MICROBIOCIDAL THIAZOLE DERIVATIVES
20220017501 · 2022-01-20
Assignee
Inventors
- David BURNS (Bracknell, Berkshire, GB)
- Mattia Riccardo MONACO (Stein, CH)
- Stefano Rendine (Stein, CH)
- Clemens Lamberth (Stein, CH)
- Andrew Edmunds (Stein, CH)
- Mathias Blum (Stein, CH)
Cpc classification
C07D411/12
CHEMISTRY; METALLURGY
A01N47/40
HUMAN NECESSITIES
C07D413/12
CHEMISTRY; METALLURGY
C07D277/56
CHEMISTRY; METALLURGY
International classification
Abstract
Compounds of the formula (I), wherein the substituents are as defined in claim 1, useful as pesticides, and especially fungicides.
##STR00001##
Claims
1. A compound of formula (I): ##STR00066## wherein, Y is C—F, C—H or N; R.sup.1 is hydrogen, halogen, cyano, C.sub.1-C.sub.4alkyl, C.sub.1-C.sub.4alkoxy, C.sub.1-C.sub.4haloalkyl, or HC(O)NH—; R.sup.2 is C.sub.1-C.sub.8alkyl, C.sub.1-C.sub.8haloalkyl, C.sub.1-C.sub.8alkoxy, C.sub.3-C.sub.8cycloalkyl, C.sub.3-C.sub.8cycloalkylC.sub.1-C.sub.2alkyl (wherein the cycloalkyl groups are optionally substituted with 1 to 3 groups represented by R.sup.3), phenyl, phenylC.sub.1-C.sub.2alkyl (wherein the phenyl rings are optionally substituted with 1 to 3 groups represented by R.sup.3), heteroaryl, heteroarylC.sub.1-C.sub.2alkyl, wherein the heteroaryl is a 5- or 6-membered aromatic monocyclic ring comprising 1, 2, 3 or 4 heteroatoms individually selected from nitrogen, oxygen and sulfur, heterocyclyl, heterocyclylC.sub.1-C.sub.2alkyl, wherein the heterocyclyl is a 4-, 5- or 6-membered non-aromatic monocyclic ring comprising 1, 2 or 3 heteroatoms individually selected from nitrogen, oxygen and sulfur, or a 5- to 12-membered non-aromatic annulated or spirocyclic carbobi- or carbotri-cyclyl ring system optionally comprising 1, 2, 3, 4 or 5 heteroatoms individually selected from nitrogen, oxygen and sulfur, and wherein the spirocyclic carbobi- or carbotri-cyclyl ring systems are each optionally bonded to the rest of the molecule through a C.sub.1-C.sub.2alkylene linker; R.sup.3 is halogen, C.sub.1-C.sub.4alkyl, C.sub.1-C.sub.4alkoxy, C.sub.1-C.sub.4haloalkyl, C.sub.3-C.sub.8cycloalkyl or C.sub.3-C.sub.8cycloalkylC.sub.1-C.sub.2alkyl; X is N or C—H; or a salt or an N-oxide thereof.
2. The compound according to claim 1, wherein R.sup.1 is chloro, bromo, methyl, methoxy, or HC(O)NH—.
3. The compound according to claim 1, wherein R.sup.1 is methyl or HC(O)NH—.
4. The compound according to claim 1, wherein R.sup.1 is methyl.
5. The compound according to claim 1, wherein R.sup.2 is C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.3alkoxy, C.sub.3-C.sub.6cycloalkyl, C.sub.3-C.sub.6cycloalkylC.sub.1-C.sub.2alkyl (wherein the cycloalkyl groups are optionally substituted with 1 to 3 groups represented by R.sup.3), phenyl, phenylC.sub.1-C.sub.2alkyl (wherein the phenyl rings are optionally substituted with 1 to 3 groups represented by R.sup.3), heteroaryl wherein the heteroaryl is a 5- or 6-membered aromatic monocyclic ring comprising 1, 2 or 3 heteroatoms individually selected from nitrogen, oxygen and sulfur, heterocyclyl wherein the heterocyclyl is a 4-, 5- or 6-membered non-aromatic monocyclic ring comprising 1, 2 or 3 heteroatoms individually selected from nitrogen, oxygen and sulfur, or a 5- to 12-membered non-aromatic spirocyclic carbobi- or carbotri-cyclyl ring system optionally comprising a single heteroatom selected from nitrogen, oxygen and sulfur.
6. The compound according to claim 1, wherein R.sup.2 is C.sub.1-C.sub.6alkyl, C.sub.3-C.sub.6cycloalkyl, C.sub.3-C.sub.6cycloalkylC.sub.1-C.sub.2alkyl (wherein the cycloalkyl groups are optionally substituted with 1 or 2 groups represented by R.sup.3), phenylC.sub.1-C.sub.2alkyl (wherein the phenyl rings are optionally substituted with 1 or 2 groups represented by R.sup.3), or a 5- to 12-membered non-aromatic spirocyclic carbobi- or carbotri-cyclyl ring system.
7. The compound according to claim 1, wherein R.sup.2 is n-butyl, isobutyl, n-pentyl, isopentyl, 2,2-dimethylpropyl, n-hexyl, 1-(cyclopropylmethyl)cyclopropylmethyl, cyclobutyl, 2,2-dimethylcyclobutyl, 1-methylcyclopentyl, benzyl, 1-phenylethyl, 3,5-bis(trifluoromethyl)phenylmethyl, spiro[3.3]heptanyl, spiro[3.4]octanyl or spiro[cyclobutane-1,2′-indanyl].
8. The compound according to claim 1, wherein R.sup.3 is C.sub.1-C.sub.3alkyl, C.sub.1-C.sub.3haloalkyl, or C.sub.3-C.sub.6cycloalkylC.sub.1-C.sub.2alkyl.
9. The compound according to claim 1, wherein X is N.
10. The compound according to claim 1, wherein Y is C—F.
11. The compound according to claim 1, wherein the compound of formula (I) is 2-[cyano-(2,6-difluoro-4-pyridyl)amino]-N-(2,2-dimethylcyclobutyl)-5-formamido-thiazole-4-carboxamide (1.b.25), 2-[cyano-(2,6-difluoro-4-pyridyl)amino]-N-[[1-(cyclopropylmethyl)cyclopropyl]methyl]-5-methyl-thiazole-4-carboxamide (1f.23), 2-[cyano-(2,6-difluoro-4-pyridyl)amino]-5-methyl-N-spiro[3.3]heptan-3-yl-thiazole-4-carboxamide (1.e.23), 2-[cyano-(2,6-difluoro-4-pyridyl)amino]-5-methyl-N-spiro[3.4]octan-3-yl-thiazole-4-carboxamide (1.c.23), 2-[cyano-(2,6-difluoro-4-pyridyl)amino]-5-methyl-N-spiro[cyclobutane-2,2′-indane]-1-yl-thiazole-4-carboxamide (1.d.23) or 2-[cyano-(2,6-difluoro-4-pyridyl)amino]-N-(2,2-dimethylcyclobutyl)-5-methyl-thiazole-4-carboxamide (1.b.23).
12. An agrochemical composition comprising a fungicidally effective amount of a compound of formula (I) according to claim 1.
13. The composition according to claim 12, further comprising at least one additional active ingredient and/or an agrochemically-acceptable diluent or carrier.
14. A method of controlling or preventing infestation of useful plants by phytopathogenic microorganisms, applying a fungicidally effective amount of a compound of formula (I) according to claim 1 to the plants, to parts thereof or the locus thereof.
15. Use of a compound of formula (I) according to any claim 1 as a fungicide.
Description
FORMULATION EXAMPLES
[0193]
TABLE-US-00002 Wettable powders a) b) c) active ingredient [compound of formula (I)] 25% 50% 75% sodium lignosulfonate 5% 5% — sodium lauryl sulfate 3% — 5% sodium diisobutylnaphthalenesulfonate — 6% 10% phenol polyethylene glycol ether — 2% — (7-8 mol of ethylene oxide) highly dispersed silicic acid 5% 10% 10% Kaolin 62% 27% —
[0194] The active ingredient is thoroughly mixed with the adjuvants and the mixture is thoroughly ground in a suitable mill, affording wettable powders that can be diluted with water to give suspensions of the desired concentration.
TABLE-US-00003 Powders for dry seed treatment a) b) c) active ingredient [compound of formula (I)] 25% 50% 75% light mineral oil 5% 5% 5% highly dispersed silicic acid 5% 5% — Kaolin 65% 40% — Talcum — 20%
[0195] The active ingredient is thoroughly mixed with the adjuvants and the mixture is thoroughly ground in a suitable mill, affording powders that can be used directly for seed treatment.
TABLE-US-00004 Emulsifiable concentrate active ingredient [compound of formula (I)] 10% octylphenol polyethylene glycol ether 3% (4-5 mol of ethylene oxide) calcium dodecylbenzenesulfonate 3% castor oil polyglycol ether (35 mol of ethylene oxide) 4% Cyclohexanone 30% xylene mixture 50%
[0196] Emulsions of any required dilution, which can be used in plant protection, can be obtained from this concentrate by dilution with water.
TABLE-US-00005 Dusts a) b) c) Active ingredient [compound of formula (I)] 5% 6% 4% talcum 95% — — Kaolin — 94% — mineral filler — — 96%
[0197] Ready-for-use dusts are obtained by mixing the active ingredient with the carrier and grinding the mixture in a suitable mill. Such powders can also be used for dry dressings for seed.
TABLE-US-00006 Extruder granules Active ingredient [compound of formula (I)] 15% sodium lignosulfonate 2% carboxymethylcellulose 1% Kaolin 82%
[0198] The active ingredient is mixed and ground with the adjuvants, and the mixture is moistened with water. The mixture is extruded and then dried in a stream of air.
TABLE-US-00007 Coated granules Active ingredient [compound of formula (I)] 8% polyethylene glycol (mol. wt. 200) 3% Kaolin 89%
[0199] The finely ground active ingredient is uniformly applied, in a mixer, to the kaolin moistened with polyethylene glycol. Non-dusty coated granules are obtained in this manner.
TABLE-US-00008 Suspension concentrate active ingredient [compound of formula (I)] 40% propylene glycol 10% nonylphenol polyethylene glycol ether (15 mol of ethylene oxide) 6% Sodium lignosulfonate 10% carboxymethylcellulose 1% silicone oil (in the form of a 75% emulsion in water) 1% Water 32%
[0200] The finely ground active ingredient is intimately mixed with the adjuvants, giving a suspension concentrate from which suspensions of any desired dilution can be obtained by dilution with water. Using such dilutions, living plants as well as plant propagation material can be treated and protected against infestation by microorganisms, by spraying, pouring or immersion.
TABLE-US-00009 Flowable concentrate for seed treatment active ingredient [compound of formula (I)] 40% propylene glycol 5% copolymer butanol PO/EO 2% tristyrenephenole with 10-20 moles EO 2% 1,2-benzisothiazolin-3-one (in the form of a 20% solution in 0.5%.sup. water) monoazo-pigment calcium salt 5% Silicone oil (in the form of a 75% emulsion in water) 0.2%.sup. Water 45.3%
[0201] The finely ground active ingredient is intimately mixed with the adjuvants, giving a suspension concentrate from which suspensions of any desired dilution can be obtained by dilution with water. Using such dilutions, living plants as well as plant propagation material can be treated and protected against infestation by microorganisms, by spraying, pouring or immersion.
Slow Release Capsule Suspension
[0202] 28 parts of a combination of the compound of formula (I) are mixed with 2 parts of an aromatic solvent and 7 parts of toluene diisocyanate/polymethylene-polyphenylisocyanate-mixture (8:1). This mixture is emulsified in a mixture of 1.2 parts of polyvinyl alcohol, 0.05 parts of a defoamer and 51.6 parts of water until the desired particle size is achieved. To this emulsion a mixture of 2.8 parts 1,6-diaminohexane in 5.3 parts of water is added. The mixture is agitated until the polymerization reaction is completed.
[0203] The obtained capsule suspension is stabilized by adding 0.25 parts of a thickener and 3 parts of a dispersing agent. The capsule suspension formulation contains 28% of the active ingredients. The medium capsule diameter is 8-15 microns.
[0204] The resulting formulation is applied to seeds as an aqueous suspension in an apparatus suitable for that purpose.
EXAMPLES
[0205] The Examples which follow serve to illustrate the invention. The compounds of the invention can be distinguished from known compounds by virtue of greater efficacy at low application rates, which can be verified by the person skilled in the art using the experimental procedures outlined in the Examples, using lower application rates if necessary, for example 50 ppm, 12.5 ppm, 6 ppm, 3 ppm, 1.5 ppm, 0.8 ppm or 0.2 ppm.
[0206] Compounds of formula (I) may possess any number of benefits including, inter alia, advantageous levels of biological activity for protecting plants against diseases that are caused by fungi or superior properties for use as agrochemical active ingredients (for example, greater biological activity, an advantageous spectrum of activity, an increased safety profile (including improved crop tolerance), improved physico-chemical properties, or increased biodegradability).
LIST OF ABBREVIATIONS
[0207] bs=broad singlet
° C.=degrees Celsius
CDCl.sub.3=chloroform-d
d=doublet
Pd.sub.2(dba).sub.3=Tris(dibenzylideneacetone)dipalladium(0)
DIPEA=N,N-diisopropylethylamine
[0208] DMF=dimethylformamide
h=hours
HATU=1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate
m=multiplet
MHz=mega hertz
mp=melting point
N=normal
ppm=parts per million
s=singlet
THF=tetrahydrofuran
Xantphos=4,5-Bis(diphenylphosphino)-9,9-dimethylxanthene
Example 1: This example illustrates the preparation of 2-[cyano-(2,6-difluoro-4-pyridyl)amino]-N-(2,2-dimethylcyclobutyl)-5-methyl-thiazole-4-carboxamide (Compound I.b.23)
[0209] ##STR00035##
a) Preparation of methyl 2-[(2,6-difluoro-4-pyridyl)amino]-5-methyl-thiazole-4-carboxylate
[0210] ##STR00036##
[0211] Under Argon atmosphere, Xantphos (0.2 equiv.), Pd.sub.2(dba).sub.3 (0.1 equiv.) and cesium carbonate (2 equiv.) were added to a degassed, stirred mixture of methyl 2-bromo-5-methyl-thiazole-4-carboxylate (4.6 g, 18.5 mmol, 1 equiv.) and 2,6-difluoropyridin-4-amine (1 equiv.) in 1,4-dioxane (660 mL). The reaction was heated to reflux and stirred for 4 h before allowing the temperature to cool to room temperature. The mixture was diluted with ethyl acetate and filtered over Celite, and the resulting filtrate was concentrated using a rotatory evaporator. Purification by column chromatography on silica gel (eluent mixtures cyclohexane/ethyl acetate) afforded the desired methyl 2-[(2,6-difluoro-4-pyridyl)amino]-5-methyl-thiazole-4-carboxylate (1.8 g, 6.31 mmol). .sup.1H-NMR (400 MHz, CDCl.sub.3): δ=2.73 (s, 3H), 3.94 (s, 3H), 6.75 (s, 1H).
b) Preparation of 2-[(2,6-difluoro-4-pyridyl)amino]-5-methyl-thiazole-4-carboxylic acid
[0212] ##STR00037##
[0213] Lithium hydroxide monohydrate (4 equiv.) was added to a solution of 2-[(2,6-difluoro-4-pyridyl)amino]-5-methyl-thiazole-4-carboxylic acid (1.8 g, 6.31 mmol) in a mixture of tetrahydrofuran (35 mL) and water (12 mL). The reaction mixture was stirred 16 h at room temperature, then the solvents were removed in vacuo. The residue was diluted with ethyl acetate and water, then 2 N hydrochloric acid was slowly added until a pH of 3-4 was reached. The formed precipitate was isolated by filtration and washed twice with water, giving the desired product 2-[(2,6-difluoro-4-pyridyl)amino]-5-methyl-thiazole-4-carboxylic acid (1.55 g, 5.71 mmol). .sup.1H-NMR (400 MHz, (CD.sub.3).sub.2SO): δ=2.69 (s, 3H), 7.30 (s, 2H), 11.35 (bs, 1H), 12.90 (bs, 1H).
c) Preparation of 2-[(2,6-difluoro-4-pyridyl)amino]-N-(2,2-dimethylcyclobutyl)-5-methyl-thiazole-4-carboxamide
[0214] ##STR00038##
[0215] (2,2-dimethylcyclobutyl) ammonium chloride (1.1 equiv.), HATU (1.1. equiv.), and DIPEA (2.6 equiv.) were added in sequence to a DMF solution (9.2 mL) of 2-[(2,6-difluoro-4-pyridyl)amino]-5-methyl-thiazole-4-carboxylic acid (250 mg, 0.92 mmol, 1 equiv.). The resulting solution was stirred at room temperature for 1 h until consumption of starting material (LCMS control). Then a saturated NaHCO.sub.3 solution was added to the mixture and the solution extracted three times with ethyl acetate. The organic phases were combined, dried over sodium sulphate and the volatiles removed by rotatory evaporator. Purification by column chromatography on silica gel (eluent: mixtures of cyclohexane/ethyl acetate) gave the desired product 2-[(2,6-difluoro-4-pyridyl)amino]-N-(2,2-dimethylcyclobutyl)-5-methyl-thiazole-4-carboxamide (280 mg, 86% yield). .sup.1H-NMR (400 MHz, CDCl.sub.3): δ=1.17 (s, 3H), 1.20 (s, 3H), 1.50-1.75 (m, 2H), 1.86-1.92 (m, 1H), 2.29-2.36 (m, 1H), 2.79 (s, 3H), 4.25-4.31 (m, 1H), 6.87 (s, 2H), 7.32 (d, 1H), 7.67 (s, 1H).
d) 2-[cyano-(2,6-difluoro-4-pyridyl)amino]-N-(2,2-dimethylcyclobutyl)-5-methyl-thiazole-4-carboxamide (Compound I.b.23)
[0216] ##STR00039##
[0217] Buthyllithium (2.5 M solution in hexane, 1.25 equiv.) was added at −78° C. to a stirred solution of 2-[(2,6-difluoro-4-pyridyl)amino]-N-(2,2-dimethylcyclobutyl)-5-methyl-thiazole-4-carboxamide (300 mg, 0.85 mmol, 1 equiv.) in THF (4.3 mL). After 30 min, cyanogen bromide was added to the solution, the reaction was allowed to reach room temperature and stirred for 2 h. Then the reaction was quenched with a NaHCO.sub.3 saturated aqueous solution and the aqueous phase was extracted three times with ethyl acetate. The combined organic phases were dried over sodium sulphate and the volatiles removed using a rotatory evaporator. Purification by column chromatography on silica gel (eluent: mixtures of cyclohexane/ethyl acetate) gave the desired product 2-[cyano-(2,6-difluoro-4-pyridyl)amino]-N-(2,2-dimethylcyclobutyl)-5-methyl-thiazole-4-carboxamide (190 mg, 0.50 mmol, 59% yield). .sup.1H-NMR (400 MHz, CDCl.sub.3): δ=1.10 (s, 3H), 1.21 (s, 3H), 1.55-1.69 (m, 2H), 1.80-1.93 (m, 1H), 2.25-2-35 (m, 1H), 2.90 (s, 3H), 4.25-4.31 (m, 1H), 6.92 (s, 2H), 7.12 (d, 1H).
[0218] Throughout this description, temperatures are given in degrees Celsius (° C.) and “m.p.” means melting point. LC/MS means Liquid Chromatography Mass Spectrometry and the description of the apparatus and the method is:
Method A: ACQUITY UPLC from Waters, Waters UPLC HSS T3, 1.8 μm particle size, 30×2.1 mm column, 0.85 ml/min., 60° C., H.sub.2O/MeOH 95:5+0.05% HCOOH (90%)/CH.sub.3CN+0.05% HCOOH (10%) −1.2 min. —CH.sub.3CN+0.05% HCOOH (100%) 0.30 min., ACQUITY SQD Mass Spectrometer from Waters, ionization method: electrospray (ESI), Polarity: positive ions, Capillary (kV) 3.00, Cone (V) 30.00, Extractor (V) 2.00, Source Temperature (° C.) 150, Desolvation Temperature (° C.) 350, Cone Gas Flow (L/Hr) 0, Desolvation Gas Flow (L/Hr) 650).
Method B: ACQUITY UPLC from Waters, Waters UPLC HSS T3, 1.8 μm particle size, 30×2.1 mm column, 0.85 ml/min., 60° C., H.sub.2O/MeOH 95:5+0.05% HCOOH (90%)/CH.sub.3CN+0.05% HCOOH (10%)−2.7 min.−CH.sub.3CN+0.05% HCOOH (100%)−0.30 min., ACQUITY SQD Mass Spectrometer from Waters, ionization method: electrospray (ESI), Polarity: positive ions, Capillary (kV) 3.00, Cone (V) 30.00, Extractor (V) 2.00, Source Temperature (° C.) 150, Desolvation Temperature (° C.) 350, Cone Gas Flow (L/Hr) 0, Desolvation Gas Flow (L/Hr) 650)).
Method C: MS: ZQ Mass Spectrometer from Waters (Single quadrupole mass spectrometer) Instrument Parameter: Ionisation method: Electrospray Polarity: positive (negative) ions Capillary (kV) 3.00, Cone (V) 30.00, Extractor (V) 2.00, Gas Temperature (° C.) 350, Drying Gas Flow (mL/min) 9.8, Neb press 45 psig, Mass range: 90 to 1000 Da.
[0219] HPLC: HP 1100 HPLC from Agilent: solvent degasser, quaternary pump (ZCQ)/binary pump (ZDQ), heated column compartment and diode-array detector. Column: porpshell 120 C18, 2.7 μm particle size, 120 Angstrom, 4.6×50 mm, Temp: 30° C. DAD Wavelength range (nm): 190 to 400 Solvent Gradient:. A=water+0.1% HCOOH. B=Acetonitril+0.08% HCOOH
TABLE-US-00010 Time (min) A % B % Flow (ml/min) 0.00 85.0 15.0 0.6 4.00 5.00 95.00 0.6 10.00 5.00 95.00 0.6
TABLE-US-00011 TABLE 2 Melting point and LC/MS data (R.sub.t = Retention time) for selected compounds of Table 1. Compound Mp No. Name Structure (°C.) LC/MS I.a.18 2-[cyano-(5-fluoro-3- pyridyl)amino]-N- cyclobutyl-5-methyl- thiazole-4-carboxamide
[0220] Surprisingly, it has been found that the novel compounds of formula (I) may show improved solubility properties (in particular in non-polar solvents), and/or photostability properties when compared to their corresponding free amine (i.e. where the nitrile group on nitrogen is replaced with a hydrogen), which are known for example from WO 2017/207362.
[0221] Throughout the following description, LogP means logarithm of the partition coefficient, ppm means parts per million, and T.sub.50 represents the half-time of the compound under irradiation conditions.
[0222] The methods used for these measurements are presented below.
Partition Coefficient
[0223] Octanol-water partition coefficients (presented as LogP) were measured by an HPLC method using reverse phase mini-columns coated with octanol. The partition coefficient P is directly proportional to the HPLC retention factor. The general principles of this method have been described for example in J. Pharm. Sci., 67 (1978) 1364-7.
[0224] A Waters HPLC system (model 1525 binary pump; 2707 autosampler with thermostat and model 2298 photodiode array detector) was used with Hichrom mini-columns and an aqueous mobile phase containing 20mM phosphate buffers adjusted to pH7, saturated with 1-octanol (Aldrich, HPLC grade). Mini-columns used were HiRPB stationary phase, either 4.6 mm internal diameter by 4 mm length or 2 mm internal diameter×10 mm length. Anisole (Aldrich, 99%+purity, LogP 2.11) was used as the primary reference to calibrate the system.
Photostability
[0225] Photostability tests were carried out by irradiation of thin-film deposits of compounds and 20 formulations on glass surfaces, using a filtered xenon lamp system (Atlas Suntest) which reproduces the spectrum and intensity of sunlight. The spectral output power of the Suntest was set to 750 W/m2, which is the typical daily maximum irradiance level at noon (UK, midsummer).
[0226] Test compounds were typically dissolved in HPLC grade methanol to give 1 g/L stock solutions. Alternatively, formulated compounds were suspended in water at the same concentration. 2 μL droplets of test solutions were spotted onto microscope cover slips in a 3D printed holder, allowed to dry then irradiated in the Suntest for varying times. Cover slips were then removed from the Suntest and placed in 4 dram vials; 1mL of wash solvent (typically 30:70 acetonitrile: 0.2% aqueous formic acid) was added, and the vials shaken to extract compounds into solution. Solutions were analysed by reverse phase HPLC, typically using a Waters UPLC system with Photodiode Array (PDA) and Waters columns (BEH C18, 100×2.1mm×1.7 μm) using mixed aqueous:acetonitrile mobile phase, acidifed with 0.2% formic acid. Peak detection was at the optimum wavelength for each candidate compound and PDA peak areas were used for quantification. Plots of % loss versus time were used to estimate T.sub.50 values, being the time taken for first 50% loss of test compound.
Solubility
[0227] Saturated solutions of test compounds were prepared in either aqueous buffer solutions (10 mM mixed phosphate, pH 7.20) or in heptane. Typically 1 mg of test compound in a 2 dram vial with 1 mL of buffer or heptane was left overnight (20 hours) on a roller shaker after an initial 20 minute period in a sonic bath. Saturated samples were then filtered through Millex-HV 0.45 micron syringe driven filters (aqueous or non-aqueous version dependent on solvent). Aqueous samples were then analysed by direct injection on LCMS, and peak areas using PDA detection were compared with standards of known concentration; heptane samples were first dried and redissolved in an LC compatible solvent, typically 30:70 acetonitrile: 0.2% formic acid. Protocol variations included pre-saturation of the filters for compounds expected to have very low solubility, and centrifugation of the saturated samples for oils.
[0228] Table 3 below illustrates surprising physical chemistry properties (partition coefficient LogP, Solubility in heptane and/or photostability) with respect to the prior art compounds of WO2017/207362.
TABLE-US-00012 TABLE 3 Solubility Solubility Photo- Compound in water in heptane stability No. Name Structure LogP (ppm) (ppm) T.sub.50 (h) E-0 2-[(2,6-difluoro-4- pyridyl)amino]-N- (2,2-dimethyl- cyclobutyl)-5-methyl- thiazole-4- carboxamide
BIOLOGICAL EXAMPLES
[0229] Example B1: Alternaria Solani/Tomato/Leaf Disc (Early Blight)
[0230] Tomato leaf disks cv. Baby are placed on agar in multiwell plates (24-well format) and sprayed with the formulated test compound diluted in water. The leaf disks are inoculated with a spore suspension of the fungus 2 days after application. The inoculated leaf disks are incubated at 23° C./21° C. (day/night) and 80% rh under a light regime of 12/12 h (light/dark) in a climate cabinet and the activity of a compound is assessed as percent disease control compared to untreated when an appropriate level of disease damage appears on untreated check disk leaf disks (5-7 days after application). The following compounds gave at least 80% control of Alternaria solani at 200 ppm when compared to untreated control under the same conditions, which showed extensive disease development: I.a.23, I.b.08, I.b.23, I.c.23, I.g.23, I.i.23, I.k.18, I.m.18.
Example B2: Botryotinia Fuckeliana (Botrytis Cinerea)/Liquid Culture (Gray Mould)
[0231] Conidia of the fungus from cryogenic storage are directly mixed into nutrient broth (Vogels broth). After placing a (DMSO) solution of test compound into a microtiter plate (96-well format), the nutrient broth containing the fungal spores is added. The test plates are incubated at 24° C. and the inhibition of growth is determined photometrically 3-4 days after application. The following compounds gave at least 80% control of Botryotinia fuckeliana at 20 ppm when compared to untreated control under the same conditions, which showed extensive disease development: I.b.23, I.c.23, I.h.23, I.k.23, I.o.23.
Example B3: Glomerella Lagenarium (Colletotrichum Lagenarium)/Liquid Culture (Anthracnose)
[0232] Conidia of the fungus from cryogenic storage are directly mixed into nutrient broth (PDB potato dextrose broth). After placing a (DMSO) solution of test compound into a microtiter plate (96-well format), the nutrient broth containing the fungal spores is added. The test plates are incubated at 24° C. and the inhibition of growth is measured photometrically 3-4 days after application.
[0233] The following compounds gave at least 80% control of Glomerella lagenarium at 20 ppm when compared to untreated control under the same conditions, which showed extensive disease development: I.a.18, I.b.08, I.b.18, I.b.23, I.b.25, I.c.23, I.d.23, I.e.23, I.f.23, I.g.08, I.g.23, I.h.18, I.h.23, I.i.23, I.j.23, I.k.18, I.k.23, 1.1.18, 1.1.23, I.m.18, I.o.23, I.p.23.
Example B4: Blumeria Graminis f. sp. Tritici (Erysiphe Graminis f. sp. Tritici)/Wheat/Leaf Disc Preventative (Powdery Mildew on Wheat)
[0234] Wheat leaf segments cv. Kanzler are placed on agar in a multiwell plate (24-well format) and sprayed with the formulated test compound diluted in water. The leaf disks are inoculated by shaking powdery mildew infected plants above the test plates 1 day after application. The inoculated leaf disks are incubated at 20° C. and 60% rh under a light regime of 24 h darkness followed by 12 h light/12 h darkness in a climate chamber and the activity of a compound is assessed as percent disease control compared to untreated when an appropriate level of disease damage appears on untreated check leaf segments (6-8 days after application).
[0235] The following compounds gave at least 80% control of Blumeria graminis f. sp. tritici at 200 ppm when compared to untreated control under the same conditions, which showed extensive disease development: I.a.23, I.b.08, I.b.18, I.b.23, I.c.23, I.d.23, I.e.23, I.f.23, I.g.08, I.g.23, I.h.23, I.i.23,I.j.23, I.k.18, I.k.23, I.l.23, I.m.18, I.o.23, I.p.23.
Example B5: Fusarium Culmorum/Wheat/Spikelet Preventative (Head Blight)
[0236] Wheat spikelets cv. Monsun are placed on agar in multiwell plates (24-well format) and sprayed with the formulated test compound diluted in water. The spikelets are inoculated with a spore suspension of the fungus 1 day after application. The inoculated spikelets are incubated at 20° C. and 60% rh under a light regime of 72 h semi darkness followed by 12 h light/12 h darkness in a climate chamber and the activity of a compound is assessed as percent disease control compared to untreated when an appropriate level of disease damage appears on untreated check spikelets (6-8 days after application). The following compounds gave at least 80% control of Fusarium culmorum at 200 ppm when compared to untreated control under the same conditions, which showed extensive disease development: I.b.23.
Example B6: Phaeosphaeria Nodorum (Septoria Nodorum)/Wheat/Leaf Disc Preventative (Glume Blotch)
[0237] Wheat leaf segments cv. Kanzler are placed on agar in a multiwell plate (24-well format) and sprayed with the formulated test compound diluted in water. The leaf disks are inoculated with a spore suspension of the fungus 2 days after application. The inoculated test leaf disks are incubated at 20° C. and 75% rh under a light regime of 12 h light/12 h darkness in a climate cabinet and the activity of a compound is assessed as percent disease control compared to untreated when an appropriate level of disease damage appears in untreated check leaf disks (5-7 days after application). The following compounds gave at least 80% control of Phaeosphaeria nodorum at 200 ppm when compared to untreated control under the same conditions, which showed extensive disease development: I.a.23, I.b.23, I.d.23, I.f.23, I.g.23, 1.1.23, I.k.18, I.k.23.
Example B7: Monographella Nivalis (Microdochium Nivale)/Liquid Culture (Foot Rot Cereals)
[0238] Conidia of the fungus from cryogenic storage are directly mixed into nutrient broth (PDB potato dextrose broth). After placing a (DMSO) solution of test compound into a microtiter plate (96-well format), the nutrient broth containing the fungal spores is added. The test plates are incubated at 24° C. and the inhibition of growth is determined photometrically 4-5 days after application.
[0239] The following compounds gave at least 80% control of Monographella nivalis at 20 ppm when compared to untreated control under the same conditions, which showed extensive disease development: I.a.18, I.a.23, I.b.08, I.b.18, I.b.23, I.b.25, I.c.23, I.d.23, I.e.23, I.f.23, I.g.08, I.g.23, I.h.18, I.h.23, I.i.23, I.j.23, I.k.18, I.k.23, I.l.18, I.l.23, I.m.18, I.o.23, I.p.23.
Example B8: Mycosphaerella Arachidis (Cercospora Arachidicola)/Liquid Culture (Early Leaf Spot)
[0240] Conidia of the fungus from cryogenic storage are directly mixed into nutrient broth (PDB potato dextrose broth). After placing a (DMSO) solution of test compound into a microtiter plate (96-well format), the nutrient broth containing the fungal spores is added. The test plates are incubated at 24° C. and the inhibition of growth is determined photometrically 4-5 days after application.
[0241] The following compounds gave at least 80% control of Mycosphaerella arachidis at 20 ppm when compared to untreated control under the same conditions, which showed extensive disease development: I.a.18, I.b.08, I.b.18, I.b.23, I.c.23, I.d.23, I.e.23, I.f.23, I.g.08, I.g.23, I.h.18, I.h.23, 1.1.23, I.k.18, 25 I.k.23, I.l.18, I.l.23, I.m.18, I.o.23, I.p.23.
Example B9: Phakopsora Lachyrhizi/Soybean/Preventative (Soybean Rust)
[0242] Soybean leaf disks are placed on water agar in multiwell plates (24-well format) and sprayed with the formulated test compound diluted in water. One day after application leaf discs are inoculated by spraying a spore suspension on the lower leaf surface. After an incubation period in a climate cabinet of 24-36 hours in darkness at 20° C. and 75% rh leaf disc are kept at 20° C. with 12 h light/day and 75% rh. The activity of a compound is assessed as percent disease control compared to untreated when an appropriate level of disease damage appears in untreated check leaf disks (12-14 days after application).
[0243] The following compounds gave at least 80% control of Phakopsora pachyrhizi at 200 ppm when compared to untreated control under the same conditions, which showed extensive disease development: I.b.23, I.f.23, I.g.23, I.h.23, I.i.23, I.l..23, I.p.23.
Example B10: Plasmopara Viticola/Grape/Leaf Disc Preventative (Late Blight)
[0244] Grape vine leaf disks are placed on water agar in multiwell plates (24-well format) and sprayed with the formulated test compound diluted in water. The leaf disks are inoculated with a spore suspension of the fungus 1 day after application. The inoculated leaf disks are incubated at 19° C. and 80% rh under a light regime of 12 h light/12 h darkness in a climate cabinet and the activity of a compound is assessed as percent disease control compared to untreated when an appropriate level of disease damage appears in untreated check leaf disks (6-8 days after application).
[0245] The following compounds gave at least 80% control of Plasmopara viticola at 200 ppm when compared to untreated control under the same conditions, which showed extensive disease development: I.g.08, I.l.18.
Example B11: Puccinia Recondita f. sp. Tritici/Wheat/Leaf Disc Curative (Brown Rust)
[0246] Wheat leaf segments cv. Kanzler are placed on agar in multiwell plates (24-well format). The leaf segments are inoculated with a spore suspension of the fungus. Plates are stored in darkness at 19° C. and 75% rh. The formulated test compound diluted in water is applied 1 day after inoculation. The leaf segments are incubated at 19° C. and 75% rh under a light regime of 12 h light/12 h darkness in a climate cabinet and the activity of a compound is assessed as percent disease control compared to untreated when an appropriate level of disease damage appears in untreated check leaf segments (6-8 days after application).
[0247] The following compounds gave at least 80% control of Puccinia recondite f. sp. tritici at 200 ppm when compared to untreated control under the same conditions, which showed extensive disease development: I.b.23, I.c.23, I.h.23, I.i.23, I.k.23, I.l.23, I.p.23.
Example B12: Puccinia Recondita f. sp. Tritici/Wheat/Leaf Disc Preventative (Brown Rust)
[0248] Wheat leaf segments cv. Kanzler are placed on agar in multiwell plates (24-well format) and sprayed with the formulated test compound diluted in water. The leaf disks are inoculated with a spore suspension of the fungus 1 day after application. The inoculated leaf segments are incubated at 19° C. and 75% rh under a light regime of 12 h light/12 h darkness in a climate cabinet and the activity of a compound is assessed as percent disease control compared to untreated when an appropriate level of disease damage appears in untreated check leaf segments (7-9 days after application).
[0249] The following compounds gave at least 80% control of Puccinia recondite f. sp. tritici at 200 ppm when compared to untreated control under the same conditions, which showed extensive disease development: I.a.18, I.a.23, I.b.08, I.b.18, I.b.23, I.c.23, I.d.23, I.e.23, I.f.23, I.g.08, I.g.23, I.h.18, I.h.23, I.i.23, I.j.23, I.k.23, I.l.18, I.l.23, I.m.18, I.o.23, I.p.23.
Example B13: Magnaporthe Grisea (Pyricularia Oryzae)/Rice/Leaf Disc Preventative (Rice Blast)
[0250] Rice leaf segments cv. Ballila are placed on agar in a multiwell plate (24-well format) and sprayed with the formulated test compound diluted in water. The leaf segments are inoculated with a spore suspension of the fungus 2 days after application. The inoculated leaf segments are incubated at 22° C. and 80% rh under a light regime of 24 h darkness followed by 12 h light/12 h darkness in a climate cabinet and the activity of a compound is assessed as percent disease control compared to untreated when an appropriate level of disease damage appears in untreated check leaf segments (5-7 days after application).
[0251] The following compounds gave at least 80% control of Magnaporthe grisea at 200 ppm when compared to untreated control under the same conditions, which showed extensive disease development: I.a.18, I.a.23, I.b.08, I.b.18, I.b.23, I.c.23, I.d.23, I.e.23, I.f.23, I.g.08, I.g.23, I.h.23, I.i.23, I.j.23, I.k.18, I.k.23, I.l.18, I.l.23, I.m.18, I.o.23, I.p.23.
Example B14: Pyrenophora Teres/Barley/Leaf Disc Preventative (Net Blotch)
[0252] Barley leaf segments cv. Hasso are placed on agar in a multiwell plate (24-well format) and sprayed with the formulated test compound diluted in water. The leaf segmens are inoculated with a spore suspension of the fungus 2 days after application. The inoculated leaf segments are incubated at 20° C. and 65% rh under a light regime of 12 h light/12 h darkness in a climate cabinet and the activity of a compound is assessed as disease control compared to untreated when an appropriate level of disease damage appears in untreated check leaf segments (5-7 days after application).
[0253] The following compounds gave at least 80% control of Pyrenophora teres at 200 ppm when compared to untreated control under the same conditions, which showed extensive disease development: I.a.23, I.b.23, I.c.23, I.d.23, I.g.23, I.i.23, I.k.23, I.p.23.
Example B15: Sclerotinia Sclerotiorum/Liquid Culture (Cottony Rot)
[0254] Mycelia fragments of a newly grown liquid culture of the fungus are directly mixed into nutrient broth (PDB potato dextrose broth). After placing a (DMSO) solution of test compound into a microtiter plate (96-well format) the nutrient broth containing the fungal material is added. The test plates are incubated at 24° C. and the inhibition of growth is determined photometrically 3-4 days after application. The following compounds gave at least 80% control of Sclerotinia sclerotiorum at 20 ppm when compared to untreated control under the same conditions, which showed extensive disease development: I.b.18, I.b.23, I.c.23, I.f.23, I.g.23, I.h.23, I.i.23.
Example B16: Mycosphaerella Graminicola (Septoria Tritici)/Liquid Culture (Septoria Blotch)
[0255] Conidia of the fungus from cryogenic storage are directly mixed into nutrient broth (PDB potato dextrose broth). After placing a (DMSO) solution of test compound into a microtiter plate (96-well format), the nutrient broth containing the fungal spores is added. The test plates are incubated at 24° C. and the inhibition of growth is determined photometrically 4-5 days after application.
[0256] The following compounds gave at least 80% control of Mycosphaerella graminicola at 20 ppm when compared to untreated control under the same conditions, which showed extensive disease development: I.a.23, I.b.08, I.b.18, I.b.23, I.b.25, I.c.23, I.d.23, I.e.23, I.f.23, I.g.23, I.h.18, I.h.23, I.i.23, I.j.23, I.k.18, I.k.23, I.l.18, I.l.23, I.m.18, I.o.23, I.p.23.