U.S. patent application number 15/332364 was filed with the patent office on 2017-02-09 for methods and compositions for controlling pests.
The applicant listed for this patent is Bio-Gene Technology Ltd.. Invention is credited to Albert Habib Basta, Robert Neil Spooner-Hart.
Application Number | 20170035048 15/332364 |
Document ID | / |
Family ID | 3828836 |
Filed Date | 2017-02-09 |
United States Patent
Application |
20170035048 |
Kind Code |
A1 |
Spooner-Hart; Robert Neil ;
et al. |
February 9, 2017 |
METHODS AND COMPOSITIONS FOR CONTROLLING PESTS
Abstract
Pest-controlling compositions include one or more .beta.-diones,
particularly .beta.-diketones and .beta.-triketones, and are used
inter alia for preventing, eradicating, destroying, repelling or
mitigating harmful, annoying or undesired pests including insects,
arachnids, helminths, molluscs, protozoa and viruses. .beta.-diones
can be prepared by de novo synthesis or from natural sources such
as volatile oil-bearing plants from families including Alliaceae,
Apiaceae, Asteraceae, Cannabinaceae, Lamiaceae, Pteridaceae,
Myrtaceae, Myoporaceae, Proteaceae, Rutaceae and Zingiberaceae.
Inventors: |
Spooner-Hart; Robert Neil;
(Kurrajong, AU) ; Basta; Albert Habib; (Glenwood,
AU) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Bio-Gene Technology Ltd. |
Hillarys |
|
AU |
|
|
Family ID: |
3828836 |
Appl. No.: |
15/332364 |
Filed: |
October 24, 2016 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14250187 |
Apr 10, 2014 |
9474270 |
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15332364 |
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13431726 |
Mar 27, 2012 |
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14250187 |
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12782125 |
May 18, 2010 |
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13431726 |
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12134035 |
Jun 5, 2008 |
7820209 |
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12782125 |
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10477057 |
Sep 23, 2004 |
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PCT/AU02/00569 |
May 8, 2002 |
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12134035 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A01N 37/18 20130101;
A61K 36/61 20130101; A01N 35/10 20130101; A01N 65/28 20130101; A01N
65/12 20130101; A01N 35/06 20130101; A01N 65/22 20130101; A01N
65/48 20130101; A01N 65/08 20130101; A01N 65/10 20130101; A01N
65/42 20130101; A01N 65/36 20130101; A01N 65/04 20130101 |
International
Class: |
A01N 35/06 20060101
A01N035/06; A01N 65/28 20060101 A01N065/28 |
Foreign Application Data
Date |
Code |
Application Number |
May 8, 2001 |
AU |
PR 4842 |
Claims
1. A method for controlling insects, arachnids, molluscs, protozoa
and helminths, said method comprising exposing the insects,
arachnids, molluscs, protozoa and helminths to a pest-controlling
effective amount of the compound flavesone
(1-isobutyroyl-3,3,5,5-tetramethylcyclohexan-2,4,6-trione).
2. The method of claim 1, wherein the compound is obtainable from a
volatile oil-bearing organism.
3. The method of claim 2, wherein the volatile oil-bearing organism
is selected from volatile oil-bearing plants.
4. The method of claim 2, wherein the volatile oil-bearing organism
is selected from genera of the Myrtaceae family.
5. The method of claim 2, wherein the volatile oil-bearing organism
belongs to a genus selected from Angophora, Austromyrtus,
Backhousia, Baeckea, Callistemon, Corymbia, Darwinia, Eucalyptus,
Kunzea, Leptospermum, Melaleuca, Syzygium and Xanthostemon
6. The method of claim 1, wherein the pest that is controlled is
selected from insects, arachnids and molluscs.
7. The method of claim 1, wherein the helminth is a nematode.
8. The method of claim 1, wherein the compound is used in the form
of a pest-controlling composition which comprises from about
0.00005% to about 90% by weight of said compound.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of U.S. application Ser.
No. 14/250,187, filed Apr. 10, 2014, which is a continuation of
U.S. application Ser. No. 13/431,726, filed Mar. 27, 2012, which is
a continuation of U.S. application Ser. No. 12/782,125, filed May
18, 2010, which is a continuation of U.S. application Ser. No.
12/134,035, filed Jun. 5, 2008, and granted Oct. 26, 2010 as U.S.
Pat. No. 7,820,209, which is a continuation of U.S. application
Ser. No. 10/477,057, filed Sep. 23, 2004. U.S. application Ser. No.
10/477,057 is the U.S. National Phase under 35 U.S.C. .sctn.371 of
International Application No. PCT/AU02/00569, filed May 8, 2002
designating the U.S. and published in English as WO 02/089587,
which claims priority to Australian Patent Application PR 4842,
filed May 8, 2001. This application incorporates herein by
reference U.S. application Ser. Nos. 13/431,726, 12/782,125,
12/134,035, and 10/477,057, U.S. Pat. No. 7,820,209, International
Application No. PCT/AU02/00569 including the International
Publication No. WO 02/089587, and Australian Patent Application PR
4842 in their entireties.
FIELD OF THE INVENTION
[0002] This invention relates generally to methods and compositions
for controlling pests. More particularly, the present invention
relates to pest-controlling compositions comprising as active
ingredients one or more .beta.-diones, particularly
.beta.-diketones and .beta.-triketones, and to the use of these
compositions inter alia for preventing, eradicating, destroying,
repelling or mitigating harmful, annoying or undesired pests
including insects, arachnids, helminths, molluscs, protozoa and
viruses. The present invention further relates to processes of
preparing .beta.-diones by de novo synthesis or from natural
sources such as volatile oil-bearing plants from families including
Alliaceae, Apiaceae, Asteraceae, Cannabinaceae, Lamiaceae,
Pteridaceae, Myrtaceae, Myoporaceae, Proteaceae, Rutaceae and
Zingiberaceae. Bibliographic details of various publications
referred to in this specification are collected at the end of the
description.
BACKGROUND OF THE INVENTION
[0003] Triketones have been used for many years as herbicides for
the control of undesired vegetation. Herbicidal triketones have
been described, for example, in EP-A-338992, EP-A-336898, U.S. Pat.
No. 4,869,748, EP-A-186118, EP-A-186119, EP-A-186120, U.S. Pat. No.
4,202,840, U.S. Pat. No. 4,695,673, U.S. Pat. No. 4,780,127, U.S.
Pat. No. 4,921,526, U.S. Pat. No. 5,006,150, U.S. Pat. No.
5,545,607, U.S. Pat. No. 5,925,795, U.S. Pat. No. 5,990,046, U.S.
Pat. No. 6,218,579, EP-A-249150, EP-A-137963, EP-A-394889,
EP-A-506907 or EP-B-135191. Examples of herbicidal triketones are
inter alia Sulcotrione (MIKADO.RTM.) whose chemical designation is
2-(2-chloro-4-methanesulfonylbenzoyl)-1,3-cyclohexandione,
2-(4-methylsulfonyloxy-2-nitrobenzoyl)-4,4,6,6-tetramethyl-1,3-cyclohexan-
e dione;
3-(4-methylsulfonyloxy-2-nitrobenzoyl)-bicyclo-[3,2,1]octane-2,4--
dione;
3-(4-methylsulfonyl-2-nitrobenzoyl)-bicyclo-[3,2,1]octane-2,4-dione-
;
4-(4-chloro-2-nitrobenzoyl)-2,6,6-trimethyl-2H-1,2-oxazine-3,5(4H,
6H)-dione;
3-(4-methylthio-2-nitrobenzoyl)-bicyclo[3,2,1]octane-2,4-dione;
4-(2-nitro-4-trifluoromethoxybenzoyl)-2,6,6-trimethyl-2H-1,2-oxazine-3,5(-
4H, 6H)-dione.
SUMMARY OF THE INVENTION
[0004] The instant invention is predicated in part on the discovery
that .beta.-diones, particularly .beta.-diketones and
.beta.-triketones, such as those obtainable from volatile
oil-bearing plants including plants from the families Alliaceae,
Apiaceae, Asteraceae, Cannabinaceae, Lamiaceae, Pteridaceae,
Myrtaceae, Myoporaceae, Proteaceae, Rutaceae and Zingiberaceae,
exhibit significant pesticidal, especially insecticidal,
arachnicidal, helminthicidal and/or molluscicidal activity. This
discovery has been reduced to practice in novel pest-controlling
compositions and methods for their preparation and use, as
described hereinafter.
BRIEF DESCRIPTION OF THE DRAWINGS
[0005] FIG. 1 shows the structures relating to the major
constituents of the published Myrtaceae essential oils.
[0006] FIG. 2 is a representation of a GC-MS trace of E. cloeziana
oil.
[0007] FIG. 3 is a tabular and graphical representation showing
.sup.1H NMR data recorded on a fraction (F4) obtained from silica
gel chromatography of E. cloeziana oil and the structure of the
major and minor isomers of the compound deduced from these
data.
[0008] FIG. 4 is a diagrammatic representation showing various
tautomeric forms of an isolated .beta.-triketone compound in
solution (CDCl.sub.3).
DETAILED DESCRIPTION OF THE INVENTION
[0009] One aspect of the present invention contemplates the use of
a .beta.-dione compound, particularly a .beta.-diketone or a
.beta.-triketone compound, in the preparation of a composition for
controlling harmful, annoying or undesired pests, said compound
being represented by the general formula (I)
##STR00001##
wherein
[0010] A is (C.dbd.O)R.sub.1, (C.dbd.S)R.sub.1, OR.sub.2, SR.sub.2,
(CR.sub.3NR.sub.4R.sub.5), C(R.sub.3).sub.2OR.sub.2,
NR.sub.4R.sub.5, (C.dbd.N--R.sub.4)R.sub.1, N.dbd.O,
N(.dbd.O).sub.2, NR.sub.4OR.sub.2 or SO.sub.4R.sub.2;
[0011] R.sub.1 is selected from H, C.sub.1-C.sub.10 alkyl,
C.sub.2-C.sub.10 arylalkyl, C.sub.3-C.sub.6 cycloalkyl,
C.sub.2-C.sub.10 alkenyl, C.sub.2-C.sub.10 heteroarylalkyl,
C.sub.1-C.sub.10 haloalkyl, C.sub.1-C.sub.10 dihaloalkyl,
C.sub.2-C.sub.10 trihaloalkyl, C.sub.2-C.sub.10 haloalkoxy,
C.sub.1-C.sub.10 hydroxyalkyl, C.sub.1-C.sub.10 thioalkyl and
C.sub.1-C.sub.10 nitroalkyl, OR.sub.2, SR.sub.2,
(CR.sub.3NR.sub.4R.sub.5), NR.sub.4R.sub.5,
(C.dbd.N--R.sub.4)R.sub.6, N.dbd.O, N(.dbd.O).sub.2,
NR.sub.4OR.sub.7 or SO.sub.4R.sub.7;
[0012] R.sub.2 is selected from H, C.sub.1-C.sub.10 alkyl,
C.sub.2-C.sub.10 arylalkyl, C.sub.3-C.sub.6 cycloalkyl,
C.sub.2-C.sub.10 alkenyl, C.sub.2-C.sub.10 heteroarylalkyl,
C.sub.2-C.sub.10 haloalkyl, C.sub.2-C.sub.10 dihaloalkyl,
C.sub.2-C.sub.10 trihaloalkyl, (CR.sub.3NR.sub.4R.sub.5),
NR.sub.4R.sub.5, (C.dbd.N--R.sub.4)R.sub.6, N.dbd.O,
N(.dbd.O).sub.2 or NR.sub.4OR.sub.7;
[0013] R.sub.3 is selected from H, C.sub.1-C.sub.10 alkyl,
C.sub.2-C.sub.10 arylalkyl, C.sub.3-C.sub.6 cycloalkyl,
C.sub.2-C.sub.10 alkenyl, C.sub.2-C.sub.10 heteroarylalkyl,
C.sub.2-C.sub.10 haloalkyl, C.sub.2-C.sub.10 dihaloalkyl,
C.sub.2-C.sub.10 trihaloalkyl, C.sub.2-C.sub.10 haloalkoxy,
OR.sub.7, SR.sub.7, (CR.sub.8NR.sub.4R.sub.5), NR.sub.4R.sub.5,
(C.dbd.N--R.sub.4)R.sub.6, N.dbd.O, N(.dbd.O).sub.2,
NR.sub.4OR.sub.7 or SO.sub.4R.sub.7;
[0014] R.sub.4 and R.sub.5 are independently selected from H,
C.sub.1-C.sub.10 alkyl, C.sub.2-C.sub.10 arylalkyl, C.sub.3-C.sub.6
cycloalkyl, C.sub.2-C.sub.10 alkenyl, C.sub.2-C.sub.10
heteroarylalkyl, C.sub.2-C.sub.10 haloalkyl, C.sub.2-C.sub.10
dihaloalkyl, C.sub.2-C.sub.10 trihaloalkyl, OR.sub.7 or
SR.sub.7;
[0015] R.sub.6 is selected from H, C.sub.1-C.sub.10 alkyl,
C.sub.2-C.sub.10 arylalkyl, C.sub.3-C.sub.6 cycloalkyl,
C.sub.2-C.sub.10 alkenyl, C.sub.2-C.sub.10 heteroarylalkyl,
C.sub.2-C.sub.10 haloalkyl, C.sub.2-C.sub.10 dihaloalkyl,
C.sub.2-C.sub.10 trihaloalkyl, C.sub.2-C.sub.10 haloalkoxy,
OR.sub.7, SR.sub.7, (CR.sub.8NR.sub.9R.sub.10), NR.sub.9R.sub.10 or
NR.sub.9OR.sub.7; R.sub.7 is selected from H, C.sub.1-C.sub.10
alkyl, C.sub.2-C.sub.10 arylalkyl, C.sub.3-C.sub.6 cycloalkyl,
C.sub.2-C.sub.10 alkenyl, C.sub.2-C.sub.10 heteroarylalkyl,
C.sub.2-C.sub.10 haloalkyl, C.sub.2-C.sub.10 dihaloalkyl,
C.sub.2-C.sub.10 trihaloalkyl;
[0016] R.sub.8 is selected from H, C.sub.1-C.sub.10 alkyl,
C.sub.2-C.sub.10 arylalkyl, C.sub.3-C.sub.6 cycloalkyl,
C.sub.2-C.sub.10 alkenyl, C.sub.2-C.sub.10 heteroarylalkyl,
C.sub.2-C.sub.10 haloalkyl, C.sub.2-C.sub.10 dihaloalkyl,
C.sub.2-C.sub.10 trihaloalkyl, OR.sub.11, SR.sub.11 or
NR.sub.9R.sub.10;
[0017] R.sub.9 and R.sub.10 are independently selected from H,
C.sub.1-C.sub.10 alkyl, C.sub.2-C.sub.10 arylalkyl, C.sub.3-C.sub.6
cycloalkyl, C.sub.2-C.sub.10 alkenyl, C.sub.2-C.sub.10
heteroarylalkyl, C.sub.2-C.sub.10 haloalkyl, C.sub.2-C.sub.10
dihaloalkyl, C.sub.2-C.sub.10 trihaloalkyl, OR.sub.12 or
SR.sub.12;
[0018] R.sub.11 is selected from H, C.sub.1-C.sub.10 alkyl,
C.sub.2-C.sub.10 arylalkyl, C.sub.3-C.sub.6 cycloalkyl,
C.sub.2-C.sub.10 alkenyl, C.sub.2-C.sub.10 heteroarylalkyl,
C.sub.2-C.sub.10 haloalkyl, C.sub.2-C.sub.10 dihaloalkyl,
C.sub.2-C.sub.10 trihaloalkyl;
[0019] R.sub.12 is selected from H, C.sub.1-C.sub.10 alkyl,
C.sub.2-C.sub.10 arylalkyl, C.sub.3-C.sub.6 cycloalkyl,
C.sub.2-C.sub.10 alkenyl, C.sub.2-C.sub.10 heteroarylalkyl,
C.sub.2-C.sub.10 haloalkyl, C.sub.2-C.sub.10 dihaloalkyl,
C.sub.2-C.sub.10 trihaloalkyl;
[0020] B is H, C.sub.1-C.sub.10 alkyl, C.sub.2-C.sub.10 alkenyl,
aryl or heteroaryl;
[0021] X and Y are independently selected from oxygen, sulfur,
--N--R.sub.4; and
[0022] Q completes a 5-8-member saturated or unsaturated
carbocyclic or heterocyclic ring in which optionally one or more
members comprise --C(.dbd.X)--; and wherein Q is optionally
substituted with one or more substituents selected from
C.sub.1-C.sub.10 alkyl, C.sub.3-C.sub.6 cycloalkyl,
C.sub.2-C.sub.10 alkenyl, C.sub.2-C.sub.10 haloalkyl,
C.sub.2-C.sub.10 dihaloalkyl, C.sub.2-C.sub.10 trihaloalkyl,
C.sub.2-C.sub.10 haloalkoxy, OR.sub.2, SR.sub.2,
(CR.sub.3NR.sub.4R.sub.5), NR.sub.4R.sub.5,
(C.dbd.N--R.sub.4)R.sub.1, N.dbd.O, N(.dbd.O).sub.2,
NR.sub.4OR.sub.2, SO.sub.4R.sub.2, C.sub.2-C.sub.10 1-arylalkyl,
C.sub.2-C.sub.10 2-arylalkyl or (C.dbd.X)R.sub.1.
[0023] Heterocyclic systems can be optionally attached to a moiety
other than those set forth above via a carbon atom or a heteroatom
of R.sub.1 to R.sub.11.
[0024] Preferred compounds represented by formula (I) are
.beta.-diketones and especially preferred are
.beta.-triketones.
[0025] As used herein, the term "alkyl" refers to linear or
branched chains. The term "haloalkyl" refers to an alkyl group
substituted by at least one halogen. Similarly the term
"haloalkoxy" refers to an alkoxy group substituted by at least one
halogen. As used herein the term "halogen" refers to fluorine,
chlorine, bromine and iodine.
[0026] As used herein the term "aryl" refers to aromatic
carbocyclic ring systems such as phenyl or naphthyl, anthracenyl,
especially phenyl. Suitably, aryl is C.sub.6-C.sub.14 with mono,
di, tri, tetra and penta substitution containing OR.sub.2, F, Cl,
Br, I, NO.sub.2, CF.sub.3, COR.sub.1, NR.sub.4R.sub.5,
SO.sub.2R.sub.2, SR.sub.2.
[0027] As used herein the terms "heterocycle", "heterocyclic",
"heterocyclic systems" and the like refer to a saturated,
unsaturated, or aromatic carbocyclic group having a single ring,
multiple fused rings (for example, bicyclic, tricyclic, or other
similar bridged ring systems or substituents), or multiple
condensed rings, and having at least one heteroatom such as
nitrogen, oxygen, or sulfur within at least one of the rings. This
term also includes "heteroaryl" which refers to a heterocycle in
which at least one ring is aromatic. Any heterocyclic or heteroaryl
group can be unsubstituted or optionally substituted with one or
more groups, as defined above. Further, bi- or tricyclic heteroaryl
moieties may comprise at least one ring, which is either
completely, or partially, saturated. Suitable heteroaryl moieties
include, but are not limited to oxazolyl, thiazaoyl, thienyl,
furyl, 1-isobenzofuranyl, 2H-pyrrolyl, N-pyrrolyl, imidazolyl,
pyrazolyl, isothiazolyl, isooxazolyl, pyridyl, pyrazinyl,
pyrimidinyl, pyradazinyl, indolizinyl, isoindolyl, indoyl, indolyl,
purinyl, phthalazinyl.
[0028] The articles "a" and "an" are used herein to refer to one or
to more than one (i.e., to at least one) of the grammatical object
of the article. By way of example, "an element" means one element
or more than one element.
[0029] A preferred carbocyclic ring formed by Q is an optionally
substituted cyclohexanedione.
[0030] A preferred subgroup of compounds of formula (I) is
represented by formula (II)
##STR00002##
[0031] Such compounds may exist in a number of tautomeric forms.
For example, in the case wherein X and Y are each oxygen, and B is
hydrogen, then the compounds of formula II may exist as one or more
of the structural formulae shown below.
##STR00003##
[0032] It is intended that all such tautomeric structures are
included within the scope of the present invention.
[0033] It should also be appreciated that some of the compounds of
formula (I) are capable of existing as different geometric isomers
and diastereomers. The invention thus includes both the individual
isomers and mixtures of such isomers.
[0034] Another preferred subgroup of compounds of formula (I) is
represented by formula (III)
##STR00004##
wherein
[0035] X, Y and Z are each independently selected from oxygen,
sulfur, --N--R.sub.4 or one of C.dbd.X, C.dbd.Y or C.dbd.Z is
CH.sub.2;
[0036] A is (C.dbd.O)R.sub.1, (C.dbd.S)R.sub.1, OR.sub.2, SR.sub.2,
(CR.sub.3NR.sub.4R.sub.5), C(R.sub.3).sub.2OR.sub.2,
NR.sub.4R.sub.5, (C.dbd.N--R.sub.4)R.sub.1, N.dbd.O,
N(.dbd.O).sub.2, NR.sub.4OR.sub.2 or SO.sub.4R.sub.2;
[0037] B is as defined above;
[0038] C, D, E and F are each independently selected from H,
C.sub.1-C.sub.10 alkyl, C.sub.3-C.sub.6 cycloalkyl,
C.sub.2-C.sub.10 alkenyl, C.sub.2-C.sub.10 haloalkyl,
C.sub.2-C.sub.10 dihaloalkyl, C.sub.2-C.sub.10 trihaloalkyl,
OR.sub.2, SR.sub.2, (CR.sub.3NR.sub.4R.sub.5), NR.sub.4R.sub.5,
(C.dbd.N--R.sub.4)R.sub.1, N.dbd.O, N(.dbd.O).sub.2,
NR.sub.4OR.sub.2, SO.sub.4R.sub.2, C.sub.2-C.sub.10 1-arylalkyl,
C.sub.2-C.sub.10 2-arylalkyl or (C.dbd.X)R.sub.1; and
[0039] R.sub.1, R.sub.2, R.sub.2, R.sub.4 and R.sub.5 are as
defined above.
[0040] Preferred .beta.-diones represented by formula (III) are
flavesone
(1-isobutyroyl-3,3,5,5-tetramethylcyclohexan-2,4,6-trione),
isoleptospermone
(1-isovaleroyl-3,3,5,5-tetramethylcyclohexan-2,4,6-trione),
leptospermone
(1-valeroyl-3,3,5,5-tetramethylcyclohexan-2,4,6-trione), papuanone
(1-pentoyl-3,3,5,5-tetramethylcyclohexan-2,4,6-trione),
grandiflorone
(1-(2-phenylethyl)-3,3,5,5-tetramethylcyclohexan-2,4,6-trione) and
jensenone (1-valeroyl-3,5-dicarbonylcyclohexan-2,4,6-trione),
including analogues and derivatives thereof.
[0041] By way of example, flavesone analogues contemplated by the
present invention include, but are not restricted to, compounds
having the following structural formulae, wherein the structural
formula of flavesone is shown for comparative purposes:
##STR00005##
[0042] Non-limiting examples of isoleptospermone analogues
contemplated by the present invention include, but are not
restricted to, compounds having the following structural formulae,
wherein the structural formula of isoleptospermone is shown for
comparative purposes:
##STR00006## ##STR00007##
[0043] Non-limiting examples of leptospermone analogues
contemplated by the present invention include, but are not
restricted to, compounds having the following structural formulae,
wherein the structural formula of leptospermone is shown for
comparative purposes:
##STR00008## ##STR00009##
[0044] Non-limiting examples of jensenone analogues contemplated by
the present invention include, but are not restricted to, compounds
having the following structural formulae, wherein the structural
formula of jensenone is shown for comparative purposes:
##STR00010## ##STR00011##
[0045] Another preferred subgroup of compounds of formula (I) is
represented by formula (IV)
##STR00012##
wherein
[0046] X and Y are each independently selected from oxygen, sulfur
--N--R.sub.4 or one of C.dbd.X or C.dbd.Y is CH.sub.2;
[0047] A is (C.dbd.O)R.sub.1, (C.dbd.S)R.sub.1, OR.sub.2, SR.sub.2,
(CR.sub.3NR.sub.4R.sub.5), C(R.sub.3).sub.2OR.sub.2,
NR.sub.4R.sub.5, (C.dbd.N--R.sub.4)R.sub.1, N.dbd.O,
N(.dbd.O).sub.2, NR.sub.4OR.sub.2 or SO.sub.4R.sub.2; [0048] B is
as defined above; [0049] C, D, E and F are each independently
selected from H, C.sub.1-C.sub.10 alkyl, C.sub.2-C.sub.10
arylalkyl, C.sub.3-C.sub.6 cycloalkyl, C.sub.2-C.sub.10 alkenyl,
C.sub.2-C.sub.10 heteroarylalkyl, C.sub.2-C.sub.10 haloalkyl,
C.sub.2-C.sub.10 dihaloalkyl, C.sub.2-C.sub.10 trihaloalkyl,
C.sub.2-C.sub.10 haloalkoxy, OR.sub.2, SR.sub.2,
(CR.sub.3NR.sub.4R.sub.5), NR.sub.4R.sub.5,
(C.dbd.N--R.sub.4)R.sub.1, N.dbd.O, N(.dbd.O).sub.2,
NR.sub.4OR.sub.2, SO.sub.4R.sub.2; and R.sub.1, R.sub.2, R.sub.2,
R.sub.4 and R.sub.5 are as defined above.
[0050] Preferred .beta.-diones represented by formula (IV) are
tasmanone
(1-isobutroyl-4-methoxy-3,5,5-trimethylcyclohex-3-en-2,6-dione),
agglomerone
(1-isobutroyl-4-methoxy-5,5-dimethylcyclohex-3-en-2,6-dione),
lateriticone
(1-valeroyl-4-methoxy-3,5,5-trimethylcyclohex-3-en-2,6-dione),
isolateriticone
(1-isovaleroyl-4-methoxy-3,5,5-trimethylcyclohex-3-en-2,6-dione and
platyphyllol
(6,6-dimethyl-2-acetyl-5-methoxycyclohex-4-ene-1,3-dione),
including analogues and derivatives thereof.
[0051] Non-limiting examples of tasmanone analogues contemplated by
the present invention include, but are not restricted to, compounds
having the following structural formulae, wherein the structural
formula of tasmanone is shown for comparative purposes:
##STR00013##
[0052] Another preferred subgroup of compounds of formula (I) is
represented by formula (V)
##STR00014##
wherein
[0053] X and Y are independently selected from oxygen, sulfur or
--N--R.sub.4; and
[0054] A is (C.dbd.O)R.sub.1, (C.dbd.S)R.sub.1, OR.sub.2, SR.sub.2,
(CR.sub.3NR.sub.4R.sub.5), C(R.sub.3).sub.2OR.sub.2,
NR.sub.4R.sub.5, (C.dbd.N--R.sub.4)R.sub.1,
[0055] N.dbd.O, N(.dbd.O).sub.2, NR.sub.4OR.sub.2 or
SO.sub.4R.sub.2;
[0056] B is as defined above;
[0057] C, D, E, F, G and H are each independently selected from H,
C.sub.1-C.sub.10 alkyl, C.sub.2-C.sub.10 arylalkyl, C.sub.3-C.sub.6
cycloalkyl, C.sub.2-C.sub.10 alkenyl, C.sub.2-C.sub.10
heteroarylalkyl, C.sub.2-C.sub.10 haloalkyl, C.sub.2-C.sub.10
dihaloalkyl, C.sub.2-C.sub.10 trihaloalkyl, C.sub.2-C.sub.10
haloalkoxy, OR.sub.2, SR.sub.2, (CR.sub.3NR.sub.4R.sub.5),
NR.sub.4R.sub.5, (C.dbd.N--R.sub.4)R.sub.1, N.dbd.O,
N(.dbd.O).sub.2, NR.sub.4OR.sub.2 or SO.sub.4R.sub.2; and
[0058] R.sub.1, R.sub.2, R.sub.2, R.sub.4 and R.sub.5 are as
defined above.
[0059] More specifically unsaturation, epoxides and thioexpoxides
may exist at positions designated by H (or G) connected to F (or E)
or F (or E) connected to C (or D). A four-membered ring forming a
part of a bicyclic structure may exist at positions designated by H
(or G) connected to C (or D).
[0060] Preferred .beta.-diones represented by formula (V) are
angustione (1-acetyl-3,5,5-trimethylcyclohex-2,6-dione),
dehydroangustione (1-acetyl-3,5,5-trimethylcyclohex-3-en-2,6-dione)
and xanthostemone
(1-isobutroyl-5,5-dimethylcyclohex-3-en-2,6-dione), including their
analogues and derivatives.
[0061] By way of example, angustione analogues contemplated by the
present invention include, but are not restricted to, compounds
having the following structural formulae, wherein the structural
formula of angustione is shown for comparative purposes:
##STR00015##
[0062] Non-limiting examples of dehydroangustione analogues
include, but are not restricted to, compounds having the following
structural formulae, wherein the structural formula of
dehydroangustione is shown for comparative purposes:
##STR00016##
[0063] Non-limiting examples of xanthostemone analogues include,
but are not restricted to, compounds having the following
structural formulae, wherein the structural formula of
xanthostemone is shown for comparative purposes:
##STR00017##
[0064] Derivatives of the above compounds include, but are not
restricted to, ethoxylate derivatives, propoxylate derivatives,
hydrates, aldehyde derivatives, ester derivatives, ether
derivatives, alcohol derivatives, phenol derivatives, amine
derivatives, other biologically or chemically equivalent
substances, and any combination of two or more of the
foregoing.
[0065] Similarly effective as pesticides are salts of the above
compounds, including mono-valent salts (e.g., sodium, potassium)
and di-valent metal salts (e.g., calcium, magnesium, iron or
copper) and ammonium salts (e.g., isopropyl ammonium, trialkyl and
tetraalkylammonium salts).
[0066] The compounds according to any one of formulae (I)-(V) can
be prepared according to methods analogous to those known in the
art for the preparation of .beta.-diones. Exemplary methods are
disclosed for example in EP-A-338992, EP-A-336898, U.S. Pat. No.
4,202,840, U.S. Pat. No. 4,869,748, EP-A-186118, EP-A-186119,
EP-A-186120, U.S. Pat. No. 4,695,673, U.S. Pat. No. 4,780,127, U.S.
Pat. No. 4,921,526, U.S. Pat. No. 5,006,150, U.S. Pat. No.
5,545,607, U.S. Pat. No. 5,925,795, U.S. Pat. No. 5,990,046, U.S.
Pat. No. 6,218,579, EP-A-249150, EP-A-137963, EP-A-394889
EP-A-506907 or EP-B-135191.
[0067] More particularly, compounds according to formulae (III)-(V)
can be synthesised using the representative procedures outlined
below.
[0068] For compounds according to formula (III),
1,3,5-trihydroxybenzene 1 is reacted with CH.sub.3CN in the
presence of zinc chloride and hydrochloric acid, according to A. H.
Blatt (1943, Org. Synth. Col. II, 522-523), affording
1-acetyl-2,4,6-trihydroxybenzene 2 (phloroacetophenone) (R=Me)
(Scheme 1).
##STR00018##
[0069] R groups other than methyl are depicted above. Reaction of
1-acetyl-2,4,6-trihydroxybenzene 2 affording
1-acetyl-3,3,5,5-tetramethylcyclohexan-2,4,6-trione 3 is a
representative procedure for all compounds according to formula
(III) (R. A. Gray et al., U.S. Pat. No. 4,202,840) (Scheme 2).
Anhydrous MeI (6 eq) is slowly added, at room temperature under an
atmosphere of nitrogen, to a mechanically stirred solution of
1-acetyl-2,4,6-trihydroxybenzene 2 (1 eq) and sodium ethoxide (6
eq) in anhydrous methanol. The mixture is refluxed for 4 hours. On
cooling the mixture is concentrated under vacuo, providing a
residue, which is diluted with water and acidified with 2 M
hydrochloric acid. Diethylether extracts are washed with saturated
sodium sulfite solution, water and then dried (Na.sub.2SO.sub.4).
Evaporation of the diethylether provides the desired product 3.
##STR00019##
[0070] For mono, di, tri and tetra B, C, D, E, F substitution
patterns, reactions between one and seven mole equivalents of R-I
and sodium ethoxide are used. Lawasson's reagent is used for
conversion of oxygen into sulfur groups and sodium borohydride or
sodium cyanoborohydride is used to reduce ketone, thioketone and
imino groups. When additional carbonyl groups are introduced into
the cyclohexane ring system the procedure of Crow is utilised (M.
L. Bolte et al., 1985, Agric. Biol. Chem., 49, 761).
[0071] Compounds of formula (IV) can be prepared according to a
first representative procedure, as follows:
3-methoxy-2,4,4-trimethylcyclohex-2-en-1,5-dione 4 (1 mole eq),
prepared according to Herzig (J. Herzig, and F. Wenzel, Monatsh,
1903, 24, 101), is dissolved in anhydrous diethylether and
hexamethylphosphoramide (solvent ratio, 20:1 respectively) under an
atmosphere of nitrogen. The mixture is cooled to 0.degree. C. and
lithium hydride (1.1 mole eq) (60% in mineral oil) is added in
portions. After addition the mixture is stirred for a further 10
mins before the addition of benzoyl cyanide 5 [R--CO--CN, R is
depicted above] (1.1 mole eq). The mixture is allowed to warm to
room temperature over 12 h at which time the reaction is quenched
with water and partitioned. The ether layer is dried
(Na.sub.2SO.sub.4) and evaporated affording crude
1-benzoyl-3-methoxy-2,4,4-trimethylcyclohex-2-en-1,5-dione 6 which
is purified by SiO.sub.2 column chromatography (hexane/ethyl
acetate, gradient) (Scheme 3).
##STR00020##
[0072] Alternatively, compounds of formula (IV) can be prepared
according to a second representative procedure, as follows:
3-methoxy-2,4,4-trimethylcyclohex-2-en-1,5-dione 4 (1 mole eq)
(commercially available) and benzoyl cyanide are dissolved in
anhydrous dichloromethane and cooled to 0.degree. C. under an
atmosphere of nitrogen. To the cooled solution is added anhydrous
finely powdered zinc chloride (1.1 mole eq.) followed by slow
addition of triethylamine (1.2 mole eq). The reaction mixture is
stirred at room temperature for 5-6 h and then poured into 2 M
hydrochloric acid. The mixture is partitioned and the
dichloromethane layer is washed with 5% sodium carbonate. The
aqueous carbonate phase is then acidified with hydrochloric acid
and extracted with methylene chloride and dried (Na.sub.2SO.sub.4).
The solvent is removed and the residue subjected to SiO.sub.2
column chromatography (hexane/ethyl acetate) affording
1-benzoyl-3-methoxy-2,4,4-trimethylcyclohex-2-en-1,5-dione 6 (W. J.
Michaely and G. W. Kraatz, EP-B-135191).
[0073] Compounds of formula (V) can be prepared according to a
first representative procedure, as follows:
4,4-dimethylcyclohexane-1,3-dione 7 (1 mole eq) (commercially
available) is dissolved in anhydrous diethylether and
hexamethylphosphoramide (solvent ratio, 20:1 respectively) under an
atmosphere of nitrogen. The mixture is cooled to 0.degree. C. and
lithium diisopropylamide (2.1 mole eq) is added dropwise over 40
mins. The mixture is then stirred for a further 10 mins before the
addition of methyl iodide (1 mole eq). The mixture is stirred for
12 h and then benzoyl cyanide (2 mole eq) is added and the mixture
stirred for a further 24 h. The reaction was quenched with water
and the ether layer partitioned and dried (Na.sub.2SO.sub.4). The
solvent was removed and the residue subjected to SiO.sub.2 column
chromatography (hexane/ethyl acetate) affording
1-benzoyl-3,3,5-trimethylcyclohexan-2,6-dione 8 (Scheme 4).
##STR00021##
[0074] Alternatively, the compounds of formula (V) can be prepared
according to a second representative procedure, as follows:
4,4-dimethyl-1,3-cyclohexanedione 7 (1 mole eq) (commercially
available) and benzoyl cyanide are dissolved in anhydrous
dichloromethane and cooled to 0.degree. C. under an atmosphere of
nitrogen. To the cooled solution is added anhydrous finely powdered
zinc chloride (1.1 mole eq) followed by slow addition of
triethylamine (1.2 mole eq). The reaction mixture is stirred at
room temperature for 5-6 h and then poured into 2 M hydrochloric
acid. The mixture is partitioned and the dichloromethane layer
washed with 5% sodium carbonate. The aqueous carbonate phase is
then acidified with hydrochloric acid and extracted with methylene
chloride and dried (Na.sub.2SO.sub.4). The solvent is removed and
the residue subjected to SiO.sub.2 column chromatography
(hexane/ethyl acetate) affording
1-benzoyl-3,3,5-trimethylcyclohexan-2,6-dione 8. (W. J. Michaely
and G. W. Kraatz, EP-B-135191).
[0075] Dehydroangustione and xanthostemone derivatives are simply
derived from dehydrogenation of angustione derivatives, for
example, by treatment of
1-benzoyl-3,3,5-trimethylcyclohexan-2,6-dione 8 with palladium on
charcoal in methanol, which thereby affords
1-benzoyl-3,5,5-trimethylcyclohex-3-en-2,6-dione 9 (Scheme 5).
##STR00022##
[0076] Metal salts (enolates) of the above compounds can be
prepared by the reaction of triketone derivatives with the
corresponding metal hydroxides suspended in methanol or ethanol.
Trialkylammonium salts can be prepared by the reaction of triketone
derivatives (e.g., 3) with trialkylamines in a chlorinated solvent
such as dichloromethane. Tetraalkylammonium salts can be prepared
by adding a halogenated tetraalkylammonium salt to a metal salt in
dichloromethane, which precipitates the metal halide removed by
filtration. The pure material is obtained by evaporation of the
filtrate.
[0077] The present inventors have discovered that the .beta.-diones
of the invention can be obtained from natural sources and, in
particular, from volatile oil-bearing organisms. Accordingly, in
another aspect, the present invention encompasses the use of a
.beta.-dione compound, particularly a .beta.-diketone or a
.beta.-triketone compound, obtainable from a volatile oil-bearing
organism, including an analogue or derivative thereof, in the
preparation of a pesticidal composition for controlling harmful,
annoying or undesired pests.
[0078] The present invention contemplates the use of any volatile
oil-bearing organism that produces .beta.-diones, preferably the
.beta.-diones according to any one of formulae (I)-(V), and
especially (3-diketones and/or .beta.-triketones, for the
preparation of the pesticidal compositions of the invention.
Preferred volatile oil-bearing organisms are volatile oil-bearing
plants including, but not restricted to, plants from the families
Alliaceae, Apiaceae, Asteraceae, Cannabinaceae, Lamiaceae,
Pteridaceae, Myrtaceae, Myoporaceae, Proteaceae, Rutaceae and
Zingiberaceae. Preferably, the volatile oil-bearing plant is
selected from genera of the Myrtaceae family including, but not
limited to, Angophora, Austromyrtus, Backhousia, Baeckea,
Callistemon, Corymbia, Darwinia, Eucalyptus, Kunzea, Leptospermum,
Melaleuca, Syzygium and Xanthostemon.
[0079] Thus, the compositions of the present invention may contain
as active ingredients substantially purified .beta.-diones or crude
.beta.-dione-containing extracts obtained from a volatile
oil-bearing organism, preferably a volatile oil-bearing plant.
Volatile oils, also known in the art as essential oils, typically
comprise a volatile mixture of esters, aldehydes, alcohols, ketones
and terpenes, which can be prepared from botanical materials or
plant cell biomass from cell culture. Volatile oils can be prepared
by subjecting botanical materials to a distillation process, for
example. A number of different procedures can be used for
distillation. For example, plant matter (e.g., foliage, stems,
roots, seeds, bark etc) of a volatile oil-bearing plant is placed
in a suitable still and steam distillation is used to break down
the cells of the plant to release the oil. The steam is then
condensed and the oil phase is separated from the aqueous phase to
obtain the volatile oil. It will be appreciated that other methods
of volatile oil extraction (e.g., solvent extraction) are known to
those of skill in the art and it will be understood, in this
regard, that the present invention is not limited to the use or
practice of any one particular method of extracting volatile
oils.
[0080] Suitably, the compositions comprise naturally-occurring
compounds derived from a volatile oil-bearing organism. Thus, in a
preferred embodiment, the pesticidal composition of the invention
comprises one or more .beta.-dione active compounds, particularly
.beta.-diketone- and/or .beta.-triketone-active compounds, that are
derived from the volatile oil of a volatile oil-bearing organism.
In this embodiment, the composition may optionally contain a
naturally-occurring carrier and/or other naturally-occurring
additives.
[0081] Naturally-occurring additives contemplated by the present
invention include natural antioxidants, which can be used
advantageously to reduce the effect of oxidation of the compounds
of the invention. An example of a suitable naturally-occurring
antioxidant is .alpha.-tocopherol. Other additives, such as
naturally-occurring stabilisers, are also contemplated, which may
desirably be added to improve the stability and shelf life of the
composition. Examples of suitable natural stabilisers include gum
arabic, guar gum, sodium caseinate, polyvinyl alcohol, locust bean
gum, xanthan gum, kelgum, and mixtures thereof.
[0082] In an alternate embodiment, the naturally-occurring
compounds derived from a volatile oil may be modified or
derivatised to improve, for instance, their shelf-life, stability,
activity and/or bioavailability.
[0083] The compounds of the present invention are useful for
controlling harmful, annoying or undesired pests. They may be used
singularly or in combination with other pest-controlling compounds
of the invention. By "controlling" is meant preventing, combating,
eradicating, destroying, repelling, or mitigating pests or
increasing the mortality or inhibiting the growth and/or
development of pests. The term "pest" is used herein in its
broadest sense and includes within its scope insects, arachnids
(e.g., acari, spiders), helminths (e.g., nematodes), molluscs,
protozoa (e.g., Plasmodium sp. Paramecium sp.), viruses (e.g.,
herpesviruses) and the like. Suitable applications for such control
include, but are not limited to, combating and/or eradicating
infestations by pests in animals (including humans) and/or plants
(including trees) and/or stored products, which includes the
administration to the animal or site of an effective quantity of a
compound of the invention.
[0084] By "effective amount" is meant the administration or
application of that amount of active compound, either in a single
dose or as part of a series, that is effective for controlling a
significant number of pests. Thus, for example, a
"pesticidally-effective" amount is the amount of active compound
that is effective for increasing the mortality or decreasing the
growth of a significant number of pests. Alternatively, a
"pest-repelling" effective amount is the amount of active compound
that is noxious to, and/or induces behavioural changes in, a
significant number of pests. The effective amount will vary
depending upon the taxonomic group of pest exposed to the active
compound, the formulation of the composition, and other relevant
factors. It is expected that the amount will fall in a relatively
broad range that can be determined through routine trials.
[0085] Accordingly, the compounds of the present invention can be
used as pesticides, such as but not limited to insecticides,
arachnicides, anti-helminthics, molluscicides antivirals,
antiprotozoals and the like, or as pest repellents including
repellents of insects, arachnids, helminths, molluscs, protozoa and
viruses. In especially preferred embodiments, the compounds of the
present invention are used in the control of insects, arachnids,
helminths or molluscs. In practice, the compounds can be applied as
formulations containing the various adjuvants and carriers known to
or used in the industry for facilitating bioavailability, stability
and dispersion. The choice of formulation and mode of application
for any given compound may affect its activity, and selection will
be made accordingly.
[0086] In general, a pest-controlling compound of the invention can
be compounded with appropriate inert carriers and additives in an
appropriate ratio by means of dissolving, separating, suspending,
mixing, impregnating, adsorbing or precipitating operation to
formulate into oil formulations, emulsifiable concentrates,
wettable powders, flowables, granules, powders, dusts, solutions,
suspensions, emulsions, controlled-release forms such as
microcapsules, aerosols or fumigants. Typically, the compounds of
the present invention can be mixed with a solid carrier, liquid
carrier or gas carrier, optionally together with a surfactant and
other adjuvants useful for such formulations.
[0087] The compounds of the invention can be used in an amount from
about 0.00005% to about 90% by weight as contained in these
formulations as their active component. As used herein, the term
"about" refers to a quantity, level, value or amount that varies by
as much as 30%, preferably by as much as 20%, and more preferably
by as much as 10% to a reference quantity, level, value or
amount.
[0088] Where the compounds are in the form of
.beta.-dione-containing extracts, the formulations will usually
comprise as their principal active ingredient from about 0.0001% to
about 90%, preferably from about 0.0001% to about 50%, more
preferably from about 0.0005% to about 10%, even more preferably
from about 0.0005% to about 5%, even more preferably from about
0.0005% to about 1% and still even more preferably from about
0.001% to about 0.5% by weight of the extract.
[0089] Alternatively, where the compounds are in the form of
substantially purified preparation of .beta.-diones, the
formulations will usually comprise as their principal active
ingredient from about 0.00005% to about 90%, preferably from about
0.0001% to about 50%, more preferably from about 0.0005% to about
10%, even more preferably from about 0.001% to about 5% and still
even more preferably from about 0.001% to about 1% by weight of the
substantially purified .beta.-dione.
[0090] By "substantially purified" is meant a compound which has
been separated from components that naturally accompany it.
Typically, a compound is substantially pure when at least 60%, more
preferably at least 75%, more preferably at least 90%, and most
preferably at least 99% of the total material (by volume, by wet or
dry weight, or by mole percent or mole fraction) in a sample is the
compound of interest. Purity can be measured by any appropriate
method, e.g., by chromatography or HPLC analysis.
[0091] Examples of solid carriers useful in preparing the
formulations are clays including kaolin clay, diatomite,
water-containing synthetic silicon oxide, bentonite, Fubasami clay,
and acid clay; talcs; ceramics; inorganic minerals such as Celite,
quartz, sulfur, active carbon, calcium carbonate and hydrated
silica; and chemical fertilisers such as ammonium sulfate, ammonium
phosphate, ammonium nitrate, urea and ammonium chloride, these
solid carriers being finely divided or granular. Examples of useful
liquid carriers are water, alcohols such as methanol and ethanol,
ketones such as acetone and methyl ethyl ketone, aromatic
hydrocarbons such as benzene, toluene, xylene, ethylbenzene and
methylnaphthalene, aliphatic hydrocarbons such as hexane,
cyclohexane, kerosene and light oil, esters such as ethyl acetate
and butyl acetate, nitriles such as acetonitrile and
isobutyronitrile, ethers such as diisopropyl and dioxane, acid
amides such as N,N-dimethylformamide and N,N-dimethylacetamide,
halogenated hydrocarbons such as dichloromethane, trichloroethane
and carbon tetrachloride, dimethyl sulfoxide, and fish oils,
mineral oils, plant derived oils such as canola oil, cotton-seed
oil, soybean oil and sesame oil as well as essential oils such as
lavender oil, eucalyptus oil, tea tree oil, citrus oil etc. Solid
or liquid carriers can be used alone or in combination. Examples of
gas carriers, i.e., those of propellants, are butane gas, LPG
(liquefied petroleum gas), dimethyl ether, fluorocarbons and carbon
dioxide gas.
[0092] Examples of surfactants are alkylsulfuric acid esters,
alkylsulfonic acid salts, alkylarylsulfonic acid salts, alkyl aryl
ethers and polyoxyethylene adducts thereof, polyethylene glycol
ethers, polyhydric alcohol esters, sugar alcohol derivatives,
sorbitane monolaurate, alkylallyl sorbitane monolaurate,
alkylbenzene sulfonate, alkylnaphthalene sulfonate, lignin
sulfonate, and sulfuric acid ester salts of higher alcohols. These
surfactants may be used alone or in combination.
[0093] Examples of adjuvants for the formulations, such as binders
and dispersants, are casein, gelatin, polysaccharides such as
starch, gum arabic, cellulose derivatives and alginic acid, lignin
derivatives, bentonite, sugars and water-soluble synthetic
high-molecular-weight substances such as polyvinyl alcohol,
polyvinyl pyrrolidone and polyacrylic acids. Examples of
stabilisers are PAP (acid isopropyl phosphate), BHT
(2,6-di-tert-butyl-4-methylphenol), BHA (mixture of
2-tert-butyl-4-methoxyphenol and 3-tert-butyl-4-methoxyphenol),
synergists such as piperonyl butoxide, vegetable oils, mineral
oils, fish oils, surfactants and fatty acids or esters thereof.
[0094] Emulsifying agents that may be used are suitably one or more
of those selected from non-ionic or anionic emulsifying agents.
Examples of non-ionic emulsifying agents include, but are not
restricted to, polyoxyethylenealkylphenylether,
polyoxyethylenealkylether, polyethyleneglycol fatty ester, sorbitan
fatty ester, polyoxyethylene sorbitan fatty ester,
polyoxyethylenesorbitol fatty ester,
polyoxyethylenepolyoxypropylenealkylether. Examples of anionic
emulsifying agents include alkyl sulphates,
polyoxyethylenealkylether sulphates, sulfosuccinates, taurine
derivatives, sarcosine derivatives, phosphoric esters,
alkylbenzenesulfonates and the like. A mixture consisting of
polyoxyethylenestyrylphenylether and calcium allylbenzenesulfonate
is preferred. These emulsifying agents may be used in an amount of
5 to 20 weight parts per 100 weight parts of the compositions of
the present invention.
[0095] Formulation thus obtained can be used solus or diluted, for
example, with water or other diluent. The formulations can be used
also as admixtures with other pesticides such as insecticides,
arachnids, anti-helminthics, molluscicides, herbicides, plant
growth regulators, synergists, soil improvers, baits and the like,
or can be used simultaneously with such agents without mixing. For
example, the pest-controlling compounds of the invention can be
combined with other naturally derived bioactive compounds or
extracts such as neem or its components, derris, pyrethrum;
microbial extracts such as avermectins or streptomycins; with
synthetic insecticides, acaricides, molluscicides,
anti-helminthics; antiprotozoals, antivirals or with microorganisms
having insecticidal, acaricidal, molluscicidal, anti-helminthic
anti-protozoal or antiviral activity e.g., bacteria such as
Bacillus thuringiensis, Bacillus popillae, entomogenous fungi such
as Metarhizium spp., Verticillium lecanii, nematodes such as
Steinernema spp and Heterorhabditis. Alternatively, or in addition,
the pest-controlling compounds of the invention can be combined
with synergists such as piperonyl butoxide, and with ultraviolet
screening compounds of natural or synthetic origin.
[0096] When used as an agricultural pesticide, the compound of the
invention is preferably applied usually in an amount of 0.01 to 500
g/100 m.sup.2. When an emulsifiable concentrate, wettable powder or
flowables are used as diluted with water, the compound is applied
usually at a concentration of 0.1 to 1000 ppm, preferably 1 to 500
ppm. The granular or dust can be applied without dilution.
[0097] The amount or concentration of application, although
exemplified above, can be suitably increased or reduced according
to the type of preparation, time, place, method of application,
kind of pest and extent of harm or annoyance suffered.
[0098] The invention also contemplates the use of the above
described .beta.-dione compounds in pest repellent, particularly
insect repellent, compositions. Repellent compositions contemplated
by the present invention include those that are noxious to, and/or
induce behavioural changes in, a pest. The latter compositions
suitably comprise an activity including, but not restricted to, an
antifeedant activity, an oviposition deterrent activity and an
insect growth regulatory activity. Insect repellent compositions in
various dosage forms can be prepared by blending the
above-described .beta.-dione compounds as active ingredients with a
base of cosmetics or pharmaceuticals, which are usually applied to
human bodies or animals. They can be formulated in, for example,
lotions, aerosols, milky lotions, creams or the like. These
compounds can be further incorporated with other insect repellents,
antioxidants, UV-absorbers, humectants or other additives.
[0099] The above compounds or the above-prepared compositions of
the present invention can be applied directly to human bodies or
animals. Besides, substrates, such as sheets, films, nets, timber
or the like, which have preliminarily been treated with the above
compounds or compositions by means of application, impregnation or
blending, can also be used.
[0100] The quantity of the above compounds to be formulated in the
noxious-insect repellents depends upon the dosage form, usage or
other conditions. Suitable dosages may be selected from about 0.1%
to about 90% by weight.
[0101] Thus, in another aspect of the present invention there is
provided a method for controlling harmful, annoying or undesired
pests, said method comprising exposing said pests to a
pest-controlling effective amount of a composition comprising a
.beta.-dione compound as broadly described above. Preferred
embodiments of this type include exposing said pests to a
pesticidally effective amount or a pest-repelling effective amount
of said composition.
[0102] The terms "comprise", "comprises" and "comprising" and the
like refer, unless the context requires otherwise, to the inclusion
of a stated step or element or group of steps or elements but not
the exclusion of any other step or element or group of steps or
elements.
[0103] The compositions and methods of the present invention may be
applied to pests including, but not restricted to, insects,
arachnids, helminths, molluscs, protozoa and viruses. For example,
suitable insects that fall within the scope of the present
invention include those:
[0104] (a) from the order of the lepidopterans (Lepidoptera), for
example, Adoxophyes orana, Agrotis ipsilon, Agrotis segetum,
Alabama argillacea, Anticarsia gemmatalis, Argyresthia conjugella,
Autographa gamma, Cacoecia murinana, Capua reticulana,
Choristoneura fumiferana, Chilo partellus, Choristoneura
occidentalis, Cirphis unipuncta, Cnaphalocrocis medinalis,
Crocidolomia binotalis, Cydia pomonella, Dendrolimus pini,
Diaphania nitidalis, Diatraea grandiosella, Earias insulana,
Elasmopalpus lignosellus, Eupoecilia ambiguella, Feltia
subterranea, Grapholitha funebrana, Grapholitha molesta, Heliothis
armigera, Heliothis virescens, Heliothis zea, Hellula undalis,
Hibernia defoliaria, Hyphantria cunea, Hyponomeuta malinellus,
Keiferia lycopersicella, Lambdina fiscellaria, Laphygma exigua,
Leucoptera scitella, Lithocolletis blancardella, Lobesia botrana,
Loxostege sticticalis, Lymantria dispar, Lymantria monacha,
Lyonetia clerkella, Manduca sexta, Malacosoma neustria, Mamestra
brassicae, Mocis repanda, Operophthera brumata, Orgyia
pseudotsugata, Ostrinia nubilalis, Pandemis heparana, Panolis
flammea, Pectinophora gossypiella, Phthorimaea operculella,
Phyllocnistis citrella, Pieris brassicae, Plathypena scabra,
Platynota stultana, Plutella xylostella, Prays citri, Prays oleae,
Prodenia sunia, Prodenia ornithogalli, Pseudoplusia includens,
Rhyacionia frustrana, Scrobipalpula absoluta, Sesamia inferens,
Sparganothis pilleriana, Spodoptera frugiperda, Spodoptera
littoralis, Spodoptera litura, Syllepta derogata, Synanthedon
myopaeformis, Thaumatopoea pityocampa, Tortrix viridana,
Trichoplusia ni, Tryporyza incertulas, Zeiraphera canadensis,
[0105] (b) furthermore Galleria mellonella and Sitotroga
cerealella, Ephestia cautella, Tineola bisselliella;
[0106] (c) from the order of the beetles (Coleoptera), for example,
Anthonomus grandis, Anthonomus pomorum, Apion vorax, Atomaria
linearis, Blastophagus piniperda, Cassida nebulosa, Cerotoma
trifurcata, Ceuthorhynchus assimilis, Ceuthorhynchus napi,
Chaetocnema tibialis, Conoderus vespertinus, Crioceris asparagi,
Dendroctonus refipennis, Diabrotica longicornis, Diabrotica
12-punctata, Diabrotica virgifera, Epilachna varivestis, Epitrix
hirtipennis, Eutinobothrus brasiliensis, Hylobius abietis, Hypera
brunneipennis, Hypera postica, Ips typographus, Lema bilineata,
Lema melanopus, Leptinotarsa decemlineata, Limonius californicus,
Lissorhoptrus oryzophilus, Melanotus communis, Meligethes aeneus,
Melolontha hippocastani, Melolontha melolontha, Oulema oryzae,
Ortiorrhynchus sulcatus, Otiorrhynchus ovatus, Phaedon cochleariae,
Phyllopertha horticola, Phyllophaga sp., Phyllotreta chrysocephala,
Phyllotreta nemorum, Phyllotreta striolata, Popillia japonica,
Psylliodes napi, Scolytus intricatus, Sitona lineatus,
[0107] (d) furthermore Bruchus rufimanus, Bruchus pisorum, Bruchus
lentis, Sitophilus granaria, Lasioderma serricorne, Oryzaephilus
surinamensis, Rhyzopertha dominica, Sitophilus oryzae, Tribolium
castaneum, Trogoderma granarium, Zabrotes subfasciatus;
[0108] (e) from the order of the dipterans (Diptera), for example,
Anastrepha ludens, Ceratitis capitata, Contarinia sorghicola, Dacus
cucurbitae, Dacus oleae, Dasineura brassicae, Delia coarctata,
Delia radicum, Hydrellia griseola, Hylemyia platura, Liriomyza
sativae, Liriomyza trifolii, Mayetiola destructor, Orseolia oryzae,
Oscinella frit, Pegomya hyoscyami, Phorbia antiqua, Phorbia
brassicae, Phorbia coarctata, Rhagoletis cerasi, Rhagoletis
pomonella,
[0109] (f) furthermore Aedes aegypti, Aedes vexans, Anopheles
maculipennis, Chrysomya bezziana, Chrysomya hominivorax, Chrysomya
macellaria, Cordylobia anthropophaga, Culex pipiens, Fannia
canicularis, Gasterophilus intestinalis, Glossina morsitans,
Haematobia irritans, Haplodiplosis equestris, Hypoderma lineata,
Lucilia caprina, Lucilia cuprina, Lucilia sericata, Musca
domestica, Muscina stabulans, Oestrus ovis, Tabanus bovinus,
Simulium damnosum;
[0110] (g) from the order of the thrips (Thysanoptera), for
example, Frankliniella fusca, Frankliniella occidentalis,
Frankliniella tritici, Haplothrips tritici, Heliothrips
haemorrhoidalis, Scirtothrips citri, Thrips oryzae, Thrips palmi,
Thrips tabaci;
[0111] (h) from the order of the hymenopterans (Hymenoptera), for
example, Athalia rosae, Atta cephalotes, Atta sexdens, Atta texana,
Hoplocampa minuta, Hoplocampa testudinea, Iridomyrmes humilis,
Iridomyrmex purpureus, Monomorium pharaonis, Solenopsis geminata,
Solenopsis invicta, Solenopsis richteri, Technomyrmex albipes;
[0112] (i) from the order of the heteropterans (Heteroptera), for
example, Acrosternum hilare, Blissus leucopterus, Cyrtopeltis
notatus, Dysdercus cingulatus, Dysdercus intermedius, Eurygaster
integriceps, Euschistus impictiventris, Leptoglossus phyllopus,
Lygus hesperus, Lygus lineolaris, Lygus pratensis, Nezara viridula,
Piesma quadrata, Solubea insularis, Thyanta perditor;
[0113] (j) from the order of the homopterans (Homoptera), for
example, Acyrthosiphon onobrychis, Acyrthosiphon pisum, Adelges
laricis, Aonidiella aurantii, Aphidula nasturtii, Aphis fabae,
Aphis gossypii, Aphis pomi, Aulacorthum solani, Bemisia tabaci,
Brachycaudus cardui, Brevicoryne brassicae, Dalbulus maidis,
Dreyfusia nordmannianae, Dreyfusia piceae, Dysaphis radicola,
Empoasca fabae, Eriosoma lanigerum, Laodelphax striatella,
Macrosiphum avenae, Macrosiphum euphorbiae, Macrosiphon rosae,
Megoura viciae, Metopolophium dirhodum, Myzus persicae, Myzus
cerasi, Nephotettix cincticeps, Nilaparvata lugens, Perkinsiella
saccharicida, Phorodon humuli, Psylla mali, Psylla piri, Psylla
pyricola, Rhopalosiphum maidis, Schizaphis graminum, Sitobion
avenae, Sogatella furcifera, Toxoptera citricida, Trialeurodes
abutilonea, Trialeurodes vaporariorum, Viteus vitifolii;
[0114] (k) from the order of the termites (Isoptera), for example,
Calotermes flavicollis, Coptotermes spp, Leucotermes flavipes,
Macrotermes subhyalinus, Nasutitermes spp such as Nasutitermes
walkeri, Odontotermes formosanus, Reticulitermes lucifugus, Termes
natalensis;
[0115] (l) from the order of the orthopterans (Orthoptera), for
example, Gryllotalpa gryllotalpa, Locusta migratoria, Melanoplus
bivittatus, Melanoplus femur-rubrum, Melanoplus mexicanus,
Melanoplus sanguinipes, Melanoplus spretus, Nomadacris
septemfasciata, Schistocerca americana, Schistocerca peregrina,
Stauronotus maroccanus, Schistocerca gregaria,
[0116] (m) furthermore Acheta domestica, Blatta orientalis,
Blattella germanica, Periplaneta americana;
[0117] (n) from the order of the phthirapterans (Phthiraptera), for
example, Mallophaga, such as Damalina spp., and Anoplura such as
Linognathus and Haematopinus spp.;
[0118] (o) from the order of the hemipterans (Hemiptera), for
example, Aphis, Bemisia, Phorodon, Aeneolamia, Empoasca,
Parkinsiella, Pyrilla, Aonidiella, Coccus, Pseudococcus,
Helopeltis, Lygus, Dysdercus, Oxycarenus, Nezara, Aleyrodes,
Triatoma, Psylla, Myzus, Megoura, Phylloxera, Adelges, Nilaparvata,
Nephotettix or Cimwx spp.;
[0119] (p) from the order of the siphonapterans (Siphonaptera), for
example, Ctenocephalides or Pulex spp.;
[0120] (q) from the order of the thysanurans (Thysanura), for
example, Lepisma spp.;
[0121] (r) from the order of the dermapterans (Dermaptera), for
example, Forficula spp.; and
[0122] (s) from the order of the psocopterans (Psocoptera), for
example, Peripsocus spp.
[0123] Arachnids contemplated by the present invention include, but
are not limited to, spiders and scorpions and especially mites such
as phytophagous mites (Acari), such as Aculops lycopersicae,
Aculops pelekassi, Aculus schlechtendali, Brevipalpus phoenicis,
Bryobia praetiosa, Eotetranychus carpini, Eutetranychus banksii,
Eriophyes sheldoni, Oligonychus pratensis, Panonychus ulmi,
Panonychus citri, Phyllocoptruta oleivora, Polyphagotarsonemus
latus, Tarsonemus pallidus, Tetranychus cinnabarinus, Tetranychus
kanzawai, Tetranchus pacificus, Tetranychus urticae, ticks, such as
Amblyomma americanum, Amblyomma variegatum, Argas persicus,
Boophilus annulatus, Boophilus decoloratus, Boophilus microplus,
Dermacentor silvarum, Hyalomma truncatum, Ixodes ricinus, Ixodes
rubicundus, Ornithodorus moubata, Otobius megnini, Rhipicephalus
appendiculatus and Rhipicephalus evertsi, and animal-parasitic
mites, such as Dermanyssus gallinae, Psoroptes ovis and Sarcoptes
scabiei.
[0124] Helminths falling within the scope of the present invention
may be selected from cestodes such as fish tapeworm, pork tapeworm,
beef tapeworm, and dwarf tapeworm; trematodes such as from the
genera Metagonimus and Heterophyes; and nematodes such as but not
limited to filariid, ascarid, strongyle and trichostrongyle
nematodes of the genera Acanthocheilonema, Aelurostrongylus,
Ancylostoma, Angiostrongylus, Ascaris, Brugia, Bunostomum,
Dictyocaulus, Dioctophyme, Dipetalonema, Dirofilaria, Dracunculus,
Filaroides, Lagochilascaris, Loa, Mansonella, Muellerius, Necator,
Onchocerca, Parafilaria, Parascaris, Protostrongylus, Setaria,
Stephanofilaria, Strongyloides, Strongylus, Thelazia, Toxascaris,
Toxocara, Trichinella, Uncinaria and Wuchereria.
[0125] Suitable molluscs include those of the Gastropoda class
examples of which include snails, slugs, conchs, and whelks.
[0126] Protozoa may be selected for example from Plasmodia,
Toxoplasma, Leishmania, Trypanosoma, Giardia, Entamoeba,
Acanthamoeba, Nagleria, Hartmanella, Balantidium, Babesia,
Cryptosporidium, Isospora, Microsporidium, Trichomonas or
Pneumocystis species.
[0127] Viruses may be selected from RNA viruses or DNA viruses,
which include but are not limited to Human Immunodeficiency Virus
(HIV), Poliovirus, Influenza virus, Rous Sarcoma virus,
Flaviviruses such as Japanese encephalitis, Influenza virus,
Respiratory Syncytial Virus, Hepatitis virus, Parvovirus,
Rotavirus, Coronavirus, Adenovirus and Herpesviruses such as
Papillomavirus and Epstein-Barr virus.
[0128] The present invention also extends to methods for producing
resistance in plants to pests including, but not limited to,
insects, arachnids, helminths, molluscs, protozoa and viruses by
crossing a plant expressing a .beta.-dione compound according to
the invention with pest susceptible lines. Crossing a
.beta.-dione-producing plant into a pest susceptible background
would produce a resistant plant with a high level of pest
resistance. Plants that could be made pest resistant include, but
are not limited to, dicotyledonous plants, especially trees and
more especially members of the Myrtaceae family. For example E.
cloeziana commonly known as Gympie Messmate is one of the many
Eucalyptus species grown for hard wood production. However the oil
present in this chemotype does not contain .beta.-diones and hence
an intra species cross with the unique North Queensland tasmanone
chemotype would introduce this phenotypic trait. Such a process
would be readily applicable to other Eucalyptus species of
commercial interest. Interspecific crossing within the Myrtaceae
family is well established to those skilled in the art and
inclusion of .beta.-dione as an additional trait into formal
breeding programs is acknowledged.
[0129] Suitable .beta.-dione-producing plants may be selected from
the families Alliaceae, Apiaceae, Asteraceae, Cannabinaceae,
Lamiaceae, Pteridaceae, Myrtaceae, Myoporaceae, Proteaceae,
Rutaceae and Zingiberaceae. Preferably, the volatile oil-bearing
plant is selected from genera of the Myrtaceae family including,
but not limited to, Angophora, Austromyrtus, Backhousia, Baeckea,
Callistemon, Corymbia, Darwinia, Eucalyptus, Kunzea, Leptospermum,
Melaleuca, Syzygium and Xanthostemon. Preferred 3-dione-producing
plants are Leptospermum morrisonii, Eucalyptus bensonii, Eucalyptus
megacornuta, Eucalyptus pilularis, Eucalyptus cornuta, Eucalyptus
baxteri, Eucalyptus macrorhyncha, Eucalyptus cloeziana, Melaleuca
cajuputi, Eucalyptus jensenii, Backhousia angustifolia and
Leptospermum scoparium. A particularly preferred
.beta.-dione-producing plant is Eucalyptus cloeziana.
[0130] As used herein, the term "plant" includes reference to whole
plants, plant organs (e.g., leaves, stems, roots, etc.), seeds and
plant cells and progeny of same. Plant cell, as used herein
includes, without limitation, seeds suspension cultures, embryos,
meristematic regions, callus tissue, leaves, roots, shoots,
gametophytes, sporophytes, pollen, and microspores. The class of
plants which can be used in the methods of the invention is
generally as broad as the class of higher plants amenable to
transformation techniques, including both monocotyledonous and
dicotyledonous plants.
[0131] Thus, the present invention contemplates conventional plant
breeding methods to transfer the genetic material associated with
.beta.-dione production via crossing and backcrossing. Such methods
will comprise the steps of: (1) sexually crossing the
.beta.-dione-producing plant with a plant from a pest susceptible
taxon; (2) recovering reproductive material from the progeny of the
cross; and (3) growing .beta.-dione-producing/pest-resistant plants
from the reproductive material. Where desirable or necessary, the
agronomic characteristics of the susceptible taxon can be
substantially preserved by expanding this method to include the
further steps of repetitively: (1) backcrossing the pest-resistant
progeny with pest-susceptible plants from the susceptible taxon;
and (2) selecting for expression of a .beta.-dione (or an
associated marker gene) among the progeny of the backcross, until
the desired percentage of the characteristics of the susceptible
taxon are present in the progeny along with the gene or genes
imparting .beta.-dione activity.
[0132] By the term "taxon" herein is meant a unit of botanical
classification. It thus includes, genus, species, cultivars,
varieties, variants and other minor taxonomic groups which lack a
consistent nomenclature.
[0133] In order that the invention may be readily understood and
put into practical effect, particular preferred embodiments will
now be described by way of the following non-limiting examples.
EXAMPLES
Example 1
.beta.-Triketone-Containing Oils Obtained from Australian Myrtaceae
Species
[0134] Australia has an extensive number of volatile oils from
species of the Myrtaceae that are rich in a diversity of
structurally related constituents known as 3-triketones. These oils
often show not only a high yield of oil, but also a high degree of
biosynthetic selectivity that produces .beta.-triketones in a high
proportion of the oil composition. The major constituents of the
published Myrtaceae essential oils (Hellyer, 1968; Boland and
Brophy, 1990, 1993; Brophy, et al., 1995; Bignall et al., 1997;
Southwell and Brophy 2000) are listed in Table 1 and their
structures are included in FIG. 1.
TABLE-US-00001 TABLE 1 .beta.-Triketone Profiles of Australian
Essential Oils Species Yield .beta.-Triketone (%) Distribn
Backhousia angustifolia 2.5 Angustione (85) QLD Backhousia
angustifolia 2.5 Dehyroangustione (80) QLD Eucalyptus cloeziana 3.0
Tasmanone (95) QLD Eucalyptus suberea 1.4 Tasmanone (94) WA
Eucalyptus lateritica 0.9 Tasmanone (37), lateriticone (14) WA
Eucalyptus camfieldii Tasmanone (40) NSW Leptospermum scoparium 0.4
Leptospermone (19), Flavesone (8), NSW, NZ Isoleptospermone (5)
Eucalyptus grandis 0.6 Leptospermone (20), Flavesone (13), NSW
Isoleptospermone (3) Eucalyptus agglomerata Agglomerone (40) NSW
Eucalyptus mckieana Agglomerone (60) NSW Eucalyptus bensonii 2.5
Agglomerone (72) Eucalyptus insularis 1 Agglomerone (19) WA
Eucalyptus jensenii 0.3 Jensenone (70) NT Eucalyptus papuana 0.7
Papuanone (40) Nth Aus Leptospermum morrisonii 1.8 Grandiflorone
(58) NSW Melaleuca cajeputi Platyphyllol Nth Aus Xanthostemon
Xanthostemone oppositifolius
[0135] The .beta.-triketones obtained from selected Myrtaceous
volatile oils were shown to have significant insecticidal and/or
acaricidal activity.
Example 2
Insecticidal Activity
[0136] Initial insecticidal screening against two important
arthropod species, two-spotted mite (Tetranychus urticae) and
diamond back moth (Plutella xylostella) lavae highlighted three
oils on the basis of efficacy, oil yield, oil profile and ease of
recollection (Table 2). Where feasible, the LD.sub.50 and LD.sub.95
values were determined.
TABLE-US-00002 TABLE 2 Percentage Mortality of Three Efficacious
Oils Two Spotted Mite Diamond Back Moth % Mortality % Mortality
Species (0.5%/1.0%) (0.5%/1.0%) Backhousia angustifolia 98/98
100/100 Backhousia angustifolia 100/100 100/100 Eucalyptus
cloeziana 100/100 70/100
[0137] Insecticidal tests using the oil from fresh plant
recollections and steam distillations varied occasionally and, in
this regard, it is believed that improving storage conditions
including temperature, light and exposure to air and inclusion of a
dessicant can enhance the stability of the active fraction of such
oils.
[0138] E. cloeziana oil continued to show high potency against both
insect tests. This oil exhibited an LD.sub.95 between 0.04-0.20%
(depending on formulation and treatment) against two-spotted mite.
An LD.sub.95 of 0.10% was observed against 1.sup.st instar lavae of
diamond back moth and this rose to 0.78% when tested against
3.sup.rd instar lavae. In additional preliminary investigations
with greenhouse thrips (Heliothrips haemorrhoidalis), a 0.1%
concentration of E. cloeziana oil induced 100% mortality.
[0139] Additional work was carried out on the E. cloeziana oil to
explore the contribution the various components make to the overall
efficacy of this oil. Fractions 1, 3, 4, and 5 outlined in Example
3 were screened against two-spotted mite. Fractions 1 and 5 showed
no significant insecticidal effect. Fractions 3 and 4, comprising
98% and 99% tasmanone were active and showed little difference to
the activity of the whole oil.
[0140] This suggests that not only is tasmanone the principle
component in the oil, but it is also the principle bioactive
constituent. It is also reasonable to assume that as the activity
of E. cloeziana oil has been demonstrated against a number of
different arthropod species, namely a mite (T. urticae), a
caterpillar (P. xylostella) and a thrips (H. haemorrhoidalis), its
insecticidal activity is broad in nature.
Example 3
Chemistry
[0141] Chemical analysis (GC-MS) of the steam-distilled oils from
the collected plants in this work are summarised in Table 3.
TABLE-US-00003 TABLE 3 .beta.-Triketone Profiles of Selected Oils
Plant Source Principle Component* (%) Backhousia angustifolia -I
Dehydroangustione 85% Backhousia angustifolia -II
Dehydroangustinone (80%) Backhousia angustifolia -III Angustinone
(65%) Backhousia angustifolia -IV Angustinone (28%) Eucalyptus
cloeziana Tasmanone (84-96%) Melaleuca cajuputi subsp platyphylla
Platyphyllol (64-71%) * As determined by gas chromatography
[0142] The most promising oils were derived from B. angustifolia,
M. cajuputi subsp platyphylla and E. cloeziana and these were then
subjected to additional chemical fractionation. The lower
insecticidal activity observed in the recollected B. angustifolia
(IV) was in part attributed to the lower levels of
.beta.-triketone. The level of .beta.-triketone was elevated by the
removal of the more volatile monoterpenes using vacuum
distillation.
[0143] One of the most efficacious oils was from E. cloeziana, a
tasmanone (84-96%) rich oil with additional terpenes and
.beta.-triketones (FIG. 2, Table 4), which displayed consistent
activity at every stage of processing and formulation. This oil
was, therefore, fractionated using column chromatography on silica
gel with a hexane-diethyl ether gradient and a final methanol
elution. The profiles of the fractions are summarised in Table
5.
TABLE-US-00004 TABLE 4 Chemical Profile of E. cloeziana Oil Peak No
Compound Composition (%) 1 a-pinene 0.5-1.9 2 B-pinene 1.5-5.7 3
Limonene 0.1-0.6 4 a-terpineol 0.7-2.0 5 Globulol 0.01-0.5 6
Agglomerone 0.01-0.6 7 Tasmanone 84-96 8 Lateriticone 0.2-0.7 9
Isolateriticone 0.3-1.2
TABLE-US-00005 TABLE 5 Fractions Cut From E. cloeziana Oil Fraction
No Solvent System Composition Amount F1 Hexane Hydrocarbons 73 mg
F2 Hex:Et2O (9:1) 80% Tasmanone 4 mg F3 Hex:Et2O (1:1) 98%
Tasmanone 3.66 g F4 Hex:Et2O (9:1) +99% Tasmanone 694 mg F5 MeOH
Terpene alcohols 64 mg
[0144] .sup.1H NMR data (FIG. 3) were recorded on F4 and confirmed
the structure of tasmanone. Moreover the compound exists in
solution (CDCl.sub.3) in tautomeric forms (FIG. 4) in the ratio
2:0:1 (A:B:C).
[0145] Another efficacious oils was from M. cajuputi subsp
platyphylla, a platyphyllol (64-71%) rich oil (Table 6), which also
displayed consistent activity at every stage of processing and
formulation.
TABLE-US-00006 TABLE 6 Typical Chemical Profile of M. cajuputi
subsp platyphylla Oil Peak No Compound Composition (%) 1 a-pinene
Tr-0.8 2 1,8-cineole Tr-0.7 3 B-caryophyllene 0.6-3.2 4 Humulene
0.6-1.3 5 Spathulenol 4.0-9.0 6 Caryophyllene oxide Tr-3.6 6
Platyphyllol 64-71 7 MW 234 - unknown 4.3 *As determined by gas
chromatography
Example 4
Phytotoxicity
[0146] Initial investigations using leaves and leaf discs of
several plant species including French bean (Phaseolus vulgaris)
lemon (Citrus limon) Orange (Citrus sinensis) and Cabbage (Brassica
oleracea) indicated that phytotoxicity did not occur for most oils
below 0.5%. More detailed investigations using soft intact leaves
of young greenhouse-grown French bean subsequently showed E.
cloeziana oil applied as a spray caused some phytotoxicity at
concentrations of 0.5% and above.
Example 5
Toxicity of E. cloeziana Oil
Bacterial Reverse Mutation Assay
[0147] This study investigated the potential of E. cloeziana oil to
induce reverse mutations at the histidine locus in the genome of
one strain of Salmonella typhimurium TA100 in the presence and
absence of a metabolic activation system (mammalian microsomal
enzymes, S9 mix). The test sample was dissolved in dimethyl
sulfoxide (DMSO). In this assay, an E. cloeziana oil test sample
did not induce an appropriate-fold increase (a 2-fold increase for
TA100) in the mean revertants per plate in the tester strain TA100
over the mean revertants per plate of the appropriate vehicle
control. Accordingly, the test sample was considered to be
non-mutagenic under the conditions of the assay.
Acute Oral
Sighting
[0148] The acute oral toxicity of E. cloeziana oil was investigated
in ten (10) Sprague Dawley Specific Pathogen Free female rats
(groups of 2) at doses of 500, 250, 125 and 50 mg/kg. The
experimental procedure was based on OECD guidelines for the testing
of chemicals, No. 420.
[0149] Clinical signs of toxicity occurred between one (1) and
twenty-four (24) hours after dosing. Both animals in the 500 mg/kg
group exhibited subdued behaviour, partial eye closure, slow
breathing, reduced motor activity, ataxia followed by death within
24 hours. The animals in the 250 mg/kg group exhibited subdued
behaviour, partial eye closure, slow breathing, social isolation,
and reduced motor activity, and had returned to normal by 24 hours
after dosing. The animals in the 125 mg/kg group exhibited subdued
behaviour, partial eye closure, slow breathing, piloerection and
reduced motor activity, and had returned to normal by 24 hours
after dosing. The animals in the 50 mg/kg group did not show any
signs of toxicity during the seven day experimental period.
[0150] There were no other clinical abnormalities in any animal
throughout the seven (7) day observation period. The stomach of one
animal in the 250 mg/kg group (5F) had a single ulcer. There were
no other gross abnormalities in the major organs of any animal at
autopsy.
Based on the results obtained from this study 50, 100 and 200 mg/kg
can be selected for a main study as the maximum non-toxic dose,
intermediate dose and high dose, respectively.
Full Study
[0151] The acute oral toxicity of E. cloeziana oil was investigated
in thirty (30) Sprague Dawley Specific Pathogen Fee rats (15 males
and 15 females) at doses of 50, 100 and 200 mg/kg. These doses were
chosen following a dose range finding in the above study. The
experimental procedure was based on OECD guidelines for the testing
of chemicals, No. 420.
[0152] The acute NOAEL of E. cloeziana oil was determined to be 50
mg/kg, and the MTD was 200 mg/kg under the conditions of this
study.
Acute Dermal
[0153] The acute dermal toxicity of E. cloeziana oil was
investigated in ten (10) Sprague Dawley Specific Pathogen Free rats
(5 males and 5 females) at a dose of 2000 mg/kg. A preliminary
study (SIGHTING) indicated no signs of toxicity at this dose. The
experimental procedure was based on OECD guidelines for the testing
of chemicals, No. 420.
[0154] No clinical abnormalities, skin irritations or body weight
losses were observed in any animal throughout the fourteen (14) day
observation period. No deaths occurred. No abnormalities were seen
in the major organs at necropsy. The rat acute dermal LD50 of E.
cloeziana oil was determined to be greater than 2000 mg/kg under
the conditions of this study.
Skin Irritancy/Corrosion
[0155] The potential of a test sample of E. cloeziana oil to
provoke skin irritation/corrosion reactions was investigated using
a primary dermal irritation/corrosion test in three (3) New Zealand
White albino rabbits (OECD Guidelines for the Testing of Chemicals,
No. 404). The results obtained from this study indicated that E.
cloeziana oil is a non-irritant according to the National
Occupational Health and Safety Commission (NOHSC) "Approved
Criteria for Classifying Hazardous Substances [NOHSC:1008
(1999)]".
Example 6
Isolation and Purification of .beta.-Triketones from Australian
Myrtaceae Species
[0156] Plant collections were commissioned at the Mt. Annan Botanic
Garden, Sydney, Australia, and the Darwin Botanic Garden, Northern
Territory, Australia to provide potential sources of a range of
.beta.-triketone isolation. The plants investigated for
.beta.-triketone exploration are listed in Table 6.
TABLE-US-00007 TABLE 6 Plants Sourced for .beta.-Triketone
Exploration Accession .beta.-triketone # Source Plant Plant
Location # anticipated 1 Leptospermum morrisonii Mt. Annan, NSW
873247 grandiflorone 2 Eucalyptus bensonii Mt. Annan, NSW 881045
agglomerone 3 Eucalyptus megacornuta Mt. Annan, NSW 831078
jensenone .sup.+ 4 Eucalyptus pilularis Mt. Annan, NSW 873166
torquatone 5 Eucalyptus cornuta Mt. Annan, NSW 852618 jensenone
.sup.+ 6 Eucalyptus baxteri Mt. Annan, NSW 860999 agglomerone/
tasmanone 7 Eucalyptus macrorhyncha Mt. Annan, NSW 860896
conglomerone 8 Eucalyptus cloeziana Lappa, QLD PF 2513 tasmanone 9
Melaleuca cajuputi ssp Bensbach, WP, PNG KW16-19 platyphyllol
platyphyla 10 Eucalyptus jensenii NT Botanic Gardens, RK114-
jensenone Darwin, NT LUS 11 Backhousia angustifolia Wilgavale,
Texas, PF1712 dehydroangustione QLD 12 Backhousia angustifolia
Didcott Creek, QLD PF 1708 angustione 13 Leptospermum scoparium NA
commercial oil NA flavesone, sample isoleptospermone, leptospermone
14 Eucalyptus conjuncta Mt Annan 854097 conglomerone .sup.+ Oil
contains other .beta.-triketone constituents
[0157] All plants, apart from 9 and 13, were steam distilled to
yield various quantities of essential oil. All oils were analysed
by Gas Chromatography Mass Spectrometry (GCMS) to determine the
presence and abundance of .beta.-triketones. Based on this
information, particular oils were targeted for isolation and
purification of .beta.-triketones using wet chemistry and
preparative HPLC techniques.
[0158] The .beta.-triketones listed in Table 7 were isolated in
quantities adequate for insecticidal screening. A minimum of 50 mg
of each compound was required. Isoleptospermone and leptospermone
were difficult to separate due to their structural similarity as
were angustione and dehydroangustione. Consequently, mixtures of
these compounds, where one isomer was significantly more abundant,
were provided for insecticidal screening as this will still allow
for differentiation in observed activity.
TABLE-US-00008 TABLE 7 .beta.-Triketones Isolated for Insecticidal
Screening Amount sent Sample Purity % by for screening
.beta.-triketones Plant source GCMS (mg) grandiflorone Leptospermum
morrisonii 100.00 99.5 jensenone Eucalyptus jensenii 100.00 67.9
dehydroangustione Backhousia angustifolia 95.02 165.02 PF1712
angustione Backhousia angustifolia 66.30 (33.70% 107.1 PF1708
dehydroangustione) agglomerone Eucalyptus bensonii 99.28 109.2
flavesone Leptospermum scoparium 99.32 101.8 isoleptospermone
Leptospermum scoparium 33.10 (66.90% 113.4 leptospermone)
leptospermone Leptospermum scoparium 95.33 77.0 tasmanone
Eucalyptus cloeziana 99.92 110.1 platyphyllol Melaleuca cajuputi
subsp. 99.62 255.6 platyphylla
[0159] The chemical structures and identities of the
.beta.-triketones isolated were confirmed by GCMS and Nuclear
Magnetic Resonance (NMR) analysis.
Example 7
Efficacy of E. cloeziana Oil on Target Organisms
Target Organisms
[0160] Two spotted mite (TSM) Tetranychus urticae Koch [Acarina:
Tetranychidae] were collected from a mass culture maintained at the
University of Western Sydney's Hawkesbury Campus in Richmond, NSW,
Australia. They were reared on potted French beans (Phaseolus
vulgaris L [Fabales: Fabaceae] in a glasshouse maintained at
25+5.degree. C., 65.+-.5% RH and 14 h D:L photoperiod. Only young
females were selected for bioassay.
[0161] Adult parthenogenetic female greenhouse thrips (GHT),
Heliothrips haemorrhoidalis Bouche (Thysanoptera: Thripidae) of
similar age were obtained from a colony reared on orange fruits and
maintained in an insectary at UWS Hawkesbury under conditions of
25.+-.3.degree. C., 65% RH and 16 h D:L photoperiod
[0162] Young nymphs of citrus aphids Toxoptera citricida (Kirkaldy)
(Hemiptera: Aphididae) were collected from lemon seedlings grown
under glasshouse conditions at UWS Hawkesbury.
[0163] Workers of the termite Nasutitermes walkeri Hill (Isoptera:
Termitidae) were collected from a laboratory culture at UWS
Richmond which was initiated from soil, termitaria and wood on
which termites were feeding were field collected at Richmond NSW,
and maintained in a darkened container under conditions of
25+2.degree. C., 35-68% RH. Termites were fed on wood collected
from near the original nest. Moistened soil from the nest together
with paper towel were placed on top of the nest and made it
possible to maintain this culture for several months in the
laboratory at UWS Hawkesbury.
[0164] Workers of the whitefooted house ant Technomyrmex albipes
(F. Smith) (Hymenoptera: Formicidae) were field collected at UWS
Hawkesbury by baiting in an empty glass jar containing sugar
granules.
[0165] Pupae of housefly, Musca domestica L (Diptera: Muscidae) and
different stages of American Cockroach, Periplaneta americana L
(Blattodea: Blattidae) were initially supplied by C.E.R.I.T and
maintained in laboratory culture at UWS Hawkesbury.
[0166] Tomato russet mites (TRM) Aculops lycopersici (Massee)
(Acarina: Eriophyidae) were collected from infested tomato plants
near Riverstone, NSW.
[0167] Mixed sex adults of the mosquito Culex quinquefaciatus
(Diptera: Culicidae) were supplied by C.E.R.I.T, held at the in the
Centre for Horticulture & Plant Science, University of Western
Sydney, Richmond NSW and treated one day after arrival.
[0168] Workers of the honeybee Apis mellifera (Hymenoptera: Apidae)
were collected from several field hives maintained at the apiary in
the Centre for Horticulture & Plant Sciences, University of
Western Sydney, Richmond NSW
[0169] Adults of ash-white-flies Aleaurocanthus woglumi (Homoptera:
Aleyrodidae) were field collected from an ornamental pear (Prunus
sp) in the Centre for Horticulture & Plant Sciences, University
of Western Sydney, Richmond NSW.
[0170] Drug store beetle Sitodrepa panicea (Coleoptera: Anobiidae)
were reared on curry powder under laboratory conditions of
25.+-.1.degree. C. and 65.+-.5% RH.
[0171] Mixed age groups of snails Helix apersa (Mollusca:
Gastropoda) were collected from infested plants in the Centre for
Horticulture & Plant Science, University of Western Sydney,
Richmond NSW.
Bioassays
TSM
[0172] From an E. cloeziana oil extract (containing 85% tasmanone),
1.1765 g was dissolved in 5 mL ethyl alcohol and distilled water
containing 200 ppm Triton X-100.TM. was added to prepare a 1% stock
solution. From this homogenised stock solution, further serial
dilutions of 0.0125, 0.025, 0.05, 0.10, and 0.12% were prepared by
mixing the required amount of stock solution in distilled water and
Triton X-100 solution. Each treatment was conducted on 60-80 TSM,
which were evenly distributed on four French bean leaf discs (25 mm
diam) contained in 90 mm diam. petri dishes. The leaf discs were
placed with their underside uppermost on moist absorbent cotton
wool covered with muslin netting. Water was added to the dishes
daily to prevent desiccation of the leaf discs. Five mL aliquots
(unless otherwise stated) were applied to each petri dish with a
Potter precision spray tower as described by Herron et al (1995).
The average mass of solution applied to each dish was calculated to
be 3.95 mg/cm.sup.2. Mortality was recorded 24 h after treatment.
Death was recognised by the absence of movement when the test
organisms were mechanically stimulated. Data were analysed using
SPSS for Windows.TM. Version 7. Probit analysis was carried out for
dose-mortality data and heterogeneity of regressions was determined
by the Pearson chi-squared characteristic.
[0173] In addition, TSM was treated with E. cloeziana oil extract
in combination with paraffin oil. In particular, E. cloeziana oil
extract at levels of 0.1, 0.2, 0.3, 0.4 and 0.5 g were weighed out
and each was made up to a weight of 10.0 g with a formulated
paraffin oil (BioPest.RTM.), which was then sonicated for 10 min. A
1.0% v/v of each blend was prepared by mixing 1.0 mL with distilled
water in a 100.0 mL volumetric flask. TSM were transferred to the
petri dish following the same standard method. Five-mL aliquots
were applied to each petri dish and mortality was recorded 24 h
after treatment. All blends of E. cloeziana oil extract with
paraffin oil produced 100.0% mortality, compared with Biopest.RTM.
alone which caused only 35.5.+-.6.9 mortality. The lowest
concentration of E. cloeziana oil extract tested in this
combination (i.e. 0.01%) resulted in 100% mortality in TSM, which
is significantly lower than that reported earlier in this document
to produce 100% mortality with E. cloeziana oil extract alone
(>0.06%).
Greenhouse Thrips
[0174] The same experimental procedure used for TSM was repeated
for GHT except that lemon leaf discs were used instead of French
beans. The required number of adult thrips for each treatment was
transferred with a fine brush to the underside of a lemon disc of
the same diameter (6 cm) as the base of a petri dish. The lemon
disc was mounted on agar with its adaxial side uppermost.
Immediately after treatment the petri dish was covered with
perforated plastic wrap. Mortality was assessed 24 h after
treatment.
Tomato Russet Mite
[0175] The same experimental procedure for TSM was repeated except
that tomato leaf discs were used instead of French bean.
Brown Citrus Aphid
[0176] Lemon leaf discs 2.5 cm diam. were cut from tender young
leaves and mounted on moistened absorbent cotton wool in 90 mm
petri dish with their adaxial surface uppermost. Each petri dish
contained four leaf discs. Uniform early instar nymphs were then
transferred with a fine brush to the leaf discs (each containing
8-10 nymphs). A Potter tower was used to apply 5 mL aliquots to
each petri dish. A control (solvent and surfactant only) was also
included in the assessment. Mortality was assessed 24 h after
treatment.
Termites
[0177] Twenty uniform termite workers were transferred to 90 mm
petri dishes lined with the same diameter moistened filter paper
(Whatman No 2). A preliminary trial was carried out using a Potter
tower to apply 5 mL aliquots of each concentration. Using this
method, all termite workers died 4 h after treatment in all
concentrations, including the lowest concentration of 0.015%. There
was no mortality recorded in the blank control treatment, and all
workers remained alive and active for >48 h after treatment.
Identical results were obtained from these investigations, whether
the petri dishes were covered or uncovered after application of the
E. cloeziana oil extract.
[0178] Subsequent investigations further assessed efficacy of E.
cloeziana oil extract by releasing termite workers on fresh dried
residues. This was carried out by uniformly distributing one mL of
each concentration over the entire surface area of a 90 mm diam
filter paper. When the paper was air dry, 20 termite workers were
placed in each petri dish. One hundred percent mortality was
recorded in all four replicates even at the lowest concentration
applied (0.015% w/v=150 ppm). This suggests that the plant extract
is a highly toxic contact poison to termites.
Ants
[0179] The same experimental procedure for termites was repeated
for ants, with the required number of worker ants for each
treatment transferred with a fine brush to the filter paper
containing a fresh dried residue of the E. cloeziana oil extract.
Immediately after release of the ants the petri dish was covered
with perforated plastic wrap, which enabled any excess vapours to
escape while retaining the ants. Mortality was assessed 4 h after
treatment. In all 4 replicates 100% mortality was obtained at
concentrations as low as 0.0075% w/v ai when applied at a rate of
1.0 mL to a 90 mm diam. filter paper.
Houseflies
[0180] One percent E. cloeziana oil extract was prepared using pure
acetone as a diluent. From this solution, further serial dilutions
were prepared by adding the required amount in acetone. Five mL
aliquots of each concentration of E. cloeziana oil extract were
dispensed into 500 mL kilner jars. The kilner jars were immediately
rotated until dryness to coat the inner surface uniformly with the
E. cloeziana oil residue. After complete dryness, 30-50 pupae were
transferred to a series of clean uncovered petri dishes (45 mm
diam.), one of which was placed inside each jar. The jar mouth was
then covered with nylon netting supported by a rubber band. All
adult houseflies started to emerge from pupae after 48 h and most
emerged within a 3 h period. Flies were fed 5% sugar solution
soaked in absorbent cotton wool. Jars were maintained in an
incubator at 29.degree. C.
[0181] Mortality was assessed at the end of the third day (i.e.,
approx. 72 h) after application of the E. cloeziana oil residues in
the kilner jar and the placement of pupae inside the jars. (This
comprised 48 h for pupae to emerge and 24 h exposure to E.
cloeziana oil residues which were now 48 h old). Flies were
observed to die within a few hours after emergence, whereas in the
control they remained alive for >48 h after emergence. The total
number of adult houseflies that emerged in each kilner jar was
counted and their mortality was recorded.
American Cockroaches
[0182] Tests were conducted on 10-20 three months old nymphs (mean
individual mass 0.2-0.3 g) and replicated three times. One mL of
1.0% E. cloeziana oil extract in acetone was uniformly distributed
on 90 mm diam. Whatman No 2 filter paper. A control treatment was
also carried out using 1 mL acetone only (i.e., minus E. cloeziana
oil extract). After complete dryness of the filter paper, the
required number of cockroaches was transferred inside the kilner
jars, and were fed dry dog food. Kilner jar necks were covered with
muslin netting supported by rubber bands. Mortality was assessed 24
h after releasing the cockroaches, and death was recognised by the
absence of movement when the test animals were mechanically
stimulated.
Adult Mosquitoes
[0183] A 0.3576 g of 85% ai of E. cloeziana oil extract was
dissolve in pure acetone as a diluent to give approximately 0.304%
concentration stock solution. From this solution, further serial
dilutions 0.152, 0.076, 0.0043 and 0.00215 were prepared by adding
the required amount in acetone. Aliquots (2.5 mL) of each
concentration of E. cloeziana oil extract were dispensed into 500
mL kilner jars with total internal surface area as 286.53 cm.sup.2.
The kilner jars were immediately rotated to coat the inner surface
uniformly with the E. cloeziana oil extract residue, until dry.
Once completely dry, 10-25 mixed sex adult mosquitos were released
into each kilner jar by allowing the mosquitoes to fly from a
darkened cage into the naturally lit jars. The jar mouth was
subsequently placed on 110 mm diam. filter paper onto which had
been placed a yellow sponge soaked in 7.0% sugar solution. Treated
jars were kept under laboratory temperature and humidity
conditions, (viz. 24.+-.1.degree. C. and 65.+-.5% RH respectively).
Mortality was assessed 24 h after releasing the mosquitoes in the
jars.
Honey Bees
[0184] Ten worker honey bees, anaesthetised with carbon dioxide,
were transferred to 90 mm diam. petri dishes lined with moistened
filter paper. Five mL aliquots were applied using a Potter Spray
Tower while the bees were still anaesthetised. Yellow sponges
soaked in 7% sugar solution were then placed inside petri dishes
for bee sustenance, and the bees after treatment were placed in
these dishes. The lids that were perforated and covered with muslin
netting were placed on the dishes, and mortality was assessed 24
hours after treatment.
Ash White Flies
[0185] For each treatment, a 2.5 cm diam. leaf disc was mounted on
a moistened Whatman #2 filter paper lining the bottom of petri
dishes. Adult white flies were anaesthetised using carbon dioxide
and 30-50 adults were transferred onto leaf discs. Serial dilutions
of 0.00546, 0.0220, 0.0894 & 0.1788% concentration of E.
cloeziana oil extract were prepared using distilled water
containing 200 ppm Triton X100. Three replicates were treated for
each concentration. Five-mL aliquots were applied to each petri
dish using a Potter Spray Tower. After treatment, petri dishes were
left to dry and then covered with muslin netting. Mortality
assessment was assessed 24 h after white flies had been transferred
to petri dishes.
Drug Store Beetle
[0186] One mL of each concentration of E. cloeziana oil extract in
pure acetone was uniformly dispensed on 90 mm diam. Whatman No 2
filter paper. The latter was left for 1 h to air dry before being
used to line the lid of a 90 mm diam. petri dish. Between 10-15
mixed adults were then transferred into the petri dish, which was
sealed with Parafilm.TM. Mortality assessment was carried out 24 h
after sealing the petri dish.
Snails
[0187] Two methods were used to assess the efficacy of E. cloeziana
oil extract against snails. In the first method different age
groups of adult snails were dipped directly into a solution of 0.5%
concentration of E. cloeziana oil extract in distilled water
containing 200 ppm Triton X10. Snails were dipped for 10 seconds
and thereafter immediately filtered in a sieve. The treated snails
were divided in three 500 mL kilner jars containing French bean
leaves as a food source. The control was carried out using 0% E.
cloeziana oil extract. Mortality was assessed 24 h after
treatment.
[0188] Allowing the snails to crawl on a E. cloeziana oil extract
contaminated surface carried out the second method. Two mL of 0.08%
concentration of E. cloeziana oil extract in pure acetone were
dispensed in 500 mL kilner jars which were rotated to uniformly
cover all inner surfaces until dry. Six different age groups of
snails were put into each jar along with plant material for food
and covered with muslin netting held by a rubber band. Three
replicates of each treatment (0.0 & 0.08% concentration) were
carried out. Mortality was assessed 24 h after releasing the snails
inside the jars.
[0189] The above bioassay results are summarised in Table 8.
TABLE-US-00009 TABLE 8 Summary of E. cloeziana Oil Extract Efficacy
Against Various Target Pests Method of Target Pest LD.sub.50 (95%
CL) LD.sub.95 (95% CL) application Remarks TSM 0.07 0.14 Potter
spray Knockdown effect (0.06-0.08) (0.12-0.16) tower; Aliquot
observed 2 h after applied 2.5 mL treatment TSM <0.01 E.
cloeziana oil 100% mortality at extract + paraffin 0.01% oil TRM
0.02 0.04 Potter spray tower GHT 0.10 0.125 Potter spray tower
Aphid 0.08 0.30 Potter spray tower American 157.27 .mu.g/cm.sup.2
E. cloeziana oil cockroach residues on filter nymph paper Ant
<0.0075 E. cloeziana oil 100% mortality at residues on filter
0.0075% when 1 mL paper applied as residue on filter paper Termite
<0.0075 E. cloeziana oil 100% mortality at residues on filter
0.0075% when 1 mL paper applied as residue on filter paper DBM
1.sup.st 0.09 0.20 5 mL Potter spray instar larva tower Housefly
69.8 .mu.g/cm.sup.2 130.87 .mu.g/cm.sup.2 E. cloeziana oil adult
residues on kilner jar wall Adult 0.00387 0.00694 Residues on
kilner Apparently very mosquitoes (0.00328-0.00451)
(0.00596-0.01051) jar wall rapid knock down. Ash white 0.01773
0.0.03728 Potter spray Assessed 4 h after fly (0.01179-0.03446)
(0.02657-0.08337) tower, 2.5 mL treatment aliquot Drug store
0.03548 0.37275 Self Mortality assessed beetle contaminating by 24
h after releasing walking on filter adults. paper Honeybees 0.12
0.40 Potter spray Knockdown effect tower observed 2 h after Aliquot
applied 5 treatment mL Snails ~0.084 E. cloeziana oil Three mL
acetone residues on kilner solution for each of jar wall 3
replicates. Snails crawled on the wall on dried residues. Snails
100% mortality Dipping method Three replicates in 0.5%
Potted Plant Investigations
[0190] The efficacy of E. cloeziana oil extract against TSM was
further assessed under greenhouse conditions. French bean plants
were grown in 15 cm diam. plastic pots in an insecticide-free
glasshouse maintained at 27.degree. C., RH 65% and natural light.
Plants were used when they reached the two true-leaf stage (i.e.,
before trifoliate leaves appeared). One plant was maintained in
each pot (i.e., two leaves/pot). Twenty gravid TSM females were
then transferred with a fine brush to the upper surface of each
leaf. Mites were left to settle for 4 h before treatment, during
which time they usually settled on the lower leaf surfaces. E.
cloeziana oil extract at a concentration of 0.07% as well as a
blank control were prepared using the same procedures described
above for the laboratory bioassays. A 400 mL hand sprayer was used
to apply the pesticide evenly to all aerial surfaces of the plants,
to run-off. Each treatment was replicated four times. The mortality
was assessed 24 h after treatment. The results were recorded as
mean percent mortality with standard deviation. These results
revealed that the mean percent mortality in E. cloeziana oil
extract treatment and control were 92.19.+-.6.39 and 0.25.+-.0.46,
respectively.
[0191] In summary, E. cloeziana oil extract was efficacious against
all pests tested namely TSM, TRM, GHT, aphids, termites,
houseflies, American cockroaches, whitefooted ants adult
mosquitoes, ash whitefly, drug store beetle and snails. It was also
toxic to honey bees.
Example 8
E. cloeziana Oil Extract as a Fumigant
[0192] An investigation was undertaken to determine the fumigation
action of E. cloeziana oil extract against arthropods. The test
organisms used were termites, Nasutitermes walkeri. A Whatman No 2
filter paper (90 mm diam.) was immersed in distilled water five
seconds and left to drain excess water before placing it on the
bottom of a 90 mm diam. petri dish, to provide moisture for termite
workers during the experimental period. A 1.0% E. cloeziana oil
solution was prepared in pure acetone. One mL was uniformly spread
on a second filter paper, which was allowed to air dry on a sheet
of aluminium foil. It was then placed under the lid of the petri
dishes containing the moist filter paper on their base. Twenty
uniform worker termites were transferred to the moist base of each
test petri dish, which was subsequently covered with its lid
containing the treated filter paper, and the dishes were then
sealed with Parafilm.TM.. A similar series of control treatments
was also prepared, using acetone only, for comparison. Five
replicates were used in each treatment.
[0193] The termites did not move to the dry top surface and
remained on the water-moistened filter paper lining the base of the
petri dishes throughout the experimental period. Mortality was
assessed 5 h after termite release.
[0194] The results revealed that E. cloeziana oil extract has
highly significant fumigant effects on termites. One hundred
percent mortality was recorded in all replicates of the E.
cloeziana oil treatment whereas no mortality occurred in any of the
control (acetone only) replicates.
Example 9
Efficacy of Purified .beta.-Triketones Against TSM
[0195] The efficacy of purified .beta.-triketones against TSM was
investigated using the TSM bioassay described in Example 8. All
.beta.-triketones tested demonstrated a high level of activity
against TSM (see Table 9).
TABLE-US-00010 TABLE 9 Efficacy of .beta.-Triketones Against TSM
Sample No. wt (mg) Chemical Name LD.sub.50 and 95% CL LD.sub.95 and
95% CL 1 109.2 Agglomerone 0.15 (0.11-021) 0.33 (0.22-1.06) 2 99.5
Grandiflorone 0.04 0.13 3 165.0 Dehydroangustione 0.36 (0.33-0.41)
0.69 (0.61-0.81) 4 107.1 Angustione 0.22 (0.21-0.24) 0.35
(0.31-0.40) 5 67.9 Jensenone No direct mortality occurred in any
concentration tested (0.05-0.4%) within 24 h. However, all treated
TSM were unable to move normally and continued to convulse until
they commenced to die 72 h after treatment. There was no recovery.
6 110.1 Tasmanone 0.055 0.150 7 101.8 Flavesone 0.020 (0.006-0.043)
0.0876 (0.076-0.114) 8 77.0 Leptospermone 0.037 0.169 9 113.4
Isoleptospermone 0.043 (0.027-0.057) 0.071 (0.058-0.109) 10 141.1
Platyphyllol 0.070 0.23
Example 10
Efficacy of Purified .beta.-Triketones Against GHT
[0196] The efficacy of purified .beta.-triketones against GHT was
investigated using the GHT bioassay described in Example 8. All
.beta.-triketones tested demonstrated a high level of activity
against GHT (see Table 9).
[0197] All .beta.-triketones except jensenone caused 100% mortality
on GHT at a concentration of 0.3% when 5 mL aliquots were applied
with a Potter spray tower. The latter .beta.-triketone did not
cause direct mortality within 24 h, but caused behavioural effects
at all concentrations tested. Convulsion and lack of movement were
consistently observed and 60.0% mortality was recorded 72 h after
application in the 0.4% treatment.
Examples 11
TABLE-US-00011 [0198] Ready-to-use miticide spray-I Ingredient
Parts E. cloeziana extract 0.1 Emulsifier: e.g.
t-octylphenoxypolyethoxyethanol, 1.0 polyoxyethylenesorbitan,
organosilicate Solvent: e.g. ethyl alcohol, isopropyl alcohol etc 5
Tannic acid 1.0 Carrier e.g. water 92.9
Examples 12
TABLE-US-00012 [0199] Concentrated natural emulsifiable concentrate
spray (4.4%)-I Ingredient Parts E. cloeziana extract 4.4 Pyrethrins
7.4 Emulsifier: e.g. t-octylphenoxypolyethoxyethanol, 14.7
polyoxyethylenesorbitan, organosilicate Solvent: e.g. ethyl
alcohol, isopropyl alcohol etc 73.5
Example 12
TABLE-US-00013 [0200] Concentrated emulsifiable concentrate spray
(44%) Ingredient Parts E. cloeziana extract 22.0 Platyphyllol
(natural or synthetic) 22.0 Emulsifier: e.g.
t-octylphenoxypolyethoxyethanol, 34.0 polyoxyethylenesorbitan,
organosilicate Solvent: e.g. ethyl alcohol, isopropyl alcohol etc
22
Example 13
TABLE-US-00014 [0201] Natural ready to use insecticide spray
Ingredient Parts E. cloeziana extract 0.3 Lavender oil 1.0
Emulsifier: e.g. t-octylphenoxypolyethoxyethanol, 1.0
polyoxyethylenesorbitan, organosilicate Solvent e.g. ethyl alcohol,
isopropyl alcohol etc 40 Carrier: Water 57.7
Example 14
TABLE-US-00015 [0202] Oil-based natural spray concentrate
Ingredient Parts E. cloeziana extract 10.0 Petroleum oil 89.0
Emulsifier: e.g. t-octylphenoxypolyethoxyethanol, 1.0
polyoxyethylenesorbitan, organosilicate
Example 15
TABLE-US-00016 [0203] Concentrated emulsifiable concentrate spray
(10%) Ingredient Parts E. cloeziana extract 10.0 Permethrin 10.0
Piperonyl butoxide 28.0 Emulsifier: e.g.
t-octylphenoxypolyethoxyethanol, 31.0 polyoxyethylenesorbitan,
organosilicate Solvent e.g. ethyl alcohol, isopropyl alcohol etc
21
Example 16
TABLE-US-00017 [0204] Molluscidal dust Ingredient Parts E.
cloeziana extract 2 Anti-caking agent (e.g. silica gel) 2
Emulsifier: e.g. t-octylphenoxypolyethoxyethanol, 3
polyoxyethylenesorbitan, organosilicate Inert carrier (talc,
kaolin, diatomaceous earth) 93
Example 17
TABLE-US-00018 [0205] Aerosol insecticidal and acaricidal spray
Ingredient Parts E. cloeziana extract 1.0 Piperonyl butoxide 0.9
Propellent hydrocarbon 98.1
Example 18
TABLE-US-00019 [0206] Repellent Ingredient Parts E. cloeziana
extract 19.5 Citronella oil 29.1 Phthalic acid dibutyl ester 29.1
N-octyl bicycloheptene dicarboxamide 22.3
Examples 19
TABLE-US-00020 [0207] Ready-to-use miticide spray-II Ingredient
Parts Melaleuca cajeputi extract 0.2 Emulsifier: e.g.
t-octylphenoxypolyethoxyethanol, 1.0 polyoxyethylenesorbitan,
organosilicate Solvent: e.g. ethyl alcohol, isopropyl alcohol etc
5.0 Tannic acid 1.0 Carrier e.g. water 92.8
Examples 20
TABLE-US-00021 [0208] Concentrated natural emulsifiable concentrate
spray (4.4%)-II Ingredient Parts Melaleuca cajeputi extract 8.8
Pyrethrins 7.4 Emulsifier: e.g. t-octylphenoxypolyethoxyethanol,
14.7 polyoxyethylenesorbitan, organosilicate Solvent: e.g. ethyl
alcohol, isopropyl alcohol etc 69.1
Examples 21
TABLE-US-00022 [0209] Ready-to-use miticide spray-III Ingredient
Parts 99% Tasmanone 0.09 Emulsifier: e.g.
t-octylphenoxypolyethoxyethanol, 1.0 polyoxyethylenesorbitan,
organosilicate Solvent: e.g. ethyl alcohol, isopropyl alcohol etc 5
Tannic acid 1.0 Carrier e.g. water 92.91
Examples 22
TABLE-US-00023 [0210] Concentrated natural emulsifiable concentrate
spray (4.4%)-III Ingredient Parts 99% Tasmanone 4.0 Pyrethrins 7.4
Emulsifier: e.g. t-octylphenoxypolyethoxyethanol, 14.7
polyoxyethylenesorbitan, organosilicate Solvent: e.g. ethyl
alcohol, isopropyl alcohol etc 73.9
Examples 23
TABLE-US-00024 [0211] Ready-to-use miticide spray-IV Ingredient
Parts 99% Platyphyllol 0.09 Emulsifier: e.g.
t-octylphenoxypolyethoxyethanol, 1.0 polyoxyethylenesorbitan,
organosilicate Solvent: e.g. ethyl alcohol, isopropyl alcohol etc 5
Tannic acid 1.0 Carrier e.g. water 92.91
Examples 24
TABLE-US-00025 [0212] Concentrated natural emulsifiable concentrate
spray (4.4%)-IV Ingredient Parts 99% Platyphyllol 4.0 Pyrethrins
7.4 Emulsifier: e.g. t-octylphenoxypolyethoxyethanol, 14.7
polyoxyethylenesorbitan, organosilicate Solvent: e.g. ethyl
alcohol, isopropyl alcohol etc 73.9
Example 25
Intra-Specific Crosses for Imparting Pest Resistance to a Pest
Susceptible Plant
[0213] Controlled and wild pollination within Eucalyptus and other
important commercial Myrtaceae are thoroughly addressed in the CRC
for Sustainable Production Forestry Symposium on Hybrid Breeding
and Genetics--Controlled Pollination of Eucalypts on 12th April,
2000 and published as the proceedings of that symposium. Genetic
Pollution from Farm Forestry (Potts et al., 2001) deals more
specifically with intra and inter species crosses occurring within
the Myrtaceae.
[0214] Using the protocols described in the above publications, an
intra-specific Eucalyptus cross breeding technique will employ the
following protocol. This begins with the selection of supreme
individuals as parent stock. Within a selected parent stock having
superior pest resistant characteristics (e.g., E. cloeziana) male
and female trees are identified for cross-pollination experiments.
Pollen is harvested from the male trees and either stored or
directly transferred to the female trees if flowering is
synchronous. Emasculation is undertaken to preclude extraneous
pollination occurring and flowers are often bagged as a further
precaution. Seed set and subsequent embryo development proceeds
over the ensuing 12-24 months and F1 seeds are collected at full
maturation.
[0215] Seeds are then germinated to produce seedlings that are
subjected to detailed analysis to assess the transfer of traits
from parent to progeny. If the F1 progeny show a desirable mix of
phenotypic traits these progeny can be used to vegetatively
propagate the new variety. If the F1 progeny show some improvement
in the selected phenotypic traits, but further improvement is
required selected F1 progeny can be back-crossed either within the
F1 progeny or with one of the parent trees to produce an F2 progeny
using the methodology outlined above. This iterative process can be
continued ad infinitum until the desired characteristics are
achieved.
Example 26
Inter-Specific Crosses for Imparting Pest Resistance to a Pest
Susceptible Plant
[0216] Inter-species hybridisation in the wild, which is a common
phenomenon within the subgenera of the Myrtaceae, have been
recorded at almost 40% in the Eucalypts, 33% in Angophora 19% in
Corymbia and 19% in Symphomyrtus. Hybridisation between the major
subgenera may also occur (Potts et al., 2001). For example,
Eucalyptus camaldulensis displays 14 natural hybrid crosses
including E. camaldulensis.times.E. robusta, E.
camaldulensis.times.E. alba, E. camaldulensis.times.E. cladocalyx,
E. camaldulensis.times.E. bigalaterita, E. camaldulensis.times.E.
tereticornis, E. camaldulensis.times.E. blakelyi, E.
camaldulensis.times.E. dwyeri, E. camaldulensis.times.E. rudis, E.
camaldulensis.times.E. ovata, E. camaldulensis.times.E.
bridgesiana, E. camaldulensis.times.E. viminalis, E.
camaldulensis.times.E. largiflorens, E. camaldulensis.times.E.
melliodora, E. camaldulensis.times.E. leucoxylon; 15 manipulated
hybrids including E. camaldulensis.times.E. diversicolor, E.
camaldulensis.times.E. grandis, E. camaldulensis.times.E.
botryoides, E. camaldulensis.times.E. cladocalyx, E.
camaldulensis.times.E. tereticornis, E. camaldulensis.times.E.
blakelyi, E. camaldulensis.times.E. urophylla, E.
camaldulensis.times.E. macarthurii, E. camaldulensis.times.E.
exerta, E. camaldulensis.times.E. maidenii, E.
camaldulensis.times.E. viminalis, E. camaldulensis.times.E.
globulus, E. camaldulensis.times.E. gunnii, E.
camaldulensis.times.E. laevopinea, E. camaldulensis.times.E.
fastigata. In another example Eucalyptus globulus displays 15
natural hybrids including E. globulus.times.E. barberi, E.
globulus.times.E. brookeriana, E. globulus.times.E. ovata, E.
globulus.times.E. kitsoniana, E. globulus.times.E. goniocalyx, E.
globulus.times.E. nortonii, E. globulus.times.E. cypellocarpa, E.
globulus.times.E. pseudoglobulus, E. globulus.times.E. bicostata,
E. globulus.times.E. johnstonii, E. globulus.times.E. viminalis, E.
globulus.times.E. cordata, E. globulus.times.E. rubida, E.
globulus.times.E. urnigera, E. globulus.times.E. perriniana and 13
successful manipulated hybrids including E. globulus.times.E.
urophylla, E. globulus.times.E. grandis, E. globulus.times.E.
robusta, E. globulus.times.E. pellita, E. globulus.times.E.
longifolia, E. globulus.times.E. loxophloeba, E. globulus.times.E.
camaldulensis, E. globulus.times.E. dunnii, E. globulus.times.E.
nitens, E. globulus.times.E. maidenii, E. globulus.times.E.
bicostata, E. globulus.times.E. viminalis, E. globulus.times.E.
gunnii. In another example Eucalyptus grandis displays 4 natural
hybrids including E. grandis.times.E. urophylla, E.
grandis.times.E. robusta E. grandis.times.E. pellita E.
grandis.times.E. terreticornis and 14 manipulated hybrids including
E. grandis.times.E. urophylla E. grandis.times.E. botryoides E.
grandis.times.E. pellita E. grandis.times.E. alba E.
grandis.times.E. terreticornis E. grandis.times.E. camaldulensis E.
grandis.times.E. dunnii E. nitens.times.E. maidenii E.
grandis.times.E. globulus E. grandis.times.E. gunnii E.
grandis.times.E. pulverulenta E. grandis.times.E. leucoxylon, E.
grandis.times.E. resinifera. In another example the Corymbia
henryi/variegata/maculata/citriodora complex displays 14 natural
hybrids including C. citriodora.times.C. catenaria, C.
citriodora.times.C. variegata, C. citriodora.times.C. maculata, C.
maculata.times.C. gummifrea, C. maculata.times.C. intermedia, C.
maculata.times.C. citriodora, C. maculata.times.C. variegata, C.
variegata.times.C. bloxsomeri, C. variegata.times.C. watsoniana, C.
variegata.times.C. citriodora, C. variegata.times.C. maculata, C.
Henryi.times.C. torelliana, C. henryi.times.C. variegata and
onemanipulated hybrid C. torelliana.times.c. In one further example
Eucalyptus cloeziana displays only 1 natural hybrid E.
cloeziana.times.E. acmenoides.
[0217] The disclosure of every patent, patent application, and
publication cited herein is hereby incorporated herein by reference
in its entirety.
[0218] The citation of any reference herein should not be construed
as an admission that such reference is available as "Prior Art" to
the instant application
[0219] Throughout the specification the aim has been to describe
the preferred embodiments of the invention without limiting the
invention to any one embodiment or specific collection of features.
Those of skill in the art will therefore appreciate that, in light
of the instant disclosure, various modifications and changes can be
made in the particular embodiments exemplified without departing
from the scope of the present invention. All such modifications and
changes are intended to be included within the scope of the
appended claims. All figures, tables, and appendices, as well as
publications, patents, and patent applications, cited herein are
hereby incorporated by reference in their entirety for all
purposes.
I. BIBLIOGRAPHY
[0220] Bignall, C. M., Dunlop, P. J., Brophy, J. J. and Fookes, C.
J. R. (1997). Volatile Leaf Oils of some South-western and Southern
Australian Species of the Genus Eucalyptus (Series I). Part XIV.
Subgenus Monocalyptus. Flav Frag. Journal, 12, 177-183. [0221]
Boland, D. and J. Brophy (1993). Essential Oils of the Eucalyptus
and Related Genera: Search for Chemical Trends. Bioactive Volatile
Compounds from Plants. R. Teranishi, R. G. Buttery and H. Sugisawa.
Washington D. C., American Chemical Society. 525: 72-87. [0222]
Brophy, J. J. and D. J. Boland (1990). "Leaf Essential Oil of Two
Chemotypes of Eucalyptus cloeziana F. Muell." Journal of Essential
Oil Research 2 (March/April): 87-90. [0223] Brophy, J. J., R. J.
Goldsack, et al. (1995). "Leaf Oils of the Genus Backhousia
(Myrtaceae)." Journal of Essential Oil Research 7 (May/June):
237-254. [0224] CRC for Sustainable Production Forestry. Symposium
on Hybrid Breeding and Genetics--Controlled Pollination of
Eucalypts, Noosa Australia, 12th April, 2000 [0225] Hellyer, R.
(1968). "The Occurrence of 3-Triketones in the Steam-Volatile Oils
of some Myrtaceous Australian Plants." Aust. J. Chem. 21(11):
2825-2828. [0226] Herron G A, Beatie G A C, Parkes R A &
Barchia I. 1995. Potter spray tower bioassay of selected citrus
pests to petroleum spray oil. Journal of Australian Entomological
Society 34: 225-263. [0227] Potts, B M, Barbour, R C and Hingston,
A H, 2001. Genetic Pollution from Farm Forestry. Rural Industries
Research and Development Corporation Publication (No 01/114).
[0228] Southwell, I. A. and J. J. Brophy (2000). "Essential oil
isolates from the Australian Flora. Part 3." Journal of Essential
Oil Research 12: 267-278.
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