THEIR USE IN INSECT CONTROL

TECHNICAL FIELD

This invention relates to insect control. More specifically the invention relates to methods and compositions for control of insects, including the cabbage looper, Trichoplusia ni.

BACKGROUND

Insecticides are chemicals used against insects. They include ovicides and larvicides used against the eggs and larvae of insects, respectively, but many insecticides kill all life stages of target insects. Insecticides are used in agriculture, medicine, industry, and general home use. The use of insecticides is believed to be one of the major factors behind the increase in agricultural productivity in the 20 ^th century.

Synthetic insecticides however may cause acute or delayed health affects in humans, and other non-target organisms, including wildlife and beneficial insects, who can be exposed. Insecticide exposure can cause a variety of adverse health effects. Furthermore certain synthetic insecticides are persistent organic pollutants that contribute to environmental contamination.

Widely distributed, the cabbage looper Trichoplusia ni (Γ. Ni) is considered an important field and greenhouse pest in vegetable crop production. This species is a generalist and attacks a variety of crops including lettuce, beets, turnip, spinach, brussel sprouts, peas, celery, tomatoes, rape, tobacco, certain ornamentals, many weedy plants, as well as cruciferous plants.

The mated females deposit dome-shaped, pale green eggs singly on the host-plants, chiefly at night. After hatching, the destructive larval stage reaches full development in two to four weeks; pupation then occurs and in almost 10 days the new adults emerge. In general, the larval stages damage the crop. The first two larval stages feed on the lower side of the leaf, eating through the upper epidermis, leaving "windows" in the leaf. Older larvae chew larger holes in the leaves, often resulting in extensive damage to leaves. Although this pest generally damages leaves, damage has been reported on fruits and flowers of various host plants. Three or more generations can be produced each season, depending on the latitude ² ^.

The loopers overwinter in the pupal stage, the pupae enclosed in flimsy silken cocoons attached to the food plants or to nearby objects. Cabbage loopers do not generally overwinter in Canada and migrate in from the south. However, they can overwinter in or around greenhouses.

Chemosensory input from contact chemosensilla present on the tarsi, antennae, and other parts of the body, such as the ovipositor, affects feeding and oviposition behaviors in cabbage looper as well as other phytophagous insects. Based on the sensory information received, an insect can choose a proper feeding or an oviposition site.

T. ni has developed resistance to a number of commercial insecticides, including early generation insecticides such as DDT, carbaryl, parathion3°, as well as more modern insecticides such as methomyl and Bt (Bacillus thuringiensis toxin), a widely used benign and specific insecticide against moth pests that are in the larval stagess ¹ .

Plants may harbor many epiphytic fungi that live on their surfaces or inside their cells. ¹ - ² A fine balance exists between a stable mutualistic plant fungal relationship, where both partners benefit from the association, and a pathogenic relationship, where the fungus overwhelms the plant's defenses leading to a disease state that can result in localized plant tissue damage or even death.3 Fungal secondary metabolites play a variety of important roles in epiphytic relationships.^ For example, it has been shown that loline alkaloids produced by the fungal epiphyte Epichloe festucae protect host Festuca and Lolium spp. grasses from insect predators such as aphids. ⁶

Epiphytic fungi are often uniquely adapted to the niche habitats of their specific plant hosts.7.8.9 Sri Lanka has a high rate of endemic speciation in both plants and microbes, ¹⁰ which offers great potential for the discovery of new epiphytic relationships that might generate novel secondary metabolites with medicinal or agricultural utility. ¹¹ - ¹² _' ^ BRIEF DESCRIPTION OF THE DRAWINGS

FIGURE l shows the structures of dhiloride compounds isolated from Penicilium purpurogenum.

FIGURE 2 shows the compounds obtained by modification of isolated dhilirolides.

FIGURE 3 shows a proposed biogenetic pathway for the dhilirolides showing representative examples of each of the new dhilirane, isodhilirane, 14,15-dinordhilirane, and 23,24-dinorisodhilirane skeletons, wherein the labeling pattern indicating intact acetate units shown for dhilirolide A (1) was determined from the [i,2- ¹ 3C2]acetate feeding study and an incredible natural abundance double quantum transfer

experiment. (INADEQUATE) experiment and is consistent with the proposed biogenesis as illustrated.

SUMMARY OF THE INVENTION

The present invention is based, in part, on the fortuitous discovery that certain meroterpenoid compounds showed significant feeding inhibition and sublethal developmental disruption in the cabbage looper, Trichoplusia ni. Furthermore, it was found that the compounds described herein may be useful in controlling insect infestations. Such compounds may therefore have important utility as an insecticide or as an insect repellent. Such compounds may have particular utility against the cabbage looper and other crop pests of the insect family Noctuidae.

In accordance with a first embodiment, there is provided a method of insect control, the method including, applying an effective amount of a compound of Formula I, Formula II and/or Formula III to an edible or non-edible plant or its surroundings, wherein the compound of Formula I has the following structure: FORMULA II;

In accordance with a further embodiment, there is provided a method of insect control, the method including, applying an effective amount of a compound of Formula I, to an edible or non-edible plant or its surroundings, wherein the compound of Formula I has the following structure:

Formula I.

Wherein: A may be C(Me)(OH), C(R ^a ) ₂ or C(Me), wherein when A is C(Me) there is a double bond between carbon 14 and carbon 3. Alternatively, A may be absent wherein carbon 3 and carbon 13 are directly bonded to another to form a six membered ring. R ¹ may be selected from H, OH, OR ^b , NR ^b ₂ , SR ^b , SiR ^b ₃ , F, Cl, Br, I, =O, =S. R ² may be H, OH, R ^c , or may be absent if R ¹ is =O or on

13) and R4 (carbon 9) through bridging moiety 1: bri ging moiety 1, wherein Xi may be selected from O, S, and CH ₂ , X ₂ may be selected from O, S, and CH ₂ and R ^! 7 may be H, CH ₃ , or R ^d . R3 may be H, OH, C(O)CH _3l OC(O)CH ₃ , R ^e , =O, or =S. R may be H, OH, C(O)CH ₃ , OC(O)CH ₃ , R ^f or may be absent if R3 is =O or =S.

Alternatively, R4 may be linked to R ² (carbon 10) and Rs (carbon 13) through bridging moiety 1. Alternatively, R4 may be linked to R ¹¹ (carbon 5) by X ₃ , wherein X ₃ may be O, S, or CH ₂ . Alternatively, R3 may be linked to R7 (carbon 8) throu h bridging moiety 2:

or =S, R ¹ * may be H, OH, R ^h or may be absent if is =O or =S, R*s may be H, OH, R =0, or =S, R ¹⁶ may be H, OH, R> or may be absent if R*s is =O or =S, X may be O, S, or CH ₂ . Rs may be H, C(O)CH ₃ , R ^k , or may be linked to R ² (carbon 10) and R4 (carbon 9) through bridging moiety 1. Alternatively, Rs may be absent when there is a double bond between carbon

13 and carbon 12. R ⁶ may be =CH ₂ or an epoxide ring as follows: . R7 may be H, OH, R ^M , C(O)OCH ₃ or may be linked to R3 (carbon 9) through bridging moiety 2. R ⁸ may be H or R ^N . R9 may be H, OH, R°, or may be absent if there is a double bond between carbon 2 and carbon 3. R ¹⁰ is H, RP or is absent if R ¹¹ is =O or =S or if there is a double bond between carbon 5 and carbon 6. R ¹¹ may be H, OH, =O, =S or R ^Q .

Alternatively, R ¹¹ may be linked to R ¹² (carbon 4) by X4, wherein X ₄ may be O, S, or CH ₂ , or may be linked to R4 (carbon 9) by X ₃ . Alternatively, R ¹¹ may be absent if there is a double bond between carbon 4 and carbon 5 or between carbon 5 and carbon 6. R ¹² may be absent if there is a double bond between carbon 3 and carbon 4 or between carbon 4 and carbon 5. Alternatively, R ¹² may be linked to R ¹¹ (carbon 5) via X4, wherein X4 may be O, S, or CH ₂ , forming a three membered ring or R ¹² may be linked to * (carbon 9) via X ₅ , wherein X ₅ may be O, S, or CH ₂ . R ^L8 may be H or R ^R . R ¹⁸ may be absent if there is a double bond between carbon 5 and carbon 6. R ¹ ? may be H or R ^S . R ² ° may be H or R*. Alternatively, R ²⁰ may be absent if there is a double bond between carbon 13 and carbon 12. R ²¹ may be H or R ^U .

R ^A may be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted 1 to 20 carbon linear, branched or cyclic, saturated or unsaturated alkyl, wherein alkyl carbon atoms may be replaced by o to 10 oxygen, o to 10 sulfur, and o to 10 nitrogen atoms. R ^B may be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted 1 to 20 carbon linear, branched or cyclic, saturated or unsaturated alkyl, wherein alkyl carbon atoms may be replaced by o to 10 oxygen, o to 10 sulfur, and o to 10 nitrogen atoms. R ^C may be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted 1 to 20 carbon linear, branched or cyclic, saturated or unsaturated alkyl, wherein alkyl carbon atoms may be replaced by o to 10 oxygen, o to 10 sulfur, and o to 10 nitrogen atoms. R ^D may be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted 1 to 20 carbon linear, branched or cyclic, saturated or unsaturated alkyl, wherein alkyl carbon atoms may be replaced by O to 10 oxygen, o to 10 sulfur, and o to 10 nitrogen atoms. R ^e may be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted l to 20 carbon linear, branched or cyclic, saturated or unsaturated alkyl, wherein alkyl carbon atoms may be replaced by o to 10 oxygen, o to 10 sulfur, and o to 10 nitrogen atoms. R ^f may be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted 1 to 20 carbon linear, branched or cyclic, saturated or unsaturated alkyl, wherein alkyl carbon atoms may be replaced by o to 10 oxygen, o to 10 sulfur, and o to 10 nitrogen atoms. Rs may be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted 1 to 20 carbon linear, branched or cyclic, saturated or unsaturated alkyl, wherein alkyl carbon atoms may be replaced by o to 10 oxygen, o to 10 sulfur, and o to 10 nitrogen atoms. R ^h may be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted 1 to 20 carbon linear, branched or cyclic, saturated or unsaturated alkyl, wherein alkyl carbon atoms may be replaced by O to 10 oxygen, o to 10 sulfur, and o to 10 nitrogen atoms. R ^j may be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted 1 to 20 carbon linear, branched or cyclic, saturated or unsaturated alkyl, wherein alkyl carbon atoms may be replaced by o to 10 oxygen, o to 10 sulfur, and o to 10 nitrogen atoms. Ri may be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted 1 to 20 carbon linear, branched or cyclic, saturated or unsaturated alkyl, wherein alkyl carbon atoms may be replaced by o to 10 oxygen, o to 10 sulfur, and o to 10 nitrogen atoms. R ^k may be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted 1 to 20 carbon linear, branched or cyclic, saturated or unsaturated alkyl, wherein alkyl carbon atoms may be replaced by 0 to 10 oxygen, o to 10 sulfur, and o to 10 nitrogen atoms. R ^m may be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted 1 to 20 carbon linear, branched or cyclic, saturated or unsaturated alkyl, wherein alkyl carbon atoms may be replaced by o to 10 oxygen, o to 10 sulfur, and o to 10 nitrogen atoms. R ⁿ may be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted 1 to 20 carbon linear, branched or cyclic, saturated or unsaturated alkyl, wherein alkyl carbon atoms may be replaced by o to 10 oxygen, o to 10 sulfur, and o to 10 nitrogen atoms. R° may be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted 1 to 20 carbon linear, branched or cyclic, saturated or unsaturated alkyl, wherein alkyl carbon atoms may be replaced by o to 10 oxygen, o to 10 sulfur, and o to 10 nitrogen atoms. RP may be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted l to 20 carbon linear, branched or cyclic, saturated or unsaturated alkyl, wherein alkyl carbon atoms may be replaced by o to 10 oxygen, o to 10 sulfur, and o to 10 nitrogen atoms. R¾ may be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted 1 to 20 carbon linear, branched or cyclic, saturated or unsaturated alkyl, wherein alkyl carbon atoms may be replaced by o to 10 oxygen, o to 10 sulfur, and o to 10 nitrogen atoms. R ^r may be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted 1 to 20 carbon linear, branched or cyclic, saturated or unsaturated alkyl, wherein alkyl carbon atoms may be replaced by o to 10 oxygen, o to 10 sulfur, and o to 10 nitrogen atoms. R ^s may be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted 1 to 20 carbon linear, branched or cyclic, saturated or unsaturated alkyl, wherein alkyl carbon atoms may be replaced by o to 10 oxygen, o to 10 sulfur, and o to 10 nitrogen atoms. R ¹ may be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted 1 to 20 carbon linear, branched or cyclic, saturated or unsaturated alkyl, wherein alkyl carbon atoms may be replaced by o to 10 oxygen, o to 10 sulfur, and o to 10 nitrogen atoms. R ^u may be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted 1 to 20 carbon linear, branched or cyclic, saturated or unsaturated alkyl, wherein alkyl carbon atoms may be replaced by o to 10 oxygen, o to 10 sulfur, and o to 10 nitrogen atoms. R may be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted 1 to 20 carbon linear, branched or cyclic, saturated or unsaturated alkyl, wherein alkyl carbon atoms may be replaced by O to 10 oxygen, o to 10 sulfur, and o to 10 nitrogen atoms.

R ¹ may be -OH and R ² is H. R ¹ may be =0 and R ² may be absent. R ¹ may be OH and R ² ma be linked to Rs (carbon 13) and R4 (carbon 9) though bridging moiety 3:

C l3 bridging moiety 3. R3 may be OC(O)(CH ₃ ) or C(O)CH ₃ . R3 may be =O and R ma be absent. Rs may be

linked to R? (carbon 8) through bridging moiety 4: ridging moiety 4.

R4 may be H, OH or OC(O)(CH ₃ ). R4 and R ¹¹ together may be O, with O bridging carbon 5 and carbon 9. R4 and R ¹² together may be O, with O bridging carbon 4 and carbon 9. R ⁶ is CH ₂ or an epoxide ring of form -CH2-O-. R? may be C(O)(OMe). R ⁸ may be H. R may be H or OH. R9 may be absent and there is a double bond between carbon 3 and carbon 2. R ¹⁰ may be H. R ¹¹ may be OH. R ¹¹ and R ¹² may be an O that bridges carbon 5 and carbon 4. R ¹¹ and R ¹² may be absent and there may be a double bond between carbon 5 and carbon 4.

The compound may be independently selected from the following:

The compound may be a feeding deterrent. The compound may be toxic to insects. The toxicity or the feeding deterrence may be selective for insects. The toxicity or the feeding deterrence may be selective for T. ni. The toxicity or the feeding deterrence may be selective for other crop pests of the insect family Noctuidae. The compounds may be applied simultaneously or sequentially. The compound may be applied in combination with another compound or treatment. The other compound may be a feeding deterrent, a toxicant or both. The insect may be a larva or an adult. The compound may form a composition with an agriculturally acceptable carrier. The compound may be provided in a formulation selected from one or more of the group consisting of a spray, aerosol, solid, or liquid. In certain circumstances the compound may be not one of the

In accordance with a further embodiment, there is provided a method of insect control, the method comprising, applying an effective amount of an extract from a culture of P. purpurogenum to an edible or non-edible plant or its surroundings.

In accordance with a further embodiment, there is provided a compound of Formula I, Formula II and/or Formula III as described herein. In accordance with a further embodiment, there is provided a compound of Formula I, as described herein.

A may be C(Me)(OH), C(R ^a ) ₂ or C(Me). A may be C(Me)(OH). A may be C(Me)(OH) or C(Me). A may be C(Me). When A is C(Me) there may be a double bond between carbon 14 and carbon 3.

Alternatively, A may be absent wherein carbon 3 and carbon 13 are directly bonded to another to form a six membered ring as shown in Formula II.

R ¹ may be selected from H, OH, OR, NR ^b ₂ , SR ^b , SiR ^b ₃ , F, CI, Br, I, =O and =S. R ¹ may be selected from H, OH, F, Cl, Br, I, =O and =S. R ¹ may be selected from H, OH, OR ^b , NR ^b ₂ , SR ^b , SiR ^b ₃ , F, Cl, Br and I. R ¹ may be selected from H, OH, F, Cl, Br and I. R ¹ may be selected from H and OH. R ² may be H, OH, R ^c , or may be absent if R ¹ is =O or =S.

Alternatively, R ² may be linked to Rs (carbon 13) and R4 (carbon 9) through bridging

Xi may be selected from O, S, and CH ₂ ;

X ₂ may be selected from O, S, and CH ₂ ; R ¹⁷ may be H, CH ₃ , or R ^d . Xi is selected from O, S, and CH ₂ . Xi may be selected from O and CH ₂ ; X ₂ may be selected from O and CH ₂ ; R ¹⁷ may be H or CH ₃ . Xi may be selected from O, S, and CH ₂ . Xi may be O. X ₂ may be O. R ¹⁷ may be H or CH ₃ . Xi may be selected from O and S. Xi may be selected from O and CH ₂ . X ₂ may be selected from O and S. X ₂ may be selected from O and CH ₂ .

R3 may be H, OH, C(O)CH ₃₎ OC(O)CH ₃ , R ^e , =O, or =S. R3 may be H, OH, C(O)CH ₃ , OC(O)CH ₃ , =O, or =S. R3 may be H, OH, C(O)CH _{3> or} OC(O)CH ₃ . Rs may be H, OH, R ^e , =O, or =S. R3 may be H, OH, C(O)CH ₃₎ OC(O)CH ₃ , R* or =O.

R4 may be H, OH, C(O)CH ₃ , OC(O)CH ₃ , or R ^f . R may be absent if Rs is =O or =S. R4 may be H, OH, C(O)CH ₃ or OC(O)CH ₃ . R may be H or OH. R may be C(O)CH ₃ or OC(O)CH ₃ . R4 may be H, C(O)CH ₃ or OC(O)CH ₃ . R4 may be OH, C(O)CH ₃ or

OC(O)CH ₃ . Alternatively, R4 may be linked to R ² (carbon 10) and Rs (carbon 13) through bridging moiety 1. Alternatively, R4 may be linked to R ¹¹ (carbon 5) by X ₃ , wherein X ₃ is O, S, natively, R3 may be linked to R7 (carbon 8) through

bridging moiety 2: wherein may be H, OH, Rs, =O, or =S, R ¹ may be

H, OH, R ^h or may be absent if R*3 is =O or =S; R^ may be H, OH, R =0, or =S, R ¹⁶ may be H, OH, R ^j or may be absent if R*s is =O or =S; and X may be O, S, or CH ₂ . R^ may be H, OH, =O, or =S. R! may be H, OH, or may be absent if R^ is =O or =S. R¾ may be H, OH, =O, or =S. R ¹⁶ may be H, OH, or may be absent if R¾ is =O or =S. X may be O, S, or CH ₂ . X maybe O. X maybe S. X may be CH ₂ .

Rs may be H, C(O)CH ₃ , R ^k , or is linked to R ² (carbon 10) and R4 (carbon 9) through bridging moiety 1. Rs may be H, C(O)CH ₃ , or is linked to R ² (carbon 10) and R4 (carbon 9) through bridging moiety 1. Rs may be H or C(O)CH ₃ . Alternatively, Rs may be absent when there is a double bond between carbon 13 and carbon 12.

R ⁶ may be =CH ₂ or an epoxide ring as follows: . R6 _ma y be =CH ₂ . R ⁶ may be an

21 epoxide ring as follows: O

R7 may be H, OH, R ^m , C(O)OCH ₃ or is linked to R3 (carbon 9) through bridging moiety

2. R? may be H, OH, C(O)OCH ₃ or is linked to R3 (carbon 9) through bridging moiety 2. R7 may be H, OH or C(O)OCH ₃ .

R ⁸ may be H or R ⁿ . R ⁸ may be H. R ⁸ maybe R ⁿ .

R9 may be H, OH, R°, or is absent if there is a double bond between carbon 2 and carbon

3. R9 may be H, OH, or is absent if there is a double bond between carbon 2 and carbon 3. R9 may be H or OH. R ¹⁰ may be H, RP or may be absent if R ¹¹ is =O or =S or if there is a double bond between carbon 5 and carbon 6. R ¹⁰ may be H or may be absent if R ¹¹ is =0 or =S or if there is a double bond between carbon 5 and carbon 6. R ¹⁰ may be H.

R ¹¹ may be H, OH, =O, =S or R ^q . R" may be H, OH, =O, or =S. R ¹¹ may be H, OH, or =O. R ¹¹ may be H or OH. Alternatively, R ¹¹ may be linked to R ¹² (carbon 4) by X4, wherein X4 may be O, S, or CH _2> or may be linked to R4 (carbon 9) by X ₃ . Alternatively, R ¹¹ may be absent if there is a double bond between carbon 4 and carbon 5 or between carbon 5 and carbon 6. X4 may be O, S, or CH ₂ . X ₄ may be O or S. X4 may be O or CH ₂ . X4 may be O. X4 may be S. X4 may be CH ₂ .

R ¹² may be absent if there is a double bond between carbon 3 and carbon 4 or between carbon 4 and carbon 5. Alternatively, R ¹² may be linked to R ¹¹ (carbon 5) via X4, wherein X4 may be O, S, or CH ₂ , forming a three membered ring or R ¹² is linked to R4 (carbon 9) via X ₅ , wherein X ₅ is O, S, or CH ₂ . X ₅ may be O or S. X ₅ may be O or CH ₂ . X ₅ may be O. X ₅ may be S. X ₅ may be CH ₂ .

R ¹⁸ may be H or R ^r or R ^l8 is absent if there is a double bond between carbon 5 and carbon 6. R ¹⁸ may be H or R ^r . R ^l8 maybe H. R ^l8 maybe R ^r . R ¹⁸ may be absent if there is a double bond between carbon 5 and carbon 6.

R^ maybe H or R ⁵ . R^ maybe H. R^ maybe R ⁸ .

R ²⁰ may be H or R ^l or may be absent if there is a double bond between carbon 13 and carbon 12. R ² ° may be H or may be absent if there is a double bond between carbon 13 and carbon 12. R ² ° may be R ^l or may be absent if there is a double bond between carbon 13 and carbon 12. R ² ° may be absent if there is a double bond between carbon 13 and carbon 12. R ² ° maybe H. R^ maybe R*.

R ¹ maybe H or R ^u . R ²¹ may be H. R ²¹ maybe R ^u .

Provided that the compound is not one of the following:

l6

In accordance with a further embodiment, there is provided an agricultural chemical formulation comprising a compound described herein and an agriculturally acceptable carrier.

In accordance with a further embodiment, there is provided a use of the agricultural chemical formulation described herein, for the control of insects. In accordance with a further embodiment, there is provided a use of a compound described herein, for the control of insects.

In accordance with a further embodiment, there is provided a use of a compound described herein, for the preparation of an agricultural chemical for the control of insects.

The insect control may be via a deterrent to feeding. The insect control may be selective for insects. The insect control may be selective for T. ni. The insect control may be via developmental disruption.

In a further embodiment compounds of Formulas II and III are shown where A in Formula I is absent (i.e. Formula II) and present (Formula III):

FORMULA III; wherein,

R ²² may be CH ₃ , OH, R ^a or may absent when there is a double bond between carbon 14 and carbon 3. R¾ may be CH ₃ , OH, or R ^a . R ² 3 maybe CH ₃ . R ² 3 may be OH. R 3 may be Ra.

In alternative embodiments, two or more compounds described herein may be applied simultaneously or sequentially. In alternative embodiments, a compound of Formula I may be applied in combination with another compound or treatment, such as one or more of an oviposition deterrent, an oviposition stimulant, a feeding deterrent, a feeding stimulant, an attractant, or a toxicant.

In alternative embodiments, the insect may be a larva or may be an adult, e.g. a female adult or a male adult. In alternative embodiments, the site of interest may be a plant or part thereof such as a cultivated plant within the host range of a pest insect, such as T. ni.

R ^{a u} may independently be H, a substituted or an unsubstituted aryl, or a substituted or an unsubstituted l to 20 carbon linear, branched or cyclic, saturated or unsaturated alkyl, wherein alkyl carbon atoms may be replaced by O to 10 oxygen, o to 10 sulfur, and O to 10 nitrogen atoms. For example, a substituted or an unsubstituted 1 to 20 carbon linear, branched or cyclic, saturated or unsaturated alkyl, wherein alkyl carbon atoms may be replaced by o to 10 oxygen atoms may include -C(0)OCH ₃ , -OC(0)CH ₃ , -OAc, - CH ₃ , =CH ₂ , =0, or epoxide ring. R ^{a u} may independently be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted 1 to 15 carbon linear, branched or cyclic, saturated or unsaturated alkyl, wherein alkyl carbon atoms may be replaced by o to 10 oxygen, o to 10 sulfur, and o to 10 nitrogen atoms. R ^{a u} may independently be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted 1 to 14 carbon linear, branched or cyclic, saturated or unsaturated alkyl, wherein alkyl carbon atoms may be replaced by o to 10 oxygen, o to 10 sulfur, and o to 10 nitrogen atoms. R ^{a u} may independently be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted 1 to 13 carbon linear, branched or cyclic, saturated or unsaturated alkyl, wherein alkyl carbon atoms may be replaced by o to 10 oxygen, o to 10 sulfur, and o to 10 nitrogen atoms. R ^{a u} may independently be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted 1 to 12 carbon linear, branched or cyclic, saturated or unsaturated alkyl, wherein alkyl carbon atoms may be replaced by o to 10 oxygen, o to 10 sulfur, and o to 10 nitrogen atoms. R ^{a u} may independently be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted 1 to 11 carbon linear, branched or cyclic, saturated or unsaturated alkyl, wherein alkyl carbon atoms may be replaced by o to 10 oxygen, o to 10 sulfur, and o to 10 nitrogen atoms. R ^{a u} may independently be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted 1 to 10 carbon linear, branched or cyclic, saturated or unsaturated alkyl, wherein alkyl carbon atoms may be replaced by o to 10 oxygen, o to 10 sulfur, and o to 10 nitrogen atoms. R ^{a u} may independently be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted 1 to 10 carbon linear, branched or cyclic, saturated or unsaturated alkyl, wherein alkyl carbon atoms may be replaced by o to 3 oxygen, o to 3 sulfur, and o to 3 nitrogen atoms. R ^{a u} may independently be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted l to 10 carbon linear, branched or cyclic, saturated or unsaturated alkyl, wherein alkyl carbon atoms may be replaced by o to 10 oxygen atoms. R ^{a u} may independently be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted l to 10 carbon linear, branched or cyclic, saturated or unsaturated alkyl. R ^{a u} may independently be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted l to 9 carbon linear, branched or cyclic, saturated or unsaturated alkyl. R ^{a u} may independently be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted 1 to 7 carbon linear, branched or cyclic, saturated or unsaturated alkyl. R ^{a u} may independently be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted 1 to 6 carbon linear, branched or cyclic, saturated or unsaturated alkyl. R ^{a u} may independently be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted 1 to 9 carbon linear, branched or cyclic, saturated or unsaturated alkyl. R ^{a u} may independently be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted 1 to 5 carbon linear, branched or cyclic, saturated or unsaturated alkyl. R ^{a u} may independently be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted 1 to 4 carbon linear, branched or cyclic, saturated or unsaturated alkyl. R ^{a u} may independently be H, a substituted or an unsubstituted aryl, a substituted or an unsubstituted 1 to 3 carbon linear, branched or cyclic, saturated or unsaturated alkyl. R ^{a u} may independently be H or a substituted or an unsubstituted 1 to 10 carbon linear, branched or cyclic, saturated or unsaturated alkyl.

Extracts of laboratory cultures of the fungus Penicilium purpurogenum obtained from rotting fruit of the tree Averrhoa bilimbi growing in Sri Lanka have yielded the meroterpenoids, dhilirolides A (1) - N (14), as described in FIGURE 1. The structures of the dhilirolides have been elucidated by analysis of spectroscopic data and a single crystal x-ray diffraction analysis of dhilirolide L (12). Dhilirolides A (1) to N (14) represent the four unprecedented and rearranged dhilirane, isodhilirane, bisnordhilirane, and bisnorisodhilirane meroterpenoid carbon skeletons. Stable isotope feeding studies have confirmed the meroterpenoid biogenetic origin of the dhilirolides and provided support for a proposed genesis of the new carbon skeletons. Dhilirolide L (12) showed significant feeding inhibition and sublethal developmental disruption in the cabbage looper Trichoplusia ni, an important agricultural pest, at low concentrations. The compound may have the following structure:

Various alternative embodiments and examples are described herein. These embodiments and examples are illustrative and should not be construed as limiting the scope of the invention.

DETAILED DESCRIPTION OF THE INVENTION

As part of an ongoing program aimed at exploring the secondary metabolites of epiphytic fungi found on Sri Lankan plants, we have investigated the fungus Penicilium purpurogenum isolated from fruits of the tree Averrhoa bilimbi. ^ The mature fruit of A. bilimbi is susceptible to infection by P. purpurogenum, which can overwhelm the plant's defenses and lead to complete decay of individual pieces of fruit.

We recently reported that cultures of P. purpurogenum produce dhilirolides A (l) to D (4), four meroterpenoids representing the unprecedented dhilirane and isodhilirane carbon skeletons. ¹ Continued chemical investigation of laboratory cultures of P. purpurogenum has resulted in the isolation of new meroterpenoids dhilirolides E-N (5- 14), which have either the dhilirane (A) and isodhilirane (B) carbon skeletons or the unprecedented bisnordhilarane (C) and bisnorisodhilarane (D) carbon skeletons.

The dhilirolides were found to have promising feeding inhibition and developmental disruption activities against the cabbage looper Trichoplusia ni (Lepidoptera: Noctuidae), an economically important agricultural pest. Details of the isolation and structure elucidation of the new dhilirolides E-N (5-14), a stable isotope feeding study that has provided insight into the biogenesis of the new carbon skeletons, and the insecticidal activities of the dhilirolides are presented below.

As used herein applying an effective amount of a compound of Formula I to an edible or non-edible plant or its surroundings, is meant to include spraying of insect pests directly, treating plants potentially infested with the insect pests, or treating surrounding soil or cultivation medium of the plants, in an amount sufficient to deter insect habitation on the plants or surroundings.

The compositions described herein, in various embodiments, are meant to be sprayable and may be sprayed onto a desired location, or formulated as a concentrate that is sprayable on addition of agriculturally acceptable carriers and/or spray adjuvants. Preferred compositions of the invention are shelf stable. The term "shelf stable" is intended to mean that a composition of the invention does not separate out into separate phases or experience significant reduction in activity.

The term "agriculturally acceptable carrier" intended to include any material that facilitates application of a composition of the invention to the desired location (for example, plants, soil, insects etc.), which may for example be a plant, plant material or equipment, or that facilitates storage, transport or handling. Carriers used in compositions for application to plants and plant material are preferably non-phytotoxic or only mildly phytotoxic. A suitable carrier may be a solid, liquid or gas depending on the desired formulation. In one embodiment preferred carriers include polar liquid carriers such as water, mineral oils and vegetable oils. Pest insects are insects that may damage or kill agricultural crops, ornamental plants, or native plants, or may consume or damage harvested food, or may cause illness or unproductivity in agricultural animals or vector human diseases or cause pain. Some insects may be beneficial at one stage of life and a pest at another stage.

Pest insects of concern in agricultural applications include: Acalymma, Acyrthosiphon pisum, African armyworm, Africanized bee, Agromyzidae, Agrotis munda, Agrotis porphyricollis, Aleurocanthus woglumi, Aleyrodes proletella, Alphitobius diaperinus, Altica chalybea, Anasa tristis, Anisoplia austriaca, Anthonomus pomorum, Anthonomus signatus, Aonidiella aurantii, Apamea apamiformis, Apamea niveivenosa, Aphid, Aphis gossypii, Apple maggot, Argentine ant, Army cutworm, Arotrophora arcuatalis, Asparagus miner, Asterolecanium coffeae, Athous haemorrhoidalis, Aulacophora, Australian plague locust, Bactericera cockerelli, Bactrocera, Bactrocera correcta, Bagrada hilaris, Banded hickory borer, Banksia, Boring Moth, Beet armyworm, Black bean aphid, Blepharidopterus chlorionis, Bogong moth, Boll weevil, Brassica pod midge, Brevicoryne brassicae, Brown locust, Brown marmorated stink bug, Brown planthopper, Cabbage Moth, Cabbage worm, Callosobruchus maculatus, Cane beetle, Carrot fly, Cecidomyiidae, Ceratitis capitata, Cereal leaf beetle Chlorops pumilionis, Citrus long- horned beetle, Coccus viridis, Codling moth, Coffee borer beetle, Colorado potato beetle, Confused flour beetle, Crambus, Cucumber beetle, Curculio nucum, Curculio occidentis, Cutworm, Cyclocephala borealis, Dark Sword-grass, Date stone beetle, Delia (genus), Delia antiqua, Delia floralis, Delia radicum, Desert locust, Diabrotica, Diabrotica balteata, Diabrotica speciosa, Diamondback moth, Diaphania indica, Diaphania nitidalis, Diaphorina citri, Diaprepes abbreviatus, Diatraea saccharalis, Differential grasshopper, Dociostaurus maroccanus, Drosophila suzukii, Dryocosmus kuriphilus, Earias perhuegeli, Epicauta vittata, Epilachna varivestis, Erionota, thrax, Eriosomatinae, Euleia heraclei, Eumetopina flavipes, Eupoecilia ambiguella, European Corn Borer, Eurydema oleracea Eurygaster integriceps, Forest bug, Frankliniella tritici, Galleria mellonella, Garden Dart, Glassy-winged sharpshooter, Greenhouse whitefly, Gryllotalpa orientalis, Gryllus pennsylvanicus, Gypsy moths, Helicoverpa armigera, Helicoverpa gelotopoeon, Helicoverpa punctigera, Helicoverpa zea, Heliothis virescens, Henosepilachna vigintioctopunctata, Hessian fly, Jacobiasca formosana, Japanese beetle, Khapra beetle, Lampides boeticus, Leaf miner, Lepidiota consobrina, Lepidosaphes beckii, Lepidosaphes ulmi, Leptoglossus zonatus, Leptopterna dolabrata, Lesser wax moth, Leucoptera (moth), Leucoptera caffeina, Light brown apple moth, Lissorhoptrus oryzophilus, Long-tailed Skipper, Lygus, Lygus hesperus, Maconellicoccus hirsutus, Macrodactylus subspinosus, Macrosiphum euphorbiae Maize weevil, Manduca sexta, Mayetiola hordei, Mealybug, Megacopta cribraria, Metcalfa pruinosa, Moth, Leek moth, Myzus persicae, Nezara viridula, Oak Processionary, Olive fruit fly, Opisina arenosella, Opomyza Opomyza florum, Opomyzidae, Oscinella frit, Ostrinia furnacalis, Oxycarenus hyalinipennis, Papaya mealy bug, Papilio demodocus, Paracoccus marginatus, Paratachardina pseudolobata, Pentatomoidea, Phthorimaea operculella, Phyllophaga (genus), Phylloxera, Phylloxeridae, Phylloxeroidea, Pieris brassicae, Pink boUworm, Planococcus citri, Platynota idaeusalis, Plum curculio, Prionus californicus, Pseudococcus viburni, Pyralis farinalis, Red imported fire ant, Red locust, Rhagoletis cerasi, Rhagoletis indifferens, Rhagoletis mendax, Rhopalosiphum maidis, Rhyacionia frustrana, Rhynchophorus, ferrugineus, Rhynchophorus palmarum, Rhyzopertha, Rice Moth, Rice stink bug, Russian wheat aphid, San Jose scale, Scale insect, Schistocerca americana, Sciaridae Scirtothrips dorsalis, Scutelleridae, Serpentine leaf miner, Setaceous Hebrew character, Silver Y, Silverleaf whitefly, Sipha flava, Small hive beetle, Southwestern corn borer, Soybean aphid, Spodoptera cilium, Spodoptera litura, Spotted cucumber beetle, Squash vine borer, Stenotus binotatus, Strauzia longipennis, Striped flea beetle, Sunn pest, Sweetpotato bug, Tarnished plant bug, Thrips, Thrips angusticeps, Thrips palmi, Toxoptera citricida, Trioza erytreae, Turnip Moth, Tuta absoluta, Varied carpet beetle, Virachola isocrates, Waxworm, Western corn rootworm, Western flower thrips, Wheat fly, Wheat weevil, Whitefly, Winter Moth, and Xyleborus glabratus.

Pest insects of concern in ornamental plant applications include: Acleris variegana, Acyrthosiphon pisum, Aphid, Bird-cherry Ermine, Grapeleaf Skeletonizer, Gypsy moths, Japanese beetle, Macrodactylus subspinosus, Mealybug, Otiorhynchus sulcatus, Paratachardina pseudolobata, Paysandisia archon, Sawfly, Scale insect, Scarlet lily beetle, Sciaridae, Spodoptera cilium, Stephanitis takeyai, Tenthredo scrophulariae, Yponomeuta malinellus, and Yponomeuta padella. Pest insects that act as vectors of plant pathogens include: Acyrthosiphon pisum, Agromyzidae, Anthomyiidae, Aphid, Beet leafhopper, Brevicoryne brassicae, Cacopsylla melanoneura, Cicadulina, Cicadulina mbila, Common brown leafhopper, Curculionidae, Diabrotica balteata, Empoasca decedens, Eumetopina flavipes, Euscelis plebejus, Frankliniella tritici, Glassy-winged sharpshooter, Haplaxius crudus, Hyalesthes obsoletus, Hylastes ater, Jumping plant louse, Leaf beetle, Leafhopper, Mealybug, Melon fly, Molytinae ,Pegomya hyoscyami, Pissodes, Pissodes strobi, Pissodini, Planthopper, Pseudococcus viburni, Psylla pyri, Rhabdophaga rosaria, Rhynchophorus palmarum, Scaphoideus titanus, Scirtothrips dorsalis, Silverleaf whitefly, Tephritidae, Thripidae, Thrips palmi, Tomicus piniperda, Toxoptera citricida, Treehopper, Triozidae, and Western flower thrips.

Pest insect that act as vectors of animal pathogens include: Alphitobius diaperinus, Calliphoridae, Cat flea, Culicoides imicola, Hippelates ,Lutzomyia shannoni, Musca autumnalis, Oriental rat flea.

Pest insects that act as vectors of human pathogens include: Aedes aegypti, Aedes albopictus, Anopheles, Anopheles barberi, Anopheles crucians, Anopheles culicifacies, Anopheles dirus, Anopheles earlei, Anopheles gambiae, Anopheles punctipennis, Black fly, Calliphoridae, Culex, Culex tritaeniorhynchus, Flea, Haemagogus, Hippelates, Housefly, Lutzomyia, Mosquito, Oriental rat flea, Phlebotomus, Rhodnius prolixus, Sarcophaga peregrina, Simuliinae, Simuliini, Simulium, Simulium yahense, Triatoma, Triatoma brasiliensis, Triatoma carrioni, Triatoma dimidiata, Triatoma gerstaeckeri, Triatoma Indictiva, Triatoma infestans, Triatoma juazeirensis, Triatoma melanica, Triatoma nigromaculata, Triatoma protracta, Triatoma sanguisuga, Triatominae, and Tsetse fly.

A feeding deterrent is a compound that once probed by the insect, may cause it to stop feeding and starve to death. MATERIALS AND METHODS General Experimental Procedures.

For compounds 1-14, melting points were taken using a Fisher-Johns apparatus and the reported values are uncorrected. Optical rotations were measured using a Jasco P-1010 Polarimeter™ with sodium light (589 nm). UV spectra were recorded with a Waters 2487 Dual λ Absorbance Detector™. The lH and 13C NMR spectra were recorded on a Bruker AV-600 spectrometer™ with a 5 mm CPTCI cryoprobe. lH chemical shifts are referenced to the residual DMSO-d6 (δ 2.50 ppm) and 13C chemical shifts are referenced to the DMSO-d6 (639.5 ppm) solvent peak. Low and high resolution ESI- QIT-MS were recorded on a Bruker-Hewlett Packard 1100 Esquire-LC™ system mass spectrometer. Merck Type 5554™ silica gel plates and Whatman MKC18F™ plates were used for analytical thin layer chromatography. Reversed-phase HPLC purifications were performed on either a Waters 1525 Binary HPLC Pump™ attached to a Waters 2487 Dual λ Absorbance Detector™ or a Waters 600E System Controller™ liquid chromatograph attached to a Waters 996 Photodiode Array Detector™. All solvents used for HPLC were Fisher™ HPLC grade and were filtered through a 0.45 μπι filter (Osmonics Inc.™) prior to use.

For Compounds 15-17, the Ή, ^X 3C, and 2D NMR spectra were recorded using a Bruker AV-600™ spectrometer with a 5 mm cryoprobe. Ή chemical shifts are referenced to the residual DMSO-cfe signal (δ 2.49 ppm). ^C chemical shifts are referenced to the DMSO-cfe signal (δ 39.5 ppm). High-resolution ESI-MS (HRESIMS) data were recorded on a Bruker-Hewlett Packard™ 1100 Esquire-LC™ system mass spectrometer. Sephadex LH-20™ (3 cm x 95 cm) was used for size exclusion column chromatography. Analytical thin layer chromatography was performed on Merck Type 5554™ silica gel plates, viewed under UV-light (254 nm), and stained with p-anisaldehyde followed by heating. Reversed-phase HPLC purifications were carried out on a Waters 1525 binary HPLC™ pump attached to a Waters 486 Tunable Absorbance Detector™. An InertSustain™ C18 HPLC column or a CSC-Inertsil 150A/ODS2, 5μπι 25 x 0.95cm column, was used for HPLC purifications. A linear gradient of 40-45% MeCN/H ₂ O over 90 minutes following with 45-40% MeCN/H ₂ O over 5 minutes at a flow rate of 2 mL/min was used during all runs. All solvents used for HPLC were Fisher™ HPLC grade and were filtered before use.

Fungal Material. The fungus Penicillium purpurogenum was isolated from infected fruits oiAverrhoa bilimbi (Averrhoa bilimbi, L., (Oxalidaceae) as described previously. ¹

EXAMPLES

EXAMPLE l: Extraction of Penicillium perpurogenum and Isolation of Dhilirolides A-N (1-14).

P. purpurogenum was cultured on potato dextrose agar (50 Petri dishes) and the culture medium cut into small pieces and extracted with EtOAc. ¹ The EtOAc extract was concentrated in vacuo and chromatographed on Sephadex LH20™ (3 cm x 95 cm) using 4:1 MeOH/CH ₂ Cl ₂ . The fractions containing the compounds of interest were combined and subjected to reversed-phase Ce and ds HPLC chromatography using a CSC- Inertsil™ 150A/ODS2, 5μπι 25 x 0.94 cm column, with a linear gradient of 40-45% MeCN/H ₂ 0 over 60 min at a flow rate of 2 mL/min. Pure samples of dhilirolides A (1) (28.9 mg), B (2) (7.6 mg), C (3) (17.7 mg), D (4) (8.7 mg), F (10) (0.4 mg), J (6) (2.5 mg), L (12) (14.8 mg), M (13) (1.0 mg) and N (14) (0.5 mg) were obtained. Additional fractions containing impure samples of dhilirolides E (5), G (7), H (8), I (9) and K (11) were fractionated via isocratic Cs reversed-phase HPLC using a Phenomenex Luna™ 100 A, 5μπι 25 x l.o cm column. Compounds are listed with eluent systems and yields: 5 (29:71 MeCN/H ₂ 0, 0.9 mg), 7 (9:16 MeCN/H ₂ 0, 0.5 mg), 8 (7:13 MeCN/H ₂ 0, 1.0 mg), 9 (9:16 MeCN/H ₂ 0, 0.9 mg) and 11 (33:67 MeCN/H ₂ 0, 1.4 mg).

Dhilirolide A (1) was obtained as optically active colorless crystals (mp 267-269 °C) that gave an [M+H] ⁺ ion in the HRESIMS at m/z 473.1796 appropriate for a molecular formula of C ₂₅ H ₂ 80g, requiring 12 sites of unsaturation. The NMR spectrum obtained for 1 contained 25 resolved resonances in agreement with the HRESIMS data (not shown). A detailed analysis of the ^! H/ ^! sC/gCOSY/gHSQC/gHMBC NMR data identified five methyl singlets [δ 0.74 (Me-25), 1.18 (Me-17), 1.25 (Me-19), 1.50 (Me-18), 1.51 (Me-15)], one methyl doublet [δ 1.42 J=7.i Hz (Me-24)], a trisubstituted olefin [5 6.42 (H-2), 125.0 (C-2); 152.2 (C-3)], two ester/lactone carbonyls [8 162.0 (C-i); 170.3 (C-20)], a trisubstituted epoxide [δ 55.6 (C-4); 3.68 (H-5), 55.9 (C-5)]; a 1,1 disubstituted epoxide [δ 64.0 (C-2i); 2.36/2.90 (Η-22β/Η-22α), 45.6 (C-22)], two oxygenated tertiary carbons [δ 91.0 (C-9); 81.5 (C-16)], an oxygenated methine carbon [δ 4-73 (H-23), 80.9 (C-23)], two ketals [δ 105.7 (C-io); 107.7 (C-14)], four other quaternary carbons [δ 40.5 (C-7); 54.2 (C-8); 44.9 (C-11); 47.9 (C-13)], and two methylenes [δ 1.77/2.46 (H-6ax/H-6eq), 31 5 (C-6); 1.74/2.11 (H-i2ax/H-i2eq), 36.0 (C- 12)]. The alkene, carbonyl, and epoxide functionalities described above accounted for 5 of the 12 sites of unsaturation indicated by the molecular formula, requiring that dhilirolide A had to contain seven additional rings.

Dhilirolide B (2) was isolated as an optically active amorphous solid that gave an [M + Na] ⁺ ion at m/z 479.1725 in the HRESIMS appropriate for the molecular formula of C25H28O8, that differs from that of 1 simply by the loss of an oxygen atom. Although the Ή and NMR spectra of 2 were similar to those of 1, the UV spectrum was markedly different. In dhilirolide A (1), a Amax typical of a trisubstituted α,β-unsaturated lactone was observed at 235 nm, while, in 2, the Ama was shifted to 280 nm. Replacing the C- 4/C-5 epoxide in 1 with a double bond in 2 would account for the loss of an oxygen and extend the conjugation from the enone in 1 to a dienone in 2, satisfying the Amax observed for 2. Examination of the lD and 2DNMR data (not shown) obtained for 2 revealed that the epoxide H-5 resonance (δ 3-68) in 1 had been replaced by an olefinic resonance at δ 6.15, that showed COSY correlations to the methylene resonances at δ 1.94 and 2.92 assigned to H-6eq/H-6ax. HMBC correlations between C-4 (δ 133.4) and H-2 (δ 5.83), H-6eq/H-6ax (δ 1.94/2.92), Me-17 (δ 1.50), and Me-18 (δ 1.54), and between H-5 (δ 6.15) and both C-3 (δ 150.0) and C-16 (δ 82.o), were consistent with the proposed structure.

Dhilirolide C (3) was isolated as an optically active amorphous solid that gave an [M + Na] ⁺ ion at m/z 479.1746 in the HRESIMS consistent with the molecular formula C25H28O8, identical to the molecular formula of dhilirolide B (2). The UV spectrum observed for 3 (Amax 239 nm) was similar to that of dhilirolide A (1) (Amax 235 nm) indicating that it did not have the dienone moiety found in 2. Comparison of the lHand ^! SCNMR spectrum of dhilirolide C (3) with the corresponding spectra recorded for dhilirolide A (l) (not shown) revealed that the resonances assigned to the 1,1- disubstituted C-21/C-22 epoxide in 1 were absent. Instead, the lH NMR spectrum of 3 contained two new singlet resonances at δ 5.04 (Η-22β) and δ 5.11 (Η-22α), which both correlated to an olefinic carbon at δ 107.7 (C-22) in the HSQC spectrum and to a second olefinic carbon at δ 151.7 (C-2i) in the HMBC spectrum. These observations showed that l and 3 differed simply by the replacement of theC-2i/C-22 epoxide in 1 with a Δ ^2ΐ,22 exocyclic alkene in 3. HMBC correlations observed between the Η-22β/Η-22α olefinic methylene resonances at δ 5.04 and 5.11 and themethine carbons at δ 6ΐ·3 and 50.4, assigned toC-8 and C-11, respectively, supported this assignment. Interestingly, although all the other structural features of 1 and 3 were identical, it was found that when the lH NMR spectrum of 3 was recorded inDMSO-d6 many resonances were doubled or broadened and significantly shifted. This phenomenon was attributed to a slow conformational equilibrium. A single set of well-resolved resonances could be obtained when the NMR spectra for 3 were recorded in MeOH-d ₄ .

Dhilirolide D (4) was isolated as an optically active amorphous solid that gave an [M - H]- ion at m/z 441.1967 in the HRESIMS appropriate for a molecular formula of C ₂ 5H ₃ o0 ₇ . The molecular formula of 4 differed from the molecular formula of 1 by the addition of two hydrogen atoms and the loss of two oxygen atoms, and it required only 11 sites of unsaturation. Examination of the Ή and ^C NMR spectra recorded for dhilirolide D (4) revealed a close relationship to dhilirolides A-C (1-3), but also several significant structural and functional group differences. The UV (Ama 276 nm) and ^! H/ ^! sC/COSY/HSQC/HMBCNMR data (not shown) obtained for 4 identified the C-i to C-5 dienone substructure present in 2 and the Δ ²¹²² exocyclic alkene present in 3.

Dhilirolide E (5): Isolated as a amorphous white powder; [α] ² ¾ +3.23 (c 0.09, MeOH); UV (MeCN) (ε) 2θ6 (3103), 248 (2348) nm; Ή NMR, see Table 1; «C NMR, see Table 3; positive ion HRESIMS [M+H] ⁺ m/z 443.1956 (calcd for C2 ₅ H ₃ i0 ₇ , 443.2070).

Dhilirolide F (6): Isolated as a amorphous white solid; [0J20D +18.1 (c 0.03, MeOH); UV (MeCN) Amax (ε) 204 (7859), 240 (4037) nm; Ή NMR, see Table 1; 13C NMR, see Table 3; positive ion HRESIMS [M+Na]+ m/z 479.2233 (calcd for C ₂ 6H ₃ 20 ₇ Na, 479.2046).

Dhilirolide G (7): Isolated as a amorphous white solid; [a] ²⁰ D +8.9 (c 0.05, MeOH); UV (MeCN) Amax (ε) 204 (5575), 269 (4786) nm; Ή NMR, see Table 1; 13C NMR, see Table 3; positive ion HRESIMS [M+Na]+ m/z 495.1982 (calcd for C ₂ 6H ₃₂ 0 ₈ Na, 495-1995).

Dhilirolide H (8): Isolated as a amorphous white solid; [0J20 D +2.92 (c 0.10, MeOH); UV (MeCN) Amax (ε) 201 (1847), 240 (941) nm; Ή NMR, see Table 1; «C NMR, see Table 3; negative ion HRESIMS [M-H]- m/z 471.2014 (calcd for C26H31O8, 471.2019).

Dhilirolide I (9): Isolated as a amorphous white solid; [a] ² °D +2.89 (c 0.09, MeOH); UV (MeCN) Amax (ε) 203 (2700), 272 (2067) nm; Ή NMR, see Table 1; ¾C NMR, see Table 3; positive ion HRESIMS [M+Na]+ m/z 453.1920 (calcd for C ₂ 4H ₃ oO ₇ Na, 453.1889).

Dhilirolide J (10): Isolated as a clear crystalline solid; mp decomposed at 223-225X5 [a] ²⁰ D +5-94 (c 0.16, MeOH); UV (MeCN) Amax (ε) 2θ6 (4349), 279 (5346) nm; Ή NMR, see Table 2; «c NMR, see Table 3; positive ion HRESIMS [M+H]+ m/z 441.1918 (calcd for C ₂ 5H ₂ 9O7, 441.1913).

Dhilirolide K (11): Isolated as a amorphous white solid; [a] ²⁰ D -2.26 (c 0.09, MeOH); UV (MeCN) Amax (ε) 2θ6 (883), 249 (978) nm; Ή NMR, see Table 2; «c NMR, see Table 3; positive ion HRESIMS [M+Na]+ m/z 479.1829 (calcd for C ₂₅ H ₂ 8O8N , 479.1682).

Dhilirolide L (12): Isolated as clear prism-like crystals; mp 27i-273°C [a] ²⁰ D +182 (co.07, MeCN) UV (MeCN) Amax (ε) 2i8 (2315) nm; Ή NMR, see Table 2; «C NMR, see Table 3; positive ion HRESIMS [M+Na]+ m/z 463.1745 (calcd for C ₂₅ H ₂ 8O ₇ Na, 463.1733)·

Dhilirolide M (13): Isolated as a amorphous white solid; [α] ² ¾ +59 (c 0.02, MeOH); UV (MeCN) Amax (ε) 197 (4526), 273 (132) nm; Ή NMR, see Table 2; «C NMR, see Table 3; positive ion HRESIMS [M+Na]+ m/z 463.1740 (calcd for C2 ₅ H ₂ 8O ₇ Na, 463.1733). Dhilirolide N (14): Isolated as a amorphous white solid; [a] ² °D -1.4 (c 0.05, MeOH); UV (MeCN) Xmax (ε) 215 (2285) nm; Ή NMR, see Table 2; «C NMR, see Table 3; positive ion HRESIMS [M+Na]+ m/z 477.1570 (calcd for C ₂₅ H ₂ 6O8Na, 477.1525).

EXAMPLE 2: Modification of isolated dhilirolides to produce compounds 15-17

Various reactions conditions were utilized in an attempt to open the epoxides in dhilirolide A (1) as shown in Scheme 1. No reaction occurred when CeCl ₃ »7H ₂ O or Bi(OTf)3 was added to dhilirolide A (1). NaN ₃ reacted with dhilirolide A (1), but did not afford the expected product. Instead of opening either of the two epoxides, NaN ₃ initiated a retro-claisen type cleavage, resulting in elimination product 15. To test whether this elimination was specific to NaN ₃ , a small, hard nucleophile, NaOH was tested. Dhilirolide A (1) was completely destroyed by NaOH when the reaction mixture was stirred at either room temperature or under heat for 18 hours. Further tests would need to be made with lower concentrations of NaOH or with shorter reaction times to determine whether NaOH catalysis can give the same product. Upon treatment with perchloric acid, dhilirolide A (1) produced products 16 and 17.

No reaction material

Scheme l: Attempted epoxide ring opening reactions with dhilirolide A (1).

Scheme 2: Reaction of dhilirolide A (1) with perchloric acid. Reaction products 16 and 17 were isolated.

EXAMPLE 3: Structure elucidation of modified dhilirolide reaction product (15)

The product 15 was isolated from the reaction of dhilirolide A (1) and NaN ₃ with an HPLC retention time of 22 minutes. 15 was obtained as an amorphous white solid that gave an [M + H] ⁺ ion in the HRESIMS at m/z 431.1712 appropriate for a molecular formula of C2 ₃ H2 ₆ O ₈ that requires 11 sites of unsaturation instead of the 12 found in 1. This molecular formula differs from that of 1 by the loss of two carbons, two protons, and one oxygen. A detailed analysis of the ^! H/^C/gCOSYeo/gHSQC/gHMBC NMR data and comparison with the NMR data for dhilirolides A-E (1-5), revealed that ring A is identical to that found in 1 and ring D were the same as seen in dhilirolides D (4) and E (5). Rings B and C were the same as seen in 1, except the quaternary carbon at C-13 was changed to a tertiary carbon in 15.

Although the Ή and ¾C NMR spectra of 15 were similar to those of 1, there were several striking differences. Only five methyl peaks were present compared to the six seen with the Ή NMR of 1. Four methyl singlets [δ 0.82 (Me-25); 1.14 (Me-17); 1.23 (Me-19); 1.49 (Me-18)] and one methyl doublet [δ 1.12 J = 7.3 Hz (Me-24)] were identified, showing a missing methyl singlet (Me-15). Also, the ketal signals at C-10 and C-14 were missing in the NMR spectrum. The C- 10 ketal resonance at 6 105.7 i 1 was replaced by a ketone resonance at δ 212. o (C-10). The C- 14 ketal resonance at 5 107.7 ^m 1 was missing, which was supported by the addition of the H-13 proton resonance (δ 3.13 ddd, J = 13.2, 4.3, 1.9). The H-13 resonance had showed gHMBC correlations to C-ll (δ 50.2) and the C-19 methyl (δ 18.4), but correlated to no carbon atoms in HSQC. gCOSY6o couplings were observed between the H-13 methine proton and the olefinic H- 2 resonance (δ 6.oi) and a pair of germinal H-i2ax/H-i2eq methylene protons (δι.97/ 1.87). gHSQC correlations between the H-i2ax/H-i2eq methylene protons and C-12 (634.8) demonstrated that the H-12 protons were germinal, which suggests that an elimination had occurred to expel a fragment that included C-14 and the C-15 methyl. The proton singlet resonance at δ 7.46 did not show a gHSQC correlation to a carbon atom, so this could be assigned to a tertiary alcohol at C-9. Supporting this assignment, the OH-9 resonance showed gHMBC correlations to C-8 (δ 57.4), C-9 (δ 89.0), and the C-23 oxygenated methine carbon (δ 83.6). Analysis of the gCOSY6o and gHMBC data as illustrated in Scheme 3 established the constitution of 15. tROESY correlations were used to facilitate the assignment of the relative configuration of 15 as illustrated in Scheme 4. tROESY correlations between the H-13 methine resonance (δ 3.13) and the H-6a methylene (δ 2.96), H-i2ex methylene (δι.97), and OH-9 tertiary alcohol (δ 7.46) resonances confirmed the configuration of the key changes in this modified dhilirolide. Additional tROESY correlations between the OH-9 and H-6a and H-23 oxygenated methine proton (δ 4.6i) further confirmed the structure of 15. The absolute configuration configuration for 15 was assigned as 4S, 5R, 7S, 8R, 9R, 11R, 13R, 21R, and 23S.

Scheme 4: Selected tROESY correlations of reaction product 15 recorded at 600 MHz in DMSO-d ₆ . Reaction product (15) from reaction with dhilirolide A (1) with NaN ₃ :

Isolated as an amorphous white solid; Ή NMR, see Table 4; NMR, see Table 4; positive ion HRESIMS [M+H] ⁺ m/z 431.1712 (calcd for C23H27O8, 431.1706).

Reaction product (16) from reaction with dhilirolide A (1) with perchloric acid: Isolated as an amorphous yellow solid; Ή NMR, see Table 5; NMR, see Table 5; positive ion HRESIMS [M+H] ⁺ m/z 455.1078 (calcd for C25H2 ₇ O8, 455.1706).

Reaction intermediate (17) from reaction with dhilirolide A (1) with

perchloric acid: Isolated as an amorphous white solid; Ή NMR, see Table 5;

NMR, see Table 5; positive ion HRESIMS [M+H] ⁺ m/z 472.

Table 1. Ή NMR Data for Dhilirolides E-I (5-9) Recorded in DMSO-d ₆ .

a Multiplicity not determined due to overlapping signals/chemical shifts determined from 2D data. Table 2. Ή NMR Data for Dhilirolides J- N (10-14) Recorded in DMSO-de.

aAssignments within a column maybe interchanged

Table 3. ¾C NMR Data for Dhilirolides E-G (5-14) Recorded in DMSO-cfe.

Table . *H and ¾C NMR Data for Reaction Product 15 Recorded in DMSO-de.

Table 5. Ή and «C NMR Data for Modified Dhilirolide Reaction Product (16) and Reaction Intermediate (1 ) Recorded in DMSO-dS.

Note: The fraction containing the intermediate (17) was very dilute due to the small amounts present in the resulting reaction mixture so the carbon signals were very weak.

EXAMPLE 4: Stable Isotope Feeding Study.

For the labeling study l g of doubly labeled acetate was dissolved in 20 mL of sterile distilled H ₂ 0 and filtered through a 0.25 μπι filter under sterile conditions. This solution was then added to 250 ml of autoclaved potato dextrose agar media, which was then divided evenly between 16 Petri dishes and the P. purpurogenum cultured as described above. The resulting culture medium was extracted with EtOAc and dhilirolides A (l) (6.2 mg), B (2) (1.9 mg), C (3) (1.8 mg), D (4) (1.7 mg), L (12) (3.3 mg) and M (13) (2.3 mg) were isolated as described above.

EXAMPLE 5: Insecticidal Activity Assays.

Feeding Deterrent Bioassay: Freshly molted third instar larvae were starved for 4-5I1 prior to each bioassay. Cabbage leaf discs of 1.5 cm diameter were painted with 10 μΐ of the methanolic solution of fungal extract or the compounds on each side. Control leaf discs were painted with the carrier solvent alone. One treated and one control disc (after being dried) were placed 0.7 cm apart in each compartment [4.2/3.0 cm (length/ width)] of a plastic assay tray. A starved larva was introduced gently into the centre of each compartment using forceps and allowed to feed. The trays were covered with plastic lids. When approximately 50% of the control leaf discs had been eaten, larvae were removed from the assay trays. Areas of control and treated leaf discs eaten were measured using Scion Image software™. A feeding deterrence index (FDI) was calculated using the formula FDI = {(C-T)/(C+T)*ioo} where C and T are the control and treated leaf areas consumed by the larvae. ²⁸

The crude fungal extract as well as the major constituents dhilirolides A (1), D (4), and L (14) were evaluated for bioactivity against the cabbage looper Trichoplusia ni (Lepidoptera: Noctuidae), an economically important agricultural pest. The crude extract was a modest feeding inhibitor to third instar T. ni larvae, but an LH ₂ 0 fraction enriched in dhilirolides A-N (1-14), dhilirolide D (4), and dhilirolide L (12) was considerably more active in a two-choice feeding bioassay. The DC ₅₀ (concentration reducing feeding by 50%) for the LH ₂ o fraction was 25 μg/cm ² , comparable to a number of natural and synthetic anti-feedants previously tested against this pest. 4- ² 7 Dhilirolide L (12) showed the highest activity of the pure compounds, giving a DC50 of 5.9 g/cm ² .

When sprayed directly on second instar larvae as a 1% aqueous emulsion, the crude extract caused 47% mortality, while the previously mentioned LH ₂ o fraction caused 63% mortality. Addition of either the crude extract or LH ₂ o fraction to the insect's diet at 1000 ppm fresh weight (fwt) resulted in significant sub-lethal effects. The crude extract reduced larval growth by 20% and adult emergence by 56%, while the dhilirolide enriched LH ₂ o fraction reduced larval growth by 70% and adult emergence by 87.5%. EXAMPLE 6: Proposed Biogeneic Pathway

Dhilirolide A (l) was isolated in sufficient quantities to acquire a 2D-INADEQUATE spectrum, which allowed the unambiguous assignment of intact acetate units as summarized in FIGURE 3. A proposed biogenetic pathway to the dhilirolides that accounts for the [i,2- ¹ 3C ₂ ] acetate incorporation data is shown in FIGURE 3. The pathway provides additional support for the earlier labeling work of the Simpson and Vederas groups.19,20,23

Although various embodiments of the invention are disclosed herein, many adaptations and modifications may be made within the scope of the invention in accordance with the common general knowledge of those skilled in this art. Such modifications include the substitution of known equivalents for any aspect of the invention in order to achieve the same result in substantially the same way. Numeric ranges are inclusive of the numbers defining the range. The word "comprising" is used herein as an open-ended term, substantially equivalent to the phrase "including, but not limited to", and the word "comprises" has a corresponding meaning. As used herein, the singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a thing" includes more than one such thing. Citation of references herein is not an admission that such references are prior art to an embodiment of the present invention. Any priority document(s) and all publications, including but not limited to patents and patent applications, cited in this specification are incorporated herein by reference as if each individual publication were specifically and individually indicated to be incorporated by reference herein and as though fully set forth herein. The invention includes all embodiments and variations substantially as hereinbefore described and with reference to the examples and drawings.

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