyrone, 8-methylpodopyrone, 8-methyl-1'-oxo-nor-podopyrone, 1'-oxo-8-methylpodopyrone, podopyrone, norpodopyrone, homopodopyrone, 10'-hydroxy-8-methylpodopyrone, 10'-acetoxy-8-methylpodopyrone, 10'-acetoxypodopyrone, and combinations thereof.

[0065] In some embodiments, the extract containing γ-pyrone is from the genera Podolepis, Gonistilus, and combinations thereof.

[0066] In one embodiment, the Podolepis-derived extract is Podolepis labill, Podolepis acuminata, Podolepis affinis Sond., Podolepis aristata, Podolepis arachnoidea, Podolepis auriculata, Podolepis basalt plain, Podolepis canescens, Podolepis capillaris, Podolepis carnarvon, Podolepis centauroides, Podolepis chrysantha, Podolepis contorta, Podolepis cupulata, Podolepis davisiana, Podolepis decipiens, Podolepis divaricata, Podolepis sect.Doratolepis, Podolepis eremaea, Podolepis ferruginea, Podolepis filiformis, Podolepis gardneri, Podolepis georgei, Podolepis gracilis, Podolepis gibertii, Podolepis gnaphalioides, Podolepis gracilis, Podolepis great victoria desert, Podolepis hieracioides, Podolepis inundata, Podolepis jaceoides, Podolepis kendallii, Podolepis laciniata, Podolepis laevigata, Podolepis lessonii, Podolepis linearifolia, Podolepis longipedata, Podolepis lucaeana, Podolepis macrocephala, Podolepis microcephala, Podolepis mitchellii, Podolepis monticola, Podolepis muelleri, Podolepis neglecta, Podolepis nutans, Podolepis omissa, Podolepis pallida, Podolepis papillosa, Podolepis remota, Podolepis rhytidochlamys, Podolepis robusta, Podolepis rosea, Podolepis rosmarinifolia, Podolepis rubida, Podolepis rugata, Podolepis rutidoclamys, Podolepis scalia, Podolepis siemssenia, Podolepis siemssenii, Podolepis simplicicaulis, Podolepis spenceri, Podolepis tepperi, Podolepis sp. aff. robusta, Podolepis NEAlps, Podolepis Warrabah, Podolepis Wollunga Well, Podolepis stylolepis, Podolepis subulata, Podolepis tepperi, Podolepis tetrachaeta, and combinations thereof.

[0067] In certain embodiments, the extract from the genus Gonystylus is Gonystylus acuminatus, Gonystylus affinis, Gonystylus areolatus, Gonystylus augescens, Gonystylus bancanus, Gonystylus borneensis, Gonystylus brunnescens, Gonystylus calophylloides, Gonystylus calophyllus, Gonystylus confusus, Gonystylus consanguineus, Gonystylus costalis, Gonystylus decipiens, Gonystylus eximius, Gonystylus forbesii, Gonystylus glaucescens, Gonystylus keithii, Gonystylus lucidulus, Gonystylus macrocarpus, Gonystylus macrophyllus, Gonystylus maingayi, Gonystylus micranthus, Gonystylus nervosus, Gonystylus nobilis, Gonystylus othmanii, Gonystylus pendulus, Gonystylus punctatus, Gonystylus reticulatus, Gonystylus spectabilis, Gonystylus stenosepalus, Gonystylus velutinus, Gonystylus xylocarpus and combinations thereof.

[0068] In a preferred embodiment, the extract containing γ-pyrone is derived from Podolepis jaceoides, Gonystylus keithii, and combinations thereof.

[0069] As noted above, the present invention provides a method for controlling one or more pests by exerting insecticidal activity and / or repelling the pests and / or inhibiting pest oviposition and / or affecting oviposition sites and / or inhibiting pest feeding on plants, comprising treating a locus with an insecticidally effective amount of a compound or insecticidal composition of the present invention, or an insecticidal composition comprising one or more extracts from the genera Podoplepis, Gonistilus, and combinations thereof, comprising one or more secondary plant compounds (SPCs) as described herein.

[0070] The present inventors have surprisingly discovered that the use of extracts containing low concentrations of γ-pyrone (e.g., less than 2 w / v % active ingredient, preferably less than 1 w / v % active ingredient) has significant insecticidal activity against pests.

[0071] In certain embodiments, the method comprises treating the habitat with a γ-pyrone-containing compound, insecticidal composition or extract of the present invention at a concentration of less than about 20% w / v, less than about 15% w / v, less than about 10% w / v, less than about 5% w / v, less than about 3% w / v, less than about 2% w / v, less than about 1% w / v, less than about 0.5% w / v and less than about 0.25% w / v.

[0072] In some embodiments, the method comprises treating the habitat with a γ-pyrone-containing compound, insecticidal composition, or extract of the present invention at a concentration of about 0.25 to about 20 w / v%, about 0.25 to about 15 w / v%, about 0.25 to about 10 w / v%, about 0.25 to about 5 w / v%, about 0.25 to about 3 w / v%, preferably about 0.25 to about 2 w / v%, about 0.25 to about 1 w / v%, or about 0.5 to about 1 w / v%.

[0073] In certain embodiments, the method comprises treating the habitat with an SPC (i.e., active ingredient) at a concentration of less than about 20 w / v%, less than about 15 w / v%, less than about 10 w / v%, less than about 5 w / v%, less than about 3 w / v%, less than about 2 w / v%, less than about 1 w / v%, less than about 0.5 w / v%, and less than about 0.25 w / v%.

[0074] In certain embodiments, the method comprises treating the habitat with SPC (i.e., active ingredient) at a concentration of about 0.001 to about 20 w / v%, about 0.001 to about 15 w / v%, about 0.001 to about 10 w / v%, about 0.001 to about 5 w / v%, about 0.001 to 3 w / v%, about 0.001 to 2 w / v%, about 0.001 to 1 w / v%, about 0.005 to 0.5 w / v%, and about 0.001 to 0.1 w / v%.

[0075] In certain embodiments, the method comprises treating the habitat with a concentration of SPC (i.e., active ingredient) from about 1 to about 30,000 ppm, from about 30 to about 30,000 ppm, from about 50 to about 30,000 ppm, from about 100 to about 30,000 ppm, from about 150 to about 30,000 ppm, from about 200 to about 30,000 ppm, from about 300 to about 30,000 ppm, from about 300 to about 25,000 ppm, from about 300 to about 16,000 ppm, from about 300 to about 15,000 ppm, from about 300 to about 12,000 ppm, or from about 3,000 to about 12,000 ppm.

[0076] In some embodiments, the pest is a plant pest and the method comprises applying a compound, pesticidal composition or extract comprising a γ-pyrone of the present invention to the plant or its vicinity.

[0077] definition In describing and claiming the present invention, the following terminology will be used in accordance with the definitions set out below. It will also be understood that the terminology used herein is for the purpose of describing particular embodiments of the present invention only, and is not intended to be limiting. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0078] Unless the context otherwise requires, throughout the specification and claims, the terms "comprises," "including," and the like, are to be construed in an inclusive sense, rather than an exclusive or exhaustive sense; in other words, "including, but not limited to."

[0079] As used herein, the term "consisting of" excludes any element, step, or ingredient not specified in the claim. When the term "consisting of" (or variations thereof) appears in a clause in the body of a claim rather than immediately following the preamble, it limits only the elements set forth in that clause; other elements are not excluded from the claim as a whole. As used herein, the term "consisting essentially of" limits a claim to only the specified elements or method steps, plus those that do not materially affect the basic characteristics of the claimed subject matter.

[0080] With respect to the terms "comprising," "consisting of," and "consisting essentially of," when any of these three terms are used herein, the presently disclosed and claimed subject matter may include the use of either of the other two terms. Thus, unless expressly stated otherwise in certain embodiments, all instances of "comprising" may be replaced with "consisting of" or "consisting essentially of."

[0081] Except in the operating examples or unless otherwise indicated, all numbers expressing quantities of ingredients or reaction conditions should be interpreted in all cases as modified by the term "about," taking into account the normal tolerances in the art. The examples are not intended to limit the scope of the invention. Unless below or otherwise indicated, "%" means "% by weight," "ratio" means "weight ratio," and "parts" means "parts by weight."

[0082] As used herein, the term "substantially" means including more than 50% by weight, where appropriate, unless otherwise specified.

[0083] The recitation of numerical ranges using endpoints includes all numbers subsumed within that range (e.g., 1 to 5 includes 1, 1.5, 2, 2.75, 3, 3.80, 4, 5, etc.).

[0084] The terms "preferred" and "preferably" refer to embodiments of the invention that may offer certain advantages, under certain circumstances. However, other embodiments may also be preferred, under the same or other circumstances. Furthermore, the recitation of one or more preferred embodiments does not imply that other embodiments are not useful, and is not intended to exclude other embodiments from the scope of the invention.

[0085] It should also be noted that as used in this specification and the appended claims, the singular forms "a," "the," and "the" include the plural forms unless the context clearly dictates otherwise.

[0086] Although exemplary embodiments of the disclosed technology have been described in detail herein, it should be understood that other embodiments are contemplated. Accordingly, the disclosed technology is not intended to be limited in scope to the construction and arrangement of components set forth in the following description or illustrated in the drawings. The disclosed technology is capable of other embodiments and of being practiced or carried out in various ways.

[0087] As used throughout the specification, the term "optionally substituted" means that a group may or may not be further substituted or fused (to form a fused polycyclic system) with one or more non-hydrogen substituents. In certain embodiments, the substituents are selected from the group consisting of halogen, ═O, ═S, —CN, —NO, —CF, —OCF, alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, aryl, heteroaryl, cycloalkylalkyl, heterocycloalkylalkyl, heteroarylalkyl, arylalkyl, cycloalkylalkenyl, heterocycloalkylalkenyl, arylalkenyl, heteroarylalkenyl, cycloalkylheteroalkyl, heterocycloalkylheteroalkyl, arylheteroalkyl, heteroarylheteroalkyl, hydroxy, hydroxyalkyl, alkyloxy, alkyloxyalkyl, alkyl, Oxycycloalkyl, alkyloxyheterocycloalkyl, alkyloxyaryl, alkyloxyheteroaryl, alkyloxycarbonyl, alkylaminocarbonyl, alkenyloxy, alkynyloxy, cycloalkyloxy, cycloalkenyloxy, heterocycloalkyloxy, heterocycloalkenyloxy, aryloxy, phenoxy, benzyloxy, heteroaryloxy, arylalkyloxy, amino, alkylamino, acylamino, aminoalkyl, arylamino, sulfonylamino, sulfinylamino, sulfonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl, sulfinyl, alkylsulfinyl, arylsulfinyl, aminosulfinylaminoalkyl, -C(=O)OH, -C(=O)R e , -C(=O)OR e , C(=O)NR e R f , C(=NOH)R e , C(=NR e )NR f R g , N.R. e R f , N.R. e C(=O)R f , N.R.e C(=O)OR f , N.R. e C(=O)NR f R g , N.R. e C(=NR f )NR g R h , N.R. e SO2R f , -SR e , SO2NR e R f , -OR e , OC(=O)NR e R f , OC(=O)R e and acyl;

[0088] where R e , R f , R g and R h are each independently H, C1-C 12 Alkyl, C1-C 12 Haloalkyl, C2-C 12 Alkenyl, C2-C 12 Alkynyl, C1-C 10 Heteroalkyl, C3-C 12 Cycloalkyl, C3-C 12 Cycloalkenyl, C1-C 12 Heterocycloalkyl, C1-C 12 Heterocycloalkenyl, C6-C 18 Aryl, C1-C 18 selected from the group consisting of heteroaryl and acyl, or R a , R b , R c and R d Any two or more of these, together with the atom to which they are attached, form a heterocyclic ring system having 3 to 12 ring atoms.

[0089] In certain embodiments, each optional substituent is independently selected from the group consisting of halogen, =0, =S, -CN, -NO2, -CF3, -OCF3, alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, aryl, heteroaryl, hydroxy, hydroxyalkyl, alkyloxy, alkyloxyalkyl, alkyloxyaryl, alkyloxyheteroaryl, alkenyloxy, alkynyloxy, cycloalkyloxy, cycloalkenyloxy, heterocycloalkyloxy, heterocycloalkenyloxy, aryloxy, heteroaryloxy, arylalkyl, heteroarylalkyl, arylalkyloxy, amino, alkylamino, acylamino, aminoalkyl, arylamino, sulfonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl, aminoalkyl, -COOH, -SH, and acyl.

[0090] Examples of particularly suitable optional substituents include F, Cl, Br, I, CH3, CH2CH3, OH, OCH3, CF3, OCF3, NO2, NH2 and CN.

[0091] As used herein, the term "secondary plant compounds" (SPCs) refers to chemical compounds synthesized by plants that are not essential for plant survival. The SPCs of the present invention have insecticidal activity and / or repel pests and / or deter pest oviposition and / or influence oviposition sites and / or deter pests from feeding on the plant.

[0092] For the purposes of simplicity, the term "insect" or its synonyms or derivatives such as "insecticide" will be used herein; however, it should be understood that the term "insect" includes not only insects but also their immature forms and larvae.

[0093] As used herein, the term "pesticidal activity" refers to the killing of pests, i.e., death of the pests. [Brief explanation of the drawings]

[0094] Certain preferred embodiments of the invention will now be described, by way of example only, with reference to the accompanying drawings, in which:

[0095] [Figure 1] Net ion flux of K+- is shown under control conditions (0–15 min) and after treatment with negative control (A) DMSO 1.0% w / v and positive controls (B) pyrethrum 0.01% w / v and (C) tasmanone 0.01% w / v.

[0096] [Figure 2] Net ion flux of Na+ is shown under control conditions (0–15 min) and after treatment with negative control (A) DMSO 1.0% w / v and positive controls (B) pyrethrum 0.01% w / v and (C) tasmanone 0.01% w / v. [Figure 3] Net ion flux of Cl- is shown under control conditions (0–15 min) and after treatment with negative control (A) DMSO 1.0% w / v and positive controls (B) pyrethrum 0.01% w / v and (C) tasmanone 0.01% w / v.

[0097] [Figure 4] (A) Net ion flux traces under control conditions (0-15 min) and after treatment with a negative K+ control; and (B) the relative response of each treatment to its respective control, comparing 0.01% w / v of the inventive extract with DMSO 1.0% w / v, pyrethrum 0.01% w / v, and tasmanone 0.01% w / v.

[0098] [Figure 5] (A) Net ion flux traces under control conditions (0-15 min) and after treatment with a negative control of Cl-; and (B) the relative response of each treatment to its respective control, comparing 0.01% w / v of the inventive extract with DMSO 1.0% w / v, pyrethrum 0.01% w / v, and tasmanone 0.01% w / v.

[0099] [Figure 6](A) Net ion flux traces under control conditions (0-15 min) and after treatment with a negative control of Na+; and (B) the relative response of each treatment to its respective control, comparing 0.01% w / v inventive extract with DMSO 1.0% w / v, pyrethrum 0.01% w / v, and tasmanone 0.01% w / v.

[0100] [Figure 7]

[0023] A Markush structure corresponding to a selected γ-pyrone of the present invention. In particular, R1, R2, R3, and R4 are each independently selected from optionally substituted C1-C12 alkyl, preferably C1-C6 alkyl, more preferably C1-C2 alkyl; H; -COOH; -OH; -OCH3 or -R5(CH2)nR6R7CH3; R5, R6, and R7 are each independently selected from optionally substituted C1-C12 alkyl, preferably C1-C6 alkyl, more preferably C1-C2 alkyl; -C=O; -COO-, N, S, or O; and n is 1 to 18, or a salt, solvate, dimer, or isomer thereof. DETAILED DESCRIPTION OF THE INVENTION

[0101] Detailed Description of the Invention Those skilled in the art will appreciate that the present invention includes the embodiments and features disclosed herein and all combinations and / or permutations of the disclosed embodiments and features.

[0102] The present inventors have developed insecticides based on γ-pyrone compound derivatives that have insecticidal activity against pests.

[0103] Compounds and Compositions As noted above, the present invention provides a γ-pyrone compound for use as an insecticide. In some embodiments, the γ-pyrone is podopyrone.

[0104] As known to those skilled in the art, pyrones are heterocyclic compounds containing an unsaturated six-membered ring containing one oxygen atom and a ketone functional group. γ-pyrones (also known as 4-pyrones) can be functionalized at the C2, C3, C5, and C6 carbons.

[0105] In some embodiments, the γ-pyrone compound has the general formula (1): [ka]

[0106] [During the ceremony,

[0107] R 1 , R 2 , R 3 and R 4 are each independently optionally substituted C-C 12 Alkyl, preferably C1-C6 alkyl, more preferably C1-C2 alkyl; H; -COOH; -OH; -OCH3 or -R 5 (CH2) n R 6 R 7 Selected from CH3;

[0108] R 5 , R 6 and R 7 are each independently optionally substituted C-C 12 alkyl, preferably C1-C6 alkyl, more preferably C1-C2 alkyl; -C=O; -COO-, N, S or O; and

[0109] n is 1 to 18.

[0110] or a salt, solvate, dimer or isomer thereof.

[0111] In some embodiments, the γ-pyrone compound has the general formula (1): [ka]

[0112] [During the ceremony,

[0113] R 1 , R 2 , R 3 and R 4 are each independently C1-C 12 alkyl, preferably C1-C6 alkyl, more preferably C1-C2 alkyl, wherein the C1-C 12 Alkyl, C1-C6 alkyl and C1-C2 alkyl are optionally epoxide, glycoside, acetoxy, halogen, cyano, amino, alcohol, phenyl, heteroaryl; H; -COOH; -OH; -OCH3 or -R 5 (CH2) n R 6 R 7 optionally substituted with CH3;

[0114] R 5 , R 6 and R 7 are each independently C1-C 12 alkyl, preferably C1-C6 alkyl, more preferably C1-C2 alkyl, wherein the C1-C 12 Alkyl, C1-C6 alkyl and C1-C2 alkyl are optionally C1-C 12 alkyl, preferably C1-C6 alkyl, more preferably C1-C2 alkyl, epoxide, glycoside, acetoxy, halogen, cyano, amino, alcohol, phenyl, heteroaryl; optionally substituted with -C=O; -COO-, N, S, or O; and

[0115] n is 1 to 18.

[0116] or a salt, solvate, dimer or isomer thereof.

[0117] In some embodiments, the γ-pyrone has the general formula (1): [ka]

[0118] [During the ceremony,

[0119] R 1 , R 2 , R 3 and R 4 are each independently C1-C 12 Alkyl, preferably C1-C6 alkyl, more preferably C1-C2 alkyl; H; -COOH; -OH; -OCH3; or -R 5 (CH2) n R 6 R 7 Selected from CH3;

[0120] R 5 , R 6 and R 7 are each independently C1-C 12 alkyl, preferably C1-C6 alkyl, more preferably C1-C2 alkyl, wherein the C1-C 12 Alkyl, C1-C6 alkyl and C1-C2 alkyl are optionally C1-C 12 alkyl, preferably C1-C6 alkyl, more preferably C1-C2 alkyl, epoxide, glycoside, acetoxy, halogen, cyano, amino, alcohol, phenyl, heteroaryl; optionally substituted with -C=O; -COO-, N, S, or O; and

[0121] n is 1 to 18.

[0122] a salt, solvate, dimer or isomer thereof.

[0123] In some embodiments, the γ-pyrone has the general formula (1): [ka]

[0124] [During the ceremony,

[0125] R 1 , R 2 , R 3 and R4 are each independently C1-C 12 Alkyl, preferably C1-C6 alkyl, more preferably C1-C2 alkyl; H; -COOH; -OH; -OCH3; or -R 5 (CH2) n R 6 R 7 Selected from CH3;

[0126] R 5 , R 6 and R 7 are each independently C1-C 12 alkyl, preferably C1-C6 alkyl, more preferably C1-C2 alkyl, wherein the C1-C 12 Alkyl, C1-C6 alkyl and C1-C2 alkyl are optionally C1-C 12 alkyl, preferably C1-C6 alkyl, more preferably C1-C2 alkyl, epoxide, glycoside, acetoxy, halogen, cyano, amino, alcohol, phenyl, heteroaryl; optionally substituted with -C=O; -COO-, N, S, or O; and

[0127] n is 1 to 18,

[0128] However, R 1 , R 2 , R 3 and R 4 At least one of 5 (CH2) n R 6 R 7 It is CH3.

[0129] or a salt, solvate, dimer or isomer thereof.

[0130] In some embodiments, the γ-pyrone has the general formula (2): [ka]

[0131] [During the ceremony,

[0132] R 1 , R 2 , R 3 are each independently C1-C 12 alkyl, preferably C1-C6 alkyl, more preferably C1-C2 alkyl, wherein the C1-C 12 Alkyl, C1-C6 alkyl and C1-C2 alkyl may be optionally substituted with epoxide, glycoside, acetoxy, halogen, cyano, amino, phenyl, heteroaryl; H; -COOH; -OH; or -OCH3;

[0133] R 5 , R 6 and R 7 are each independently C1-C 12 alkyl, preferably C1-C6 alkyl, more preferably C1-C2 alkyl, wherein the C1-C 12 Alkyl, C1-C6 alkyl and C1-C2 alkyl are optionally C1-C 12 alkyl, preferably C1-C6 alkyl, more preferably C1-C2 alkyl, epoxide, glycoside, acetoxy, halogen, cyano, amino, alcohol, phenyl, heteroaryl; optionally substituted with -C=O; -COO-, N, S, or O; and

[0134] n is 1 to 18.

[0135] or a salt, solvate, dimer or isomer thereof.

[0136] In some embodiments, the γ-pyrone compound has the general formula (2): [ka]

[0137] [During the ceremony,

[0138] R 1 , R 2 , R 3 are each independently C1-C 12alkyl, preferably C1-C6 alkyl, more preferably C1-C2 alkyl; H; -COOH; -OH; or -OCH3;

[0139] R 5 , R 6 and R 7 are each independently C1-C 12 alkyl, preferably C1-C6 alkyl, more preferably C1-C2 alkyl, wherein the C1-C 12 Alkyl, C1-C6 alkyl and C1-C2 alkyl are optionally C1-C 12 alkyl, preferably C1-C6 alkyl, more preferably C1-C2 alkyl, epoxide, glycoside, acetoxy, halogen, cyano, amino, alcohol, phenyl, heteroaryl; optionally substituted with -C=O; -COO-, N, S, or O; and

[0140] n is 1 to 18.

[0141] or a salt, solvate, dimer or isomer thereof.

[0142] In some embodiments, the γ-pyrone compound has the general formula (2): [ka]

[0143] [During the ceremony,

[0144] R 1 , R 2 , R 3 are each C1-C2 alkyl; H; -COOH; -OH; or -OCH3;

[0145] R 5 , R 6 and R 7 are each independently C1-C 12 alkyl, preferably C1-C6 alkyl, more preferably C1-C2 alkyl; selected from -C=O, -COO-, N, S or O; and

[0146] n is 1 to 18.

[0147] or a salt, solvate, dimer or isomer thereof.

[0148] In some embodiments, the γ-pyrone has the general formula (3): [ka]

[0149] [During the ceremony,

[0150] n is 6, 7, or 8;

[0151] R 1 is C1-C 12 alkyl, preferably C1-C6 alkyl, more preferably C1-C2 alkyl; and

[0152] R 5 , R 6 and R 7 are each independently selected from —C═O and —CH—;

[0153] However, R 5 , R 6 and R 7 If one of the groups is -C=O, the remaining group is -CH2-. It is of the type.

[0154] In a preferred embodiment, the γ-pyrone is selected from the group consisting of compounds (1a) to (1q) or a salt, solvate, dimer or isomer thereof. [Table 1] [Table 2]

[0155] In a preferred embodiment, the γ-pyrone compound of general formula (1) is 10′-oxopodopyrone (1c).

[0156] In a preferred embodiment, the γ-pyrone compound of general formula (1) is 10′-oxo-8-methylpodopyrone (11).

[0157] In another aspect, the present invention provides pesticidal compositions comprising the γ-pyrone compounds described herein.

[0158] In certain embodiments, the podopyrone is selected from the group consisting of 10'-oxopodopyrone, 10'-oxo-8-methylpodopyrone, 9'-oxopodopyrone, 9'-oxo-8-methylpodopyrone, 1'-oxo-nor-podopyrone, 1'-oxopodopyrone, 1'-deoxo-8-methyl-1'-oxonorpodopyrone, 8-methylpodopyrone, 8-methyl-1'-oxo-nor-podopyrone, 1'-oxo-8-methylpodopyrone, podopyrone, norpodopyrone, homopodopyrone, 10'-hydroxy-8-methylpodopyrone, 10'-acetoxy-8-methylpodopyrone, 10'-acetoxypodopyrone, and combinations thereof.

[0159] In some embodiments, an extract comprising gamma-pyrone from the genera Podorepis, Gonistilus, and combinations thereof is provided.

[0160] In one embodiment, the Podolepis-derived extract is Podolepis labill, Podolepis acuminata, Podolepis affinis Sond., Podolepis aristata, Podolepis arachnoidea, Podolepis auriculata, Podolepis basalt plain, Podolepis canescens, Podolepis capillaris, Podolepis carnarvon, Podolepis centauroides, Podolepis chrysantha, Podolepis contorta, Podolepis cupulata, Podolepis davisiana, Podolepis decipiens, Podolepis divaricata, Podolepis sect.Doratolepis, Podolepis eremaea, Podolepis ferruginea, Podolepis filiformis, Podolepis gardneri, Podolepis georgei, Podolepis gracilis, Podolepis gibertii, Podolepis gnaphalioides, Podolepis gracilis, Podolepis great victoria desert, Podolepis hieracioides, Podolepis inundata, Podolepis jaceoides, Podolepis kendallii, Podolepis laciniata, Podolepis laevigata, Podolepis lessonii, Podolepis linearifolia, Podolepis longipedata, Podolepis lucaeana, Podolepis macrocephala, Podolepis microcephala, Podolepis mitchellii, Podolepis monticola, Podolepis muelleri, Podolepis neglecta, Podolepis nutans, Podolepis omissa, Podolepis pallida, Podolepis papillosa, Podolepis remota, Podolepis rhytidochlamys, Podolepis robusta, Podolepis rosea, Podolepis rosmarinifolia, Podolepis rubida, Podolepis rugata, Podolepis rutidoclamys, Podolepis scalia, Podolepis siemssenia, Podolepis siemssenii, Podolepis simplicicaulis, Podolepis spenceri, Podolepis tepperi, Podolepis sp. aff. robusta, Podolepis NEAlps, Podolepis Warrabah, Podolepis Wollunga Well, Podolepis stylolepis, Podolepis subulata, Podolepis tepperi, Podolepis tetrachaeta, and combinations thereof.

[0161] In certain embodiments, the extract from the genus Gonystylus is Gonystylus acuminatus, Gonystylus affinis, Gonystylus areolatus, Gonystylus augescens, Gonystylus bancanus, Gonystylus borneensis, Gonystylus brunnescens, Gonystylus calophylloides, Gonystylus calophyllus, Gonystylus confusus, Gonystylus consanguineus, Gonystylus costalis, Gonystylus decipiens, Gonystylus eximius, Gonystylus forbesii, Gonystylus glaucescens, Gonystylus keithii, Gonystylus lucidulus, Gonystylus macrocarpus, Gonystylus macrophyllus, Gonystylus maingayi, Gonystylus micranthus, Gonystylus nervosus, Gonystylus nobilis, Gonystylus othmanii, Gonystylus pendulus, Gonystylus punctatus, Gonystylus reticulatus, Gonystylus spectabilis, Gonystylus stenosepalus, Gonystylus velutinus, Gonystylus xylocarpus and combinations thereof.

[0162] In a preferred embodiment, the extract containing γ-pyrone is derived from Podolepis jaceoides, Gonystylus keithii, and combinations thereof.

[0163] Embodiments of the present invention may be used to treat crops to limit or prevent insect infestation. The present invention is particularly suitable for plants of agricultural importance, which refers to plants that are harvested or grown on a commercial scale.

[0164] An example of an agriculturally important crop is cotton. Other examples of such agricultural plants (or crops) are cereals such as wheat, barley, rye, oats, rice, maize or sorghum; beets such as sugar beets or fodder beets; fruits, including pome fruits, stone fruits and soft fruits, such as apples, pears, plums, prunes, peaches, almonds, cherries or berries, e.g. strawberries, raspberries or blackberries; legumes such as beans, lentils, peas or soybeans; oilseed rape, mustard, poppy, olives, sunflowers, coconuts, castor beans, cocoa or peanuts. These include oil crops, Cucurbitaceae such as pumpkin, cucumber or melon, fibrous plants such as cotton, flax, hemp or jute, citrus fruits such as orange, lemon, grapefruit or tangerine, vegetables such as spinach, lettuce, asparagus, cabbage seeds, carrot, onion, chili pepper, tomato, potato or capsicum, Lauraceae such as avocado, Cinnamon or camphor, and tobacco, nuts (e.g. walnuts), coffee, eggplant, cane, tea, mustard, grapes, hops, Musaceae, latex plants and ornamental plants. Also important are fodder crops such as grasses and legumes.

[0165] In some embodiments, the plants are fibrous plants, cereal crops, legume crops, cereals, vegetables and fruits, more particularly cotton, corn, sorghum, sunflower, lucerne, various legumes, especially soybean, pigeon pea, mung bean and chickpea, tomato, okra and similar plants.

[0166] In some embodiments, the plants include ornamental plants. By way of example, these ornamental plants can be orchids, roses, tulips, trees, shrubs, herbs, lawns and grasses, bulbs, vines, perennials, succulents, and foliage plants.

[0167] The present invention involves applying a compound, insecticidal composition, or extract containing γ-pyrone in a carrier (e.g., a non-polar solvent, a polar solvent, oil, water, or any carrier product) to a plant affected by a pest or its surroundings, or to an animal affected by a pest. Treatment can include the use of a non-polar solvent-based formulation, an oil-based formulation, a water-based formulation, a residual formulation, a wettable powder, a dry powder, or the like. In some embodiments, the compound, insecticidal composition, or extract containing γ-pyrone can be applied directly to plant material, such as dried, powdered plant material, as a powder. In some embodiments, a combination of formulations can be used to achieve the benefits of different formulation types. The compound, insecticidal composition, or extract containing γ-pyrone can be added to a carrier, or in the case of a liquid formulation, a carrier such as chloroform, methanol, water, and combinations thereof, may be used to extract SPC.

[0168] In certain embodiments, the formulation is an oil-based formulation that may further comprise a surfactant.

[0169] In some embodiments, the formulation is an oil-based formulation, and the oil is C 19 -C 27 It is a hydrocarbon.

[0170] In some embodiments, the formulation comprises γ-pyrone in an organic solvent such as an alcohol, ketone, aldehyde, or sulfoxide. In some embodiments, the suitable organic solvent can be selected from the group consisting of pentane, cyclopentane, hexane, cyclohexane, benzene, toluene, 1,4-dioxane, chloroform, diethyl ether, dichloromethane, tetrahydrofuran, ethyl acetate, acetone, dimethylformamide, acetonitrile, dimethyl sulfoxide, nitromethane, propylene carbonate, n-butanol, isopropanol, n-propanol, ethanol, methanol, formic acid, acetic acid, hexafluoroisopropanol, trifluoroacetic acid, and combinations thereof. In particular, the formulation comprises γ-pyrone in an alcohol.

[0171] In some embodiments, the formulation comprises a methanolic extract from Podolepis, Gonistylus, and combinations thereof. In some embodiments, the formulation comprises an ethanolic extract from Podolepis, Gonistylus, and combinations thereof.

[0172] In some embodiments, the formulation contains γ-pyrone in a low molecular weight oil, such as crude or refined cottonseed oil or canola oil. Other oils include white oil, DC Tron oil (nC21 and nC24 oil), canopy oil (nC27 oil), Biopest oil (nC24 oil), dormant oil, or summer oil, which are known in the horticultural industry. Most of these oils are nC 19 ~nC 27 However, other hydrocarbons with acceptable toxicological profiles may be used. A number of such products suitable for use in the present invention are commercially available. These include Sunspray® oil, tea tree oil, and Sunspray® Ultra Fine, manufactured by Sun Refining and Marketing Company.

[0173] Petroleum-based spray oils may be used in conjunction with suitable agriculturally acceptable diluents and / or carriers and other additives common in the art, such as emulsifiers, wetting agents, surfactants, stabilizers, spreading agents, and the like.

[0174] In some embodiments, the formulation comprises an aqueous solution comprising γ-pyrone.

[0175] In some embodiments, the formulation comprises immobilizing the γ-pyrone in a low hydrocarbon solvent, such as hexane.

[0176] In some embodiments, the formulation comprises a fraction of the extract that contains crude γ-pyrone.

[0177] In some embodiments, the formulation comprises a fraction of a crude extract from the genera Podolepis, Gonistylus, and combinations thereof.

[0178] In some embodiments, the formulation comprises a mixture of fractions of crude extracts from the genera Podolepis, Gonistylus, and combinations thereof.

[0179] The term "carrier" as used herein means a liquid or solid substance, which may be inorganic or organic, of synthetic or natural origin, with which the active compound is mixed or formulated, which facilitates application or facilitates storage, transport, and / or handling of the compound according to the invention, the pesticidal composition, the γ-pyrone-containing extract, or the SPC, which is applied to the locus to be treated. Generally, any of the substances customarily used in formulating pesticides are suitable.

[0180] As used herein, the term "habitat" refers to the location to which the compositions or SPCs of the present invention are applied, including application to individual plants, groups of plants such as plants and / or their surroundings, animals, either individually or in groups, and areas where plants may be planted or where animals may congregate, as well as application to one or more insects and / or their surroundings.

[0181] The γ-pyrone-containing compounds, pesticidal compositions or extracts of the present invention may be used alone or, if desired, in the form of a solid and / or liquid dispersible carrier medium and / or a mixture with other known compatible active agents such as pesticides or acaricides, nematicides, fungicides, bactericides, rodenticides, herbicides, fertilizers, growth regulators, or in the form of specific dosage formulations for specific applications prepared therefrom, such as solutions, emulsions, suspensions, dusts, pastes and granules, and thus in a ready-to-use form.

[0182] The γ-pyrone-containing compounds, pesticidal compositions or extracts of the present invention may, if desired, be formulated or mixed with conventional inert pesticide diluents or extenders of the type useful in conventional pest control agents, e.g., conventional dispersible carrier media in the form of solutions, emulsions, suspensions, emulsifiable concentrates, spray powders, pastes, soluble powders, dusts, granules or foams.

[0183] Typical emulsifiers suitable for use in the compounds, pesticidal compositions, or extracts of the present invention include, but are not limited to, low molecular weight oils (e.g., canola oil, cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil) and non-anionic, anionic, and cationic surfactants. Mixtures of any of the above emulsifiers may also be used in the compounds, pesticidal compositions, or extracts of the present invention.

[0184] Typical nonionic surfactants are ethoxylated alkanols, in particular ethoxylated fatty alcohols and ethoxylated oxo alcohols, such as ethoxylated lauryl alcohol, ethoxylated isotridecanol, ethoxylated cetyl alcohol, ethoxylated stearyl alcohol and esters thereof, such as acetate esters; ethoxylated alkylphenols, such as ethoxylated nonylphenyl, ethoxylated dodecylphenyl, ethoxylated isotridecylphenol and esters thereof, such as alkylglucoside acetate esters and alkylpolyglucosides, ethoxylated alkylglucosides; ethoxylated fatty amines, ethoxylated fatty acids, partial esters, such as mono-, di- and triesters of fatty acids with glycerin or sorbitan, such as glycerin monostearate, glycerin monooleate. These include glycerin, sorbitan monolaurate, sorbitan monopalmitate, sorbitan monostearate, sorbitan monooleate, sorbitan tristearate, sorbitan trioleate; ethoxylated esters of fatty acids with glycerin or sorbitan, such as polyoxyethylene glycerin monostearate, polyoxyethylene sorbitan monolaurate, sorbitan monopalmitate, polyoxyethylene sorbitan monostearate, polyoxyethylene sorbitan monooleate, polyoxyethylene sorbitan tristearate, polyoxyethylene sorbitan trioleate; ethoxylated vegetable oils or animal fats, such as ethoxylated corn oil, ethoxylated castor oil, ethoxylated tallow oil; ethoxylated fatty amines, fatty amides or fatty acid diethanolamides.

[0185] Typical anionic surfactants are alkyl sulfonates, such as lauryl sulfonate, isotridecyl sulfonate, alkyl sulfates, in particular fatty alcohol sulfates, such as lauryl sulfate, isotridecyl sulfate, cetyl sulfate, stearyl sulfate, aryl- and alkylaryl sulfonates, such as naphthyl sulfonate, dibutyl naphthyl sulfonate, alkyl diphenyl ether sulfonates, such as dodecyl diphenyl ether sulfonate, alkyl benzene sulfonates, such as cumyl sulfonate, nonyl benzene sulfonate and dodecyl benzene sulfonate salts, in particular sodium, potassium, calcium or ammonium salts; sulfonates of fatty acids and fatty acid esters; sulfates of fatty acids and fatty acid esters; sulfates of ethoxylated alkanols, for example, the sulfate of ethoxylated lauryl alcohol; sulfates of alkoxylated alkylphenols; alkyl phosphates and dialkyl phosphates; dialkyl esters of sulfosuccinates, for example, dioctyl sulfosuccinate, acyl alcosinates, including low molecular weight condensates of fatty acids, for example, stearates, acyl glutamates, lignin sulfonates, naphthalene sulfonic acid or phenol sulfonic acid with formaldehyde and, optionally, urea;

[0186] Typical cationic surfactants include quaternary ammonium compounds, particularly alkyltrimethylammonium salts and dialkyldimethylammonium salts, such as halides, sulfates and alkyl sulfates.

[0187] In some embodiments, the compound, insecticidal composition, or extract may be combined with one or more insecticides or pesticides. In some embodiments, the compound, insecticidal composition, or extract may be combined with one or more synthetic insecticides or pesticides. In some embodiments, the insecticide or pesticide is selected from one or more of endosulfan, dicofol, chlorpyrifos, dimethoate, disulfoton, omethoate, parathion, phorate, profenofos, sulprofos, thiometon, aldicarb, carbaryl, beta-cyfluthrin, deltamethrin, esfenvalerate, fenvalerate, fluvalinate, lambda-cyhalothrin, chlorfluazuron, piperonyl butoxide, and petroleum-based spray oils. In other embodiments, the pesticide is a biological pesticide selected from nuclear polyhedrosis viruses and / or plant extracts known to be antifeedants of pests. In still other embodiments, the insecticide or pesticide is used at a reduced labeling rate. For example, insecticides or repellents may be used at half or one-third the labeled rate.

[0188] The compounds, pesticidal compositions or extracts of the present invention can be used to control pests by treating the host directly or by treating the area in which the host is located. For example, the host can be treated directly using a spray formulation that can be applied to plants, such as agricultural plants, individually or when crowded.

[0189] The formulations of the present invention may further contain other formulation auxiliaries known in the field of agricultural chemical formulations in conventional amounts. Such auxiliaries include, but are not limited to, antifreeze agents (such as, but not limited to, glycerin, ethylene glycol, propylene glycol, monopropylene glycol, hexylene glycol, 1-methoxy-2-propanol, and cyclohexanol), buffers (such as, but not limited to, sodium hydroxide and phosphoric acid), preservatives (such as, but not limited to, derivatives of 1,2-benzisothiazolin-3-one, benzoic acid, sorbic acid, formaldehyde, and combinations of methyl and propyl parahydroxybenzoates), stabilizers (such as, but not limited to, acids, preferably organic acids, such as, but not limited to, dodecylbenzenesulfonic acid, acetic acid, propionic acid, or butylhydroxyltoluene and butylhydroxylanisole), thickeners (such as, but not limited to, heteropolysaccharides and starches), and antifoaming agents (such as, but not limited to, silicones, particularly those based on polydimethylsiloxane). Such adjuvants are commercially available and known in the art.

[0190] In one embodiment, the present invention uses fractions, SPC and crude extracts from Podolepis, Gonistylus and combinations thereof formulated for cotton pest control.

[0191] Typically, extracts can be obtained by adding a solvent to a plant sample to obtain a mixture. The mixture is then expanded, such as by sonication and / or maceration. The mixture is then filtered and transferred to a separatory funnel for solvent extraction. The two layers formed during solvent extraction are then separated, and the solvent is removed, for example, under reduced pressure, to obtain at least two fractions of extract.

[0192] method As described above, the present invention provides a method for controlling one or more pests comprising treating a locus with an insecticidally effective amount of a compound, insecticidal composition comprising one or more γ-pyrone compounds of the present invention, or an insecticidal composition comprising one or more extracts from the genera Podorepis, Gonistilus, and combinations thereof, comprising one or more secondary plant compounds (SPCs) of the present invention, thereby exerting insecticidal activity and / or repelling the pests and / or inhibiting pest oviposition and / or affecting oviposition sites and / or inhibiting pest feeding on the plant.

[0193] Those skilled in the art will recognize that an appropriate amount of compound, pesticidal composition, or extract can be applied for pest control. In some embodiments, the method comprises treating a locus with an extract comprising γ-pyrone at a concentration of less than about 20 w / v%, less than about 15 w / v%, less than about 10 w / v%, less than about 5 w / v%, less than about 3 w / v%, less than about 2 w / v%, less than about 1 w / v%, less than about 0.5 w / v%, and less than about 0.25 w / v%.

[0194] In certain embodiments, the method comprises treating the locus with a compound, insecticidal composition, or extract comprising γ-pyrone at a concentration of about 0.25 to about 20 w / v%, about 0.25 to about 15 w / v%, about 0.25 to about 10 w / v%, about 0.25 to about 5 w / v%, about 0.25 to about 3 w / v%, preferably about 0.25 to about 2 w / v%, and more preferably about 0.25 to about 1 w / v%.

[0195] Those skilled in the art will recognize that any suitable amount of SPC can be applied for pest control. In certain embodiments, the method comprises treating the locus with an SPC (i.e., active ingredient) at a concentration of less than about 20 w / v%, less than about 15 w / v%, less than about 10 w / v%, less than about 5 w / v%, less than about 3 w / v%, less than about 2 w / v%, less than about 1 w / v%, less than about 0.5 w / v%, and less than about 0.25 w / v%.

[0196] In certain embodiments, the method comprises treating the habitat with SPC (i.e., active ingredient) at a concentration of about 0.001 to about 20 w / v%, about 0.001 to about 15 w / v%, about 0.001 to about 10 w / v%, about 0.001 to about 5 w / v%, about 0.001 to 3 w / v%, about 0.001 to 2 w / v%, about 0.001 to 1 w / v%, about 0.005 to 0.5 w / v%, and about 0.001 to 0.1 w / v%.

[0197] In certain embodiments, the method comprises treating the habitat with a concentration of SPC (i.e., active ingredient) from 1 to about 30,000 ppm, from about 30 to about 30,000 ppm, from about 50 to about 30,000 ppm, from about 100 to about 30,000 ppm, from about 150 to about 30,000 ppm, from about 200 to about 30,000 ppm, from about 300 to about 30,000 ppm, from about 300 to about 25,000 ppm, from about 300 to about 16,000 ppm, from about 300 to about 15,000 ppm, from about 300 to about 12,000 ppm, or from about 3,000 to about 12,000 ppm.

[0198] Preferably, the compounds, insecticidal compositions, extracts, and formulations of the present invention are suitable for killing insects (i.e., insecticidal activity). Without being bound by theory, the inventors believe that γ-pyrone, when applied to a plant or insect, penetrates the insect's cuticle or is ingested, killing the insect. Alternatively, the extract remaining on the plant may repel insects or deter insects from laying eggs or feeding. The formulations may kill insects or deter insects from laying eggs or feeding within 3-4 days of application to the insect or target crop.

[0199] The inventors have surprisingly discovered that the γ-pyrones of the present invention can affect at least two ion channels in pest insects, in contrast to known insecticides that typically only affect a single ion channel as part of their mechanism of action, e.g., dichlorodiphenyltrichloroethane (DDT) and pyrethroids, which typically target sodium channels in pest insects.

[0200] In some embodiments, γ-pyrone affects at least two neural ion channels in pest insects, preferably selected from the group consisting of sodium, potassium, and chloride ion channels. In some embodiments, γ-pyrone causes sodium and potassium ion efflux. In some embodiments, γ-pyrone causes chloride ion influx.

[0201] Without being bound by any one theory, the inventors believe that the insecticidal activity of the γ-pyrones of the present invention is provided by having a γ-pyrone moiety "core" in combination with at least one optionally substituted aliphatic chain on any one of the C2, C3, C5 or C6 carbons of the γ-pyrone "core."

[0202] Typically, the compound, insecticidal composition, extract, fraction, crude, or SPC (i.e., active ingredient) of the present invention is incorporated into a carrier (e.g., an organic solvent or an organic solvent / water mixture) and applied to crops infested by the target insect. Those skilled in the art will recognize that an appropriate application rate of γ-pyrone can be applied for pest control. The application rate of the compound, insecticidal composition, or extract of the present invention is typically about 1 to 500 L of compound, insecticidal composition, extract, or SPC in carrier per hectare, preferably about 60 to 120 L of compound, insecticidal composition, extract, or SPC in carrier per hectare, more preferably about 100 L of compound, insecticidal composition, extract, or SPC in carrier per hectare. Treatment may optionally include repeated spraying. In some embodiments, treatment involves at least two, three, or four sprays at intervals of 3 to 28 days, 3 to 14 days, and preferably 7 days.

[0203] The present inventors have surprisingly and advantageously discovered that the gamma-pyrones of the present invention have minimal impact on beneficial insect species such as pollinators, including the Western honeybee (Apis mellifera).

[0204] Indiscriminate use of and over-reliance on broad-spectrum pesticides leads to pesticide resistance and the re-emergence of previously controlled pests, contributing to environmental contamination, food residues, and unacceptable risks to human health and biodiversity. Broad-spectrum pesticides can also kill beneficial predators and pollinators, which is undesirable.

[0205] In Australia, bollworm insecticide resistance is tightly controlled through the adoption of geographic region-specific Insecticide Resistance Management Strategies, which are promoted and continually updated by the Cotton Research & Development Corporation (CRDC).

[0206] There are currently 185 registered products across 17 chemical insecticide classes. To reduce the development of resistance among pests such as the cotton bollworm, most of these chemical classes can only be applied once or twice per growing season. Resistance to indoxacarb, avermectin, Rynaxypyr® (ry-nax-ipier), and organophosphates is relatively low; however, in recent years, resistance to bifenthrin has increased to 40% and to common pyrethroids to 90%.

[0207] The bollworm is resistant to at least 47 active ingredients individually, and in some cases this leads to cross-resistance to chemicals that act at similar sites of action, reducing the number of effective products available to growers in turn. Additionally, secondary cotton pests can develop multiple resistance to insecticides.

[0208] The present inventors have surprisingly discovered that, in certain embodiments, the γ-pyrone compounds, insecticidal compositions, extracts, and formulations of the present invention exhibit selectivity toward pests. In particular, the γ-pyrone compounds of the present invention exhibit insecticidal activity against aphids and twospotted spider mites, with minimal effect on Helicoverpa nitidae. Furthermore, as described above, the γ-pyrone compounds have minimal effect on beneficial insect species, such as pollinators, including the Western honeybee (Apis mellifera).

[0209] This allows the compounds, pesticidal compositions and extracts of the present invention to be used while avoiding the possibility of pesticide resistance by certain pests, such as the Helicoverpa gallinae, which, as noted above, are known to be a problem in the agricultural industry.

[0210] The compounds, pesticidal compositions and extracts of the present invention can be used to augment a complete pest control and resistance management program while offering growers an environmentally responsible option.

[0211] In some embodiments, the plant structures used to obtain the extract are any combination of leaves, stems, roots, pods, seeds, and plant parts. In some embodiments, the plant structures used to obtain the extract are in the pre-flowering stage. In some embodiments, the plant structures used to obtain the extract are in the late-flowering stage. In some embodiments, the plant structures used to obtain the extract are in the mid-flowering stage. These can be used as fractions or crude extracts in formulations such as methanol, water, or some other carrier to control chewing or sap-sucking pests through repellency, oviposition deterrence, anti-feeding, and direct contact activity.

[0212] Botanically, the genus Podolepis is a group of annual and perennial herbs with a wool-like appearance, often with finely septate hairs and often with finely glandular or almost hairless hairs. The plants typically have basal and cauline leaves. The inflorescences are borne in one to several terminal heads, which are solitary or contracted to form cymes. The terminal heads are bell-shaped to hemispherical, and the scale leaves at the apex of the pedicel are usually integrated into the involucre. The marginal florets are typically yellow. The genus Podolepis is distributed worldwide, with 18 species known to date, endemic to Australia.

[0213] Botanically, the genus Gonistilus is a Southeast Asian species of about 30 species of hardwood trees, also known as Ramin, Melawis (Malaysia), and Ramin Terur (Sarawak). Ramins are native to Malaysia, Singapore, Indonesia, Brunei, the Philippines, and Papua New Guinea and represent the largest species diversity in Borneo. They are related to Arnhemia, Deltaria, Lethedon, and Solmsia. Ramins are medium-sized trees, typically reaching a height of about 24 m with a straight, clean (unbranched), buttress-free trunk about 18 m long and 60 cm in diameter. The trees are slow-growing and occur primarily in swamp forests.

[0214] In some embodiments, the insecticidal compositions of the present invention are active against pests, which may include economically important pests of agriculture, forestry, greenhouses, nurseries, ornamentals, food and fiber, public and animal health, home and commercial construction, households, and stored products. The pests are insects selected from the orders Coleoptera, Diptera, Hymenoptera, Lepidoptera, Trichophora, Homoptera, Acarina, Hemiptera, Orthoptera, Thysanoptera, Dermoptera, Isoptera, Phthiraptera, Siphonaptera, Trichoptera, Coleoptera, etc., especially Hemiptera and Trichoptera.

[0215] Lepidoptera larvae include armyworms, cutworms, loopers, and heliothins of the Noctuidae family, such as Spodoptera frugiperda J.E. Smith (spodoptera fall armyworm); S. exigua Huebner (beet armyworm); S. litura Fabricius (tobacco cutworm, Spodoptera litura Fabricius); Mamestra configurata Walker (bertha armyworm); M. brassicae Linnaeus (diamond back moth); Agrotis ipsilon Hufnagel (cutworm moth); A. orthogonia Morrison (western cutworm); A. subterranea Fabricius (granulated cutworm); Alabama argillacea Huebner (cotton leafworm); Trichoplusia ni Huebner (nettle looper); Pseudoplusia includens Walker (soybean looper); Anticarsia gemmatalis Huebner (velvet bean caterpillar); Hypena scabra Fabricius (green cloverworm); Heliothis virescens Fabricius (false cotton moth); Pseudaletia unipuncta Haworth (armyworm); Athetis mindara Barnes and Mcdunnough (roughskinned cutworm); Euxoa messona Harris (dark-sided cutworm); Earias insulana Boisduval (spinil ballworm); E. vittella Fabricius (spotted bowlworm); Helicoverpa armigera Huebner (cotton bollworm); H.zea Boddie (corn earworm or cotton ball worm); Melanchra picta Harris (zebra caterpillar); Egira (xylomyges), curialis Grote (citrus cutworm); borers, cocooning insects, webworms, cornworms, and leaf-eating larvae of the Pyralidae family Ostrinia nubilalis Huebner (European corn borer); Amyelois transitella Walker (navel orange worm); Anagasta kuehniella Zeller (striped grain moth); Cadra cautella Walker (gray moth); Chilo suppressalis Walker (rice stem borer); C. partellus (sorghum borer); Corcyra cephalonica Stainton (corn weed moth); Crambus caliginosellus Clemens (corn root webworm); C. teterrellus Zincken (grass moth); Cnaphalocrocis medinalis Guenee (rice leaf borer); Desmia funeralis Huebner (grape leaf folder); Diaphania hyalinata Linnaeus (melon worm); D. nitidalis Stoll (pickleworm); Diatraea grandiosella Dyar (southwestern corn borer), D.saccharalis Fabricius (sugarcane pest); Eoreuma loftini Dyar (rice water weevil); Ephestia elutella Huebner (tobacco (cocoa) moth); Galleria mellonella Linnaeus (large wax moth); Herpetogramma licarsisalis Walker (sod webworm); Homoeosoma electellum Hulst (sunflower moth); Elasmopalpus lignosellus Zeller (corn moth); Achroia gnsella Fabricius (small wax moth); Loxostege sticticalis Linnaeus (white-striped moth); Orthaga thyrisalis Walker (tea tree web moth); Maruca testulalis Geyer (bean moth); Plodia interpunctella Huebner (Indian meal moth); Scirpophaga incertulas Walker (sunflower moth); Udea rubigalis Guenee (celery leaf tearer); and leaf rollers, caterpillars, seed worms, and fruit worms of the Tortricidae family, such as Acleris gloverana Walsingham (western black-headed budworm); A. variana Fernald (eastern black-headed budworm); Archips argyrospila Walker (fruit tree tortrix moth); A. rosana Linnaeus (European leaf roller); and other Archips species, Adoxophyes orana Fischer von Rosslerstamm (summer fruit tortrix moth); Cochylis hospes Walsingham (banded sunflower moth); Cydia latiferreana Walsingham (fibertworm); C. pomonella Linnaeus (codling moth); Platynota flavedana Clemens (variagated leaf roller); P.stultana Walsingham (omnivorous leafroller); Lobesia botrana Denis & Schiffermiiller (European grapevine moss); Spironota ocellana Denis & Schiffermiiller (ice-spotted bud moss); Endopiza viteana Clemens (grapeberry moss); Eupoecilia ambiguella Huebner (vineberry moss); Bonagota salubncola Meyrick (Brazilian apple leaf roller); Grapholita molesta Busck (pear fruit moth); Suleima helianthana Riley (sunflower bud moss); Argyrotaenia spp.; Choristoneura spp.

[0216] Other selected agricultural pests of the order Lepidoptera include Alsophila pometaria Harris (fallen oakworm); Anarsia lineatella Zeller (peach leaf moth); Anisota senatoria J.E. Smith (orange-striped oakworm); Antheraea pernyi Guerin-Meneville (Chinese oak tussock moth); Bombyx mori Linnaeus (silkworm); Bucculatnx thurbenella Busck (cotton leaf perforator); Colias eurytheme Boisduval (alfalfa caterpillar); Datana integerrima Grote & Robinson (walnut caterpillar); Dendrolimus sibiricus Tschetwerikov (Siberian silk moth), Ennomos subsignaria Huebner (elm spanworm); Erannis tiliaria Harris (linden looper); Euproctis chrysorrhoea Linnaeus (white-winged tussock moth); Harrisina americana Guerin-Meneville (grape leaf skeletonizer); Hemileuca oliviae Cockrell (range caterpillar); Hyphantria cunea Drury (fall webworm); Keiferia lycopersicella Walsingham (tomato pinworm); Lambdina fiscellaria fiscellaria Hulst (eastern hemlock looper); L. fiscellaria lugubrosa Hulst (western hemlock looper); Leucoma salicis Linnaeus (yellow-legged tussock moth); Lymantria dispar Linnaeus (gypsy moth); Manduca quinquemaculata Haworth (five-spotted hawkmoth, tomato hornworm); M.sexta Haworth (Tomato Hornworm, Tobacco Hawkmoth); Operophtera brumata Linnaeus (Winter Scale); Paleacrita vernata Peck (Spring Cankerworm); Papilio cresphontes Cramer (Giant Swallowtail, Orange Dog); Phryganidia californica Packard (California Oakworm); Phyllocnistis citrella Stainton (Citrus Leaf Miner); Phyllonorycter blancardella Fabricius (Spotted Tenchfoam Leaf Miner); Pieris brassicae Linnaeus (Large White); P. rapae Linnaeus (Cabbage White); P. napi Linnaeus (Siberian White); Platyptilia carduidactyla Riley (Artichoke Plum Moth); Plutella xylostella Linnaeus (Diamondback Moth); Pectinophora gossypiella Saunders (pink bollworm); Pontia protodice Boisduval & Leconte (southern cabbageworm); Sabulodes aegrotata Guenee (omnivorous looper); Schizura concinna J.E. Smith (red-humped caterpillar); Sitotroga cerealella Olivier (gray moth); Thaumetopoea pityocampa Schiffermuller (pine processionary caterpillar); Tineola bisselliella Hummel (webbing crow moth); Tuta absoluta Meyrick (tomato tooth moth); Yponomeuta padella Linnaeus (ermine moth); Heliothis subflexa Guenee; Malacosoma spp. and Orgyia spp.Including, but not limited to: Of interest are weevils from the families Pholiota, Bruchidae and Curculionidae (including, but not limited to, Anthonomus grandis Boheman (boll weevil); Lissorhoptrus oryzophilus Kuschel (rice water weevil); Sitophilus granarius Linnaeus (granary maize weevil); S. oryzae Linnaeus (rice borer); Hypera punctata Fabricius (cloverleaf waveville); Cylindrocopturus adspersus LeConte (sunflower stem waveville); Smicronyx fulvus LeConte (red sunflower seed waveville); S. sordidus LeConte (gray sunflower seed waveville); Sphenophorus maidis Chittenden (maysbill bug)); flea beetles, cucumber beetles, rootworms, leaf beetles, potato beetles and leaf miners from the family Chrysomelidae (Leptinotarsa decemlineata Say (Colorado potato beetle); Diabrotica virgifera virgifera LeConte (western corn rootworm); D. barberi Smith & Lawrence (northern corn rootworm); D.undecimpunctata howardi Barber (southern corn rootworm); Chaetocnema pulicaria Melsheimer (corn flare beetle); Phyllotreta cruciferae Goeze (corn flare beetle); Colaspis brunnea Fabricius (grape colaspis); Oulema melanopus Linnaeus (cereal leaf beetle); Zygogramma exclamationis Fabricius (sunflower beetle); Coccinellidae beetles (including but not limited to Epilachna vanvestis Mulsant (green ladybird)); scarab beetles and other scarab beetles (Popillia japonica Newman (bean beetle); Cyclocephala borealis Arrow (scarab beetle, scarab beetle larvae); C. immaculata Olivier (scarab beetle, scarab beetle larvae); Rhizotrogus majalis Razoumowsky (European Schaefer); Phyllophaga crinita Burmeister (scarab beetle larvae); Ligyrus gibbosus De Geer (black beetle); Dermestidae (dermestidae); Eleodes spp., Melanotus spp., Conoderus spp., Limonius spp., Agriotes spp., Ctenicera spp., Aeolus spp.; and larvae and adults of Coleoptera, including bark beetles and Tenebrionid beetles.

[0217] Adult and immature members of the Diptera order are of interest, including the leaf-mining insect Agromyza parvicornis Loew (cornblotch leaf miner); small insects (including, but not limited to, Contarinia sorghicola Coquillett (sorghum gall midge); Mayetiola destructor Say (wheat gall midge); Sitodiplosis mosellana Gehin (wheat gall midge); Neolasioptera murtfeldtiana Felt (sunflower seed midge)); fruit flies (Tephritidae), Oscinella frit Linnaeus (fruit fly), Bactrocera tryoni (Queensland fruit fly); maggots (Delia platura Meigen (seed fly); D. coarctata Fallen (wheat borer); and other Delia spp., Meromyza americana Fitch (wheat root miner); Musca domestica Linnaeus (house fly); Fannia canicularis Linnaeus, F. femoralis Stein (small house flies); Stomoxys calcitrans Linnaeus (stable flies); face flies, horn flies, blow flies, Chrysomya spp.; Phormia spp.; and other blowfly pests, horn flies Tabanus spp.; bot flies Gastrophilus spp.; Oestrus spp.; cow flies Hypoderma spp.; deer flies Chrysops spp.; Melophagus ovinus Linnaeus (sheep lice); and other Brachycera, mosquitoes Aedes spp.; Anopheles spp.; Culex spp.; black flies Prosimulium spp.; Simulium spp.; midges, sand flies, sea flies and other Nematocera.Insects of interest include: aphids of the Aphididae family, Hemiptera of the Miridae family, cicadas of the Cicadidae family, leafhoppers of the Cicadidae family, Empoasca spp.; planthoppers of the Delphacidae, Aethiopicidae, Aethiopoidea, Delphacidae and Delphacidae families, treehoppers of the Ceroplastidae family, psyllid lice of the Psyllidae family, Aleyrodes Whiteflies of the Aleyrodidae family including proletella (barley whitefly, kale whitefly, brassica whitefly), aphids of the Aphididae family, grape aphids of the Phyllophididae family, mealybugs of the Pseudococcidae family, Asterolecanidae, scale insects of the Coccidae family, cochineal scales, scale insects of the Pseudococcidae family, Pseudogidae, and Pseudococcidae families, and scale beetles of the Tingidae family, such as the olive lace bug (Froggattia oliviana), brown marmorated stink bugs, cinch bugs, and Blissus spp.; and adults and nymphs of Hemiptera and Homoptera genera such as, but not limited to, other seed bugs of the Lygaeidae family, foxtail bugs of the Scypholidae family, squash bugs of the Coreidae family, and chiggers and cotton strainers of the Pyrrhocoridae family.

[0218] Further agriculturally important members of the Homoptera genera include Acyrthisiphon pisum Harris (pea aphid); Aphis craccivora Koch (cowpea aphid); Aphis fabae Scopoli (bean black aphid); Aphis gossypii (cotton aphid, melon aphid); A. maidiradicis Forbes (corn root aphid); A. pomi De Geer (apple aphid); A. spiraecola Patch (spirea aphid); Aulacorthum solani Kaltenbach (potato aphid); Chaetosiphon fragaefolii Cockerell (strawberry aphid); Diuraphis noxia Kurdjumov / Mordvilko (Russian wheat aphid); Dysaphis plantaginea Paaserini (rosy apple aphid); Enosoma lanigerum Hausmann (apple aphid); and Brevicoryne brassicae Linnaeus (radish aphid); Hyalopterus pruni Geoffroy (peach syrup aphid); Lipaphis erysimi Kaltenbach (false radish aphid); Metopolophium dirrhodum Walker (cereal aphid); Macrosiphum euphorbiae Thomas (potato long-horn aphid); Myzus persicae Sulzer (peach aphid); Nasonovia ribisnigri Mosley (lettuce aphid); Pemphigus spp. (root and gall aphids); Rhopalosiphum maidis Fitch (corn leaf aphid); R.padi Linnaeus (bird cherry auto aphid); Schizaphis graminum Rondani (greengrass aphid); Sipha flava Forbes (yellow sugarcane aphid); Sitobion avenae Fabricius (English grain aphid); Therioaphis maculata Buckton (alfalfa aphid); Toxoptera aurantii Boyer de Fonscolombe (phylloxera aphid); and T. citricida Kirkaldy (brown citrus aphid); Adelges spp. (parasol aphids); Phylloxera devastatrix Pergande (pecan phylloxera); Bemisia tabaci Gennadius (tobacco whitefly, sweet potato whitefly); B. argentifolii Bellows & Perring (silverleaf whitefly); Dialeurodes citri Ashmead (citrus whitefly); Trialeurodes abutiloneus (banded-winged whitefly) and T. vaporariorum Westwood (greenhouse whitefly); Empoasca fabae Harris (potato leafhopper); Laodelphax striatellus Fallen (small brown planthopper); Macrolestes quadrilineatus Forbes (aster leafhopper); Nephotettix cinticeps Uhler (green leafhopper); N. nigropictus Stal (rice leafhopper); Nilaparvata lugens Stal (brown planthopper); Peregrinus maidis Ashmead (corn planthopper); Sogatella furcifera Horvath (white-backed planthopper); Sogatodes orizicola Muir (rice delphacid); Typhlocyba pomaria McAtee (white apple leafhopper); Erythroneoura spp.(Grape Leafhopper); Magicicada septendecim Linnaeus (North American Cicada); lcerya purchasi Maskell (Bollworm Scale); Quadraspidiotus perniciosus Comstock (San Jose Scale); Planococcus citri Risso (Citrus Mealybug); Pseudococcus spp. (Other Mealybug Hybrids); Cacopsylla pyricola Foerster (Pia Psylla); Trioza diospyn Ash Mead (Persimmon Psylla).

[0219] Interesting and agriculturally important species of Hemiptera include Acrosternum hilare Say (green lace bug); Anasa tristis De Geer (squash bug); Blissus leucopterus leucopterus Say (Japanese long-legged bug); Corythuca gossypii Fabricius (cotton lace bug); Cyrtopeltis modesta Distant (tomato bug); Dysdercus suturellus Herrich-Schaffer (cotton strainer); Euschistus servus Say (brown stink bug); E. vanolanus Palisot de Beauvois (one-spotted stink bug); Graptostethus spp. (seed bug hybrids); Leptoglossus corculus Say (leaf-footed pine seed bug); Lygus lineolaris Palisot de Beauvois (rusty white stink bug); and L. hesperus Knight (Western Tarnished Plant Bug); L. pratensis Linnaeus (Common Meadow Bug); L. rugulipennis Poppius (European Tarnished Plant Bug); Lygocoris pabulinus Linnaeus (Common Green Capsid); Nezara viridula Linnaeus (Southern Green Stink Bug); Oebalus pugnax Fabricius (Rice Stink Bug); Oncopeltus fasciatus Dallas (Large Milkweed Bug); Pseudatomoscelis seriatus Reuter (Cotton Flarehopper).

[0220] Further embodiments of the present invention include Calocoris norvegicus Gmelin (strawberry bug); Orthops campestris Linnaeus; Plesiocons rugicollis Fallen (apple capsid); Cyrtopeltis modestus Distant (tomato bug); Cyrtopeltis notatus Distant (sac fly); Spanagonicus albofasciatus Reuter (white-marked flare hopper); Diaphnocoris chlorionis Say (honey locust plant bug); Labopidicola allii Knight (onion plant bug); Pseudatomoscelis seriatus Reuter (cotton flare hopper); Adelphocoris rapidus Say (rapid plant bug); Poecilocapsus lineatus Fabricius (four-lined plant bug); Nysius ericae Schilling (fair's chinch bug); Nysius raphanus Howard (fair's chinch bug); Nezara viridula Linnaeus (southern green bugs); Eurygaster spp.; Coreidae spp.; Pyrrhocoridae spp.; Tinidae spp.; Blostomatidae spp.; Reduviidae spp.; and Cimicidae spp.

[0221] Also included are adult and larval members of the order Acari (mites), such as Aceria tosichella Keifer (wheat curl mite); Petrobia latens Mueller (ground spider mite); spider mites and red mites of the family Tetranychidae, Panonychus ulmi Koch (apple red mite); Tetranychus urticae Koch (two-spotted spider mite); (T. mcdanieli McGregor (McDaniel mite); T. cinnabarinus Boisduval (carmine spider mite); T. turkestani Ugarov & Nikolski (strawberry spider mite); small spider mites of the family Tenebrionidae, Brevipalpus lewisi McGregor (green house spider mite); rust and bud mites and other leaf-feeding mites of the family Eriophyidae, and mites of importance to human and animal health, i.e. house mites of the family Dermatophagoides, Demodex mites of the family Demodex mites, grain mites of the family Myristicidae, mites of the family Ixodes mites Other interesting pests include the deer tick, Dermacentor scapularis Say (deer tick), / Holocyclus Neumann (Australian paralytic tick), Dermacentor variabilis Say (American dog tick), Amblyomma americanum Linnaeus (Lone star tick), and the scabies and mange mites of the genera Pseudococcus, lice mites, and Sarcoptes mites. Other pests of interest include Lepisma saccharina Linnaeus (silver silverfish), and Thermobia domestica Packard (spotted silverfish).

[0222] Interesting and agriculturally important species of the order Thysanoptera (Thysanoptera) include, but are not limited to, Frankliniella occidentalis (Occitan flower thrips); F. occidentalis, Thrips simplex, Thrips palmi, Frankliniella tritici, and Heliothrips haemorrhoidalis (Greenhouse thrips).

[0223] In some embodiments, the pest is selected from cotton bollworms, native budworms, mirid bugs, aphids, southern green stink bugs, apple dimple bugs, thrips (plaque thrips, tobacco thrips, onion thrips, western flower thrips), whiteflies, and two-spotted spider mites.

[0224] In some embodiments, animal pests include fleas, lice, mosquitoes, flies, tsetse flies, ants, ticks, mites, silverfish, and sand fleas.

[0225] The insecticidal activity of the present invention can be tested against pests at any stage of their life cycle, for example, as early developmental stages, such as larvae or other immature forms. Insects can be reared in complete darkness or natural light at about 20°C to about 30°C and about 30% to about 70% relative humidity. Methods for rearing insect larvae and conducting bioassays are well known to those skilled in the art.

[0226] In some embodiments, the insecticidal effect is that the treatment kills at least about 10% of the pests exposed to the treatment. In some embodiments, the insecticidal effect is that the treatment kills at least about 25% of the pests. In some embodiments, the insecticidal effect is that the treatment kills at least about 50% of the pests exposed to the treatment. In some embodiments, the insecticidal effect is that the treatment kills at least about 75% of the pests exposed to the treatment. In some embodiments, the insecticidal effect is that the treatment kills at least about 90% of the pests exposed to the treatment. In some embodiments, the insecticidal effect is that the treatment kills at least about 95% of the pests exposed to the treatment. In some embodiments, the insecticidal effect is that the treatment kills at least about 99% of the pests exposed to the treatment. In some embodiments, the insecticidal effect is that the treatment kills at least about 99.5% of the pests exposed to the treatment. In some embodiments, the insecticidal effect is that the treatment kills at least about 99.9% of the pests exposed to the treatment.

[0227] Beneficial insects protected by the present invention, i.e., those not significantly harmed by exposure, include: (1) predatory beetles such as adults of Harmonia arcuata (Fabricius), Diomus notescens (Blackburn), Coccinella repanda (Thunberg), and Dicranolauis bellulus (Gaerin); (2) carnivorous insects such as adults of Geocoris lubra (Kirkaldi), Cermatulus nasalis (Westwood), and Nabis capsiformis (Germar); (3) spiders, particularly of the Salticidae family, Araneus spp., Oxypes spp., and (parasitoids) Pterocormus promissorius (Erickson), Heteropelma scaposum (Moray), and Netelia producta (Bullet) and (4) pollinators, such as honeybees (Apis genus), especially Apis mellifera (Western honeybee), Apis mellifera capensis (African honeybee), Apis koschevnikovi (Koschevnikov's honeybee), Apis nigrocincta, Apis cerana (Oriental honeybee or Asiatic honeybee), Apis cerana indica (Indian honeybee), and Apis cerana nuluensis.

[0228] Embodiments of the present invention also relate to the production of improved pest control agents by identifying one or more fractions of a combination agent, screening the one or more fractions using the methods disclosed herein, and characterizing the one or more fractions as having a positive or negative effect on the likelihood of activity against a target insect.

[0229] In some embodiments, one or more fractions in complex agents (e.g., essential oils) can be isolated by using fractionation techniques, including, for example, fractional solvent extraction, fractional distillation, fractional crystallization, fractional freezing, dry distillation, surfactant fractionation, solvent extraction, supercritical CO2 fractionation, vacuum distillation, column chromatography, reversed-phase chromatography, high performance liquid chromatography, etc. These methods are known to those skilled in the art and widely practiced.Vacuum distillation is preferred because it is relatively simple to use and does not require the use of solvent.

[0230] In some embodiments, one or more fractions of the complexing agent can be isolated by column chromatography using silica or alumina solid support. For example, organic solvents including alkanes, such as hexane and petroleum ether, toluene, methylene chloride (or other halogenated hydrocarbons), diethyl ether, ethyl acetate, acetone, alcohol, acetic acid, etc., can be used alone or in combination with column solvents or mobile phases. In some embodiments, the complexing agent is fractionated by column chromatography using increasing concentrations of polar solvents as elution solvents. Methods for performing column chromatography and common solvents for use therein are well known in the art.

[0231] In some embodiments, SPC can be isolated by solvent extraction. For example, the extract can be extracted with a solvent such as methanol, acetic acid, acetone, acetonitrile, benzene, 1-butanol, 2-butanol, 2-butanone, t-butyl alcohol, carbon tetrachloride, chlorobenzene, chloroform, cyclohexane, 1,2-dichloroethane, diethyl ether, diethylene glycol, diglyme (diethylene glycol dimethyl ether), 1,2-dimethoxyethane (glyme, DME), dimethyl ether, dimethylformamide (DMF), dimethyl sulfoxide (DMSO), dioxane, ethanol, ethanol acetate, or the like. The solvent may be mixed with an organic solvent, including organic solvents such as methyl methyl ether (MTBE), ethylene glycol, glycerin, heptane, hexamethylphosphoramide (HMPA), hexamethylphosphoramide (HMPT), hexane, methyl t-butyl ether (MTBE), methylene chloride, JV-methyl-2-pyrrolidinone (NMP), nitromethane, pentane, petroleum ether (ligroin), 1-propanol, 2-propanol, pyridine, tetrahydrofuran (THF), toluene, triethylamine, water, heavy water, o-xylene, m-xylene, and / or 1-xylene. Other organic solvents known to those skilled in the art may also be used.

[0232] The mixture of SPC and organic solvent can then be combined with an extraction solvent that is immiscible with the organic solvent, including, for example, water, ethanol, and methanol. This combination is vigorously shaken for several minutes in a glass container such as a separatory funnel, and then allowed to settle into separate phases for several minutes. The lower, darker phase is then drained from the separatory funnel. The organic phase is then repeatedly re-extracted with the extraction solvent to separate compounds that are still soluble in the extraction solvent from the organic phase. The volume of the organic phase and the extracted phase can then be reduced using rotary evaporation to obtain two separate fractions of SPC.

[0233] In some embodiments, a method for identifying improved agents against target insects can include identifying compounds present in a combination formulation or individual isolated fractions of the combination formulation and screening the component compounds for activity. Identification of the compounds can be carried out by analyzing the combination formulation or its isolated fractions by high-performance liquid chromatography (HPLC) or gas chromatography (GC) coupled with mass spectrometry (MS). Component compounds can also be identified by first concentrating or purifying the individual components to homogeneity using techniques including, for example, fractional solvent extraction, fractional distillation, vacuum distillation, fractional crystallization, fractional freezing, dry fractionation, detergent fractionation, solvent extraction, supercritical CO2 fractionation, column chromatography, reversed-phase chromatography, high-performance liquid chromatography, and the like. The enriched or purified components can be identified using spectroscopic techniques including, for example, infrared (IR) spectroscopy, Raman spectroscopy, nuclear magnetic resonance spectroscopy (NMR), and the like.

[0234] Some embodiments relate to the use of chemical derivatives or analogs of the identified chemicals to produce improved agents against target insects. Chemical derivatives of the identified chemicals can include compounds derivatized with inorganic or organic functional groups. In some embodiments, the chemical derivatives are compounds derivatized with organic functional groups. In some embodiments, the organic functional groups can be alkyl groups. In some embodiments, the organic functional groups can be methyl, ethyl, propyl, butyl, seryl, decyl, heptyl, hexyl, myricyl, myristyl, nonyl, octyl, palmityl, pentyl, stearyl, isopropyl, isobutyl, lignoceryl, pentacosyl, heptacosyl, montanyl, nonacosyl, pentan-2-yl, isopentyl, 3-methylbutan-2-yl, tert-pentyl, neopentyl, undecyl, tridecyl, pentadecyl, margaryl, nonadecyl, arachidyl, henicosyl, behenyl, tricosyl, cyclobutyl, cyclopropyl, and the like. In some embodiments, the organic functional group can be an aryl group. In some embodiments, the organic functional group can be a phenyl or biphenyl-4-yl group.

[0235] In some embodiments, the improved agents against target insects include chemical derivatives that are halogenated derivatives of the identified compounds. In some embodiments, the chemical derivatives are fluorinated, chlorinated, brominated, or iodinated derivatives.

[0236] In some embodiments, the improved agent against the target insect comprises a chemical derivative that is an alkenylated derivative of the compound. In some embodiments, the improved agent against the target insect comprises a chemical derivative that is an alkenylated derivative of the compound, such as a compound that improves the water solubility of a glycosylated derivative. In some embodiments, the chemical derivative is an oleylated, allylated, isopropenylated, vinylated, prenylated, glycosylated, or phytylated derivative.

[0237] In some embodiments, the improved agent against the target insect comprises a chemical derivative that is a hydroxylated derivative of the identified compound. In some embodiments, the improved agent against the target insect comprises a chemical derivative that is a thiolated derivative of the identified compound. In some embodiments, the improved agent against the target insect comprises a chemical derivative that is a carboxylated derivative of the identified compound. In some embodiments, the improved agent against the target insect comprises a chemical derivative that is an amidated derivative of the identified compound. In some embodiments, the improved agent against the target insect comprises a chemical derivative that is an esterified derivative of the identified compound. In some embodiments, the improved agent against the target insect comprises a chemical derivative that is an acylated derivative of the identified compound. In some embodiments, the improved agent against the target insect comprises a chemical derivative that is a sulfonated derivative of the identified compound.

[0238] In some embodiments, improved agents against target insects include chemical derivatives that are derivatized by the introduction of substituent homologs.

[0239] In some embodiments, improved agents against target insects include chemical derivatives that are derivatized by moving substituents around the ring to different positions.

[0240] In some embodiments, the effectiveness of test compositions can be determined by testing them on pests.For example, the effectiveness of test compositions for killing pests, changing feeding or egg-laying tendencies, etc. can be tested by using controlled experiments in which insects are exposed to test compositions.In some embodiments, the toxicity of test compositions to pests can be tested by using controlled experiments in which insects are exposed to test compositions.The effectiveness of test compositions can also be determined using beneficial and predatory species.

[0241] In some embodiments, the formulation consists of an emulsifier with high solvency and ability to form a stable emulsion of the total formulation in water and a "carrier" oil that may also have pesticidal properties. A preferred "carrier" oil is an esterified vegetable oil.

[0242] In some embodiments, the pest is an animal pest and the method includes applying a compound, insecticidal composition, or extract containing γ-pyrone to the animal. In some embodiments, the animal can be a dog, cat, cow, sheep, horse, goat, pig, chicken, guinea pig, donkey, duck, bird, water buffalo, camel, reindeer, goose, llama, alpaca, elephant, deer, rabbit, mink, chinchilla, hamster, fox, emu, or ostrich.

[0243] In some embodiments, the method involves treating a habitat, for example, a habitat of any one or more of the above-identified animals.

[0244] In certain embodiments, the LC of the compound, pesticidal composition or extract 50 Values ​​are less than about 3000 ppm (i.e., 0.3%), less than about 2500 ppm, less than about 2000 ppm, less than about 1500 ppm, less than about 1000 ppm, less than about 500 ppm, less than about 200 ppm, less than about 100 ppm, less than about 50 ppm and preferably less than about 30 ppm.

[0245] In certain embodiments, the LC of the compound, pesticidal composition or extract 50The value is about 10 to 3000 ppm, about 10 to 2500 ppm, about 10 to 2000 ppm, about 10 to 1500 ppm, about 10 to 1000 ppm, about 10 to 500 ppm, about 10 to 200 ppm, about 10 to 100 ppm, about 10 to 50 ppm and preferably about 10 to 30 ppm.

[0246] In certain embodiments, the LC of the compound, pesticidal composition or extract 95 Values ​​are less than about 1500 ppm (ie, 0.15%), less than about 1000 ppm, less than about 500 ppm, less than about 200 ppm, less than about 100 ppm, less than about 50 ppm, less than about 30 ppm, and preferably less than about 10 ppm.

[0247] In certain embodiments, the LC of the compound, pesticidal composition or extract 95 The value is about 10 to 1500 ppm, about 10 to 1000 ppm, about 10 to 800 ppm, about 10 to 500 ppm and preferably about 100 to 300 ppm.

[0248] In use, the method of the present invention can be carried out by spraying the dispersion solution. The spray can be applied from a spray container such as a can, bottle, or other container by means of a pump or by release from a pressurized container, for example, a pressurized aerosol spray can. Such spray compositions can take various forms, such as a spray, mist, foam, fume, or fog. Such spray compositions can therefore optionally further comprise propellants, foaming agents, etc.

[0249] In another embodiment of the present invention, a kit for sale on the shelf is provided, the kit comprising spraying means, a container means which may or may not be integral with the spraying means, predetermined amounts of the components of the spray formulation (e.g., γ-pyrone, carrier, surfactant, if applicable), and instructions for preparing the formulation for use.

[0250] Typical propellants include, but are not limited to, methane, ethane, propane, butane, isobutane, butene, pentane, isopentane, neopentane, pentene, hydrofluorocarbons, chlorofluorocarbons, dimethyl ether, and combinations thereof.

[0251] In other embodiments, the method may be practiced using creams, ointments, emollients, pastes, gels, powders, solids and combinations thereof. [Example]

[0252] Example 1: Plant Extracts Approach 1 Plant samples (aerial parts) from identified surviving plants were collected, placed in labeled paper bags, and dried in an oven at 40°C for 7 days. The material was then crushed and stored at room temperature in suitable containers. Original samples of Podolepis jaceoides flowers were obtained from the Mount Annan Botanic Garden, Mount Annan, New South Wales, Australia. Plants were then grown at the Hawkesbury Campus of Western Sydney University from seeds obtained from commercial supplier Nindethana Australian Seeds, Albany, Western Australia; and from seeds collected from this crop.

[0253] Technical grade methanol and Milli-Q (ultrapure) water were used to prepare the test extracts. Approximately 25 g of each sample was added to a 1:1 mixture of methanol and chloroform (250 mL). The mixture was sonicated for 60 minutes, softened by soaking (24 hours), suction filtered, and the filtrate was transferred to a separatory funnel. A small amount of water (approximately 25 mL) was added to the mixture to create a two-layer system consisting of a nonpolar chloroform layer (N) and a polar aqueous-methanol layer (P). These two layers were separated into pre-weighed round-bottom flasks. The fractions were either evaporated to dryness using a rotary evaporator (P) or evaporated in a fume hood (N). Each fraction was tested separately.

[0254] Once the active ingredient was determined, a suitable organic solvent such as methanol, ethanol, acetone or a binary (two-solvent) system was used.

[0255] Approach 2 An extract was prepared from dried, harvested flowers of Podolepis jaceoides (plant #68) grown at Western Sydney University (WSU). The flowers were ground to a fine powder using a Waring blender, extracted with pure methanol (20 g ground plant in 200 mL pure methanol), shaken in a Ratec platform mixer for 24 hours, sonicated for 80 minutes, and then filtered through a glass filter funnel fitted with Whatman No. 1 filter paper. The filtrate formed the "methanol extract."

[0256] A 1.0% w / v emulsion was then prepared by taking a weight of the extract to prepare a quantity sufficient for treatment of the three test organisms, two-spotted spider mite, cotton aphid, and tobacco budworm. The dried extract was first dissolved in acetone, sonicated for 5 minutes, and then diluted with 200 ppm surfactant Triton X-100. TM A 1.00% w / v emulsion was prepared from each sample by dilution with distilled water.

[0257] The stock solution was then serially diluted in Triton X-100 / water to give concentrations of 0.200%, 0.100%, 0.050%, and 0.0125% w / v. These lower concentrations were tested against TSM and cotton aphid; 1.0% was tested against Helicoverpa spp.

[0258] Example 2: Target Pests Aphis gossypii Glover (Hemiptera: Aphidoidea), commonly known as the cotton or melon aphid, is a cosmopolitan pest with an extremely wide host range (e.g., www.cabi.org / isc / datasheet / 6204). It is also a vector for numerous viruses. Aphis gossypii (susceptible species) (Hemiptera: Aphidoidea) was established as a culture from material provided by Dr. Grant Herron, NSW Department of Primary Industries, Menangle, New South Wales, Australia. Aphids were reared on potted Gossypium hirsutum L. (Malvaceae) plants grown from non-insecticide-treated conventional cotton seed provided by Dr. Robert Mensah, NSW Department of Primary Industries, Narrabri, New South Wales, Australia. Cultures were maintained in a temperature-controlled greenhouse at 27±3°C and 55±10% RH (relative humidity) with natural light. Only young adult females were selected for the bioassay.

[0259] Tetranychus urticae Koch (Acari: Tetranychidae), commonly known as the two-spotted spider mite, is a cosmopolitan, herbivorous species and a significant agricultural pest in many temperate and subtropical countries. It is highly omnivorous, feeding primarily on plant leaves. Tetranychus urticae Koch UWS 1 (organophosphate-sensitive) species was originally obtained from Grant Herron, NSW Department of Primary Industries, Menangle, New South Wales, Australia, and has been maintained at the WSU Hawkesbury campus for approximately 18 years. Potted green beans, Phaseolus vulgaris var. Redland Pioneer, were reared in a constant-temperature room maintained at 25 ± 2°C, 65 ± 5% RH, and a 14-hour D:L photoperiod. Cultures were reared on potted dwarf green bean (Phaseolus vulgaris L. var. Redland Pioneer) plants grown in a composted pine bark-based potting medium in a large (7.0 m (l) × 6.0 m (w) × 4.0 m (h)), pesticide-free, constant-temperature room (28–30°C, 50% RH, 18:6 D:L). Only young, gravid female spider mites were selected for bioassays.

[0260] Helicoverpa armigera (Huebner) (Lepidoptera: Noctuidae), commonly known as the cotton ball worm or heliodis, is a widely distributed species with a broad host range (e.g., https: / / www.cabi.org / isc / datasheet / 26757 Helicoverpa armigera are obtained as new eggs from AgBiTech Pty. Ltd, Grandvale, Queensland, Australia, or from the CSIRO Agriculture and Food, Cotton Research Institute, Myall Vale, New South Wales, Australia.

[0261] Eggs (50–75) were transferred with a fine camelhair brush to 90 mm Whatman No. 1 filter paper partially moistened with distilled water, and a thin piece of artificial medium provided by AgBiTech was placed on the filter paper as a food source for the hatched caterpillars. Eggs were incubated under laboratory conditions of 25 ± 3°C and 65 ± 10% RH. Hatching occurred within 24 hours, and bioassays were performed on newborn larvae with an average weight of 4.12 mg.

[0262] Example 3: Bioassay Screening of each sample for two-spotted spider mites was performed with 20–40 adult female mites evenly distributed across one to three green bean leaf segments (25 mm diameter) contained in a 90 mm diameter Petri dish. The leaf segments were placed underside up on moist absorbent cotton. Water was added to the dish daily to prevent the leaf segments from drying out and to prevent mites from escaping from the segments.

[0263] A 3.0 mL aliquot was applied to each petri dish with a Potter-type precision spray tower as described by Herron et al., Potter spray tower bioassay of selected citrus pests to petroleum spray oil, Journal of the Australian Entomological Society 34, 253-263, 1995.

[0264] The average weight of the solution sprayed on each dish was 5.385 mg / cm 2 or 0.05385 mg / cm 2 The active ingredient (ai) was calculated. Dishes were kept under laboratory conditions (24±2°C, 50±15% RH) for post-treatment observation. Mortality was recorded 24 and 48 hours after treatment (HAT). Death was determined by the absence of movement when the test organisms were mechanically stimulated by prodding with a brush.

[0265] For cotton aphids, parthenogenetic adult female aphids were collected from cotton plant cultures. Fifteen adults were transferred using a fine camel hair brush to the underside of each 50 mm diameter leaf piece placed on 1.0% w / v agar in a plastic insect cage measuring 50 mm diameter x 15 mm high (SPL Life Sciences, Korea).

[0266] The baskets had lids with holes (15 mm diameter) covered with fine mesh; the lids were removed immediately before spraying. Aliquots (3 mL) of 1% w / v extract were applied in a Potter-type spray tower for each treatment as described above, and the spray was allowed to partially dry before the baskets were covered and the sides of the baskets were covered with Parafilm M®.

[0267] Dishes were kept at laboratory conditions (24±2°C, 50±15% RH) for post-treatment observation. Mortality counts were performed at 24 and 48 HAT. Immobility of aphids when poked with a fine brush was taken as the criterion for mortality.

[0268] For Helicoverpa, neonate larvae 4–12 h after hatching were individually transferred to cotton leaf pieces (35 mm diameter) on 1% agar in Easy Grip Tissue Culture Dishes (35 × 10 mm, Corning Incorporated, Corning, NY, USA) with the underside facing up. Only one neonate was transferred to each dish (because initial investigations, in which small groups of neonates were treated before separation, resulted in larval damage due to cannibalism). Four of these dishes were transferred to large 90 mm diameter Petri dishes and sprayed with 3 mL volumes using a Potter-type spray tower as described above. Thus, each replicate consisted of four neonates.

[0269] After spraying, the small dishes were covered with perforated plastic sheets and then placed in the plastic cages described above for the aphid bioassay. The number of dishes sprayed per sample ranged from 4 to 10. Dishes were maintained under laboratory conditions (24 ± 2°C, 50 ± 15% RH) for post-treatment observation. Mortality counts were performed at 24 and 48 h. The immobility of caterpillars when poked with a fine brush was taken as the criterion for mortality.

[0270] For each batch of study, a control treatment was performed in the same manner as the other treatments. The control treatment solution contained 200 ppm Triton X-100 in 3 mL of distilled water and acetone or hexane at the same concentration as the test solution. If control mortality exceeded 15%, the study was abandoned. Treatment mortality was calculated and expressed as adjusted mortality to account for any control mortality (Abbott W.S., A method of computing the effectiveness of an insecticide. Journal of Economic Entomology, 18, 265-267, 1925).

[0271] Example 4: Insecticidal activity test results Table 1 presents initial experimental data demonstrating the efficacy of non-polar extracts of P. jaceiodes against cotton aphids and twospotted spider mites, but not Helicoverpa. Polar extracts were substantially less effective against these target species. Activity was observed almost immediately after application of effective concentrations, with no signs of toxicity to the target organisms within the first hour after treatment (HAT) and 100% mortality and no progeny within a few hours. However, the polar fraction was substantially less effective against the target species than the non-polar fraction, particularly against cotton aphids.

[0272] Subsequent studies showed 100% mortality of cotton aphids at 0.25% ai (Table 2).

[0273] Further evaluation of other P. jaceoides N extracts (flowers) using only methanol as the solvent resulted in extremely high mortality of cotton aphids, high mortality of twospotted spider mites, and no mortality of Helicoverpa moths (Table 3). For cotton aphids, 100% mortality of cotton aphids occurred at 0.05 and 0.20% treatments, while at 0.20% w / v there was 84.4% mortality of TSM. The estimated LC of cotton aphids 50 and L.C. 95 The values ​​were 0.014% (0.004-0.021), (i.e., 140 [40-210] ppm) and 0.051% (0.039-0.112) (510 [390-1120 ppm]), respectively. [Table 1] * Low concentrations not tested; HAT = time after treatment; N = non-polar fraction; P = polar fraction [Table 2] [Table 3] * X 2 =0.734

[0274] Heterogeneity of regression was evaluated using Pearson χ 2 The heterogeneity factor, determined by the index and with a significance level of <0.150, was used to calculate the confidence limits. This allowed the calculation of the LC value and its 95% confidence limits (CL). 50 and L.C. 95 was selected as a good representation of the potential activity of the insecticides. Analyses were performed using the program IBM SPSS Statistics 25.

[0275] Example 5: Pot trials to evaluate P. jaceoides methanol extract against cotton aphid Replicate pot experiments also demonstrated high efficacy of P. jaceoides methanol extract against cotton aphid.

[0276] Twelve potted cotton plants were grown under controlled greenhouse conditions (26-28°C and 60±10% RH) for three weeks until two true leaves were fully developed. Each pot contained one plant. They were moved between potted plants heavily infested with cotton aphids for three days to allow for leaf contamination. The newly infested pots were randomly divided into two groups of six plants each.

[0277] A 1.0% w / v concentration of P. jaceoides methanol extract in water containing 200 ppm Triton X-100 and a control containing all ingredients except the extract were prepared as described above. One group of plants was sprayed with the 1.0% w / v extract, and the other with the control emulsion. A 500 mL hand-held sprayer was used to apply 45 mL of each treatment to six plants. Each plant was sprayed separately to dripping to cover the entire upper and lower leaf surface; this was approximately 7.5 mL of treatment per plant.

[0278] Aphid counts were performed on marked leaves before and after spraying, with post-treatment counts at 24 HAT and 48 HAT. Observations were made at 2 HAT to determine any knockdown effect. Mortality reached 100% at 48 HAT, and the experiment was terminated. Plants were held for 5 DAT to assess any phytotoxicity due to the treatment.

[0279] Data were analyzed using a general linear model analysis of variance (ANOVA). Each variable was visually examined for normality using a QQ plot, and a Levene test was used to test for equality of error variances. The significance level was set at P = 0.05. In this case, however, the results were clear and there was one treatment and control, so statistical analysis was not necessary.

[0280] The results are summarized in Table 4. At 24 HAT, the mean aphid mortality was 97.37 (±SE 2.162), with 5 of 6 plants experiencing 100% mortality at 48 HAT. Other plants with >90% mortality at 48 HAT may not have been adequately sprayed. Mortality at 24 and 48 HAT in the control treatment was negligible, and the treatment results were highly significant (F = 1888.475, P < 0.001 and F = 1603.362, P < 0.001), respectively.

[0281] At 24 HAT evaluation, many aphids on the extract-treated leaves were nearly moribund, while control plants remained healthy. This study confirms the laboratory bioassay that the non-polar or methanol extract of P. jaceoides (labeled extract #68) has extremely high aphicidal activity. [Table 4]

[0282] There was no sign of phytotoxicity in any of the plants up to 5 HAT, indicating that the P. jaceoides methanol extract had no phytotoxic effect under the conditions tested.

[0283] Furthermore, the extracts did not cause phytotoxicity to cotton or green bean plants when applied at concentrations much higher than those found to be effective, indicating that crop damage is unlikely to occur with end-use insecticides derived from these plant extracts of the present invention.

[0284] Example 6: Field trial to evaluate the efficacy of Podolepis jaceoides methanol extract against cotton aphid material and method The trials were conducted on irrigated commercial Bollgard II cotton crops (Sicot 74 variety) at the Australian Cotton Research Institute (ACRI) in Narrabri, New South Wales, Australia. The cotton crops used in the trials were mature plants near cutting and infested with Aphis gossypii-infested leaves collected from a nearby commercial cotton field at Yarral, Narrabri, New South Wales, Australia.

[0285] Treatment plots were arranged in a randomized, completely block design with three replicates per treatment. Each replicate plot consisted of 2 m (two rows) wide and 1 m long with a 2 m (row) buffer between replicate plots. Three plants in each treatment replicate with aphid-infested ends were randomly selected, and the ends were tagged for pre- and post-treatment evaluation. The following treatments were evaluated against Aphis gossypii adults and nymphs: (1) 1.0% v / v P. jaceoides methanol extract, (2) 125 mL / hectare clothianidin (Shield), and (3) control.

[0286] The methanol extract of P. jaceoides was dissolved in 10 mL acetone and then diluted with 100 ppm Triton-X 100 water. The control was sprayed with only the aqueous solution of 100 ppm Triton-X 100.

[0287] Foliar applications of each treatment were performed, with the control applied first, followed by the plant extract, and finally the industrial standard. Each treatment was applied to drip with a small handheld pressurized sprayer equipped with a flat fan nozzle delivering a 200 μm droplet size (equivalent to a field application of 100 L / hectare). The decision to apply treatments was based on the IPM Guidelines and CottonLogic recommended economic threshold of 50% contamination of the plant end.

[0288] The abundance of A. gossypii and beneficial insect species (primarily predatory insects) was assessed by visual counting A. gossypii adults and nymphs and beneficial organisms on the upper and lower surfaces of tagged leaves below the terminal 3–4 nodes of the plants. Pre-treatment counts were performed 24 h before treatment, and post-treatment counts were performed 1, 2, 5, 6, 7, 8, 9, 12, and 14 days after treatment (DAT).

[0289] Data for both A. gossypii adults and nymphs and predatory insects were initially expressed as numbers per end, respectively. Data were calculated as percentage reduction in population compared to pre-treatment counts.

[0290] Population reduction data means were compared using one-way ANOVA after normality checks using SPSS v21. Significance was typically set at 5% (α = 0.05) and, when significant, means were separated using Duncan's multiple range test.

[0291] The data are presented in Table 5. At 1 DAT and 2 DAT, there is a significant reduction in cotton aphid population with P. jaceoides extract and bifenthrin treatments compared to the control (P 2,9 =0.047;P 2,9 =0.001), there was no difference between them.

[0292] At 5 DAT, there was no significant difference between treatments at the 5% level, but at the 10% level (P 2,9 = 0.087), with the plant extract being superior to the control but with no other differences. At 6, 7, and 14 DAT, there were no differences between the treatments, but there was a trend toward the P. jaceoides extract being superior.

[0293] A range of beneficial insect species was recorded throughout the trial. The most common species were spiders, ladybugs (adults and larvae), pirate bugs, and hoverflies. Comparisons between treatments were difficult. First, some data (e.g., mummified aphids in the control at 1 DAT) were the result of activity before the start of the trial. Second, the high abundance of aphid predators in the control at 5–6 DAT likely reflects the earlier abundance of aphids. In general, the abundance of beneficial insect species was low early in the trial but generally increased in all treatments (including the control) by 9 DAT.

[0294] The trial had untreated buffer rows where beneficial insect species could reside before migrating to the trial location. It was late in the season (when beneficial insect species are generally abundant) and most of the other surrounding cotton crops had been harvested, making the trial location attractive.

[0295] Therefore, data for beneficial insects and other species are somewhat variable, and there are insufficient data points to make any detailed assessment. Nevertheless, there is no evidence that any of the chemical treatments (including P. jaceoides extracts) had adverse effects on beneficial insect species. [Table 5] #Not significant at 5%, but significant at 10%

[0296] Example 7: Evaluation of the acute toxicity of P. jaceoides methanol extract to honeybees The acute toxicity of a methanol extract of P. jaceoides to honeybees was evaluated. Although other insects can be efficient pollinators, the Western honeybee, Apis mellifera, is considered the most important pollinator of agricultural crops worldwide due to its abundance and amenability to human handling.

[0297] material and method A laboratory-based bioassay was conducted to evaluate the acute toxicity of the extract to mature foraging workers of the Western honeybee Apis mellifera. A 3 mL volume of P. jaceoides extract was applied at 1.0% ai. w / v using a Potter-type spray tower, using the method previously described for the preparation and application of this plant extract to the test insect.

[0298] Bifenthrin (CAS No. 82657-04-3) 97.0% technical (Batch 50118) was provided by Dr. Ehrenstorfer GmbH, D-86190 Augsburg, Germany. A 0.05% w / v ai concentration was obtained by dissolving 0.013 g bifenthrin in 1 mL acetone, sonicating for 5 minutes, and then diluting with 200 ppm Triton X-100 in distilled water in a 25 mL volumetric flask. Application was via a Potter-type spray tower using a 3 mL volume of 0.05% ai; this concentration was based on the recommended label rate on cotton. Controls consisted of 5 mL acetone and the water-Triton X-100 mixture alone; two controls were performed.

[0299] Western honeybee forager workers for the bioassays were collected from strong, healthy honeybeehives at the Western Sydney University Apiary, Richmond, New South Wales, Australia, by blocking the hive entrance for 1 min and collecting the foragers with a net.

[0300] They were immediately transferred to mesh cages (25 × 33 × 33 cm) and maintained in the laboratory at 24°C, where they were given free access to 1:1 w / v sugar:water via cotton wool placed on the mesh above the cage until the bioassay, which was performed within 1 h of collection. Bees for the bioassay were randomly collected from the cages using 120 mL plastic sample tubes and anesthetized with pharmaceutical-grade CO2 for 30 seconds. Ten bees were then placed in a 90 mm diameter Petri dish lined with filter paper. The dish containing the bees was placed in a Potter-type spray tower platform, and each treatment was applied, starting with the control and ending with the bifenthrin treatment.

[0301] Immediately after treatment, treated bees were gently transferred to a 250 mL glass beaker, topped with a fine nylon mesh rubber band, and the bees were fed sugar:water as above and maintained at 26°C in minimal light. Bee activity was observed and recorded periodically at 10-minute intervals until 420 minutes (i.e., 7 HAT), and then less frequently until 48 HAT. By this time, bees had died in both controls, and the study was therefore terminated. Data recorded were signs of toxicity, knockdown (K), and mortality (D).

[0302] The results are shown in Table 6. P. jaceoides extract was safe to honeybees as measured by acute toxicity, far superior to the industrial control, bifenthrin. There was no mortality or knockdown at 36 HAT, at which point mortality began in one of the control replicates. In contrast, in the bifenthrin treatments, knockdown began at 10 MAT (minutes after treatment), mortality began at 90 MAT, 50% of bees were knocked down at 45-50 MAT, and 50% died at 240 MAT. By 480 MAT, 100% had died. The only treatment in which signs of bee poisoning were observed was the bifenthrin treatment. Lack of bee coordination, vomiting of stomach contents, and hyperactivity were all observed. [Table 6-1] [Table 6-2]

[0303] Although this was a single study evaluating acute dermal toxicity (not oral toxicity), P. jaceoides extract has limited acute toxicity to honeybees and is relatively safe for application to flowering crops where honeybees can feed. It should be noted that the application concentration was 1.0% w / v, which may be higher than would typically be needed in field applications to target pests, and that under test conditions, bees were directly exposed to the spray applied by a Potter-type spray tower. This level of exposure is unlikely to occur routinely in the field.

[0304] Example 8: Evaluation of the efficacy of a methanol extract of P. jaceoides against Queensland fruit fly and barley whitefly Topical application of a 1.0% w / v P. jaceoides (labeled #68) flower methanol extract in a Potter-type spray tower resulted in approximately 100% mortality of mixed-sex adult Queensland fruit flies (QFF) (Bactrocera tryoni, Diptera: Tephritidae) at 48 HAT using the method previously described. Shortly after treatment, flies became hyperactive and showed knockdown at 20 MAT. However, at 0.5% w / v, the extract did not show similar knockdown, with only 1 of 5 flies killed at 48 HAT. death All treated controls resulted in 0.0% mortality at 48 HAT.

[0305] Against the cabbage whitefly, Aleyrodes proletella (Hemiptera: Aleyrodidae), a 1.0% w / v P. jaceoides (labeled #68) flower methanol extract using a Potter-type spray tower provided 100% mortality at 30 MAT.

[0306] material and method P. jaceoides (labeled #68) flower methanol extract (batch ARL181076) was obtained from Southern Cross University.

[0307] Approximately 50 pupae of Queensland fruit flies were collected from a multigenerational inbred culture by Markus Riegler at Western Sydney University. Pupae were transferred to 30 x 30 x 30 cm greenhouse cages at 25°C and 70% RH. Adults emerged approximately 5 days later, and recently emerged mixed-sex adults were used in the bioassay.

[0308] The cabbage whitefly is distributed globally and is a pest of various Brassica species, particularly kale (Brassica oleracea var. sabellica). Bioassay specimens were collected from naturally heavily infested, healthy potted kale plants in Western Sydney. When kale plants were observed to contain large numbers of adult whiteflies, the leaves were gently cut and placed into a 1 L Perspex beaker covered with muslin netting and secured with rubber bands.

[0309] Queensland fruit fly bioassay Three- to five-day-old, mixed-sex flies were transferred to a 1-L Perspex beaker. Each fly was captured in a 20-mL glass beaker, then released into a larger beaker and transferred to the beaker by covering it with a muslin net secured with a rubber band. The flies were anesthetized with pharmaceutical-grade carbon dioxide until their movements were restricted. Batches of flies were divided into a control treatment (i.e., all components in the emulsion except for the #68 extract) and a 1.0% w / v concentration of the #68 methanol extract. Treatment sites were small 50 x 15 mm plastic Petri dishes lined with filter paper of the same diameter. A 3-mL aliquot was applied to the Petri dishes containing the anesthetized flies using a Potter-type spray tower. The uncovered treatment dishes were transferred to a 500-mL glass beaker and covered with a muslin net secured with a rubber band.

[0310] Kale Whitefly Bioassay Whiteflies from freshly cut kale leaves were transferred to a -20°C deep freezer using a 1 L Perspex beaker. Approximately 40 minutes in the deep freezer was found to restrict movement sufficiently to transfer the whiteflies and allow treatment with Potter-type tower applications of the control and #68 methanol extract treatments. Using a fine camel hair brush, adult whiteflies were transferred to filter paper placed in the bottom of small 50 x 15 mm Petri dishes. 3 mL volumes were applied with a Potter-type spray tower at control (0.0%) and 1.0% w / v concentrations (three separate dishes, replicate treatments). After treatment application, fresh kale leaf pieces approximately 25 mm in diameter were placed in the Petri dishes, which were then covered with their lids, each containing a 15 mm diameter hole covered with fine mesh. result

[0311] The results are shown in Table 7 (Queensland fruit fly, QFF) and Table 8 (Kale white fly). For QFF, 1 in 5 flies were writhing and all survived at 48 HAT in the 0.5% w / v treatment. However, in the 1.0% w / v treatment, 80% of the flies treated at 48 HAT in one container were dead and writhing; in the other container, 100% were dead. However, flies sometimes had slightly retracted appendages and otherwise death Interestingly, the flies became hyperactive for several minutes after treatment, after which they squirmed about until they fell to the bottom of the beaker, were unable to fly again, and died.

[0312] For kale whiteflies, 100% mortality was observed in all three treatment dishes, with mortality occurring within 30 MAT. Observation at 48 HAT confirmed mortality, not simply knockdown. [Table 7] [Table 8]

[0313] These results showed that the methanol extract of #68 had high efficacy against kale white fly and moderate to high efficacy against Queensland fruit fly.

[0314] Example 9: Investigation of the chemistry of Podolepis jaceoides to determine the active components of the solvent extract This example has two objectives: 1) to elucidate the chemistry of insecticidal P. jaceoides extracts and 2) to identify active fractions / compounds by performing a bioassay-guided fractionation.

[0315] Stems (including leaves) and flowers of P. jaceoides were harvested and evaluated separately. Once dried and ground, samples were extracted with methanol (1 L) by sonication (1 h) and maceration (24 h). The filtered extract was freeze-dried and partitioned in a separatory funnel with water (250 mL) and hexane (3 × 250 mL), followed by extractions with diethyl ether (3 × 250 mL) and ethyl acetate (3 × 250 mL). The resulting aqueous (AQ), hexane (Hex), diethyl ether (DE), and ethyl acetate (EA) fractions were freeze-dried to yield four fractions.

[0316] Liquid chromatography-mass spectrometry (LCMS) analysis was performed on a Waters Acquity Xevo TQ triple quadrupole mass spectrometer coupled to a binary pump, a photometric diode array (PDA) detector, and an autosampler (Waters, Milford, USA) with a 3 μL injection volume. Chromatographic separation was performed using a C 18 The column (Phenomenex kinetex 1.7 μM, 100 A, 150 × 2.10 mm) and an ACN / H2O (acetonitrile / water) gradient (both mobile phases contained 0.1% formic acid) were used, and the mobile phase gradient was 10% ACN to 95% ACN for 25 min, followed by 95% ACN for 5 min at a flow rate of 0.2 mL / min.

[0317] Mass spectra were performed in atmospheric pressure chemical ionization (APCI) positive mode using a mass range m / z (mass-to-charge ratio) of 150–800 a.m. (atomic mass units) and diode array detection (200–500 nm) at a capillary voltage of 2.5 kV, a cone voltage of 10 V, a probe temperature of 350–375 °C, and a gas flow of 300 L / h. Data were analyzed with MassLynx software (Waters, Milford, USA).

[0318] Whole plant samples were extracted with 100% methanol (MeOH), sonicated for 1 hour, then soaked overnight in a 1:10 w / v (dry weight) ratio, filtered, and analyzed. Fractions or pure compounds were analyzed at 0.5 mg / mL in methanol (MeOH).

[0319] NMR spectroscopy NMR spectra were obtained on a Bruker Avance DRX-400. Spectral data were analyzed using Mnova NMR software (Mestrelab, Santiago de Compostela, Spain). 1 H NMR spectra were measured at 400 MHz and 13 C NMR spectra were recorded at 100 MHz. Chemical shifts (δ) are expressed as δ values ​​in parts per million (ppm) and coupling constants (J) are expressed in Hertz (Hz). COSY, NOESY, HSQC and HMBC experiments were obtained using standard Bruker pulse programs. Experiments were performed in deuterated solvents and chemical shifts were recorded relative to the methanol solvent peak ( 1 H δ 3.31 and 13 C δ 49.00 ppm), DMSO solvent peak ( 1 H δ 2.50 and 13 C δ 39.52 ppm) or chloroform solvent peak ( 1 H δ 7.26 and 13 C δ 77.16 ppm).

[0320] Preparative HPLC fractionation of P. jaceoides whole extract was carried out based on LC-MS analysis, and bioassay results are from polar and non-polar fractions.

[0321] Bioassays were performed with the same three target organisms described above: twospotted spider mite, cotton aphid, and Helicoverpa, and mortality counts were taken at 24 and 48 HAT. Concentrations were 1.0% w / v unless otherwise noted in Table 9. Data are presented as adjusted mortality. In addition, any signs of phytotoxicity to the target plant material (leaf discs) were noted and recorded.

[0322] Bioassay results of the fractions are shown in Table 9. Several fractions of #68, especially those from flowers (#68F_P (polar) and #68F_N (non-polar)), showed very high activity against TSM and cotton aphid, and fraction #44 showed very high activity against both Tetranychus urticae and Helicoverpa ulcerata; other fractions showed very high activity only against spider mites.

[0323] Further data on cotton aphid were not collected on the HPLC fractions as they were further fractionated to produce podopyrone. Interestingly, the waste (column wash), which presumably contains long-chain compounds, also showed very high activity against cotton aphid.

[0324] We then compared chromatograms of methanol extracts from seeds, flower heads, and white petals (ray florets) of Podolepis jaceoides cultivated at Western Sydney University to determine the presence of podopylone in these various plant parts (not shown). Chromatograms of whole flower methanol extracts were compared with those of seeds, flower heads, and white petals. The whole flower methanol extract and the fractionated components, white petals, flower heads, and seeds showed the presence of podopylone (dominant large double UV peaks at approximately 16 and 17 min). Because the same column was used for seeds, flower heads, and white petals, comparisons were possible. The podopylone peak was present at 18–20 min in seeds, flower heads, and petals. [Table 9]

[0325] Comparing the whole plant extract with the flower head extract, podopylone appears to be more concentrated in the flower head because other compound peaks are more dominant. This was also confirmed by the total ion count (TIC) (not shown), in which the podopylone ion was detected less frequently. The TIC of the seed extract (not shown) showed a high abundance of the podopylone total ion at 18.85 and 19.14 min. The TIC of the white petals (not shown) detected numerous podopylone ions at 19.02 and 19.3 min. These results confirmed that the strategy for extracting the pappus of P. jaceoides seeds was appropriate.

[0326] Example 10: Identification of active ingredients from Podolepis jaceoides (non-polar) extract Table 9 shows the efficacy data for multiple extract fractions from these three plant species. Bioactive components were more concentrated in the stems and leaves than in the flowers. Furthermore, fraction 44 was the most active fraction.

[0327] Extraction and purification of active metabolites from Podolepis jaceoides The above-ground parts of the plants were harvested and dried. 50 g of dried plant material was ground and extracted with 1:1 chloroform:methanol (250 mL) by sonication (1 h) and maceration (24 h). The filtered extract was separated into polar (#68P) and non-polar (#68N) fractions in a separatory funnel by adding water (approximately 75 mL) until immiscible layers formed. The polar fraction was evaporated to dryness using a rotary evaporator, and the non-polar extract was evaporated in a fume hood. The dried extract was screened for insecticidal activity.

[0328] "Whole Plant Extract" for Preparative HPLC Fractionation Flowering, dried aerial parts of P. jaceoides were collected from plants grown at the Hawkesbury Campus of Western Sydney University, dried in a laboratory oven at 40°C for 7 days, then crushed and extracted (100 g dry weight) in 1:1 chloroform:methanol (1 L) by sonication (1 h) and maceration (24 h). The filtered extract (porous 4-sintered glass frit) was evaporated to dryness using a rotary evaporator to give 16.6 g (16.6% yield, w / w) of a dark green crude extract.

[0329] HPLC fractionation of active extracts Preparative HPLC fractionation was performed on a Shimadzu LC-20AP system (Kyoto, Japan) equipped with a DGU-20A3 inline vacuum degasser and a SIL-20AHT autoinjector and operated with Lab Solutions software. The crude extract was solubilized in MeOH and purified by C 18 (Alltech, Davisil C18 bonded silica, 35-75 μm, 150 Å) in a 1:2 gram ratio and packed into a stainless steel guard cartridge (Alltech, 10 × 30 mm).

[0330] Purification of podopyrone from P. jaceoides The extract (2.5 g) was fractionated on a reverse-phase preparative C18 Luna 5 μm 100A (150 × 21.2 mm) column (Phenomenex, Lane Cove, Australia) using a binary solvent system: Solvent A (MQ water, 0.01% TFA (trifluoroacetic acid)) and Solvent B (MeOH (methanol), 0.01% TFA), monitored at 210 nm (Shimadzu SPD-20A UV / Vis detector). A gradient HPLC condition of 25% B to 98% B was used over 60 min at a flow rate of 9 mL / min. Sixty fractions were collected from 1 to 60 min. Any strongly adsorbing material was subsequently eluted with 100% B to clean the column, creating a "waste" fraction for each sample. All corresponding fractions from five replicate injections were combined and evaporated to dryness using a speed vacuum concentrator (Savant SC250EXP, Thermo Scientific).

[0331] Insecticidal activity was identified across fractions 40, 42, 44, 53, and 55, with column wash fractions 42 (m / z 323) and 44 (m / z 337) containing the major metabolites in the extract, which, upon subsequent lyophilization, yielded the known podopyrones: 10'-oxopodopyrone (1c) and 10'-oxo-8-methylpodopyrone (1l).

[0332] Fraction #42, colorless oil, 30.4 mg, ≥90% pure by NMR, 1.22% yield dry weight of crude extract, unoptimized; 1 H NMR (400MHz, CDCl3): 3.94 (3H, s, 2-OMe), 2.57 (2H, t, J=7.5 Hz H-1'), 2.39 (2H, t, J=7.1 Hz H-9'), 2.11 (3H, s, 11'-Me), 1.91 (3H, S, H-7), 1.83 (3H, s, H-8), 1.62 (2H, dt, J=7.4, 6.9 Hz H-2'), 1.54 (2H, m, H-8'), 1.4 - 1.2 (10H, m, H-3'-H7'). Lit. 1 H NMR 400MHz, CDCl3, 13 C NMR 67.9MHz CDCl3(Jaensch et al., Pyrones and other constituents from Podolepis species, Phytochemistry, 28, 3497-3501, 1989; (+)-LRAPCIMS m / z (rel. int.): [M+H + ] (100%), C 19 H 31 Calculated O4, 323.222 (100) [ka]

[0333] Fraction 44, colorless oil, 38.2 mg, ≥95% purity by NMR, 1.53% yield dry weight of crude extract, unoptimized; 1H NMR (400MHz, CD3OD): 4.03 (3H, s, 2-OMe), 2.69 (2H, t, J=7.4 Hz H-1'), 2.46 (2H, t, J=7.1 Hz H-9'), 2.38 (2H, q, J=7.5, 7.4 Hz H-8), 2.12 (3H, s, 11'-Me), 1.92 (3H, S, H-7), 1.70 (2H, dt, J=7.2, 7.4 Hz H-2'), 1.55 (2H, m, H-8'), 1.3 - 1.5 (10H, m, H-3'-H7'), 1.00 (3H, t, J=7.4 Hz, H-9). Lit. 1 H NMR 400MHz, CDCl3(Zdero et al., Pyrone derivatives from Podolepis hieracioides and sesquiterpene acids from Cassinia longifolia, Phytochemistry, 26, 187-190, 1986; (+)-LRAPCIMS m / z (rel. int.): [M+H + ] C 20 H 33 Calculated O4, 337.238, 337 (100)

[0334] Samples of these three fractions of podopyrones (1c) and (1l) and #68N were subjected to bioassays against cotton aphids, as previously described.

[0335] The results (Table 10) indicate that podopyrone is the active metabolite in the methanol extract and N fraction of P. jaceoides. Mortality increased with increasing podopyrone concentration, with 100% mortality at 0.0625% w / v. The most active fraction was from 19 to 30 minutes, followed by 14 to 19 minutes. These two fractions contain podopyrone. [Table 10]

[0336] Example 11: Mechanism of Action - Ion Flux Response An electrophysiology technique known as microelectrode ion flux estimation (MIFE) was used to measure the net ion flux of a cell community (using Schneider's Drosophila cell line, D.mel-S1). Cells were measured under control conditions for 15 minutes, followed by an additional 25–35 minutes for each treatment. A chamber was used to hold a very small amount of γ-pyrone for treatment, with a known amount of cells deposited on a glass platform to measure equal population sizes for each recording.

[0337] An ion-selective microelectrode was placed near the surface of the cell population (40 μm at position 1 - first position) and slowly removed from the surface (80 μm at position 2 - second position) using a computer-driven micromanipulator without disturbing the surrounding solution. The change in ion concentration (Δ) between the two positions was used to calculate the net ion influx or efflux by the cells, providing an indication of which ion challenges were concentration-pumped, cotransported, or undergoing passive diffusion after treatment.

[0338] The results indicated that γ-pyrones induce a neurotoxic mode of action.

[0339] Figures 1-3 show MIFE traces of net potassium, sodium, and chloride fluxes for comparison: A) shows the effect of the negative control DMSO at 1.0%, B) shows the positive control pyrethrum at 0.01%, and C) shows the positive control plant extract tasmanone at 0.01%.

[0340] Figures 4-6 show the net ion flux measured with the extract of the present invention, compared to a positive control for comparison.

[0341] Using D.mel-S2 cells, the γ-pyrone-containing non-polar extract used in Examples 1 to 9 exhibited multiple neuronal activities, resulting in the efflux of both potassium and sodium ions and the influx of chloride ions.

[0342] γ-pyrone-containing non-polar extracts exhibited significant K-like activity similar to that of pyrethrum. + Effluent concentration and Cl - This causes inflow concentration and also significant Na + It induces effluent concentrations that are superior to pyrethrum in both potency and persistence with a dual mechanism of action (Figures 4-6).

[0343] The ability of this extract to induce multiple ion flux responses is surprising as it suggests multiple mechanisms of action. This is due to the persistent Na+ concentration during the stabilization phase of the assay. + Together with the efflux, this indicates that this extract cannot be readily metabolized by cells for detoxification.

[0344] Those skilled in the art will appreciate that the invention described herein is susceptible to variations and modifications other than those specifically described, and the invention includes all such variations and modifications within its spirit and scope.

Claims

1. An insecticidal composition comprising γ-pyrone, wherein the γ-pyrone is represented by the general formula (3): 【Chemistry 1】 [During the ceremony, n is 6, 7 or 8; R １ is C １ -C １２ is alkyl; and R ５ , R ６ and R ７ are each independently —C═O and —CH ２ - is selected from, However, R ５ , R ６ and R ７ If one of the groups is -C=O, the remaining group is -CH ２ - is. The insecticidal composition according to claim 1, 2. The insecticidal composition of claim 1, wherein R 1 is C 1 -C 2 alkyl.

3. 3. The insecticidal composition according to claim 1, wherein the γ-pyrone is selected from the group consisting of compounds (1a) to (1q), salts or solvates thereof. 【Chemistry 2】 【Transformation 3】

4. γ-pyrone is a compound 【Chemistry 4】 2. The insecticidal composition of claim 1, selected from the group consisting of salts or solvates thereof.

5. γ-pyrone 【Transformation 5】 2. The insecticidal composition of claim 1, which is a salt or solvate thereof.

6. γ-pyrone 【Transformation 6】 2. The insecticidal composition of claim 1, which is a salt or solvate thereof.

7. An insecticidal composition according to any one of claims 1 to 6, further comprising one or more agriculturally acceptable diluents and / or carriers and / or other additives selected from emulsifiers, wetting agents, surfactants, stabilizers and spreading agents.

8. An insecticidal composition according to any one of claims 1 to 7, wherein γ-pyrone is present as an extract.

9. The insecticidal composition of claim 8, wherein the extract is derived from the genera Podorepis, Gonistylus, and combinations thereof.

10. 10. The insecticidal composition of any one of claims 1 to 9, which is water-based, solvent-based or oil-based.

11. An insecticidal composition according to any one of claims 1 to 10, in the form of a dosage formulation of a solution, emulsion, suspension, emulsifiable concentrate, spray powder, paste, soluble powder, dusting agent, granule or foam.

12. 12. The insecticidal composition of claim 11 for controlling pests selected from cotton bollworms, native budworms, mirid bugs, aphids, southern green stink bugs, apple dimple bugs, thrips (plaque thrips, tobacco thrips, onion thrips, and western flower thrips), whiteflies, two-spotted spider mites, fleas, lice, mosquitoes, flies, tsetse flies, ants, ixodid mites, mites, silverfish, sand fleas, and the like.

13. 1. A formulation for controlling pests comprising: one or more agriculturally acceptable diluents and / or carriers and / or other additives such as emulsifiers, wetting agents, surfactants, stabilizers, spreading agents, etc.; and An insecticidal composition according to any one of claims 1 to 12 in an insecticidally effective amount. wherein, upon use, the formulation exerts insecticidal activity and / or repels pests and / or deters pest oviposition and / or influences oviposition sites and / or deters pests from feeding on the plant.

14. 13. Use of an insecticidally effective amount of an insecticidal composition according to any one of claims 1 to 12 to control pests by exerting insecticidal activity and / or repelling pests and / or inhibiting pest oviposition and / or influencing oviposition sites and / or inhibiting pest feeding on plants.

15. 13. A method for controlling one or more pests by exerting insecticidal activity and / or repelling the pests and / or inhibiting pest oviposition and / or affecting oviposition sites and / or inhibiting pest feeding on plants, the method comprising treating a locus with an insecticidally effective amount of the pesticidal composition of any of claims 1 to 12.

16. The method of claim 15, wherein the insecticidal composition of any one of claims 1 to 12 affects at least two ion channels in the pest.

17. The method of claim 15, wherein the insecticidal composition of any one of claims 1 to 12 affects at least two neural ion channels in the pest.

18. 17. The method of claim 15 or 16, wherein the at least two ion channels are selected from the group consisting of sodium, potassium and chloride ion channels.

19. The method of claim 18, wherein the insecticidal composition of any one of claims 1 to 12 effluxes sodium and potassium ions.

20. The method of claim 18, wherein the insecticidal composition of any one of claims 1 to 12 induces an influx of chloride ions.

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