WO2008070891A1

WO2008070891A1 - Ligands for the modulation of ecdysone receptors and control of insect growth

Info

Publication number: WO2008070891A1
Application number: PCT/AU2006/001905
Authority: WO
Inventors: Andris Juris Liepa; Wynona Marguerite Johnson; Kathleen Anne Turner
Original assignee: Commonwealth Scientific and Industrial Research Organization CSIRO; Australian Wool Innovation Ltd
Current assignee: Commonwealth Scientific and Industrial Research Organization CSIRO; Australian Wool Innovation Ltd
Priority date: 2006-12-15
Filing date: 2006-12-15
Publication date: 2008-06-19
Anticipated expiration: 2009-06-15
Also published as: WO2008070934A1

Abstract

The present invention relates to the use of N-substituted γ-methylene γ-lactams as non-steroidal ligands for modulating ecdysone receptors generally and, in particular, ecdysone receptors of the Australian blowfly, Lucilia cuprina. The present invention further relates to novel N-substituted γ-methylene γ-lactams. These compounds are useful as insecticides or as lead compounds for the development of insecticides. The compounds are also useful as effectors for ecdysone switches.

Description

LIGANDS FOR THE MODULATION OF ECDYSONE RECEPTORS AND CONTROL OF INSECT GROWTH

FIELD OF THE INVENTION

The present invention relates to the use of N-substituted γ-methylene γ-Iactams as non-steroidal ligands for modulating ecdysone receptors generally and, in particular, ecdysone receptors of the Australian blowfly, Lucilia cuprina. The present invention further relates to novel N-substituted γ-methylene γ-laclams. These compounds are useful as insecticides or as lead compounds for the development of insecticides.

BACKGROUND OF THE INVENTION

The management of insect pest species is a particular problem, particularly as insecticidal treatments are often damaging to the environment. Insect steroid hormones and receptors are promising targets for control agents. The ecdysone receptor is a nuclear hormone receptor found in arthropod cells. The moulting, metamorphosis and reproduction of insects are controlled by the binding of 20-hydroxyecdysone to this ligand-activated transcription factor (Riddiford et al., 2000). There is sufficient variability in ecdysone receptors between insect orders to allow development of compounds which have low toxicity to off target insect orders. Due to its specificity, ligands for this receptor should provide environmentally friendly control agents with little toxicity to other species.

Until the 1980's, chemical approaches to the development of ecdysone mimics were hampered by the structural complexity and synthetic inaccessibility of the steroids for commercial-scale field applications. However in 1988, Rohm and Haas Company scientists (Wing et al., 1988; Wing, 1988) reported that a class of bisacylhydrazine insecticides, which the company had discovered serendipitously, were acting primarily via interaction with ecdysone receptors. The binding affinity of members of this class for an ecdysone receptor correlates well with the strength of their moulting hormone activity (Minakuchi et al., 2003). Members of this class display remarkable selectivity at the level of orders within the Insecta, for example RH-5992 is some two to three orders of magnitude more effective against Lepidopteta than it is against Diptera. This difference correlates with different dissociation constants for interaction of the compounds with ecdysone receptors from the two insect orders (Dhadialla et al., 1998), Although subsequent studies (Sundaram et al., 1998) have demonstrated a contribution in some cases by active transport clearance, there is little doubt that variation in the structure of the ecdysone receptors per se between different orders plays a very significant role in underlying the selectivity of extant insecticides in this class.

The cloning of the EcR and USP subunits of the L. cuprina receptor is described in WO 01/02436 A1. WO 2005/054271 describes high-throughput fluorescent polarisation assays for discovering ecdysone receptor ligands. Use of these assays to perform high-throughput screening of compound libraries against specific ecdysone receptors or the ligand-binding portions thereof, facilitates the discovery of novel ecdysone receptor agonists and antagonists.

Furthermore, since ecdysone receptors and their functional domains are employed as components of ecdysone switches for the control of reporter and therapeutic genes in mammalian cells (Lafont & Dinan, 2003; Yang et al., 1986) and for control of transgenes more generally in agriculturally important species, both animal and plant (Lafont & Dinan, 2003; Padidam et al., 2003), the ability to screen compound libraries against selected ecdysone receptors (or the ligand-binding portions thereof) should aid in the discovery of safer and/or more effective ligands to act as effectors for such switches. Several different classes of non-steroidal compounds have previously been established to act as ecdysone receptor ligands and effectors for ecdysone switches including the bisacylhydrazines (US 6,258,603 B1 ), tetxahydroquinolines (WO 03/105849 A1) and oxadiazolines (US 2004/0171651 A1).

Modulators of ecdysone receptors would find particular application in the control of the blowfly L. cuprina. L. cuprina is indigenous to Africa but is also a major pest species in Asia and Australasia. It is a known carrier of anthrax and is suspected of transmission of other diseases. It is responsible for initiating about 90% of all cases of flystrike in Australia. A current estimate of the annual cost of prevention, treatment and losses due to flystrike in Australia alone is $280 million.

Flystrike occurs when the female lays its eggs in damp, protected areas on sheep. The larvae migrate to the skin of the animal, and where the skin is damaged they become established and feed on the flesh of the sheep. If left untreated it can lead to death in a few days. Current management practices include animal husbandry methods such as shearing, dagging, crutching, wound treatment, docking and mulesing. These have limited effectiveness. With the phasing out of mulesing in Australia by 2010 effective alternatives are required.

Chemical treatment methods available include application of organophosphales, insect growth regulators and spinosyns. Resistance to organophosphatc insecticides in L. cuprina populations has already developed. There is also continuing concern about pesticide residues on the wool, from an environmental perspective and due to health concerns in relation to shearers and other wool handlers. As a result, controls in many of the wool processing countries are becoming increasingly stringent.

The N-substituted γ-methylene γ-lactam derivatives that are the subject of this invention are related to compounds disclosed in the literature as outlined below. However, as will be seen from the following, the scope of the known compounds in this area is very limited and their use in the modulation of ecdysone receptor activity has not previously been elucidated.

JP2001294771 claims compounds of general formula A, where R¹ and R² are substituents, n is 0-1 , X is O, NSO₂R³, R³ is alkyl or aryl, * is bond location and EWG is an electron aspiration group with Hammett substituent constant σp >0.30. These are stated to be useful as dyes for printing and recording inks.

The only specific examples given are compounds of formula B and C.

JP 2001010242 claims compounds of formula D, where ring A is phenyl; R¹ is H, alkyl, alkenyl, cycloalkyl or Ph, X is O or S and R² and R³ are alkyl, alkenyl, phenyl or cycloalkyl.

These compounds are stated to be useful as ink components for printing materials,

US 92-995436 claims compounds of formula E where R¹ is an electron withdrawing group, R² is C1-20 alkyl or alkylthio, C2-20 alkenyl, aryl, aralkyl, heterocycle or cycloalkyl, any of which are C5-14, or R² is hydroxy, cyano, chloro, nitro or H; R³ is C6-14 aryl or C1-12 alkyl; L¹ -L³ are methine, n is 0-3 and D is a moiety in conjugation with the O of the pyrrolinone ring.

Similar compounds conlaining conjugated side chains are claimed in JP 07319097, JP05197079, JP 2001242584, EP0985968, JP 2005190770, and US5283165. These compounds are stated to be useful as dyes.

A related patent is JP2001.334756 which discloses compounds of the formula F

where R¹, R² and R³ are H or a substituent, O is a group consisting of non-metallic atoms which forms a visible or IR-absorbing dye, L is CH or N and X is O, S or NR³ as dyes for use in thermal-transfer recording materials.

WO2005005430 discloses a process for preparing furopyrroles from compounds of Formula G

where A¹ and A² are C1-C18 alkyl, C2- C18 alkenyl, C2-C18 alkynyl, C5-C8 cycloalkyl, C5- C8 cycloalkenyl, aryl or heteroaryl; A³ is hydrogen, C1-C18 alkyl, cyanomethyl, Ar³, -CR³⁰R³¹-(CH₂)_m-Ar³ or Y-R³², wherein R³⁰ and R³¹ independently of each other stand for hydrogen or C1-C4 alkyl, or phenyl which can be substituted up in three times with C1-C4 alkyl;

Ar³ stands for aryl, C5-C8 cycloalkyl, C5-C8 cycloalkenyl or heteroaryl, which can be substituted one to three times with C1-C8 alkyl, C1 -C8 alkoxy, halogen or phenyl, which can be substituted with C1-C8 alkyl or C1-C8 alkoxy one to three times and m stands for 0,1,2,3 or 4;

R is C1-C10 alkyl, in particular C1-C4 alkyl, aryl , in particular phenyl, or aralkyl, in particular benzyl, which can be substituted one to three times with C1-C8 alkyl, C1-C8 alkoxy, or halogen ;

Y is C(O), C(O)O, C(O)NH, SO₂NH or SO₂ and

R³² is C1-C18 alkyl, Ar³ or aralkyl.

WO9732847 discloses compounds of Formula H

where R¹ is 11 or C1-20 alkyl and R² is cyano or acetyl. These compounds are stated to be useful as protein dephosphorylase inhibitors.

WO9732579 discloses compounds of Formula J

where R¹ is C1-3 alkyl or is bonded to R² to thereby foπn methylidene; R² is OH, C1-C20 alkoxy or C1-C10 alkoxy substituted by nitrooxy or phenyl; R³ is H or C1-C20 alkyl and R⁴ is CN, C1-C3 alkoxycarbonyl or C2-C10 alkanoyl optionally substituted by hydroxy. These compounds gave a neural differentiation induction effect

US3299095, US3361626, PE2352448, GB1478643, EP1106604, WO2000028064 and JP05317068 disclose tetramic acid derivatives including magnesidin, amycomycin and vancoresmycin which are stated to be useful as antibiotics. Related compounds are also disclosed in the journal literature. GB2372986 claims tetramic acid derivatives as inhibitors of plasminogen activator inhibitor. US3306909 claims tetramic acid derivatives as mammalian diuretics.

DE4223015 discloses tetramic acid derivatives of Formula K

where X and Y are independently H, alkyl, halogen, alkoxy, haloalkyl, haloalkoxy, nitro, alkoxycarbonyl, carboxy, cyano, or S(O)_m-R ¹ ;

Z is alkyl, halo, alkoxy, haloalkyl, haloalkoxy, alkoxyalkyl or alkoxycarbonyl

n is 0-3; m is 0-2;

A is H, alkyl, haloalkyl, alkoxyalkyl, polyalkoxyalkyl, alkylthioalkyl, alkenyl, alkynyl, optionally heteroatom containing cycloalkyl, cycloalkyl, phenyl optionally substituted with halogen, alkyl, haloalkyl, alkoxy, nitro, haloalkyl, or aralkyl;

B is hydrogen, alkyl, haloalkyl, cycloalkyl, alkoxyalkyl, alkylthioalkyl, phenyl optionally substituted with alkyl, alkoxy, halogen, nitro, halogenoalkyl or aralkyl; C is H, or alkyl optionally substituted with halogen; or

A and B together with the atom to which they are bound and optionally substituted with haloalkyl or alkoxy form a saturated or unsaturated ring system that may contain oxygen or sulphur;

B and C together with the carbon atom to which they are bound form a saturated or unsaturated ring which may be substituted with halogen, haloalkyl or alkoxy and may contain oxygen or sulphur;

E is H, alkyl, haloalkyl, alkenyl, alkynyl, alkoxyalkyl, aralkyl, COR², metal or metal equivalent;

R¹ is alkyl, haloalkyl or optionally substituted phenyl;

R² is alkyl, haloalkyl, alkenyl, alkoxy, alkoxyalkyl, alkylthio, optionally substituted phenyl or aralkyl;

These compounds are stated to be useful as insecticides and herbicides.

In Adhikari et al., Aust. J. Chem, 2005, 58, 882-8900 which was published on 20 December 2005, the synthesis of a number of compounds of Formula L is disclosed.

These compounds were considered as possible fungicides. However, their use as potential pesticides or as insecticides was not considered by the authors.

The present inventors have found that a class of N-substituted γ-methylene γ-lactams act as non-steroidal ligands for the modulation of ecdysone receptors. These compounds have applications as insecticides. In particular, the present invention provides compounds which bind to the Lucilia cuprina ecdysone receptor and thereby exhibit larvicidal activity.

SUMMARY OF THE INVENTION

In a first aspect, the present invention provides a method of inhibiting invertebrate pest development in a subject or an environment, said method comprising the step of administering an effective amount of a compound of formula I, or an agriculturally or pharmaceuticalIy acceptable salt thereof, to the subject or the environment

wherein:

R¹ is selected from the group consisting of: CN, CF₃, CO₂R⁶, COR⁷, CONR⁸R⁹, S(O)_mR¹⁰, P(O)_nR¹¹R¹², halogen, C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl, and heterocyclylalkyl;

R² is selected from the group consisting of C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl, heterocyclylalkyl, CN, CO₂R⁶, COR⁷, CONR⁸R⁴, S(O)_mR¹⁰, halogen or P(O)_nR¹¹R¹²; or, R² and R⁵ may together with the carbons that connect them form an optionally substituted five-to seven-membered ring system optionally comprising one or more heteroatoms selected from N, O and S;

or R¹ and R² are joined together to form the group -C(O)-X-(CH₂)_y- wherein the -C(O)- moiety is at the R¹ position and the -(CH₂)_y- moiety is at the R² position; X is selected from CH₂, O, and NR; R is selected from H and C1-3 alkyl; each CH₂ group is optionally substituted; and y is 1 to 3;

R³ is selected from the group consisting of C2-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl, heterocyclylalkyl, CN, CO₂R¹³ or COR¹⁴, N[(CO)pR¹⁵ ](CO)_qR¹⁶, NHS(O_r)R¹⁷;

each m is independently selected from 0-2;

each n is independently selected from 0-1;

p is selected from 0-1 and q is selected from 0-2, provided that when p is 1, q is not 2;

r is selected from 0-2;

R⁴ and R⁵ are each independently selected from the group consisting of H, halogen, C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, helerocyclyl and heterocyclylalkyl; or, R⁴ and R⁵ may together with the carbon to which they are attached form an optionally substituted three-to seven-membered ring system optionally comprising one or more heteroatoms selected from N, O and S;

each R⁶, R⁷, R⁹, R¹⁰, R¹³, R¹⁶ and R¹⁷ is independently selected from the group consisting of C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl and heterocyclylalkyl;

each R⁸ and R¹⁵ is independently selected from the group consisting of H, C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, helerocyclyl and heterocyclylalkyl;

R¹¹ and R¹² are each independently selected from the group consisting of C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, C1- C10 alkoxy, C3-C8 cycloalkyloxy, C4-C8 cycloalkenyloxy, C2-C10 alkenyloxy, C2- C10 alkynyloxy, phenyl, phenoxy, biphenyl, aralkyl, aralkyloxy, heterocyclyl, heterocyclyloxy, heterocyclylalkyl and heterocyclylalkyloxy; NR¹⁵R¹⁶ can together form an optionally substituted ring system, either monocyclic or polycyclic, containing single bonds or a combination of single and double bonds and optionally containing 1 to 8 heteroatoms selected from N, O and S;

when q = 1 or 2, R¹⁶ may be C1-10alkoxy or NR¹⁵R¹⁶.

R¹⁴ is selected from the group consisting of C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl, heterocyclylalkyl and NR¹³R¹⁶.

In a second aspect, the invention provides compositions for administration of the compounds of Formula I.

In a third aspect, the present invention provides a method of modulating gene activity mediated by an ecdysone receptor or functional domain thereof, said method comprising administrating a compound of Formula I to the ecdysone receptor or functional domain thereof.

In a fourth aspect, the present invention provides a compound of formula I, or an agriculturally or pharmaceutically acceptable salt thereof,

wherein:

R¹ is selected from the group consisting of: CN, CF₃, CO₂R⁶, COR⁷, CONR⁸R⁹, S(O)_mR.¹⁰,P(O)_nR₁ ¹R¹², halogen, C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl, and heterocyclylalkyl; R² is selected from the group consisting of C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl, heterocyclylalkyl, CN, CO₂R⁶, COR⁷, CONR⁸R⁹, S(O)_mR^10, halogen or P(O)_nR¹¹R¹²; or, R² and R⁵ may together with, the carbons that connect them form an optionally substituted five-to seven-membered ring system optionally comprising one or more heteroatoms selected from N, O and S;

or R¹ and R² ate joined together to form the group -C(O)-X-(CH₂)_y- wherein the -C(O)- moiety is at the R¹ position and the -(CH₂)_y- moiety is at the R² position; X is selected from CH₂, O, and NR; R is selected from H and C1-3alkyl; each CH₂ group is optionally substituted; and y is 1 to 3;

R³ is selected from the group consisting of C2-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, substituted phenyl, biphenyl, substituted aralkyl, heterocyclyl, unsaturated heterocyclyl, heterocyclylalkyl, CN, CO₂R¹³ or COR¹⁴, N[(CO)_pR¹⁵](CO)_qR₁ ⁶, NHS(O_r)R₁ ⁷;

each m is independently selected from 0-2;

each n is independently selected from 0-1 ;

p is selected from 0-1 and q is selected from 0-2, provided that when p is 1 , q is not 2;

r is selected from 0-2;

R⁴ and R⁵ are each independently selected from the group consisting of H, halogen, C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl and heterocyclylalkyl; or, R⁴ and R⁵ may together with the carbon to which they are attached form an optionally substituted three-to seven-membered ring system optionally comprising one or more heteroatoms selected from N, O and S;

each R⁶, R⁷, R⁹, R¹⁰, R¹³, R¹⁶ and R¹⁷ is independently selected from the group consisting of C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl.

C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl and heterocyclylalkyl; each R⁸ and R¹⁵ is independently selected from the group consisting of H, C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl and heterocyclylalkyl;

R¹¹ and R¹² are each independently selected from the group consisting of C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, C1- C10 alkoxy, C3-C8 cycloalkyloxy, C4-C8 cycloalkenyloxy, C2-C10 alkenyloxy, C2- C10 alkynyloxy, phenyl, phenoxy, biphenyl, aralkyl, aralkyloxy, heterocyclyl, heterocyclyloxy, heterocyclylalkyl and heterocyclylalkyloxy;

NR¹⁵R¹⁶ can together form an optionally substituted ring system, either monocyclic or polycyclic, containing single bonds or a combination of single and double bonds and optionally containing 1 to 8 heteroatoms selected from N, Q and S;

when q = 1 or 2, R¹⁶ may be C1-10alkoxy or NR¹⁵R¹⁶.

R¹⁴ is selected from the group consisting of substituted C1 alkyl, C2-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, substituted phenyl, biphenyl, aralkyl, heterocyclyl, heterocyclylalkyl and NR¹⁵R¹⁶.

provided that:

when R⁴ and R⁵ are H, R² is methyl, R¹ is CN, C1-3 alkoxycarbonyl, or C2-10 alkanoyl, R3 is not C2-10alkyl;

when R³ is pyrid-2-yl, R⁴ or R^s may not be 1-propyl-5-(butylpropylamino)imidazol-2- yl or 5-(dibutyIamino)oxazoI-2-yl;

when R¹ is CH₃ and R⁴ and R⁵ are both H, R³ may not be n-butyl;

the compound is not the compound of formula I where:

R¹ is CO₂Me, R² is Me, R³ is n-butyl, and R⁴ and R⁵ are H

R² is Ph and R³ is 4-MeSO₂NMPh or 4-HO₂CPM; R¹ is CO₂Me, R² is CO₂Me R³ is 4-MeOPh and R⁴ and R⁵ are Ph;

R¹ is CO₂Me, R² is H, R³ is 4-MeOPh, R⁴ is Me and R⁵ is Et.

DETAILED DESCRIPTION OF THE INVENTION

In a first aspect, the present invention provides a method of inhibiting invertebrate pest development in a subject or an environment, said method comprising the step of administering an effective amount of a compound of formula I, or an agriculturally or pharmaceutically acceptable salt thereof, to the subject or the environment

wherein:

R² is selected from the group consisting of C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl, heterocyclylalkyl, CN, CO₂R⁶, COR⁷, CONR⁸R⁹, S(O)_mR¹⁰, halogen or P(O)_nR¹¹R¹²; or, R² and R⁵ may together with the carbons that connect them form an optionally substituted three- to seven-membered ring system optionally comprising one or more heteroatoms selected from N, O and S;

or R¹ and R² are joined together to form the group -C(O)-X-(CH₂)_y- wherein the -C(O)- moicty is at the R¹ position and the -(CH₂)_y- moiety is at the R² position; X is selected from CH₂, O, and NR; R is selected from H and C1-3alkyl; each CH₂ group is optionally substituted; and y is 1 to 3;

R³ is selected from the group consisting of C2-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heteroeyclyl, heterocyclylalkyl, CN, CO₂R¹³ or COR¹⁴, N[(CO)_nR¹⁵](CO)₀R¹⁶, NHS(O_r)R¹⁷;

each m is independently selected from 0-2;

each n is independently selected from 0-1 ;

r is selected from 0-2;

. each R⁶, R⁷, R⁹, R¹⁰, R¹³, R¹⁶ and R¹⁷ is independently selected from the group consisting of C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl and heterocyclylalkyl;

each R⁸ and R¹⁵ is independently selected from the group consisting of H, C1-C10 alkyl. C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl and heterocyclylalkyl;

R¹¹ and R¹² are each independently selected from the group consisting of C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyy, C2-C10 alkenyl, C2-C10 alkynyl, C1- C10 alkoxy, C3-C8 cycloalkyloxy, C4-C8 cycloalkenyloxy, C2-C10 alkenyloxy, C2- C10 alkynyloxy, phenyl, phenoxy, biphenyl, aralkyl , aralkyloxy, heterocyclyl, heterocyclyloxy, heterocyclylalkyl and heterocyclylalkyloxy; NR¹⁵R¹⁶ can together form an optionally substituted ring system, either monocyclic or polycyclic, containing single bonds or a combination of single and double bonds and optionally containing 1 to 8 heteroatoms selected from N, O and S;

when q = 1 or 2, R¹⁶ may be C1-10 alkoxy or NR¹⁵R¹⁶.

R¹⁴ is selected from the group consisting of C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl, heterocyclylalkyl and NR¹⁵R¹⁶.

The term "inhibiting invertebrate pest development" means inhibition of invertebrate pest development (including mortality) that causes significant reduction in feeding or other injury or damage caused by the invertebrate pest. The compounds of Formula I may act as agonists or antagonists of the function of ecdysone receptors. An agonist will bind to the ecdysone receptor and initially mimic the action of an ecdysteroid. This initiates the process of moulting and the pest organism ceases feeding. In normal ecdysteroid induced moulting, the hormone drops over the period of the moulting process switching on the genes which control the late stage moulting including ecdysis, pigmentation and chitin synthesis. This does not occur with a synthetic agonist and moulting is incomplete. The organism does not resume feeding and dies of starvation and desiccation (Remakaran et. al. 1997). An antagonist will bind to the receptor and may prevent or delay the process of moulting. Ecdysone receptor antagonists have been shown to affect the development of certain insects and cause weight loss and death during moulting (Blackford and Dinan 1997, Miro 1995). In either case, the compound will disrupt the life cycle and growth of the invertebrate pest, potentially killing it.

The method of the present invention finds particular application in controlling growth of insects detrimental to crops or to human and animal health by administration of an effective amount of a compound of Formula I.

Preferably, the invertebrate pest is an arthropod and/or helminth parasite. Preferably, the invertebrate pest is an insect. More preferably, the insect is Lucilia cuprina.

In other preferred forms, the parasite is Bovicola ovis (sheep body louse) or human head lice (Pedicllus humanus capitis). In a preferred form, the subject is an animal infested by the invertebrate pest. Preferably, the animal is a sheep.

In one embodiment, the present invention provides a method for controlling insect growth of Lucilia cuprina comprising contacting Lucilia cuprina or their environment with an effective amount of a compound of Formula I.

Although, particular invertebrate pests have been identified above, modulators of ecdysone activity are suitable for inhibiting the development of a large number of pests. For instance, invertebrate pests of economic importance such as arthropods, gastropods and nematodes. The term" arthropod" includes insects, mites, spiders, scorpions, centipedes, millipedes, pill bugs and symphylans. The term "gastropod" includes snails, slugs and other

Stylommatophora. The term "nematode" includes all of the helminths, such as roundworms, heartworms, and phytophagous nematodes (Nematode), flukes (Tematoda), Acanthocephala, and tapeworms (Cestoda).

Particular examples of invertebrate pests of economic importance are listed in WO 03/016304, .the subject matter of which is hereby incorporated by reference.

In a further preferred optional embodiment of the invention there are provided methods of preventing or treating invertebrate pest infestation in crop plants, stored grain or other stored plant or agricultural products, and people or animals, comprising administering a pest- suppressive or pest killing amount of at least one inventive compound or combinations thereof, into an environmental area where invertebrate pests of interest are present, or may become present. By "administering" in this context is meant contacting environmental materials or surfaces, including plants and external surfaces (e.g. skin, hair, wool, fur or hides) of animals including humans, with amounts of the inventive compound or with a selected mixture or combination of more than one of the inventive compounds that is effective to kill, suppress and/or repel one or more invertebrate pests of interest.

Compositions that include solutions, emulsions, suspensions and dry forms of the inventive compound(s) are discussed below. The process of administering such compositions in the environmental context can be achieved by methods well known in the art. These include spraying, brushing, dipping, rinsing, washing, dusting, using art-known equipment, in a selected area. The selected area to be treated optionally includes plants, e.g., crops, and/or animals. In a particular embodiment, a composition comprising a compound of the invention is placed on a minor portion of the outer surface of an animal, generally as a line or spot on the animal's back (e.g., as a pour-on application) and the compound migrates over the whole external surface of the animal to protect the animal [see, US6,492,419 B1, the contents of which are hereby incorporated by reference in their entireties].

Environmental areas contemplated to be treated in this way include, e.g., fields, orchards, gardens and the like, buildings and their environs, including landscaping; storage facilities, transport or fixed storage or analogous structures and structural components, such as walls, floors, roofs, fences, windows and window screens, and the like. Animal living spaces are also included, e.g., animal pens, chicken coops, corrals, barns and the like. Human homes and other human residential, business or commercial and educational facilities are also contemplated to be treated or contacted with the inventive compounds or. compositions thereof as described above.

The present inventors have found that compounds of formula I in which R³ is a cyclic substituent are particularly effective as ligands of the ecdysone receptor. Accordingly, in a preferred form, R³ is selected from phenyl, biphenyl, heterocyclyl, and NR¹⁵R¹⁶ wherein NR¹⁵R¹⁶ is an optionally substituted 5- to 10-membered monocyclic or bicyclic ring system optionally comprising 1-4 heteroatoms selected from N, O and S, or R¹⁵ and R¹⁶ are independently C3-C8 alkyl, C3-C8 alkenyl, C3-C7 cycloalkyl or aryl.

Modelling studies have shown that if the cyclic substituent R³ has a high tilt angle to the lactam there is improved interaction with the ecdysone receptor. Accordingly, in a further preferred form, R³ is ortho-substituted phenyl.

Preferably, R⁴ and R⁵ are each selected from H or C1-3 alkyl.

Preferably R² is C1-10 alkyl.

Preferably, R¹ is CN. Preferably, the compound of Formula I is a compound of Formula II

wherein:

each of R¹, R², R⁴ and R⁵ are as defined above:

R¹⁸ is selected from halo, CF₃, CF₃O, C1-4alkyl, CO₂C1-4alkyl, OC1-4alkyl, NO₂, OH, aryl, O-aryl;

R¹⁹ is selected from the group consisting of H, halo, O-aryl, OC1-4alkyl, CF3, NO₂, and C1-4alkyl;

R²⁰ is selected from the group consisting of H, C1-4alkyl, halo, CN, OC1-4alkyl, phenyl;

R²¹ is selected from the group consisting of H, halo, O-aryl, OC1-4alkyl, CF₃, NO₂, and C1-4alkyl;

R²² is selected from the group consisting of H, halo, C1-4alkyl, CO2C1-4alkyl, OC1- 4alkyl and NO₂. In a second aspect, the invention provides compositions for administration of the compounds of Formula 1.

The compositions of the second aspect comprise an effective amount of one or more of the compounds of Formula I together with a suitable carrier. When the compound of Formula I is employed in the field, in order to treat the ground, structures, food plants, animal care facilities, and the like, the composition will comprise a solid or liquid formulation.

Solid compositions according to the invention include, for example, a powdered carrier into which an effective amount and concentration of at least one compound of Formula I is admixed. Such solid compositions optionally further include stabilizers, preservatives, colouring agents, perfumes, additional art-known active agents selected to provide synergistic invertebrate pest killing activity, and/or agents selected to complement the pest killing spectrum of the inventive compound or compounds.

Liquid compositions according to the invenlion include, for example, one or more optional liquid solvents, diluents or carriers that are polar, e.g., based on water, alcohol, or other polar solvent, or a solvent or carrier that is non-polar, e.g., an organic solvent or the like. An effective amount and concentration of at least one compound of Formula I is admixed, dispersed, emulsified, or dissolved in the liquid carrier. Such liquid compositions optionally further include emulsifiers, detergents, anti-foaming agents, stabilizers, preservatives, colouring agents, perfumes, additional art-known active agents selected to provide synergistic invertebrate pest killing activity, and/or agents selected to complement the pest killing spectrum of the inventive compound or compounds. Such optional diluents or carriers are selected for compatibility with the selected inventive compound, as well as for environmental compatibility and safety, while allowing for administering the inventive compound or compounds into an area or location of interest, at concentrations effective for the intended purpose.

More preferably, the invention provides for a pharmaceutical composition for treatment of animals infected with parasites that comprises a therapeutically effective dosage amount of compound of and/or combinations thereof of compounds of Formula 1 and a pharmaceutically acceptable excipient. The pharmaceutical composition is contemplated to be administered to animals for in vivo treatment by any suitable art known route, including, e.g., oral, parenteral, topical, and/or rectal, routes of administration.

In a solid form, the pharmaceutical composition includes pharmaceutically acceptable excipients, and carriers, and is prepared as a powder that is optionally dispensed in soluble capsules for oral ingestion, in any art-known tableted form. A solid composition according to the invention is also optionally formulated into a patch for transdermal administration.

In a liquid form, the pharmaceutical composition is provided, together with any optional pharmaceutically acceptable excipients, and carriers, in solution and/or in suspension in a pharmaceutically acceptable liquid composition for administration orally, by infusion or injection and/or by spray or inhalation, and the like.

Ecdysone receptors and their functional domains are employed as components of ecdysone receptor gene switches for the control of reporter and therapeutic genes in mammalian cells (Lafont & Dinan, 2003; Yang et al., 1986) and for control of transgencs more generally in agriculturally important species, both animal and plant (Lafont & Dinan, 2003; Padidam et al., 2003) .

Accordingly, the present invention provides a method for activating or suppressing the transcription of one or more exogeneous genes in a cell, wherein transcription of said one or more exogenous genes is controlled by a ecdysone receptor gene switch, said method comprising administering a compound of Formula I to the cell.

The compound of Formula I may act as an agonist or an antagonist of the ecdysone receptor or functional domain thereof which forms part of the ecdysone receptor gene switch.

wherein:

R¹ is selected from (he group consisting of: CN, CF₃, CO₂R^Λ, COR⁷, CONR⁸R⁹, S(O)₁₁₁R¹⁰, P(O)_nR¹¹R¹², halogen, C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl, and heterocyclylalkyl;

R² is selected from the group consisting of C I-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl, heterocyclylalkyl, CN, CO₂R⁶, COR⁷, CONR^SR°, S(O)_mR¹⁰, halogen or P(O)_nR¹¹R¹²; or, R² and R⁵ may together with the carbons that connect them form an optionally substituted five-to sevcn-membered ring system optionally comprising one or more heteroatorns selected from N, O and S;

or R¹ and R² are joined together to form the group -C(O)-X-(CH₂)_y- wherein the -C(O)- moiety is at the R¹ position and the -(CH₂)_y- ^moiety is at the R² position; X is selected from CH₂, O, and NR; R is selected from H and Cl -3alkyl; each CH₂ group is optionally substituted; and y is 1 to 3;

R³ is selected from the group consisting of C2-C.10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, substituted phenyl, biphenyl, substituted aralkyl, licterocyclyl, unsaturated heterocyclyl, heterocyclylalkyl, CN, CO₂R¹³ or COR¹⁴, NKCO)pR^ιsKCO)_qR¹⁶, NHS(O₁)R¹⁷;

eacti m is independently selected from 0-2;

each ii is independently selected from 0- 1 ; p is selected from 0-1 and q is selected from 0-2, provided that when p is 1, q is not 2;

r is selected from 0-2;

each R⁶, R⁷, R⁹, R¹⁰, R¹³, R¹⁶ and R¹⁷ is independently selected from the group consisting of C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl and heterocyclylalkyl ;

each R⁸ and R¹⁵ is independently selected from the group consisting of H, C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl and heterocyclylalkyl;

R¹¹ and R¹² are each independently selected from the group consisting of C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, C1- C10 alkoxy, C3-C8 cycloalkyloxy, C4-C8 cycloalkenyloxy, C2-C10 alkenyloxy, C2- C10 alkynyloxy, phenyl, phenoxy, biphenyl, aralkyl, aralkyloxy, heterocyclyl, heterocyclyloxy, heLerocyclylalkyl and heterocyclylalkyloxy;

NR¹⁵R¹⁶ can together form an optionally substituted ring system, cither monocyclic or polycyclic, containing single bonds or a combination of single and double bonds and optionally containing 1 to 8 heteroatoms selected from N, O and S;

when q = 1 or 2, R¹⁶ may be C1-10 alkoxy or NR¹⁵R¹⁶.

R¹⁴ is selected from the group consisting of substituted C1 alkyl, C2-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, substituted phenyl, biphenyl, aralkyl, heterocyclyl, heterocyclylalkyl and NR¹⁵R¹⁶. provided that:

when R³ is pyrid-2-yl, R⁴ or R⁵ may not be 1-propyl-5-(butylpropylamino)imidazoI-2- yl or 5-(dibutyIamino)oxazol-2-yl;

when R¹ is CH₃ and R⁴ and R⁵ are both If, R³ may not be n-butyl;

the compound is not the compound of formula I where:

R¹ is CO₂Me, R² is Me, R³ is n-butyI, and R⁴ and R⁵ are H;

R² is Ph and R³ is 4-MeSO₂NHPh or 4-HO₂CPh;

R¹ is CO₂Me₁ R² is CO₂Me R³ is 4-MeOPh and R⁴ and R⁵ are Ph;

R¹ is CO₂Me, R² is H, R³ is 4-MeOPh, R⁴ is Me and R⁵ is Et.

In a preferred form, when R¹ is CN, R² is Me, each of R⁴ and R⁵ is H, then R³ is not 4- chlorophenyl, 2,6-diethylphenyl, 4-cyanophenyl, cyclohexyl, 4-chloronaphth-1-yl, 5- chloropyrid-2-yl, 5-methyl-1,3,4-thiadiazol-2-yl, (4-chlorobenzylidene)amino, phenethyl, or 3,4-dimethoxybenzyl

Preferably, the compound is not a compound of formula I where:

R¹ is CN, R² is Et, R³ is 2-chloro-6-methylphenyl, and R⁴ and R⁵ are H;

R¹ is CN, R² is i-Pr, R³ is 2-isopropyl-6-methylphenyl, and R⁴ and R⁵ are H;

R¹ is CN, R² is Bu, R³ is 2-chloro-6-methylphenyl, and R⁴ and R⁵ are H;

R¹ is CN, R² is phenyl, R³ is cyclohexyl, and R⁴ and R⁵ are H;

R¹ is CN, R² and R⁵ together are (CH₂)₃, R³ is cyclohexyl and R⁴ is H;

R¹ is CO₂Et, R² is Mc, R³ is 4-chlorophenyl, and R⁴ and R⁵ are H. In a preferred form, R³ is selected from phenyl, biphenyl, heterocyclyl, and NR¹⁵R¹⁶ wherein NR¹⁵R¹⁶ is an optionally substituted 5- to 10-nicmbered monocyclic or bicyclic ring system optionally comprising 1-4 heteroatoms selected from N, O and S, or R¹⁵ and R¹⁶ are independently C3-C8 alkyl, C3-C8 alkenyl, C3-C7 cycloalkyl or aryl.

In a further preferred form, R³ is ortho-substituted phenyl.

Preferably, R⁴ and R⁵ are each selected from H or C1-3 alkyl.

Preferably R² is C1-10 alkyl.

Preferably, R¹ is CN.

As used herein, the term "halo" or "halogen" refers to fluorine (fluoro), chlorine (chloro), bromine (bromo) or iodine (iodo).

As used herein, the term "alkyl" either used alone or in compound terms such as NH(alkyl) or N(alkyl)₂, refers to straight chain or branched hydrocarbon groups, having 1 to 3, 1 to 6, 1 to 10 or 1 to 21 carbon atoms. For example, suitable alkyl groups include, but are not limited to methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, nmpentyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 2-, 3- or 4-methylpentyl or 2-ethylbutyl.

The term "alkenyl" as used herein, refers to straight chain or branched hydrocarbon groups containing one or more double bonds. Suitable alkenyl groups include, but are not limited to ethenyl, propenyl, butenyl, pentenyl and hexenyl.

The term "alkynyl" as used herein, refers to straight chain or branched hydrocarbon groups containing one or more triple bonds. Suitable alkynyl groups include, but are not limited to ethynyl, propynyl, butynyl, pentynyl and hexynyl.

The term "cycloalkyl" as used herein, refers to cyclic hydrocarbon groups containing one or more rings including fused and bridged ring systems. Suitable cycloalkyl groups include, but are not limited to cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl. The term "aryl" as used herein, refers to C₆-C₁₀ aromatic hydrocarbon groups containing one or more rings including fused ring systems, for example phenyl or naphthyl.

The term "aralkyl" as used herein, refers to an aryl group as defined above directly connected lo an alkyl group as defined above.

The term "helerocycle" when used alone or in compound words includes monocyclic, polycyclic, or fused saturated, unsaturated or aromatic hydrocarbon residues, preferably C_{3 10}, wherein one or more carbon atoms (and where appropriate, hydrogen atoms attached thereto) are replaced by a heteroatom. Suitable heteroatoms include, O, N and S. Where two or more carbon atoms are replaced, this may be by two or more of the same heteroatom or by different heteroatoms. Suitable examples of heterocyclyl groups may include, but are not limited to imidazolidinyl, pyrazolidinyl, furanyl, piperidinyl, piperazinyl, morpholinyl, etc.

Each alkyl, alkenyl, alkynyl, cycloalkyl, aryl or heterocyclyl group may be optionally substituted with one or more of C_1-4 alkyl, OH, OC _1-4alkyl, O(aryl), OCOC_1-4alkyl, OCO(aryl), halo, CN, NO₂, CO₂H, CO₂C_1-4alkyl, CONH₂, CONH(C_1-4alkyl), CON(C_1-4alkyl)₂, trifluoromethyl, NH₂, NH(alkyl), N(alkyl)₂, NH(aryl), NHCO(alkyl),

N(alkyl)CO(alkyl), NHCO(aryl), N(alkyl)CO(aryl), P(O)(C_{1 4}alkoxy)₂, P(O)(C_1-4alkyl)(C_1-4 alkoxy), S(O)_n(alkyl) where n=0-2. For example, an optionally substituted aryl group may be a 4-methylphenyl or 4-hydroxyphenyl group, and an optionally substituted alkyl group may be 2-hydroxyethyl, trifluoromethyl or difluoromethyl.

The compounds of the invention can be prepared by a number of methods. Simply by way of example, and without limitation, the compounds can be prepared using one or more of the reaction schemes and methods described below.

Cyanoacetamides could be prepared by methods well known in the art. For example by heating an amine with ethyl cyanoacetate, [Schellhase el al (1984)], or by condensation with cyanoacetic acid in the presence of diisopropyl carbodiimide [Sjogren et al (1991); Papageorgiou et al (1998)].

Thus by way of a non-limiting example and with reference to Reaction Scheme 1, a cyanoacetamide 2a where R¹ is alkyl, cycloalkyl, aryl, heterocycle, aralkyl etc. may be prepared by heating the corresponding amine with ethylcyanoacetate in an appropriate solvent. Suitable solvents include THF (tetrahydrofuran), ethanol and methanol. Alternatively a cyanoacetamide 2a can be prepared by condensing the amine with cyanoacetic acid in the presence of diisopropyl carbodiimide in an appropriate solvent. A preferred solvent in this regard is DMF (dimethyl formamide).

Reaction Scheme 1

Preparation of a compound of formula 2a from a compound of formula 1 a

By way of a non-limiting example and with reference to Reaction Scheme 2, a cyanoacetamide 4a, where R¹ and R² are independently alkyl, cycloalkyl, aryl, heterocycle, aralkyl etc. or R¹ and R² together with the nitrogen to which they are attached form a ring, may be prepared by condensing a hydrazine 3a with cyanoacetic acid in the presence of diisopropyl carbodiimide in an appropriate solvent, for example DMF or THF.

Reaction Scheme 2

Preparation of a compound of formula 4a from a compound of formula 3a.

By way of a non-limiting example and with reference to Reaction Scheme 3, a diacylhydrazide 6a, where R¹ is alkyl, cycloalkyl, aryl, heterocycle, aralkyl etc. may be prepared by condensing the corresponding acid chloride 5a with cyanoacetohydrazide in the presence of a suitable base, for example pyridine or potassium carbonate.

Reaction Scheme 3

Preparation of a compound of formula 6a from a compound of formula 5a.

By way of a non-limiting example and with reference to Reaction Scheme 4, a diacyl hydrazide 9a, where R¹ is alkyl, cycloalkyl, aryl, heterocycle, aralkyl etc. and R² is alkyl, cycloalkyl, aryl, aralkyl etc. may be prepared by treatment of an acid chloride 7a with an appropriate hydrazine to give the corresponding monoacylhydrazide 8a, followed by treatment with cyanoacetic acid and phosphoryl chloride.

Reaction Scheme 4

Preparation of a compound of formula 9a from a compound of formula 7 a.

By way of a non-limiting example and with reference to Reaction Scheme 5, a malonamide 11a, where R¹ is alkyl, cycloalkyl, aryl, heterocycle, aralkyl etc. may be prepared by treatment of the appropriate amine 1a with the malonate ester 10a at elevated temperatures, which may be achieved by conventional or microwave heating. The reaction may be conducted neat or in an appropriate solvent such as xylene, chlorobenzene or toluene and may or may not contain a base, for example sodium acetate or pyridine, or an acid such as toluenesulfonic acid.

Reaction Scheme 5

Preparation of a compound of formula 11a from a compound of formula 10a and a compound of formula 1a.

By way of a non-limiting example and with reference to Reaction Scheme 6, an amide 13a, where R¹ is alkyl, cycloalkyl, aryl, heterocycle, aralkyl etc. and R² is alkyl , cycloalkyl, aryl, heterocycle, aralkyl, acyl, alkylsulfonyl etc. may be prepared from the corresponding amine 1a and an appropriately substituted acetyl chloride 12a in a suitable solvent in the presence of an appropriate base, for example pyridine, triethylamine, diisopropylethylamine, N- methylmorpholine, DMAP (4-dimethylaminopyridine), potassium carbonate, sodium hydroxide, potassium hydroxide etc. Suitable solvents may include toluene, benzene, THF, dichloromethane, diethyl ether, dioxane and acetone.

Reaction Scheme 6 Preparation of a compound of formula 13 a from a compound of formula 12a and a compound of formula 1 a.

By way of a non-litniting example and with reference to Reaction Scheme 7, a 1,2,4- oxadiazol-3-ylacetamide 15a, where R¹ is alkyl, cycloalkyl, aryl, heterocycle, aralkyl etc. and R² is alkyl, substituted alkyl, cycloalkyl, aryl, heterocycle, aralkyl etc. may be prepared from the corresponding cyanoacetamide 2a by treatment with hydroxylamine hydrochloride in the presence of base to give the amidoxime 14a, followed by reaction with the appropriate anhydride or reaction with an appropriate ester in the presence of a suitable base, for example sodium ethoxide, sodium hydroxide or sodium carbonate. Suitable solvents for the latter transformation include ethanol and aqueous ethanol.

Reaction Scheme 7

Preparation of a compound of formula 15a from a compound of formula 2a.

By way of a non-limiting example and with reference to Reaction Scheme 8, a sulfonylacetamide 17a, where R¹ is alkyl, cycloalkyl, aryl, heterocycle, aralkyl etc. and R² is aryl may be prepared by treatment of the corresponding chloroacetamide 16a with an appropriate sodium sulfinate. The chloroacetamide 16a may in turn be prepared from the corresponding amine 1a and chloroacelyl chloride in an appropriate solvent such as benzene, dichloromethane, diethyl ether, THF, dioxane or ethyl acetate and in the presence of an appropriate base such as potassium carbonate, sodium carbonate, aqueous sodium hydroxide, pyridine, triethylamine etc.

Reaction Scheme 8

Preparation of a compound of formula 17a from a compound of formula 1a.

By way of a non-limiting example and with reference to Reaction Scheme 9, a mixture of tetrazoles 18a and 19a, where R ¹ is alkyl, cycloalkyl, aryl, helerocycle, aralkyl etc. and R² is alkyl may be prepared by treatment of the appropriate cyanoacetamide 2a with sodium azide in the presence of triethylamine hydrochloride. These may then be alkylated with the appropriate orthoacetate to give the products.

Reaction Scheme 9 Preparation of a compound of formula 20a and a compound of formula 21 a from a compound of formula 2a.

By way of a non-limiting example and with reference to Reaction Scheme 10, an acyl sulfonylhydrazide 23a, where R¹ is aryl may be prepared from the corresponding sulfonyl chloride 22a by treatment, with cyanoacetohydrazidc in the presence or absence of an appropriate base, for example sodium carbonate or pyridine, and in a suitable solvent, for example acetonitrile or ethanol.

Reaction Scheme 10

Preparation of a compound of formula 23a from a compound of formula 22a.

By way of a non-limiting example and with reference to Reaction Scheme 11 , an acylhydrazone 25a, where R¹ is alkyl, cycloalkyl, aryl, heterocyclc, aralkyl etc. may be prepared from the corresponding aldehyde 24a by treatment with cyanoacetohydrazide with or without a suitable base such as pyridine or triethylamine, and in an appropriate solvent, such as methanol, ethanol or aqueous ethanol.

Reaction Scheme 11

Preparation of a compound of formula 25a from a compound of formula 24a.

Condensation of the cyanoacetamides with 1,2 diketones, followed by dehydration to give the N-substituted γ-methylene γ-lactams is described by Adhikari et al 2005 and references therein.

Thus, by way of a non-limiting example and with reference to Reaction Scheme 12, a preferred method for preparing methylenelactams 30a and 31 a, where R¹, R³ and R⁴ are is alkyl, cycloalkyl, aryl, heterocycle, aralkyl, alkylamino, arylamino, alkylcarbonyl, arylcarbonyl, etc. and R² is an electron withdrawin1 group is by condensation of a suitably substituted acetamide 26a, including 2a, 4a, 6a, 9a, 11a, 13a, 15a, 17a, 20a and 21a, with a 1,2-diketone 27a in an appropriate solvent and in the presence of a suitable base to give a single hydroxylactam 28a or 29a or a mixture of the two. Suitable bases may include for example piperidine, morpholine and DABCO (1,4-diazabicyclo[2.2.2]octane). A preferred solvent is DMF. The hydroxylactams 28a and 29a may be individually or together treated with an appropriate acid in the presence or absence of a suitable co-solvent to provide the methylenelactams 30a and 31a. Suitable acids may include formic acid, trifluoroacetic acid and polyphosphate ester. An example of a suitable co-solvent for trifluoroacetic acid is dichloromethane.

Reaction Scheme 12

Preparation of a compound of formula 30a and a compound of formula 31 a from a compound of formula 26a and a compound of formula 27a.

In order that the nature of the present invention may be more clearly understood, preferred forms thereof will now be described with reference to the following non-limiting examples. EXAMPLE 1

Preparation of 1-(2-isopropyl-6-methylphenyl)-5-methylene-2-oxo-4-isopropyl-2,5-dihydro- 1H-pyrrole-3-carbonitrile (Compound 142)

The following compounds were prepared according to reaction schemes 1 and 12.

(a) To a stirred suspension of 2-isopropyl-6-methylaniline (1 equiv.) and cyanoacetic acid ( 1. equiv.) in DMF (380 ml/mol) at 0-5°C under argon was added 1 ,3- diisopropylcarbodiimide (1.2 equiv.) dropwise. The reaction mixture was left stirring at room temperature for 2 days, poured into water. The precipitate was collected by filtration and washed with water then dichloromethane to give the cyanamide as a pale yellow solid. Trituration with hot methanol and filtration gave the product in 71% yield, m.p. 124-126°C (aqueous ethanol).

(b) Piperidine (6.5 mL/mol) was added to a mixture of 4-methylpentan-2,3-dione (1 equiv.) and the cyanamide (1 equiv.) in dry DMF (800 mL/mol) and the solution left to stand at room temperature for 7 days. Addition of water caused separation of a tarry material which was extracted into dichloromethane. The organic extract was washed with water and the solvent removed under reduced pressure to leave a gum which was used without further purification.

(c) The gum was dissolved in dichloromethane/ trifluoroacetic acid (10/1), left, to stand overnight and the reaction evaporated to dryness. The product was obtained as needles from isopropyl alcohol m.p. 149-152°C in a 61% overall yield.

EXAMPLE 2

Preparation of 4-butyl-1-(2-chloro-6-methylphenyl)-5-methylene-2-oxo-2,5-dihydro-1H- pyrrole-3-carbonitrile (Compound 151)

The following compounds were prepared according to reaction schemes 1 and 12.

(a) A mixture of ethyl cyanoacetate (1 equiv.), DMF (500 mL/mol) and 2-chloro-6- methylaniline (1 equiv.) was heated at. 100°C for 10 h, diluted with water (1.5 L/mol) and the precipitate was collected, washed with water and used in the next step without further purification.

(b) Piperidine (6.5 mL/mol) was added to a mixture of heptan-2,3-dione (1 equiv.) and the cyanoacetamide (1 equiv.) in DMF (800 mL/mol) and the solution left to stand at room temperature for 7 days. Addition of water caused separation of a tarry material which was extracted into dichloromethane. The organic extract was separated, washed with water and the solvent removed under reduced pressure to leave a gum which was used without further purification.

(c) The gum was dissolved in dichloromethane/trifluoruacetic acid (10/ϊ), left to stand overnight and the reaction evaporated to dryness. The product was obtained as prisms from (ether/light petroleum) m.p. 101-102°C in 42% overall yield.

Compounds 1 to 197 as listed in Table 1 and Compounds 233 to 290 as listed in Table 3 were prepared by the method of Example 1 or Example 2, with the following variations:

In step (a) of Example 1 the mixture may be stirred from 1-2 days.

In step (a) of Example 2 the mixture may be heated from 100 to 160°C for 1 h-3 days.

In step (b) the piperidine may be replaced by another suitable base and the mixture stood or stirred for 1-15 days.

In step (c) dichloromethane/trifluoroacetic acid may be replaced by another suitable acid. The mixture may be heated to 40-80°C for several hours and/or stood for 3h to 1 day.

In step (c) the product may be isolated from the crude mixture by chromatography on silica with a suitable eluent, for example ether/petroleum ether (b.p. 40-60°C), dichloromethane/petroleum ether or ethyl acetate/petroleum ether.

Compounds 291-330 as listed in Table 3 can be prepared by the method of Example 1 or Example 2 with the above variations and where the aniline is replaced by the appropriate hydrazine. This is illustrated by reaction schemes 2 and 12.

EXAMPLE 3

Preparation of 4-ethyl-1-(4-chlorobenzoylamino)-5-ethylidene-2-oxo-2,5-dihydro-1H- pyrrole-3-carbonitrile (Compound 337)

The following compounds were prepared according to reaction schemes 3 and 12. (a) 4-Chlorobenzoyl chloride (1 equiv.) was added dropwisc to a stirred, chilled (0-5°C) mixture of cyanoacetohydrazide (1.1 equiv.) and pyridine (1.5 equiv.) in dichloromethane (1 L/mol), The resulting mixture was allowed to warm to room temperature and stirred for 4 h, then diluted with water and dichloromethane and filtered to give the product as a white crystalline solid.

(b) Morpholine (6.5 mL/mol) was added to a mixture of heχxn-3,4-dione (1.1 equiv.) and the cyanoacetamide (1 equiv.) in dry DMF (500 mL/mol) and the solution stirred at room temperature overnight. The mixture was poured into water and filtered to give the product as a brown solid, which was dried by evaporation of the water as the benzene azeotrope, then used without further purification .

(c) The solid was dissolved in formic acid (1.3 L/mol) and heated to 90°C for 8 h then poured into water and extracted with dichloromethane. The combined organic extracts were washed with water and 10% sodium bicarbonate solution, dried (MgSO₄), filtered and concentrated under reduced pressure. The residue was subjected to radial chromatography on a silica plate (eluent 5% ethyl acetate in dichloromethane) to give the product as an amber crystalline solid, m.p. 142°C in 15% overall yield.

Compounds 332 to 338 as listed in Table 3 were prepared by the method of Example 3, with the following variations:

In step (b) the mixture may be stirred at room temperature for 18 h to 3 days

In step (c) the mixture may be heated at 85-90°C for 2-8 h

Compound 331 was prepared by the method of Example 3 with the above variations and where the benzoyl chloride/pyridine is replaced by the appropriate aldehyde and ethanol. This is illustrated by reaction scheme 11.

EXAMPLE 4

Preparation of 4-ethyl-1-[N-(3-chlorobenzoyl)-N-methylamino]-5-ethylidene-2-oxo-2,5- dihydro-1H-pyrrole-3 -carbonitrile (Compound 342) The following compounds were prepared according to reaction schemes 4 and 12.

(a) A solution of 3-chlorobenzoyl chloride (1 cquiv.) in dichloromethane (350 mL/inol) was added dropwisc to a stirred solution of methyl hydrazine (3 equiv.) in dichloromethane (350 πiL/mol) maintained below -30^DC. It was then allowed to warm to room temperature and the reaction mixture washed with water then extracted into 2 M hydrochloric acid. The aqueous solution was washed with ethyl acetate, basified willi 10% sodium hydroxide solution and extracted with dichloroiαethanc. The organic phase was dried (Na₂SO₄), filtered and concentrated under reduced pressure to give the product as an oil which was used without further purification.

(b) Phosphoryl chloride (2 cquiv.) was added to a stirred, chilled (0-5°C) mixture of cyanoacetic acid (1 equiv.) and the hydrazide (1 cquiv.) in dichloroethane (1.5 L/mol). The resulting mixLurc was then heated and stirred at S0°C for 1 h, then diluted with water and the organic layer removed. The aqueous layer was extracted with dichloromethane and the combined organic layers washed "with water, dried (Na₂SOu), filtered and concentrated under reduced pressure. The residue was recrystalliscd from dichloroethane/ dichlpromethane to give the product as a pale yellow solid.

(c) Morpholine (31 mlVinol) was added to a mixture of hexan-3,4-dκme (1.3 equiv.) and the cyanoacctamide (2.4 g, 1 equiv.) in dry DMF (1.6 L/mol) and the solution stirred at room temperature for 22 h. The mixture was poured into water, acidified with 2 M hydrochloric acid and extracted with dichloromethane. The organic phase was dried (Na₂SO₄), filtered and concentrated under reduced pressure. Purification of the crude material by radial chromatography on silica (eluent: ethyl acetate/dichloromethane/ petroleum spirit) gave the product as a colourless oil.

(d) The oil was dissolved in formic acid (1.5 iVmoI) and heated to 90°C for 3 h then poured into water and extracted with dichlorometαanc. The combined dichloromethane extracts were washed wiih wuiur, dried (Na₂SO₄), ±ilicreύ unci wMceuiraicύ unύci icducou pressure. The residue was subjected to radial chromatography on a silica plate (eluent: ethyl acctatc/dichloromethane/petroleurn spirit 1/1/2) to give the product as a crystalline solid, m.p. 126°C in 3% overall yield. Compounds 339 to 359 as listed in Table 3 were prepared by the method of Example 4.

EXAMPLE 5

Preparation of 3 -benzoyl-1-(2, 6-diisopropylphenyl)-5-methylene-2-oxo-4-methyl-2,5- dihydro-1H-pyrrole (Compound 203)

The following compounds were prepared according to reaction schemes 5 and 12.

(a) A mixture of 2,6-diisopropylaniline (1 equiv.) and ethyl benzoylacetate (1.1 equiv.) in DMF (600 mL/mol) was heated at 150°C for 4 h, then cooled to 80°C and water added till turbid. The mixture was then stirred for 30min, water and ethanol added and stirring continued for a further 1 h. The solid was then filtered off and used without further purification.

(b) DABCO (4 g/mol) was added to a mixture of butan-2,3-dione (1 equiv.) and the benzoylacetamide (1 equiv.) in dry DMF (600 mL/moI) and the solution left to stand at room temperature for 3 weeks. Trituration with water caused separation of a gummy solid which was filtered off and used without further purification.

(c) The hydroxylactam was dissolved in formic acid ( 1.3 L/mol) and stood at room temperature for 1 h then poured into water and extracted with ether. The combined organic extracts were washed with water, dried (MgSO₄), filtered and concentrated under reduced pressure. The residue was subjected to radial chromatography on a silica plate (eluent 5% ethyl acetate in petroleum spirit) to give the product as a solid.

Compounds 199-213 as listed in Table 2 and Compound 364-380, 382-395 and 397-404 as listed in Table 3 can be prepared by the method of Example 5 using the appropriate materials.

EXAMPLE 6

Preparation of 3-ethoxycarbonyl-1-{4-chlorophenyl) 5 methylene-2-oxo-4-methyl-2,5- dihydro-1H-pyrrole (Compound 198)

The following compounds were prepared according to reaction schemes 6 and 12. (a) A solution of ethyl malonylchloride (1 equiv.) in dichloromethane (560 mL/mol) was added dropwise to a stirred, chilled (0-5°C) solution of 4-chIoroaniline (1 equiv.) and pyridine (112 mL/mol) in dichloromethane. The resulting mixture was stirred at room temperature overnight then diluted with waLer and extracted with ether. The ether phase was washed with dilute hydrochloric acid and water, dried (Na₂SO₄), filtered and concentrated under reduced pressure. The residue was crystallised from ether/petroleum spirit (b.p. 40-60°C to give the product as a pink solid, m.p. 84°C.

(b) Butane-2,3-dione (1.05 equiv.) was added to a solution of ethyl malonyl-4- chlorobenzamide (1 equiv.) in DMSO (600 mL/mol). Morpholine (6.5 mL/mol) was then added and the mixture stirred at room temperature for 10 days. Water was then added and the mixture stirred. The solvent was decanted off and the gummy solid crystallised from ethyl acetate/petroleum spirit to give the product as a crystalline solid.

(c) The solid was dissolved in formic acid (290 mL/mol) and stirred for 2 days at room temperature. It was then filtered and the solid washed with formic acid and water to give the product as a white solid, m.p. 251°C in 34% overall yield..

Compounds 216 and 221-223 as listed in Table 2 and compounds 381 and 396 as listed in Table 3 can be prepared by the method of Example 6 using the appropriate materials.

EXAMPLE 7

Preparation of 3-(5-trifluoromethyl-1,2,4-oxadiazol-3-yl)-1-(4-fluorophenyl)-5-methylene-2- oxo-4-methyl-2,5-dihydro-1H-pyrrole (Compound 227)

The following compounds were prepared according to reaction schemes 7 and 12.

(a) A mixture of N-(4-fluorophenyl)cyanoacetamide (1 equiv.), hydroxylamine hydrochloride (2 equiv.) and triethylamine (2 equiv.) in DMF was heated to 50-60°C for 4 h then diluted with water and the product filtered off and dried.

(b) Trifluoroacetic anhydride (2 equiv.) was added to a chilled (0-5°C) mixture of the amidoxime (1 equiv.) and dichloromethane. The resulting mixture was allowed to warm to room temperature and stirred for 24 h then washed with water, dried (MgSO₄), filtered and concentrated under reduced pressure. The residue was stirred with aqueous ethanol and the solid product filtered off and dried.

(c) The solid was taken up in DMF (800 mL/mol) and treated with butan-2,3-dione (1.1 equiv.) and morpholine (6.5 mL/mol) at room temperature for 24 h. Addition of water caused separation of a tarry material which was extracted into dichloromethane. The organic extract was separated; washed with water and the solvent removed under reduced pressure to leave a gum which was used without further purification.

(d) The gum was dissolved in formic acid ( 1.3 L/mol) and heated to 70°C for 2 h then poured into water. The precipitate was filtered off and dried to give the product.

EXAMPLE 8

Preparation of3-{4-chlorobenzenesulphonyl)-1-{2-methyl-5-isopropylphenyl)-5-methylene-2- oxo-4-methyl-2,5-dihydro-1H-pyrrole (Compound 215)

The following compounds were prepared according to reaction schemes 8 and 12.

(a) A mixture of N-(2-methyl-5-isopropylphenyl)chloroacetamide (1 equiv.) and sodium 4-chlorobenzenesulfinate ( 1 equiv.) in DMF was heated to 60°C overnight then diluted with water and the precipitated solid filtered off.

(b) Morpholine (6.5 mL/mol) was added to a mixture of butan-2,3-dione (1.1 equiv.) and the benzenesulfonylacetamide (1 equiv.) in dry DMF (500 mL/mol) and the solution stirred at room temperature for 7 days. The mixture was poured into water and extracted with ethyl acetate. The organic phase was washed with water, dried (MgSO₄), filtered and concentrated under reduced pressure to give the product which was used without further purification.

(c) The hydroxylactam was dissolved in formic acid (290 mL/mol) and heated to 80°C for 6 h. It was then diluted with water and the solid filtered off, washed with water and dried to give the product as a solid. EXAMPLE 9

Preparation of 3-(4-nitrophenyl)-1-(4-chlorophenyl)-5-methylene-2-oxo-4-methyl-2,5- dihydro-1H-pyrrole (Compound 232)

The following compounds were prepared according to reaction schemes 6 and 12.

(a) A solution of 4-nitrophenylacctyl chloride (1 equiv.) in dichloromethane (560 mL/mol) was added dropwise to a stirred, chilled (0-5°C) solution of 4-chloroaniline (1 equiv.) and pyridine (112 mL/mol) in dichloromethane. The resulting mixture was stirred at room temperature overnight then diluted with water and extracted with ether. The ether phase was washed with dilute hydrochloric acid and water, dried (Na₂SO₄), filtered and concentrated under reduced pressure.

(b) N-(4-chlorophenyl)-4-nitrophenylacetamide (1 equiv.), butan-2,3-dione (1 equiv.) and morpholine (6.5 mL/mol) in DMF (800 mL/mol) were stirred overnight at room temperature then diluted with water and the solid product filtered off and used without further purification.

(c) The hydroxylactam was dissolved in dichloromethane/ trifluoroacelic acid ( 10/1) and left to stand overnight and the reaction evaporated to dryness. The crude solid was recrystallised from chloroform/ ethanol (1/1) to give the product as yellow crystals, m.p. 227- 230°C.

EXAMPLE 10

Preparation of 3-cyano-1-(2.4,6-trimethylphenyLsulphonylamino)-5-methylene-2-oxo-4-methyl-2,5-dihydro-1H-pyrrole (Compound 362)

The following compounds were prepared according to reaction schemes 10 and 12.

(a) A suspension of cyanoacetohydrazide (1 equiv,), mesitylsulfonyl chloride, and sodium carbonate in acetonitrile was stirred overnight at room temperature then diluted with water and the solid filtered off. (b) The sulphonylhydrazide (1 equiv.), butan-2,3-dione (1 equiv.) and morpholine (6.5 mL/mol) in DMF (800 mL/mol) were stirred overnight at room temperature then diluted with water. The mixture was extracted with ethyl acetate and the combined extracts washed with water, dried (MgSO₄), filtered and concentrated under reduced pressure to give the product which was used without further purification.

(c) The product from step (b) was dissolved in formic acid (290 mL/mol) and heated to 80°C for 24 h. It was then diluted with water and the solid filtered off, washed with water and dried to give the product.

EXAMPLE 11

Preparation of 3-(1-methyltetrazol-5-yl)-1-(2-chlorophenyl)-5-methylene-2-oxo-4-methyl-2,5- dihydro-1H-pyrrole (Compound 228) and 3-(2-methyltetrazol-5-yl)-1-(2-chlorophenyl)-5- methylene-2-oxo-4-methyl-2,5-dihydro-1H-pyrrole (229)

The following compounds were prepared according to reaction schemes 9 and 12.

(a) A mixture of ethyl cyanoacetate (1 equiv.), DMF (500 mL/mol) and 2-chloroaniline (1 equiv.) was heated at. 100°C for 10 h, diluted wilh water (1.5 L/mol) and the precipitate was collected, washed with water and used in the next step without further purification.

(b) The cyanoacetamide (1 equiv.), sodium azide (1 equiv) and triethylamine hydrochloride (1 equiv.) in DMF (500 mL/mol) were healed to 100°C for 6 h then cooled and stirred into water. Sodium bicarbonate (100 g/mol) was then added and the mixture extracted with ethyl acetate. The aqueous phase was acidified with hydrochloric acid then the product filtered off.

(c) The product from step (b) (1 equiv.) was taken up in trimethyl orthoacetate (10 equiv.) and heated to 100°C for 8 h then cooled and ethyl acetate (5 L/mol) and water (5 L/mol) added, followed by sodium bicarbonate (128 g/mol). The organic layer was removed, dried (MgSO₄), filtered and concentrated under reduced pressure to give the product.

(d) The tetrazolylacetamides from step (c) were taken up in DMF (800 mL/mol) and treated with butan-2,3-dione (1 equiv.) and morpholine (6.5 mL/mol) for 4 h at room temperature then diluted with water and extracted with ethyl acetate. The organic layer was washed with water, dried (MgSO₄), filtered and concentrated under reduced pressure to give the product.

(e) The alcohol was taken up in formic acid (290 mL/mol) and heated to 100°C for 6 h, then cooled, diluted with methanol and the solid product filtered off.

Compounds 230 and 231 as listed in Table 2 were prepared by the method of Example 11.

Listing of exemplified compounds .

Table 1

Table 2

Table 3

ECDYSONE RECEPTOR BINDING OF LIGANDS AND INHIBITION OF INSECT GROWTH

Competitive inhibition assays

The ligand binding regions from the EcR and USP proteins of the insect pest, Lucilia cuprina, were co-expressed from a baculovirus vector, purified as a recombinant heterodimer ( LcEcR_DEF/LcUSP_DEF) by immobilised metal affinity chromatography, and used in fluorescent polarisation assays (patent specifications PCT /AU2004/001701 and WO2005054271 , which are hereby incorporated by cross-reference).

A 30 mM stock solution of the test compound, i.e. the putative ligand in DMSO was used to prepare a dilution series covering the assay range of 3 nM to 3 mM (final concentration). The diluent for the test compound was DMSO. These preparative pipetting steps were performed in 0.6 ml microfuge tubes.

The standard fluorescence polarisation (FP) assay buffer was 50 mM sodium phosphate, 100 mM NaCl, pH 7.4. Each 200μI assay included 6 μl of test compound in DMSO (3% v/v final concentration DMSO in each assay), 0.5 mg/ml bovine serum albumin (BSA), 36 nM fluorescein-inokosterone A conjugate and 57 μg/ml LcEcR_DEF/LcUSP_DEF receptor in standard FP assay buffer. The assays were set up in opaque flat-bottomed 96 well plates designed for fluorescence polarisation measurements and were incubated at 4°C overnight then equilibrated at room temperature for 3 hours before reading the mP values.

The FP plate reader was a PHERAstar instrument (BMG Labtech). The standard setup used the default FP parameters programmed into the PHERAstar software. The instrument was normalised to 100 mP using a 200 μl sample containing 36 nM fluorescent conjugate, 0.5 mg/ml BSA and 3% DMSO in standard FP assay buffer. Data were plotted in Excel.

It should be noted that the FP assays (described above) are competitive inhibition assays. All compounds that modulate the ecdysone response by binding to the ecdysone receptor, agonists and antagonists alike, will give a positive result on this assay. However on the larvicidal test, an agonist would cause premature moult of the larvae, whereas an antagonist would delay the moult, process, both resulting in mortality of the larvae.

L. cuprina larvicidal test

The compounds of the invention could be tested in the following manner. The method is a modification of that of Roxburgh & Shanahan (1973). The test compound is dissolved in methanol and added to sheep serum containing 20 g/L yeast extract and 5 g/L potassium dihydrogen orthophosphate. The final concentration of the methanol is 1%. The solution is left to stand for 24 h. At the end of this time strips (12 by 3 cm) of chromatography paper are treated with the serum. This is repeated with a serial dilution of test compound in concentrations expected to elicit 0-100% mortality of the larvae. The rolled papers are placed in glass vials (4 by 1 cm). Newly hatched first-instar larvae are introduced into the tubes. Four replicates are set up for each concentration. Mortality is assessed after incubation for 12 and 24 h at 28°C. Controls consist of the scrum solution without any test compound.

L. cuprina ovicidal test

The compounds of the invention could be tested in the following manner. The method is a modification of that of Roxburgh & Shanahan ( 1973). The test compound is dissolved in methanol and added to sheep serum containing 20 g/L yeast extract and 5 g/L potassium dihydrogen orthophosphate. The final concentration of the methanol is 1%. The solution is left to stand for 24h. At the end of this time strips (12 by 3 cm) of chromatography paper are treated with the serum. This is repeated with a serial dilution of test compound in concentrations expected to elicit 0-100% ovicidal activity. The rolled papers are placed in glass vials (4 by 1 cm). Eggs of L. cuprina are introduced into the tubes. Four replicates are set up for each concentration. Hatching of the eggs is assessed after incubation for 12 and 24 h at 28°C. Controls consist of the serum solution without any test compound.

Bovicola ovis testing

Method 1

The mortality testing can be done by the method of Levot & Hughes (1990). Lice are removed from donor sheep using a vacuum pump. Two 60 x 60mm cloth squares are prepared for each insecticide dilution. Starting at the centre of the cloth, 1 mL of each dilution is pipetted onto each cloth rectangle and allowed to dry at room temperature for 24 h. Control cloths are prepared with solvent only. Cloths are placed into labelled glass tubes and live lice are placed into the tubes. Tubes are sealed and incubated at 34°C for 16 h. Lice are then removed from the tubes and their condition tabulated.

Method 2

A modification of the method of Gough et al. (2002) can be used to assess the action of the compounds of the invention on B. ovis. Lice are collected from infested sheep. Bioassays can be performed in multi-welled tissue culture plates. A diet of epidermal scrapings from sheepskin is added to the wells and lice either adults or juveniles or a mixture of both are added to the wells. Wool treated with the test compounds is then added to the well and the plates incubated in sealed humidity jars containing saturated ammonium chloride (70% RH) and maintained at 34°C. Control wells have the wool treated with the solvent only. Mortality of the louse population is assessed at 72 h and again after 5-9 days.

Throughout this specification the word "comprise", or variations such as "comprises" or "comprising", will be understood to imply the inclusion of a stated element, integer or step, or group of elements, integers or steps, but not the exclusion of any other element, integer or. step, or group of elements, integers or steps. All publications mentioned in this specification are herein incorporated by reference. Any discussion of documents, acts, materials, devices, articles or the like which has been, included in the present specification is solely for the purpose of providing a context for the present invention. It is not to be taken as an admission that any or all of these matters form part of the prior art base or were common general knowledge in the field relevant to the present invention as it existed anywhere before the priority dale of each claim of this application.

It will be appreciated by persons skilled in the art that numerous variations and/or modifications may be made to the invention as shown in the specific embodiments without departing from the spirit or scope of the invention as broadly described. The present embodiments are, therefore, to be considered in a!l respects as illustrative and not restrictive.

REFERENCES

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Dhadialla, T.S., Carlson, O.R., Lc, D.P., 1998. New Insecticides with ecdysteroidal and juvenile hormone activity. Ann. Rev. Entomol.43, 545-569.

Gough, J.M., Akhurst, RJ., Ellar, DJ., Kemp, D.H., Wijffels, G. L., 2002. New Isolates of Bacillus thuringiensis for control of livestock ectoparasites. Biological Control, 23, 179-189.

Lafont, R., Dinan, L., 2003. Practical uses for ecdysteroids in mammals including humans: an update. J. Insect Sci.3 article 7. Online at insectscience.org/3.7

Levot, G. W., Hughes, P, B., 1990. Laboratory studies on resistance to cypermethrin in Damalinia ovis (Shrank). Journal of the Australian Entomological Society, 29, 257-259.

Minakuchi, C, Nakagawa, Y.. Kamimura, M., Miyagawa, H., 2003. Binding affinity of nonsteroidal ecdysone agonists against the ecdysone receptor complex determines the strength of their molting hormonal activity. Eur. J. Biochem.270, 4095-104.

Miro, M., Cucurbitacins and their pharmacological effects. Phytother. Res. 8, 159-168.

Padidam, M., Gore, M., Lu, D.L., Smimova, O., 2003. Chemical-inducible, ecdysone receptor-based gene expression system for plants. Transgenic Res. 12, 101-109.

Retnakaran, A., MacDonald, A, Tomkins, W.L., Davis, C.N., Brownwright, AJ., Palli, S.R., 1997. Ultrastructural effects of a non-steroidal ecdysone agonist, RH-5992, on the sixth instar larva of the spruce budworm, Chorisioncura fumiferana. J. Insect Physiol. 43, 55-68. Riddiford, L.M., Cherbas, P., Truman, J.W., 2000. Rcdysone receptors and their biological actions. Vitara. Horm. 60, 1-73.

Roxburgh, N. A., Shanahan, GJ., 2003. A method for the detection and measurement of insecticide resistance in larvae of Lucilia cuprina (Wied.). Bulletin of Entomological Research. 63, 99-102.

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Sjogren, E.B., Rider, P.H., Nelson, P.M., Bingham, S. Jr., Poulton, A.L., Emanuel, M.A., Kmunieki, R., 1991. Synthesis and biological activity of a series of diaryl-substituted α- cyano-β-hydroxypropenamides, a new class of anthelmintic agents. J. Med. Chem., 34, 3295- 3301.

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Wing, K.D., 1988. A nonsteroidal ecdysone agonist: effects on a Drosophila cell line. Science 241 , 467-469.

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Yang, G., Harman, G.N., Lockett, T.J., Hill, R.J., 1986. Functional transfer of an elementary ecdysone gene regulatory system to mammalian cells: transient transfection and stable cell liees. Eur. J. Entomol.92, 379-389.

Claims

1. A method of inhibiting invertebrate pest development in a subject or an environment, said method comprising the step of administering an effective amount of a compound of formula I, or an agriculturally or pharmaceutically acceptable salt thereof, to the subject or the environment

wherein:

R¹ is selected from the group consisting of: CN, CF₃, CO₂R⁶, COR⁷, CONR⁸R⁹, S(O)_mR¹⁰, P(O)_nR¹¹R¹², halogen, C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl,- C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl, and heterocyclylalkyl;

R² is selected from the group consisting of C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl, heterocyclylalkyl, CN, CO₂R⁶, COR⁷, CONR⁸R⁹, S(O)_mR¹⁰, halogen or P(O)_nR¹¹R¹²; or, R² and R⁵ may together with the carbons that connect them form an optionally substituted five-to seven-membered ring system optionally comprising one or more heteroatoms selected from N, O and S;

or R¹ and R² are joined together to form the group -C(O)-X-(CH₂)_y- wherein the -C(O)- moiety is at the R¹ position and the -(CH₂)_y- moiety is at the R² position; X is selected from CH₂, O, and NR; R is selected from H and C1-3alkyl; each CH₂ group is optionally substituted; and y is 1 to 3; R³ is selected from the group consisting of C2-C10 alkyl, C3-C8 eycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl, heterocyclylalkyl, CN, CO₂R^rj or COR¹⁴, Nf(CO)_PR¹⁵I(CO)₁₁R¹⁶, NI fS(Or)R¹⁷;

each m is independently selected from 0-2;

each n is independently selected from 0-1 ;

p Ls selected from ϋ- 1 and q is selected from 0-2, provided that when p is 1, q is not 2;

r is selected from 0-2;

R⁴ and R⁵ are each independently selected from the group consisting of H, halogen, C1-C10 alkyl, C3-C8 eycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl and heterocyclylalkyl; or, R⁴ and R may together with the carbon to which they are attached form an optionally substituted thrcc-to seven-membered ring system optionally comprising one or more heicroatoms selected from N, O and S;

each R⁶, R⁷, R⁹, R¹⁰, R¹³, R^lή and R¹⁷ is independently selected from the group consisting of C1-C10 alkyl, C3-C8 eycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl and heterocyclylalkyl;

each R⁸ and R¹⁵ is independently selected from the group consisting of H, C1-C10 alkyl, C3-C8 eycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, helerocyclyl and heterocyclylalkyl;

R^π and R¹² are each independently selected from ihc group consisting of C1-C10 alkyl, C3-C8 eycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, C1- C10 alkoxy, C3-C8 eycloalkyloxy, C4-C8 cycloalkenyloxy, C2-C10 alkenyloxy, C2- C10 alkynyloxy, phenyl, phenoxy, biphenyl, aralkyl, aralkyloxy, heterocyclyl, heterocyclyloxy, heterocyclylalkyl and heteracyclylalkyloxy;

NR¹⁵R^l6can togctlicr form an optionally substituted ring system, either monocyclic or polycyclic, containing single bonds or a combination of single and double bonds and optionally containing 1 to 8 hcfcroatoms selected from N, O and S; when q = 1 or 2, R » 1^m6 may be C1-10 alkoxy or NR I¹S³ ₁R₃ 1¹6⁰.

2. A method according to claim 1 wherein the compound of Formula I is a compound of Formula II

R²

wherein:

R² is selected from the group consisting of C1-C10 alkyl, C3-CS cycloalkyl, C4-C8 cycloalkenyi, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl, heterocyciylalkyl, CN, CO₂R⁶, COR⁷, CONR⁸R⁹, S(O)₀₁R¹⁰, halogen or P(O)_nR¹¹R¹²; or, R² and R⁵ may together with the carbons that connect them form an optionally substituted five-to sevcn-membered ring system optionally comprising one or more heleroatoms selected from N, O and S;

or R¹ and R² are joined together to form the group -C(O)-X-(CH₂)_y- wherein the - C(O)- inoiety is at the R¹ position and the -(CH₂)_y- moiety is at the R² position; X is selected from CH₂, O, and NR; R is selected from H and C1-3alkyl; each CH₂ group is optionally substituted; and y is I to 3;

each m is independently selected from 0-2;

each n is independently selected from 0- 1 ; .

R⁴ and R⁵ are each independently selected from the group consisting of 11, halogen, C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl and heterocyclylalkyl; or, R⁴ and R⁵ may together with the carbon to which they are attached form an optionally substituted thxee-to seven-membered ring system optionally comprising one or more heteroatoms selected from N, O and S;

each R⁶, R⁷, R⁹, and R¹⁰ is independently selected from the group consisting of C1-C10 alkyl, C3-CS cyckmlkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2- C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl and heterocyclylalkyl;

each R⁸ is independently selected from the group consisting of H, C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, hiphenyl. aralkyl. heterocyclyl andheterocyclylalkyl;

R¹¹ and R¹² are each independently selected from the group consisting of C1- C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, C1-C10 alkoxy, C3-C8 cycloalkyloxy, C4-C8 cycloalkenyloxy, C2- C10 alkenyloxy, C2-C10 alkynyloxy, phenyl, phenoxy, biphenyl, aralkyl, aralkyloxy, heterocyclyl, heterocyclyloxy, heterocyclylalkyl and • heterucyclylalkyloxy; R¹⁸ is selected from halo, CF₃, CF₃O, C1-4alkyl, CO₂C1-4alkyl, pci-4alkyl, NO₂, OH, aryl, O-aryl;

R¹⁹ is selected from the group consisting of H, halo, O-aryl, OC1-4alkyl, CF), NO₂, and C1-4alkyl;

R²¹ is selected from the group consisting of H, halo, O-aryl, OC!-4alkyl, CF3, NOz, and C1-4alkyl;

R²² is selected from the group consisting of H, halo, C1-4alkyl, CO₂C1-4alkyI, OC1- 4alkyl and NO₂.

3. A method according to claim I wherein R ^f is selected from phenyl, biphenyl, helerocyclyl, and KR¹⁵R¹⁶ wherein NR¹⁵R¹⁶ is an optionally substituted 5- to 10- membered monocyclic or bicyclic ring system optionally comprising 1-4 heteroatoms selected rrom N, O and S, or. R¹⁵ and R¹⁶ are independently C3-C8 alkyl, C3-C8 alkenyl, C3-C7 cycloalkyl or aryL

4. A method according to claim 3 wherein R³ is selected lrotn heterocyclyl and NR¹⁵R¹⁶ ^■wherein NR¹ R¹⁶ is an optionally substituted 5- to 10-membered monocyclic or bicyclic ring system optionally comprising 1-4 heteroatoms selected from N, O and S, or R¹⁵ and R¹⁶ are independently C3-C8 alkyl, C3-C8 alkenyl, C3-C7 cycloalkyl or aryl..

5. A method according to claim 3 wherein R^"' is ortho-substituted phenyl.

6. A method according to .my one of claims 1 to 5 wherein R and R are each selected from H or C1-3 alkyl.

7. A method according to any one of claims I to 5 wherein R¹ is C1-10 alkyl.

8. A method according to any one of claims 1 to 7 wherein R¹ is CN.

9. A method for activating or suppressing the transcription of one or more exogeneous genes in a cell, wherein transcription of said one or more exogenous genes is controlled by a ecdysone receptor gene switch, said method comprising administering a compound of Formula T to the cell

wherein:

R¹ is selected from the group consisting of: CN, CF₃, CO₂R⁶, COR⁷, CONR⁸R⁹, S(O)_mR¹⁰, P(O)_nR¹¹R¹², halogen, C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloal kenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl, and heterocyclylalkyl;

R² is selected from the group consisting of C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, .heterocyclyl, heterocyclylaliyl, CN, CO₂R⁶, COR⁷, CONR⁸R⁹, S(O)_mR¹⁰, halogen or P(O)_nR¹¹R¹²; or, R² and R⁵ may together with the carbons that connect them form an optionally substituted five-to seven-membered ring system optionaiiy comprising one or more heteroatoms selected from N, O and S;

or R¹ and R² are joined together to form the group -C(O)-X-(CH₂)_y,- wherein the - C(O)- moiety is at the R¹ position and the -(CH₂)_y- moiety is at the R² position; X is selected from CH₂, O, and NR; R is selected from H and C1-3alkyl; each CH₂ group is optionally substituted; and y is 1 to 3; R³ is selected from the group consisting of C2-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl, heterocyclylalkyl, CN, CO₂R¹³ or COR¹⁴, N[(CO)pR¹⁵|(CO)_qR¹⁶, NHS(O_r)R¹⁷;

each m is independently selected from 0-2;

each n is independently selected from 0-1;

r is selected from 0-2;

each R⁶, R⁷, R⁹, R¹⁰, R¹³, R¹⁶ and R^J7 is independently selected from the group consisting of C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl and heterocyclylalkyl;

R¹¹ and R¹² are each, independently selected from the group consisting of C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, C1- C10 alkoxy, C3-C8 cycloalkyloxy, C4-C8 cycloalkenyloxy, C2-C10 alkenyloxy, C2- C10 alkynyloxy, phenyl, phenoxy, biphenyl, aralkyl, aralkyloxy, heterocyclyl, heterocyclyloxy, heterocyclylalkyl and heterocyclylalkyloxy; NR¹⁵R¹⁶ can together form an optionally substituted ring system, either monocyclic or polycyclic, containing single bonds or a combination of single and double bonds and optionally containing 1 to 8 heteroatoms selected from N, O and S;

when q = 1 or 2, R¹⁶ may be C1-10 alkoxy or NR¹⁵R¹⁶.

10. A method according to claim 9 wherein the compound of Formula 1 is a compound of Formula II

wherein:

R¹ is selected from the group consisting of: CN, CF₃, CO₂R⁶, COR⁷, CONR⁸R⁹, S(O)_mR¹⁰, P(O)_nR¹¹R¹², halogen, C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl, and heterocyclylalkyl; R² is selected from the group consisting of C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl, heterocyclylalkyl, CN, CO₂R⁶, COR⁷, CONR⁸R⁹, S(O)_mR¹⁰, halogen or P(O)_nR¹¹ R¹²; or, R² and R⁵ may together with the carbons that connect them form an optionally substituted five-to seven-membered ring system optionally comprising one or more heteroatoms selected from N, O and S;

or R¹ and R² are joined together to form the group -C(O)-X-(CH₂)_y- wherein the - C(O)- moiety is at the R¹ position and the -(CH₂)_y- moiety is at the R² position; X is selected from CH₂, O, and NR; R is selected from H and C1-3alkyl; each CH₂ group is optionally substituted; and y is 1 to 3;

each m is independently selected from 0-2;

each n is independently selected from 0-1;

each R⁶, R⁷, R⁹, and R¹⁰ is independently selected from the group consisting of C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 atkenyl, C2- C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl and helerocyclylalkyl;

each R⁸ is independently selected from the group consisting of H, C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl and heterocyclylalkyl;

R¹¹ and R¹² are each independently selected from the group consisting of C1- C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, C1-C10 alkoxy, C3-C8 cycloalkyloxy, C4-C8 cycloalkenyloxy, C2- C10 alkenyloxy, C2-C10 alkynyloxy, phenyl, phenoxy, biphenyl, aralkyl, aralkyloxy, heterocyclyl, heterocyclyloxy, heterocyclylalkyl and heterocyclyl alkyloxy;

R²² is selected from the group consisting of H, halo, C1-4alkyl, CO₂C1-4alkyl, OC1- 4alkyl and NO₂.

11. A method according to claim 9 wherein R³ is selected from phenyl, biphenyl, heterocyclyl, and NR¹⁵R¹⁶ wherein NR¹⁵R¹⁶ is an optionally substituted 5- to 10- membered monocyclic or bicyclic ring system optionally comprising 1-4 heteroatoms selected from N, O and S, or R¹⁵ and R¹⁶ are independently C3-C8 alkyl, C3-C8 alkenyl, C3-C7 cycloalkyl or aryl.

12. A method according to claim 11 wherein R³ is selected from heterocyclyl and NR¹⁵R¹⁶ wherein NR¹⁵R¹⁶ is an optionally substituted 5- to 10-membered monocyclic or bicyclic ring system optionally comprising 1-4 heteroatoms selected from N, O and S, or R¹⁵ and R¹⁶ are independently C3-C8 alkyl, C3-C8 alkenyl, C3-C7 cycloalkyl or aryl.

12. The method of claim 11 wherein R¹ is ortho-substituted phenyl.

13. A method according to any one of claims 9 to 13 wherein R⁴ and R⁵ are each selected from H or C1-3 alkyl.

14. A method according to anyone of claims 9 to 13 wherein R² is C1-10 alkyl.

15. A method according to any one of claims 9 to 14 wherein R¹ is CN.

16. A method according to any one of claims 9 to 15 wherein the compound of Formula I is an antagonist of the ecdysone receptor or a functional domain thereof.

17. A compound of formula I, or an agriculturally or pharmaceutically acceptable salt, thereof,

wherein:

R¹ is selected from the group consisting of: CN, CF₃, CO₂R⁶, COR⁷, CONR⁸R⁹, S(O)_mR¹⁰, P(O)_nR¹¹R¹², halogen, C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl , heterocyclyl, and helerocyclylalkyl;

R² is selected from the group consisting of C1-C10 alkyl, C3 -C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl, heterocyclylalkyl, CN, CO₂R⁶, COR⁷, CONR⁸R⁹, S(O)_mR¹⁰, halogen or P(O)_nR¹¹R¹²; or, R² and R⁵ may together with the carbons that connect them form an optionally substituted five-to seven-membered ring system optionally comprising one or more heteroatoms selected from N, O and S;

or R¹ and R² are joined together to form the group -C(O)- X-(CH₂)_y- wherein the - C(O)- moiety is at the R¹ position and the -(CH₂)_y- moiety is at the R² position; X is selected from CH₂, O, and NR; R is selected from H and C1-3alkyl; each CH₂ group is optionally substituted; and y is 1 to 3;

R³ is selected from the group consisting of C2-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, substituted phenyl, biphenyl, substituted aralkyl, heterocyclyl, unsaturated heterocyclyl, heterocyclylalkyl, CN, CO₂R¹³ or COR¹⁴, N[(CO)_pR¹⁵](CO)_qR¹⁶, NHS(O_r)R¹⁷;

each in is independently selected from 0-2;

each n is independently selected from 0-1;

p is selected from 0- 1 and q is selected from 0-2, provided that when p is 1 , q is not 2;

r is selected from 0-2;

each R⁸ and R¹⁵ is independently selected from the group consisting of H, C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl C2-C10 alkenyl, C2-C10 alkynyl, phenyl, biphenyl, aralkyl, heterocyclyl and hererocyclylalkyl;

R¹¹ and R¹² are each independently selected from the group consisting of C1-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, C1- C10 alkoxy, C3-C8 cycloalkyloxy, C4-C8 cycloalkenyloxy, C2-C10 alkenyloxy, C2- C10 alkynyloxy, phenyl, phenoxy, biphenyl, aralkyl, aralkyloxy, helerocyclyl, heterocyclyloxy, heterocyclylalkyl and heterocyclylalkyloxy;

when q = 1 or 2, R¹⁶ may be C1-10 alkoxy or NR¹⁵R¹⁶.

R¹⁴ is selected from the group consisting of substituted C1 alkyl, C2-C10 alkyl, C3-C8 cycloalkyl, C4-C8 cycloalkenyl, C2-C10 alkenyl, C2-C10 alkynyl, substituted phenyl, biphenyl, aralkyl, heterocyclyl, heterocyclylalkyl and NR¹⁵R¹⁶

provided that:

when R⁴ and R³ are H, R² is methyl, R¹ is CN, C1-3alkoxycarbonyl, or C2- 10alkanoyl, R3 is not C2-10alkyl;

when R³ is pyrid-2-yl, R⁴ or R⁵ may not be 1-propyl-5-(butylpropylamino)imidazol-2- yl or 5-(dibutylamino)oxazol-2-yl;

when R¹ is CH₃ and R⁴ and R⁵ are both H, R³ may not be n-butyl;

the compound is not the compound of formula I where:

R¹ is CO₂Me, R² is Me, R³ is n-butyl, and R⁴ and R⁵ are H;

R² is Ph and R³ is 4-McSO₂NHPh or 4-HO₂CPh;

R¹ is CO₂Me, R² is CO₂Me R³ is 4-MeOPh and R⁴ and R⁵ are Ph;

R¹ is CO₂Me, R² is H, R³ is 4-MeOPh, R⁴ is Me and R⁵ is Et.

18. A compound according to claim 12 or claim 13 wherein when R¹ is CN, R² is Me, each of R⁴ aud R³ is H, then R³ is not 4-chlorophenyl, 2,6-diethylphenyl, 4- cyanophenyl, cyclohexyl, 4-chloronaphth-1-yl, 5-chloropyrid-2-yl, 5-methyl-1,3,4- thiadiazol-2-yl, (4-chlorobenzylidene)amino, phenethyl, or 3,4-dimethoxybenzyl.

219. A compound according to any one of claims 12 to 14 wherein the coxnpound of formula I is not one of the group consisting of:

R¹ is CN, R² is Et, R³ is 2-chloro-6-methylphenyl, and R⁴ and R⁵ are H;

R¹ is CN, R² is Bu, R³ is 2-chloro-6-methylphenyl, and R⁴ and R⁵ are H;

R¹ is CN, R² is phenyl, R³ is cyclohexyl, and R⁴ and R⁵ are H;

R¹ is CO₂Et, R² is Me, R³ is 4-chlorophenyl, and R⁴ and R⁵ are H.

20. A compound according to any one of claims 17 to 19 wherein R³ is selected from phenyl, biphenyl, heterocyclyl, and NR¹³R¹⁶ wherein NR¹⁵R¹⁰ is an optionally substituted 5- to 10-membered monocyclic or bicyclic ring system optionally comprising 1-4 heteroatoms selected from N, O and S, or R ⁵ and R are independently C3-C8 alkyl, C3-C8 alkenyi, C3-C7 cycloalkyl or aryl.

21. A compound according to claim 20 wherein R³ is selected from heterocyclyl and

NR¹⁵R¹⁶ wherein NR¹⁵R¹⁶ is an optionally substituted 5- to 10-membered monocyclic or bicyclic ring system optionally comprising 1-4 heteroatoms selected from N, O and S, or R¹⁵ and R¹⁶ are independently C3-C8 alkyl, C3-C8 alkenyl, C3-C7 cycloalkyl or aryl.

22. A compound according to claim 20 wherein R³ is ortho-substituted phenyl.

23. A compound according to any one of claims 17 to 22 wherein R⁴ and R⁵ are each selected from H or C1-3 alkyl.

24. A compound according to anyone of claims 17 to 23 wherein R^" is C1-10 alkyl.

25. A compound according to any one of claims 17 to 24 wherein R¹ is CN .