Fungicidal compounds

Abstract

This invention relates to deuterated derivatives of metyltetraprole which are of use in the field of agriculture.

Description

Fungicidal Compounds

[0001] This invention relates to deuterated derivatives of metyltetraprole which are of use in the field of agriculture as fungicides.BACKGROUND

[0002] Metyltetraprole (MTP) is the first of a second generation strobilurin-class fungicide. They are designated Quinone outside Inhibitors (Qol’s) which inhibit ubiquinol oxidation in the mitochondrial respiratory chain complex III which ultimately results in the arresting of ATP synthesis.Metyltetraprole

[0003] MTP is able to control a wide range of fungal diseases, including important cereal diseases, and is being developed globally for the control of resistant Zymoseptoria tritici (Septoria Leaf Blotch) in wheat, which in the event of severe outbreaks can result in up to 50% crop losses, and Pyrenophora teres (net blotch) in barley, which can result in crop production losses between 10-40%.

[0004] The use of Qol’s has been in decline for some time in several segments and some firms ceased their research activities in this area 10 years or more ago. This decline is due to the development of resistance in certain pathogens. Whilst several different mechanisms have been reported to explain this, by far the most prevalent type of resistance is that imparted by the G143A mutation, which brings about almost complete loss of efficacy.

[0005] An aim of certain embodiments of the present invention is to provide fungicides which have activity either non-selectively, i.e. broad-spectrum activity, or which are active specifically against selective target organisms.

[0006] Certain compounds of the invention may be as active or more active than prior art compounds, e.g. Qol’s such as metyltetraprole. They may have activity against organisms that have developed a resistance to prior art compounds. However, certain embodiments of the present invention may also concern compounds which have a lower level of activity relative to prior art compounds. These lower activity compounds are still effective asfungicides but may have other advantages relative to existing compounds such as, for example, a reduced environmental impact.

[0007] Certain embodiments of the invention provide compounds that achieve one or more of the above aims. The compounds may be active in their own right or may metabolise or react in aqueous media to yield an active compound.SUMMARY OF THE INVENTION

[0008] In accordance with a first aspect of the invention there is provided a compound of Formula (I):agronomically acceptable salt or / V-oxide thereof, wherein:R1is selected from D and H;R2is independently at each occurrence selected from D and H;R3is selected from CH3, CH2D, CHD2 and CD3; andR4is selected from CH3, CH2D, CHD2 and CD3, wherein R1, R2, R3and R4are selected so that at least one of the groups selected from R1, the two R2groups, R3and R4is D or comprises D.

[0009] The inventors of the present invention have found that compounds of Formula (I) exhibit a surprising increase in fungicidal activity in comparison to metyltetraprole.

[0010] The following embodiments apply to compounds of Formula (I). These embodiments are independent and interchangeable. Any one embodiment may be combined with any other embodiment, where chemically allowed. In other words, any of the features described in the following embodiments may (where chemically allowable) be combined with the features described in one or more other embodiments. In particular, where a compound is exemplified or illustrated in this specification, any two or more of the embodiments listed below, expressed at any level of generality, which encompass that compound may be combined to provide a further embodiment which forms part of the present disclosure.

[0011] It may be that R1is H. Alternatively, R1may be D.

[0012] It may be that at least one R2is H. It may be that R2is at each occurrence H. It may be that one R2is D and one R2is H. It may be that at least one R2is D. Preferably, R2is at each occurrence D. It has been shown that compounds wherein at least one R2is D (e.g. wherein R2is at each occurrence D) display improved activity against SDHI resistant species of Zymoseptoria, such as Zymoseptoria tritici.

[0013] R3may be CH3. R3may be selected from CH2D, CH D2 and CD3. It may be that R3is CD3.

[0014] R4may be CH3. R4may be selected from CH2D, CH D2 and CD3. It may be that R4is CD3.

[0015] The compound of Formula (I) may be a compound selected from:

[0016] In an embodiment, the compound of Formula (I) is not in the form of an agronomically acceptable salt or / V-oxide. In an embodiment, the compound of Formula (I) is not in the form of an agronomically acceptable / V-oxide.

[0017] In accordance with a second aspect of the invention there is provided fungicidal formulation comprising a compound according to the first aspect of the invention. The formulation will typically comprise an agronomically acceptable auxiliary, such as an extender, solvent or carrier.

[0018] In accordance with a third aspect of the invention there is provided a method for preventing, controlling or treating a fungal disease, the method comprising applying an agronomically effective and substantially non-phytotoxic quantity of a compound according to the first aspect of the invention, or a fungicidal formulation according to the second aspect of the invention, to seeds of plants, to plants themselves, or to an area where it is intended that plants will grow or are growing.

[0019] In accordance with a fourth aspect of the inventions there is provided a use of a compound according to the first aspect of the invention, or a fungicidal formulation according to the second aspect of the invention, as a fungicide (i.e. to prevent, control or treat fungal disease in plants).BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Embodiments of the invention are further described hereinafter with reference to the accompanying drawings, in which:Figure 1 is a bar chart showing the % control of metyltetraprole and Examples 2, 3 and 5 against an SDHI resistant isolate of Zymoseptoria tritici in planta.DETAILED DESCRIPTION

[0021] In the compounds of the invention, when a position is specifically designated as “D”, this position would be understood by a skilled person to be occupied by deuterium (2H) at an isotopic abundance greater than its natural isotopic abundance (e.g. greater than about 0.015%).

[0022] For example, a position designated “D” may be occupied with deuterium at an isotopic purity of2H of greater than 50%. Isotopic purity can be determined using conventional analytical methods known to a person skilled in the art, such as mass spectrometry and nuclear magnetic resonance spectroscopy. A position designated “D” may be occupied with deuterium at an isotopic purity of2H of at least 90%, e.g. at least 95%. A position designated “D” may be occupied with deuterium at an isotopic purity of2H of at least 99%, e.g. at least 99.5%.

[0023] Where structural isomers are interconvertible via a low energy barrier, tautomeric isomerism (‘tautomerism’) can occur. This can take the form of proton tautomerism in compounds of the invention containing, for example, an imino, keto, or oxime group, or so- called valence tautomerism in compounds which contain an aromatic moiety. It follows that a single compound may exhibit more than one type of isomerism.

[0024] Included within the scope of the present invention are all stereoisomers, geometric isomers and tautomeric forms of the compounds of the invention, including compounds exhibiting more than one type of isomerism, and mixtures of one or more thereof. Also included are acid addition or base salts wherein the counter ion is optically active, for example, D-lactate or L-lysine, or racemic, for example, DL-tartrate or DL- arginine.

[0025] The compounds of the invention may be obtained, stored and / or used in the form of an agronomically acceptable salt. Suitable salts include, but are not limited to, salts of acceptable inorganic acids such as hydrochloric, sulfuric, phosphoric, nitric, carbonic, boric, sulfamic, and hydrobromic acids, or salts of agronomically acceptable organic acids such as acetic, propionic, butyric, tartaric, maleic, hydroxymaleic, fumaric, malic, citric, lactic, mucic, gluconic, benzoic, succinic, oxalic, phenylacetic, methanesulfonic, toluenesulfonic, benzenesulfonic, salicylic, sulfanilic, aspartic, glutamic, edetic, stearic, palmitic, oleic, lauric, pantothenic, tannic, ascorbic and valeric acids. Suitable salts also include salts of inorganic and organic bases, e.g. counterions such as Na, Ca, K, Li, Mg, ammonium, trimethylsulfonium.

[0026] The present invention also includes all agronomically acceptable isotopically- labelled compounds of Formulae (I) and (II), wherein one or more atoms are substituted by an isotope(s) of the same element. Examples of isotopes suitable for inclusion in the compounds of the invention include isotopes of hydrogen, such as2H and3H, carbon, such as11C,13C and14C, nitrogen, such as13N and15N, and oxygen, such as150,17O and18O.

[0027] Isotopically-labelled compounds can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described using an appropriate isotopically-labelled reagent in place of the non-labelled reagent previously employed.

[0028] The term “effective and non-phytotoxic amount” means an amount of fungicide which is sufficient to control the fungal disease, but which does not have any significant detrimental effect on any seeds, and / or plants present in the area to which the fungicide is applied. The amount should typically have a positive effect on plant vigour and yield in theabsence of the targeted pest. The amount will vary depending on the pest to be controlled, the type of plant, the climatic conditions and the compounds included in the fungicidal formulation. This amount can be determined by systematic field trials, which are within the capabilities of a person skilled in the art.

[0029] The term “plant” is intended to include plants at every stage of their life cycle postgermination. The term “plant” therefore includes plant seedlings. The term “plant” includes trees.

[0030] Throughout the description and claims of this specification, the words “comprise” and “contain” and variations of them mean “including but not limited to”, and they are not intended to (and do not) exclude other moieties, additives, components, integers or steps. Throughout the description and claims of this specification, the singular encompasses the plural unless the context otherwise requires. In particular, where the indefinite article is used, the specification is to be understood as contemplating plurality as well as singularity, unless the context requires otherwise.

[0031] Features, integers, characteristics, compounds, chemical moieties or groups described in conjunction with a particular aspect, embodiment or example of the invention are to be understood to be applicable to any other aspect, embodiment or example described herein unless incompatible therewith. All of the features disclosed in this specification (including any accompanying claims, abstract and drawings), and / or all of the steps of any method or process so disclosed, may be combined in any combination, except combinations where at least some of such features and / or steps are mutually exclusive. The invention is not restricted to the details of any foregoing embodiments. The invention extends to any novel one, or any novel combination, of the features disclosed in this specification (including any accompanying claims, abstract and drawings), or to any novel one, or any novel combination, of the steps of any method or process so disclosed.

[0032] The reader's attention is directed to all papers and documents which are filed concurrently with or previous to this specification in connection with this application and which are open to public inspection with this specification, and the contents of all such papers and documents are incorporated herein by reference.Fungicidal Formulations

[0033] In a second aspect of the invention there is provided a fungicidal formulation comprising a compound according to the first aspect of the invention.

[0034] Depending on their particular physical and / or chemical properties, the active compounds of the invention can be formulated as solutions, emulsions, suspensions, powders, foams, pastes, granules, aerosols, microencapsulations in polymeric substances and in coating materials for seed, and also as ULV cold and warm fogging formulations.

[0035] The formulation may be a ready-to-use solution, emulsion, water- or oil-based suspension, powder, wettable powder, paste, soluble powder, dust, soluble granules, granules for broadcasting, suspoemulsion concentrate, natural substance impregnated with active compound, synthetic substance impregnated with active compound, fertilizer or a microencapsulation in polymeric substances.

[0036] Formulations containing the compounds of the invention are produced in a known manner, for example by mixing the compounds with extenders (e.g. liquid solvents and / or solid carriers), optionally with the use of surfactants (e.g. emulsifiers and / or dispersants and / or foam-formers). The formulations are typically prepared either in factories / production plants. The formulations may be prepared before or during the application.

[0037] Formulations which could be used to administer compounds of the invention are outlined in the ‘Assay Procedure’, below.

[0038] Auxiliaries are substances which are suitable for imparting to the formulation itself and / or to preparations derived therefrom (for example spray liquors, seed dressings) particular properties such as certain technical properties and / or also particular biological properties. Typical suitable auxiliaries include extenders, solvents and carriers.

[0039] Suitable extenders are, for example, water, polar and nonpolar organic chemical liquids, for example from the classes of the aromatic and non-aromatic hydrocarbons (such as paraffins, alkylbenzenes, alkylnaphthalenes, chlorobenzenes), the alcohols and polyols (which, if appropriate, may also be substituted, etherified and / or esterified), the ketones (such as acetone, cyclohexanone), esters (including fats and oils) and (poly)ethers, the unsubstituted and substituted amines, amides, lactams (such as / V-alkylpyrrolidones) and lactones, the sulfones and sulfoxides (such as dimethylsulfoxide).

[0040] If the extender used is water, it is also possible to use, for example, organic solvents as auxiliary solvents. Essentially, suitable liquid solvents are: aromatics such as xylene, toluene or alkylnaphthalenes, chlorinated aromatics and chlorinated aliphatic hydrocarbons such as chlorobenzenes, chloroethylenes or methylene chloride, aliphatic hydrocarbons such as cyclohexane or paraffins, for example petroleum fractions, alcoholssuch as butanol or glycol and also their ethers and esters, ketones such as acetone, methyl ethyl ketone, methyl isobutyl ketone or cyclohexanone, strongly polar solvents such as dimethylformamide and dimethylsulfoxide.

[0041] Suitable solid carriers are: for example, ammonium salts and ground natural minerals such as kaolins, clays, talc, chalk, quartz, attapulgite, montmorillonite or diatomaceous earth, and ground synthetic minerals, such as finely divided silica, alumina and silicates; suitable solid carriers for granules are: for example, crushed and fractionated natural rocks such as calcite, marble, pumice, sepiolite and dolomite, and also synthetic granules of inorganic and organic meals, and granules of organic material such as paper, sawdust, coconut shells, maize cobs and tobacco stalks; suitable emulsifiers and / or foamformers are: for example, nonionic and anionic emulsifiers, such as polyoxyethylene fatty acid esters, polyoxyethylene fatty alcohol ethers, for example alkylaryl polyglycol ethers, alkylsulfonates, alkyl sulfates, arylsulfonates and also protein hydrolysates; suitable dispersants are nonionic and / or ionic substances, for example from the classes of the alcohol-POE and / or -POP ethers, acid and / or POP-POE esters, alkylaryl and / or POP-POE ethers, fat- and / or POP-POE adducts, POE- and / or POP-polyol derivatives, POE- and / or POP-sorbitan- or -sugar adducts, alkyl or aryl sulfates, alkyl- or arylsulfonates and alkyl or aryl phosphates or the corresponding PO-ether adducts. Furthermore, suitable oligo- or polymers, for example those derived from vinylic monomers, from acrylic acid, from EO and / or PO alone or in combination with, for example, (poly)alcohols or (poly)amines. It is also possible to employ lignin and its sulfonic acid derivatives, unmodified and modified celluloses, aromatic and / or aliphatic sulfonic acids and their adducts with formaldehyde.

[0042] Tackifiers such as carboxymethylcellulose and natural and synthetic polymers in the form of powders, granules or latices, such as gum arabic, polyvinyl alcohol and polyvinyl acetate, as well as natural phospholipids such as cephalins and lecithins, and synthetic phospholipids, can be used in the formulations.

[0043] Further additives may be mineral and vegetable oils. It is also possible to add colorants such as inorganic pigments, for example iron oxide, titanium oxide and Prussian Blue, and organic dyestuffs, such as alizarin dyestuffs, azo dyestuffs and metal phthalocyanine dyestuffs, and trace nutrients such as salts of iron, manganese, boron, copper, cobalt, molybdenum and zinc. Other possible additives are perfumes, mineral or vegetable, optionally modified oils and waxes.

[0044] The formulations may also comprise stabilizers, e.g. low-temperature stabilizers, preservatives, antioxidants, light stabilizers or other agents which improve chemical and / or physical stability.

[0045] The formulations generally comprise between 0.01 and 98% by weight of active compound, preferably between 0.1 and 95% and particularly preferably between 0.5 and 90%.

[0046] The formulation may further comprise one or more additional fungicides, for example, to improve the activity spectrum or to reduce or slow the development of resistance. A mixture with other known active compounds such as nematicides, acaricides, herbicides, bactericides or insecticides, or with fertilizers and growth regulators, safeners or semiochemicals is also possible.

[0047] The formulation may be as described in the ‘Catalogue of pesticide formulation types and international coding system’ (Technical Monograph n° 2, 8th Edition by CropLife International).Method / Use

[0048] The compounds of the invention, in combination with good plant and environmental tolerance and favourable toxicity to warm-blooded animals, are suitable for protecting plants and plant organs, for increasing the harvest yields, for improving the quality of the harvested material and for controlling diseases caused by fungi which are encountered in agriculture, in horticulture, in animal husbandry, in forests, in gardens and leisure facilities, in the protection of stored products and of materials, and in the hygiene sector. They may be preferably employed as crop protection agents.

[0049] In a third aspect of the invention there is provided a method for preventing, controlling or treating a fungal disease, the method comprising applying an agronomically effective and substantially non-phytotoxic quantity of a compound according to the first aspect of the invention, or a fungicidal formulation according to the second aspect of the invention, to seeds of plants, to plants themselves or to an area where it is intended that plants will grow.

[0050] In a fourth aspect of the inventions there is provided a use of a compound according to the first aspect of the invention, or a fungicidal formulation according to the second aspect of the invention, as a fungicide (i.e. to prevent, control or treat fungal disease in plants).

[0051] The compounds of the present invention are suitable for protecting any plant variety which is employed in agriculture, in the greenhouse, in forests or in horticulture. Typically, the plants are crop plants. Crop plants that may be protected by the compounds of the present invention include cereal grains (e.g. wheat, maize, rice, barley, rye, millet,triticale and oats); root and tuber vegetables (e.g. carrot, potato, radish, sugar beet, mangelwurzel and yam); bulb vegetables (i.e. Allium spp., e.g. onion and garlic); non- Brassica leafy Vegetables (e.g. celery, head lettuce, lettuce and spinach); Brassica leafy vegetables (e.g. broccoli, cauliflower, cabbage and mustard greens); legume vegetables (e.g. bean, pea, soybean, chickpea, peanut and lentil); fruiting vegetables (e.g. tomato, bell pepper and chilli); citrus fruits (e.g. orange, lemon and grapefruit); pome fruits (e.g. apple, pear and nectarine); cucurbit vegetables (e.g. cucumber, melon and squash); stone fruits (e.g. cherry, peach, plum and apricot); berries (e.g. strawberry, blackberry, raspberry, blueberry, blackcurrant, redcurrant and grape); tree nuts (e.g. almond, pecan, hazelnut, macadamia and pistachio); grass forage, fodder and hay (e.g. Bermuda grass, meadow- grass and Bromus spp.); non-grass animal feeds (e.g. alfalfa and clover); herbs and spices (e.g. basil, black pepper, chive and celery seed or dill seed); legume vegetables (e.g. rapeseed, sunflower and cottonseed); petiole vegetables (e.g. asparagus and rhubarb); tropical and subtropical fruits (e.g. cocoa, fig, guava, olive, avocado, banana, pomegranate, and pineapple); coffee (Coffea spp.); cotton; and sugarcane. The plants may however be ornamental plants. Typically, however, the plants will be crops. The crops may be cereal crops, e.g. wheat crops. It may be that the plants are growing or are being stored.

[0052] Fungal diseases are caused by fungi. It may be that the fungal disease is caused by a sac fungi. The fungi may be in the Mycosphaerellaceae family. For example, the fungi may be Asperisporium spp., Pseudocercospora spp., Mycosphaerella spp., Passalora spp., Pseudocercospora spp., Ramularia spp.., Sphaceloma spp., Sphaerellothecium spp., Thedgonia spp., and Zymoseptoria (Septoria) spp.. It may be that the fungal disease is caused by Zymoseptoria spp., e.g. Zymoseptoria tritici.

[0053] The fungi may be in the Pleosporaceae family. For example, the fungi may be Alternaria spp. or Pyrenophora spp.. It may be that the fungal disease is caused by Pyrenophora spp, e.g. Pyrenophora teres, Pyrenophora graminea, or Pyrenophora tritici- repentis. It may be that the fungal disease is caused by Pyrenophora teres.

[0054] The fungi may be Fusarium spp. The fungi may be Microdochium spp., e.g. Microdochium nivale.

[0055] It may be that the fungal disease is caused by fungi that has acquired antifungal resistance. Antifungal resistance may be defined as the ability to grow at antifungal drug concentrations that arrest growth and / or kill most isolates of that species.

[0056] It may be that the fungal disease is caused by fungi that has acquired resistance to SDHI (succinate dehydrogenase inhibitor) fungicides. It may be that the fungal disease is caused by SDHI-resistant Zymoseptoria spp., e.g. SDHI-resistant Zymoseptoria tritici. It may be that the fungal disease is caused by fungi (e.g. any of the aforementioned fungi) having the G143A mutation. The G143A mutation arises from the change of glycine to alanine at position 143 of the fungal cytochrome b gene.

[0057] It may be that the fungal disease is caused by fungi that has acquired resistance to a Qol. For example, it may be that the fungal disease is caused by Zymoseptoria tritici that has acquired resistance to azoxystrobin (i.e. Zymoseptoria tritici (Az reduced sensitivity)).

[0058] It may be that the disease is Septoria leaf blotch. It may be that the disease is net blotch.

[0059] Typically, the compounds of the invention will be applied as a fungicidal formulation. Application may be carried out, for example, by watering, spraying, atomizing, broadcasting, dusting, foaming, spreading, etc. It is also possible to apply the active compounds by the ultra-low volume method or to inject the preparation of active compound or the active compound itself into the soil. It is also possible to treat the seeds of the plants.

[0060] Exemplary application rates of the active compounds according to the invention are: when treating leaves: from 0.1 to 10 000 g / ha, preferably from 10 to 1000 g / ha, particularly preferably from 50 to 300 g / ha (when the application is carried out by watering or dripping, it is even possible to reduce the application rate, especially when inert substrates such as rockwool or perlite are used); when treating seed: from 2 to 200 g per 100 kg of seed, preferably from 2.5 to 150 g per 100 kg of seed, and particularly preferably from 2.5 to 25 g per 100 kg of seed, very particularly preferably from 2.5 to 12.5 g per 100 kg of seed; when treating the soil: from 0.1 to 10 000 g / ha, preferably from 1 to 5 000 g / ha.EXAMPLESGeneral Synthesis

[0061] The skilled person will appreciate that adaptation of methods known in the art could be applied in the manufacture of the compounds of the present invention.

[0062] For example, the skilled person will be immediately familiar with standard textbooks such as "Comprehensive Organic Transformations - A Guide to FunctionalGroup Transformations", RC Larock, Wiley-VCH (1999 or later editions); "March's Advanced Organic Chemistry - Reactions, Mechanisms and Structure”, MB Smith, J. March, Wiley, (5th edition or later); “Advanced Organic Chemistry, Part B, Reactions and Synthesis”, FA Carey, RJ Sundberg, Kluwer Academic / Plenum Publications, (2001 or later editions); "Organic Synthesis - The Disconnection Approach", S Warren (Wiley), (1982 or later editions); "Designing Organic Syntheses" S Warren (Wiley) (1983 or later editions); “Heterocyclic Chemistry”, J. Joule (Wiley 2010 edition or later); ("Guidebook To Organic Synthesis" RK Mackie and DM Smith (Longman) (1982 or later editions), etc., and the references therein as a guide.

[0063] The skilled person is familiar with a range of strategies for synthesising organic and particularly heterocyclic molecules and these represent common general knowledge as set out in text books such as Warren “Organic Synthesis: The Disconnection Approach”; Mackie and Smith “Guidebook to Organic Chemistry”; and Clayden, Greeves, Warren and Wothers “Organic Chemistry”.

[0064] The skilled chemist will exercise his judgement and skill as to the most efficient sequence of reactions for synthesis of a given target compound and will employ protecting groups as necessary. This will depend inter alia on factors such as the nature of other functional groups present in a particular substrate. Clearly, the type of chemistry involved will influence the choice of reagent that is used in the said synthetic steps, the need, and type, of protecting groups that are employed, and the sequence for accomplishing the protection I deprotection steps. These and other reaction parameters will be evident to the skilled person by reference to standard textbooks and to the examples provided herein.

[0065] Sensitive functional groups may need to be protected and deprotected during synthesis of a compound of the invention. This may be achieved by conventional methods, for example as described in “Protective Groups in Organic Synthesis” by TW Greene and PGM Wuts, John Wiley & Sons Inc. (1999), and references therein.

[0066] Throughout this specification these abbreviations have the following meanings: aq. aqueous POE polyoxyethyleneD deuterium POP polyoxypropyleneDMSO dimethyl sulfoxide r.h. relative humidityEO ethylene oxide sat. saturatedLC-MS liquid chromatography-iSQ single quadrupole spectrometryPDA photo diode array ULV ultra-low volumePO Propylene oxide LIPLC ultra-performance liquid chromatographyAnalytical Procedures

[0067] NMR spectra were run on either a Bruker Ultrashield 400 MHz or 500MHz NMR spectrometer. Spectra were recorded at 298K and were referenced using the solvent peak.

[0068] Mass spectra were run on LC-MS systems using electrospray ionization. These were run using a Waters Acquity Classic UPLC with PDA and SQ mass detection or a Waters Acquity H-Class UPLC with PDA and QDA mass detection. [M+H]+ refers to monoisotopic molecular weights.LC-MS-1 SQDMethod 2AColumn: Acquity UPLC BEH C18 2.1 x 50 mm 1.7 pmColumn Temp: 50 °C Flow rate: 0.8 mL / min. Eluents: A: H2O, 0.1 % formic acid, B: MeCN Gradient: 0.0-1.8 min 2-98% B, 1.8-2.1 min 98% B, 2.1-2.5 98% A.Method 2BColumn: Acquity UPLC BEH C18 2.1 x 50 mm 1.7 pmColumn Temp: 50 °C Flow rate: 0.8 mL / min. Eluents: A: H2O, 0.1 % ammonia B: MeCN Gradient: : 0.0-1.8 min 2-98% B, 1.8-2.1 min 98% B, 2.1-2.5 98% A.LC-MS-2 QDAMethod 2AColumn: Acquity UPLC BEH C18 2.1 x 50 mm 1.7 pmColumn Temp: 50 °C Flow rate: 0.8 mL / min.Eluents: A: H2O, B: MeCN, C: 50% H2O I 50% MeCN + 2.0% formic acid Gradient: 0.0 - 1.7 mins 0-95% B, 5% C; 1.7-2.1 mins 95% B, 5% C 2.1-2.5 mins 95% A, 5% C.Method 2BColumn: Acquity UPLC BEH C18 2.1 x 50 mm 1.7 pmColumn Temp: 50 °CFlow rate: 0.8 mL / min.Eluents: A: H2O, B: MeCN, C: 50% H2O I 50% MeCN + 2.0% ammonia (aq.) Gradient: 0.0 - 1.7 mins 0-95% B, 5% D; 1.7-2.1 mins 95% B, 5% D 2.1-2.5 mins 95% A, 5% D.

[0069] A solution of 3-bromo-2-methylaniline (2 g, 10.75 mmol) in toluene (6 mL) was added dropwise to a solution of triphosgene (4.8 g, 16.1 mmol) in toluene (30 mL). The reaction mixture was heated at reflux temperature for 3 h then concentrated in vacuo to give the title compound as a purple oil (2 g).

[0070] 1H N MR (400 MHz, CDCh) 6 7.40 (dd, = 7.8, 1.5 Hz, 1H), 7.13 - 6.94 (m, 2H), 2.43 (s, 3H).Intermediate X2: 1-(3-Bromo-2-tolyl)- 1 ,4-dihydro-5-tetraazolone

[0071] 2-Bromo-6-isocyanatotoluene (intermediate X1) (2.0 g, 9.43 mmol) and azidotrimethylsilane (5.87 g, 50.9 mmol) were heated in a sealed tube at 90 °C for 24 h. Upon cooling, the reaction mixture was added to sat. aq. NaHCCh (10 mL) and the crude product extracted into ethyl acetate (3 x 25 mL). The combined organics were dried over MgSCU and concentrated in vacuo. The crude product was purified by flash column chromatography on the Combiflash® Nextgen 300+ (40 g MODUS silica cartridge, eluting 0 to 50% ethyl acetate in petroleum ether) to give the title compound as a yellow solid (650 mg).

[0072] LC-MS-1 (Method 2A): Rt 1.21 mins; MS m / z = 257.0 [M+H]+(85% @ 254nm).

[0073] 1H NMR (400 MHz, DMSO) 5 14.63 (s, 1H), 7.84 (dd, J = 8.1, 1.3 Hz, 1H), 7.49 (dd, J = 8.0, 1.3 Hz, 1H), 7.35 (td, J = 7.9, 0.7 Hz, 1 H), 2.23 (s, 3H).Intermediate X3: 1-(3-Bromo-2-tolyl)-4-methyl- 1 ,4-dihydro-5-tetraazolone

[0074] Sodium hydride (60% dispersion in mineral oil, 75 mg, 1.88 mmol) was added to a solution of 1-(3-bromo-2-tolyl)-1,4-dihydro-5-tetraazolone (intermediate X2) (400 mg, 1.57 mmol) in / V, / V-dimethylformamide (7.3 mL) at 0 °C. After 5 min the cooling was removed and the reaction mixture stirred at room temperature for 1 h before being re-cooled to 0 °C. lodomethane (267 mg, 1.88 mmol) was added, the reaction mixture stirred for 2 h then added to water (75 mL) and the crude product extracted into ethyl acetate (3 x 50 mL). The combined organics were washed with brine (30 mL), dried over MgSCUand concentrated in vacuo to give the title compound as a brown solid (303 mg).

[0075] LC-MS-1 (Method 2A): Rt 1.33 mins; MS m / z = 271.0 [M+H]+(85% @ 254nm).

[0076] 1H NMR (400 MHz, CDCh) 6 7.70 (dd, J = 7.9, 1.5 Hz, 1H), 7.29 (dd, J = 8.0, 1.4 Hz, 1 H), 7.24 - 7.20 (m, 1 H), 3.72 (s, 3H), 2.32 (s, 3H).Intermediate X4: 1-[3-Bromo-2-(bromomethyl)phenyl]-4-methyl- 1 ,4-dihydro-5-tetraazolone

[0077] / V-Bromosuccinimide (143 mg, 0.80 mmol) and 2,2'-azobis(2-methylpropionitrile) (23 mg, 0.14 mmol) were added to a solution of 1-(3-bromo-2-tolyl)-4-methyl-1 ,4-dihydro- 5-tetraazolone (intermediate X3) (188 mg, 0.70 mmol) in degassed chlorobenzene (8.7 mL). The reaction mixture was heated at 165 °C for 5 h. Further / V-bromosuccinimide (143 mg, 0.80 mmol) and 2,2'-azobis(2-methylpropionitrile) (23 mg, 0.14 mmol) were added and the reaction mixture was heated at 165 °C for 24 h. Further / V-bromosuccinimide (143 mg, 0.80 mmol) and 2,2'-azobis(2-methylpropionitrile) (23 mg, 0.14 mmol) were again added and the reaction mixture was heated at 165 °C for 4 h. Upon cooling the reaction mixture was diluted with water (25 mL) and the crude product extracted into ethyl acetate (3 x 25 mL). The combined organics were washed with brine (2 x 20 mL), dried over MgSCU and concentrated in vacuo. The crude product was purified by flash column chromatography on the Combiflash® Nextgen 300+ (25 g MODUS silica cartridge, eluting 0 to 34% ethyl acetate in petroleum ether) to give the title compound as a white solid (162 mg).

[0078] LC-MS-1 (Method 2A): Rt 1.36 mins; mass ion not observed (97% @ 254nm).

[0079] 1H NMR (400 MHz, CDCh) 6 7.76 (dd, J = 7.7, 1.7 Hz, 1H), 7.43 - 7.29 (m, 2H), 4.72 (s, 2H), 3.75 (s, 3H).

[0080] 1-(4-chlorophenyl)pyrazol-3-ol (44.7 mg, 0.23 mmol) and potassium carbonate (38.13 mg, 0.275 mmol) were added to a solution of 1-[3-bromo-2-(bromomethyl)phenyl]-4- methyl-1,4-dihydro-5-tetraazolone (intermediate X4) (80 mg, 229.89 pmol) in acetonitrile (3.2 mL). The reaction mixture was heated at reflux temperature for 3 h. Upon cooling the reaction mixture was diluted with water (20 mL) and the crude product extracted into ethyl acetate (3 x 25 mL). The combined organics were washed with brine (2 x 20 mL), dried over MgSCU and concentrated in vacuo. The crude product was purified by flash column chromatography on the Combiflash® Nextgen 300+ (25 g MODUS silica cartridge, eluting 0 to 53% ethyl acetate in petroleum ether) to give the title compound as a yellow oil (87 mg).

[0081] LC-MS-1 (Method 2B): Rt 1.71 mins; MS m / z = 462.9 [M+H]+(100% @ 254nm).

[0082] 1H NMR (400 MHz, CDCh) 6 7.79 (ddd, J = 8.1, 4.3, 1.6 Hz, 1H), 7.68 - 7.62 (m, 1 H), 7.54 - 7.43 (m, 2H), 7.43 - 7.33 (m, 4H), 5.81 (d, J = 2.6 Hz, 1 H), 5.53 (s, 2H), 3.60 (s, 3H).Intermediate X6: 2-Bromo-1-[(2H3)methyl]-6-nitrotoluene

[0083] Methanol-d4 (2 mL) and sodium methoxide (250 mg, 4.63 mmol) were added to 2- bromo-6-nitrotoluene (1.0 g, 4.63 mmol) and the reaction mixture was heated in the microwave at 100 °C for 2 h. The suspension was filtered, washing with methanol-d4(3 x 1 mL). The filtrate was concentrated in vacuo then methanol-d4(2.5 mL) and sodium methoxide (250 mg, 4.63 mmol) were added. The reaction mixture was heated in the microwave at 100 °C for 2 h. The suspension was filtered, washing with methanol-d4(4 x1 mL) and the filtrate concentrated in vacuo. The reaction cycle was repeated twice more. The residue was dissolved in ethyl acetate (50 mL) and washed with deuterium oxide (2 x 5 mL), dried over MgSCU and concentrated in vacuo to give the title compound as a dark yellow oil (0.71 g).

[0084] 1H NMR (400 MHz, CDCh) 6 7.79 (dd, J = 8.0, 1.2 Hz, 1H), 7.72 (dd, J = 8.1, 1.3 Hz, 1H), 7.20 (t, J = 8.1 Hz, 1 H)Intermediate X7: 3-Bromo-2-l'(2H3)methyl]tolylamine j D3C^yNH2

[0085] Tin (II) chloride (2.64 g, 13.92 mmol) was added to a solution of 2-bromo-1- [(2H3)methyl]-6-nitrotoluene (intermediate X6) (0.61 g, 2.78 mmol) in ethanol (50 mL). The reaction mixture was heated at reflux temperature for 2.5 h then cooled to room temperature and poured into ice water (50 mL). After basification with sat. aq. NaHCCh (30 mL), ethyl acetate (250 mL) was added and the mixture was filtered through celite. The phases were separated and additional crude product was extracted from the aqueous phase with ethyl acetate (4 x 50 mL). The combined organics were washed with brine (40 mL), dried over MgSCU and concentrated in vacuo to give the title compound as a brown oil (0.51 g).

[0086] LC-MS-2 (Method 2B): Rt 1.43 min, mass ion not observed (50 % @ 254 nm).

[0087] 1H NMR (400 MHz, CDCh) 6 7.04 (dd, J = 7.9, 1.1 Hz, 1H), 6.88 (t, J = 7.9 Hz, 1 H), 6.68 (dd, J = 7.9, 1.2 Hz, 1 H). Amine NH2not observed.Intermediate X8: 2-Bromo-6-isocvanato-1-[(2H3)methylltoluene

[0088] A solution of 3-bromo-2-[(2H3)methyl]tolylamine (intermediate X7) (0.51 g, 2.70 mmol) in toluene (2.7 mL) was added dropwise to a solution of triphosgene (1.2 g, 4.05 mmol) in toluene (8.1 mL). The reaction mixture was heated at reflux temperature for 135 min then concentrated in vacuo to give the title compound as a brown oil (0.54 g).

[0089] 1H NMR (400 MHz, CDCh) 6 7.39 (dd, J = 7.7, 1.4 Hz, 1H), 7.10 - 6.94 (m, 2H).Intermediate X9: 1- / 3-Bromo-2-[(2H3)methylltolyl}-1,4-dihydro-5-tetraazolone

[0090] 2-Bromo-6-isocyanato-1-[(2H3)methyl]toluene (intermediate X8) (0.54 g, 2.51 mmol) and azidotrimethylsilane (1.56 g, 13.56 mmol) were heated in a sealed tube at 90 °C for 19 h. Upon cooling, the reaction mixture was added to sat. aq. NaHCCh (5 mL) and the crude product extracted into ethyl acetate (4 x 15 mL). The combined organics were washed with brine (10 mL), dried over MgSCU and concentrated in vacuo. The crude product was purified by flash column chromatography on the Combiflash® Nextgen 300+ (12 g MODUS silica cartridge, eluting 2 to 60% ethyl acetate in petroleum ether) to give the title compound as an off-white solid (0.44 g).

[0091] LC-MS-2 (Method 2B): Rt 0.80 mins; MS m / z 256.0 / 258.0 = [M-H]’ (97% @ 254nm).

[0092] 1H NMR (400 MHz, DMSO) 6 14.71 (s, 1H), 7.84 (dd, J = 8.1, 1.1 Hz, 1H), 7.49 (dd, J = 7.9, 1.2 Hz, 1H), 7.36 (t, J = 8.0 Hz, 1 H).Intermediate X10: 1- / 3-Bromo-2-[ f2H3) methyl ltolyl}-4-methyl- 1 ,4-dihydro-5-tetraazolone

[0093] Sodium hydride (60 % dispersion in mineral oil, 41 mg, 1.02 mmol) was added to a solution of 1-{3-bromo-2-[(2H3)methyl]tolyl}-1 ,4-dihydro-5-tetraazolone (intermediate X9) (220 mg, 0.85 mmol) in / V, / V-dimethylformamide (1.7 mL) at 0 °C. After 5 min the cooling was removed and the reaction mixture stirred at room temperature for 1 h before being recooled to 0 °C. lodomethane (64 pL, 1.02 mmol) and the reaction mixture stirred, allowing to warm to room temperature, for 20 h. The reaction mixture was diluted with water / brine (30 mL, 5:1) and the crude product extracted into ethyl acetate (3 x 25 mL). The combined organics were washed with brine (3 x 25 mL), dried over MgSCU and concentrated in vacuo to give the title compound as an off-white solid (219 mg).

[0094] LC-MS-2 (Method 2B): Rt 1.45 mins; mass ion not observed (78% @ 254nm).

[0095] 1H NMR (400 MHz, CDCh) 6 7.71 (dd, J = 8.0, 1.2 Hz, 1H), 7.30 (dd, J = 7.9, 1.3 Hz, 1H), 7.21 (t, J = 8.0 Hz, 1 H), 3.73 (s, 3H).Intermediate X11: 1- 3-Bromo-2-[bromo(2H2)methyl]phenyl}-4-methyl-1,4-dihydro-5- tetraazolone

[0096] / V-Bromosuccinimide (164 mg, 0.92 mmol) and 2,2'-zzobis(2-methylpropionitrile) (26 mg, 0.16 mmol) were added to a solution of 1-{3-bromo-2-[(2H3)methyl]tolyl}-4-methyl- 1 ,4-dihydro-5-tetraazolone (intermediate X10) (218 mg, 0.80 mmol) in degassed chlorobenzene (3.3 mL). The reaction mixture was heated at 165 °C for 5 h. Further N- bromosuccinimide (164 mg, 0.92 mmol) and 2,2'-azobis(2-methylpropionitrile) (26 mg, 0.16 mmol) were added and the reaction mixture was heated at 165 °C for 9 h. The same quantities of both reagents were added twice more over a further 9 h heating. Upon cooling the reaction mixture was diluted with water (25 mL) and the crude product was extracted into ethyl acetate (3 x 50 mL). The combined organics were washed with brine (50 mL), dried over MgSCU and concentrated in vacuo. The crude product was purified by flash column chromatography on the Combiflash® Nextgen 300+ (40 g MODUS silica cartridge, eluting 0 to 30% ethyl acetate in petroleum ether) to give the title compound as a pale orange solid (65 mg).

[0097] 1H NMR (400 MHz, CDCh) 6 7.76 (dd, J = 7.7, 1.6 Hz, 1H), 7.41 - 7.31 (m, 2H), 3.75 (s, 3H).

[0098] 1-(4-chlorophenyl)pyrazol-3-ol (36 mg, 0.19 mmol) and potassium carbonate (31 mg, 0.22 mmol) were added to a solution of 1-{3-bromo-2-[bromo(2H2)methyl]phenyl}-4- methyl-1,4-dihydro-5-tetraazolone (intermediate X11) (65 mg, 0.19 mmol) in acetonitrile (3 mL). The reaction mixture was heated at reflux temperature for 2 h. Upon cooling the reaction mixture was diluted with water (10 mL) and the crude product was extracted into ethyl acetate (3 x 20 mL). The combined organics were washed with brine (20 mL), driedover MgSCU and concentrated in vacuo. The crude product was purified by flash column chromatography on the Combiflash® Nextgen 300+ (12 g MODUS silica cartridge, eluting 0 to 60% ethyl acetate in petroleum ether) to give the title compound as an off-white solid (68 mg).

[0099] LC-MS-2 (Method 2B): Rt 1.81 mins; MS m / z 462.8 / 464.8 = [M+H]+(100% @ 254nm).

[0100] 1H NMR (400 MHz, CDCh) 6 7.81 - 7.77 (m, 1 H), 7.64 (d, = 2.6 Hz, 1 H), 7.54 - 7.49 (m, 2H), 7.42 - 7.34 (m, 4H), 5.81 (d, J = 2.6 Hz, 1 H), 3.60 (s, 3H).

[0101] Sodium hydride (60 % dispersion in mineral oil, 41 mg, 1.02 mmol) was added to a solution of 1-{3-bromo-2-[(2H3)methyl]tolyl}-1 ,4-dihydro-5-tetraazolone (intermediate X9) (220 mg, 0.85 mmol) in / V, / V-dimethylformamide (1.7 mL) at 0 °C. After 5 min the cooling was removed and the reaction mixture stirred at room temperature for 1 h before being recooled to 0 °C. Iodomethane-d3 (64 pL, 1.02 mmol) was added and the reaction mixture stirred, allowing to warm to room temperature, for 18 h. The reaction mixture was diluted with water / brine (30 mL, 5:1) and the crude product extracted into ethyl acetate (3 x 25 mL). The combined organics were washed with brine (3 x 25 mL), dried over MgSCU and concentrated in vacuo to give the title compound as an off-white solid (227 mg).

[0102] LC-MS-2 (Method 2B): Rt 1.45 mins; mass ion not observed (59% @ 254nm).

[0103] 1H NMR (400 MHz, CDCh) 6 7.64 (dd, J = 8.0, 1.2 Hz, 1 H), 7.23 (dd, J = 7.9, 1.3 Hz, 1 H), 7.14 (t, J = 8.0 Hz, 1 H).

[0104] / V-Bromosuccinimide (168 mg, 0.94 mmol)and 2,2'-azobis(2-methylpropionitrile) (27 mg, 0.16 mmol) were added to a solution of 1-{3-bromo-2-[(2H3)methyl]tolyl}-4- [(2H3)methyl]-1,4-dihydro-5-tetraazolone (intermediate X13) (226 mg, 0.82 mmol) in degassed chlorobenzene (3.3 mL). The resulting mixture was heated at 165 °C for 5 h. The same quantities of both reagents were added twice more over a further 9 h heating. Upon cooling the reaction mixture was diluted with water (25 mL) and the crude product extracted into ethyl acetate (3 x 50 mL). The combined organics were washed with brine (50 mL), dried over MgSCU and concentrated in vacuo. The crude product was purified by flash column chromatography on the Combiflash® Nextgen 300+ (40 g MODUS silica cartridge, eluting 0 to 30% ethyl acetate in petroleum ether) to give the title compound as a pale orange solid (63 mg).

[0105] 1H NMR (400 MHz, CDCh) 6 7.76 (dd, J = 7.7, 1.5 Hz, 1H), 7.41 - 7.31 (m, 2H).

[0106] 1-(4-chlorophenyl)pyrazol-3-ol (35 mg, 0.18 mmol) and potassium carbonate (30 mg, 0.21 mmol) were added to a solution of 1-{3-bromo-2-[bromo(2H2)methyl]phenyl}-4- [(2H3)methyl]-1,4-dihydro-5-tetraazolone (intermediate X14) (63 mg, 0.18 mmol) in acetonitrile (3 mL). The reaction mixture was heated at reflux temperature for 2 h. Upon cooling the reaction mixture was diluted with water (10 mL) and the crude product extracted into ethyl acetate (3 x 20 mL). The combined organics were washed with brine (20 mL), dried over MgSCU and concentrated in vacuo. The crude product was purified by flash column chromatography on the Combiflash® Nextgen 300+ (12 g MODUS silica cartridge, eluting 0 to 40% ethyl acetate in petroleum ether) to give the title compound as an off-white solid (69 mg).

[0107] LC-MS-2 (Method 2B): Rt 1.80 mins; MS m / z 465.9 / 467.9 = [M+H]+(100% @ 254nm).

[0108] 1H NMR (400 MHz, CDCh) 6 7.81 - 7.76 (m, 1H), 7.68 - 7.62 (m, 1H), 7.55 - 7.44 (m, 2H), 7.44 - 7.33 (m, 4H), 5.81 (d, J = 2.6 Hz, 1 H).Intermediate X16: 1-(p-Chlorophenyl)(5-2H)-1,2-dihydro-3-pyrazolone

[0109] Potassium terf-butoxide (778 mg, 6.94 mmol) was added to a solution of 1-(p- chlorophenyl)-1,2-dihydro-3 / 7-pyrazol-3-one (450 mg, 2.31 mmol) in dimethylsulfoxide-d6 (2.6 mL) and deuterium oxide (2.6 mL). The reaction mixture was heated at 100 °C for 17 h then diluted with water (50 mL) and neutralised by the addition of 2M HCI (aq.). The crude product was extracted into ethyl acetate (3 x 25 mL). The combined organics were dried over MgSCU and concentrated in vacuo. The crude product was purified by flash column chromatography on the Combiflash® Nextgen 300+ (25 g MODUS silica cartridge, eluting 0 to 35% ethyl acetate in petroleum ether) to give the title compound as a yellow solid (189 mg).

[0110] LC-MS-2 (Method 2A): Rt 1.26 mins; MS m / z 196.9 = [M+H]+(100% @ 254nm).

[0111] 1H NMR (400 MHz, DMSO) 5 10.30 (s, 1 H), 7.73 - 7.65 (m, 2H), 7.50 - 7.43 (m, 2H), 5.83 (s, 1 H).Intermediate X17: 4-(3-Bromo-2-{ 1-(p-chlorophenyl)(5-2H)-1H-pyrazol-3- yloxylmethyl}phenyl)-1-methyl-1,4-dihydro-1,2,3,4-tetraazol-5-one

[0112] 1-(p-Chlorophenyl)(5-2H)-1 ,2-dihydro-3-pyrazolone (intermediate X16) (35 mg, 0.18 mmol) and potassium carbonate (30 mg, 0.21 mmol) were added to a solution of 1-[3- bromo-2-(bromomethyl)phenyl]-4-methyl-1,4-dihydro-5-tetraazolone (intermediate X4) (63 mg, 0.18 mmol) in acetonitrile (2.4 mL). The reaction mixture was heated at reflux temperature for 3 h. Upon cooling the reaction mixture was diluted with water (10 mL) and the crude product extracted into ethyl acetate (3 x 20 mL). The combined organics were washed with brine (20 mL), dried over MgSCU, and concentrated in vacuo. The crude product was purified by flash column chromatography on the Combiflash® Nextgen 300+ (12 g MODUS silica cartridge, eluting 0 to 40% ethyl acetate in petroleum ether) to give the title compound as a yellow solid (44 mg).

[0113] LC-MS-1 (Method 2B): Rt 1.70 mins; MS m / z 464.0 / 466.0 = [M+H]+(81% @ 254nm).

[0114] 1H NMR (400 MHz, CDCh) 6 7.79 (ddd, J = 8.0, 4.3, 1.5 Hz, 1 H), 7.55 - 7.47 (m, 2H), 7.43 - 7.33 (m, 4H), 5.80 (s, 1 H), 5.53 (s, 2H), 3.60 (s, 3H).Intermediate X18: 1-(3-Bromo-2-tolyl)-4-[(2H3)methyl]- 4-dihydro-5-tetraazolone■X?'N\ ii N-N D3C

[0115] Sodium hydride (60% dispersion in mineral oil, 15 mg, 0.39 mmol) was added to a solution of 1-(3-bromo-2-tolyl)-1,4-dihydro-5-tetraazolone (intermediate X2) (82 mg, 0.32 mmol) in / V, / V-dimethylformamide (0.6 mL) at 0 °C. After 5 min the cooling was removed and the reaction mixture stirred at room temperature for 1 h before being re-cooled to 0 °C. lodomethane (22 pL, 0.35 mmol) was added, the reaction mixture stirred at room temperature for 16 h then added to water (10 mL) and the crude product extracted into ethyl acetate (3 x 10 mL). The combined organics were washed with brine (100 mL), dried over MgSCUand concentrated in vacuo. The crude product was purified by flash column chromatography on the Combiflash® Nextgen 300+ (12 g MODUS silica cartridge, eluting 15 to 40% ethyl acetate in petroleum ether) to give the title compound as a white solid (69 mg).

[0116] LC-MS-1 (Method 3A): Rt 1.80 mins; MS m / z = 272.1 [M+H]+(94% @ 254nm).

[0117] 1H NMR (500 MHz, Chloroform-d) 5 7.71 (dd, J = 8.0, 0.9 Hz, 1H), 7.30 (dt, J = 7.9, 0.9 Hz, 1H), 7.21 (t, J = 8.0 Hz, 1 H), 2.33 (s, 3H).Intermediate X19: 1-[3-Bromo-2-(bromomethyl)phenyll-4-[ Hsimethyll- 1,4-dihydro-5- tetraazolone

[0118] / V-Bromosuccinimide (50 mg, 0.28 mmol) and 2,2'-azobis(2-methylpropionitrile) (8 mg, 0.05 mmol) were added to a solution of 1-(3-Bromo-2-tolyl)-4-[(2H3)methyl]-1 ,4- dihydro-5-tetraazolone (intermediate X18) (67 mg, 0.25 mmol) in degassed chlorobenzene (1.0 mL). The reaction mixture was heated at 165 °C for 5 h. Further / V-bromosuccinimide (9 mg, 0.05 mmol) and 2,2'-azobis(2-methylpropionitrile) (4 mg, 0.03 mmol) were added and the reaction mixture was heated at 165 °C for 24 h. Further / V-bromosuccinimide (143mg, 0.80 mmol) and 2,2'-azobis(2-methylpropionitrile) (23 mg, 0.14 mmol) were again added and the reaction mixture was heated at 165 °C for 4 h. Upon cooling the reaction mixture was diluted with water (5 mL) and the crude product extracted into ethyl acetate (3 x 10 mL). The combined organics were washed with brine (10 mL), dried over MgSCU and concentrated in vacuo. The crude product was purified by flash column chromatography on the Combiflash® Nextgen 300+ (12 g MODUS silica cartridge, eluting 0 to 45% ethyl acetate in petroleum ether) to give the title compound as a white solid (62 mg).

[0119] LC-MS-1 (Method 3A): Rt 1.85 mins; MS m / z = 352.0 [M+H]+(100% @ 254nm).

[0120] 1H NMR (500 MHz, Chloroform-d) 5 7.76 (dd, J = 7.8, 1.5 Hz, 1H), 7.38 (dd, J = 8.0, 1.5 Hz, 1H), 7.34 (t, J = 7.9 Hz, 1 H), 4.72 (s, 2H).

[0121] 1-(4-chlorophenyl)pyrazol-3-ol (35 mg, 0.18 mmol) and potassium carbonate (29 mg, 0.21 mmol) were added to a solution of 1-[3-Bromo-2-(bromomethyl)phenyl]-4- [(2H3)methyl]-1,4-dihydro-5-tetraazolone (intermediate X19) (62 mg, 0.18 mmol) in acetonitrile (1.3 mL). The reaction mixture was heated at reflux temperature for 2 h. Upon cooling the reaction mixture was diluted with water (5 mL) and the crude product extracted into ethyl acetate (3 x 10 mL). The combined organics were washed with brine (10 mL), dried over MgSCU and concentrated in vacuo. The crude product was purified by flash column chromatography on the Combiflash® Nextgen 300+ (4 g MODUS silica cartridge, eluting 0 to 50% ethyl acetate in petroleum ether) to give the title compound as a yellow solid (69 mg).

[0122] LC-MS-1 (Method 3A): Rt 2.25 mins; MS m / z = 466.1 [M+H]+(100% @ 254nm).

[0123] 1H NMR (500 MHz, Chloroform-d) 5 7.81 - 7.76 (m, 1 H), 7.64 (d, J = 2.6 Hz, 1 H), 7.54 - 7.49 (m, 2H), 7.40 - 7.34 (m, 4H), 5.81 (d, J = 2.6 Hz, 1 H), 5.53 (s, 2H).Exemplary Compounds

[0124] Methylboronic acid-d3 (18 mg, 0.283 mmol), tripotassium phosphate (120 mg, 0.565 mmol), palladium (II) bis(diphenylphosphino)ferrocene dichloride (13.5 mg, 0.019 mmol) and water (95 pL) were added to a solution of 1-(3-bromo-2-{[1-(p-chlorophenyl)-3- pyrazolyloxy]methyl}phenyl)-4-methyl-1,4-dihydro-5-tetraazolone (intermediate X5) (87 mg, 0.188 mmol) in dioxane (1.5 mL). The reaction mixture was heated at 80 °C for 5.5 h. Upon cooling, further methylboronic acid-d3 (6 mg, 0.094 mmol) and palladium (II) bis(diphenylphosphino)ferrocene dichloride (7 mg, 0.009 mmol) were added. The reaction mixture was heated at 80 °C for a further 5 h then, upon cooling, was diluted with water (20 mL) and the crude product extracted into ethyl acetate (3 x 15 mL). The combined organics were washed with brine (10 mL), dried over MgSCU and concentrated in vacuo. The crude product was purified by flash column chromatography on the Combiflash® Nextgen 300+ (25 g MODUS silica cartridge, eluting 0 to 46% ethyl acetate in petroleum ether) and then further purified by flash column chromatography on the Interchim Puriflash ® 450 (20 g MODUS C cartridge, 5 to 60% acetonitrile in water (0.1% formic acid) followed by lyophilisation to give the title compound as a white solid (30 mg).

[0125] LC-MS-1 (Method 2B): Rt 1.64 mins; MS m / z = 400.2 [M+H]+(97% @ 254nm).

[0126] 1H NMR (400 MHz, CDCh) 6 7.65 (d, J = 9.3 Hz, 1 H), 7.56 - 7.46 (m, 2H), 7.44 - 7.33 (m, 4H), 7.24 (d, J = 3.1 Hz, 1 H), 5.82 (d, J = 2.7 Hz, 1 H), 5.33 (s, 2H), 3.63 (s, 3H).Example 2: 1-(2-ff 1-(p-Chlorophenyl)-3-pyrazolyloxy](2H2)methyl}-3-tolyl)-4-methyl-1 ,4- dihydro-5-tetraazolone

[0127] Methylboronic acid (6 mg, 0.10 mmol), tripotassium phosphate (41 mg, 0.19 mmol), palladium (II) bis(diphenylphosphino)ferrocene dichloride (5 mg, 0.006 mmol) and water (30 pL) were added to a solution of 1-(3-bromo-2-{[1-(p-chlorophenyl)-3- pyrazolyloxy](2H2)methyl}phenyl)-4-methyl-1 ,4-dihydro-5-tetraazolone (intermediate X12) (30 mg, 0.06 mmol) in dioxane (0.5 mL). The reaction mixture was heated at 80 °C for 3 h. Upon cooling, further methylboronic acid (2 mg, 0.04 mmol) and palladium (II)bis(diphenylphosphino)ferrocene dichloride (2 mg, 0.003 mmol) were added. The reaction mixture was heated at 80 °C for a further 2 h then, upon cooling, was diluted with water (5 mL) and the crude product extracted into ethyl acetate (10 mL then 3 x 5 mL). The combined organics were washed with brine (5 mL), dried over MgSCU and concentrated in vacuo. The crude product was purified by flash column chromatography on the Combiflash® Nextgen 300+ (4 g MODUS silica cartridge, eluting 2 to 40% ethyl acetate in petroleum ether) followed by lyophilisation to give the title compound as an off-white solid (19 mg).

[0128] LC-MS-2 (Method 2B): Rt 1.70 mins; MS m / z 399.0 = [M+H]+(100% @ 254nm).

[0129] 1H NMR (400 MHz, CDCh) 6 7.67 - 7.63 (m, 1H), 7.50 (d, J = 8.8 Hz, 2H), 7.44 - 7.35 (m, 5H), 5.82 (d, J = 2.7 Hz, 1 H), 3.64 (d, J = 8.4 Hz, 3H), 2.57 (d, J = 11.9 Hz, 3H).Example 3: 1-(p-Chlorophenyl)-3-pyrazolyloxy](2H2)methyl}-3-r(2H3)methyl]tolyl)-4- methyl- 1 ,4-dihydro-5-tetraazolone

[0130] Methylboronic acid-d3 (6 mg, 0.10 mmol), tripotassium phosphate (41 mg, 0.19 mmol), palladium (II) bis(diphenylphosphino)ferrocene dichloride (5 mg, 0.006 mmol) and water (30 pL) were added to a solution of 1-(3-bromo-2-{[1-(p-chlorophenyl)-3- pyrazolyloxy](2H2)methyl}phenyl)-4-methyl-1 ,4-dihydro-5-tetraazolone (intermediate X12) (30 mg, 0.06 mmol) in dioxane (0.5 mL). The reaction mixture was heated at 80 °C for 3 h. Upon cooling, further methylboronic acid-d3 (2 mg, 0.04 mmol) and palladium (II) bis(diphenylphosphino)ferrocene dichloride (2 mg, 0.003 mmol) were added. The reaction mixture was heated at 80 °C for a further 2 h. Upon cooling, further methylboronic acid-d3 (2 mg, 0.04 mmol) and palladium (II) bis(diphenylphosphino)ferrocene dichloride (2 mg, 0.003 mmol) were added. The reaction mixture was heated at 80 °C for a further 1 h then, upon cooling, was diluted with water (5 mL) and the crude product extracted into ethyl acetate (10 mL then 3 x 5 mL). The combined organics were washed with brine (5 mL), dried over MgSCU and concentrated in vacuo. The crude product was purified by flash column chromatography on the Combiflash® Nextgen 300+ (8 g MODUS silica cartridge, eluting 2 to 40% ethyl acetate in petroleum ether) followed by lyophilisation to give the title compound as an off-white solid (18 mg).

[0131] LC-MS-2 (Method 2A): Rt 1.75 mins; MS m / z 402.0 = [M+H]+(98% @ 254nm).

[0132] 1H NMR (400 MHz, CDCh) 6 7.67 - 7.62 (m, 1 H), 7.54 - 7.47 (m, 2H), 7.44 - 7.35 (m, 5H), 5.82 (d, J = 2.6 Hz, 1 H), 3.64 (d, J = 8.4 Hz, 3H).Example 4: 1-(p-Chlorophenyl)-3-pyrazolyloxyl(2H2)methyl}-3-tolyl)-4-[(2H3)methyll-1 , 4-dihydro-5-tetraazolone

[0133] Methylboronic acid (6 mg, 0.10 mmol), tripotassium phosphate (41 mg, 0.19 mmol), palladium (II) bis(diphenylphosphino)ferrocene dichloride (5 mg, 0.006 mmol) and water (30 pL) were added to a solution of 1-(3-bromo-2-{[1-(p-chlorophenyl)-3- pyrazolyloxy](2H2)methyl}phenyl)-4-[(2H3)methyl]-1 ,4-dihydro-5-tetraazolone (intermediate X15) (30 mg, 0.06 mmol) in dioxane (0.5 mL). The reaction mixture was heated at 80 °C for 3 h. Upon cooling the reaction mixture was diluted with water (5 mL) and the crude product extracted into ethyl acetate (4 x 5 mL). The combined organics were washed with brine (5 mL), dried over MgSCU and concentrated in vacuo. The crude product was purified by flash column chromatography on the Combiflash® Nextgen 300+ (4 g MODUS silica cartridge, eluting 2 to 30% ethyl acetate in petroleum ether) followed by lyophilisation to give the title compound as an off-white solid (18 mg).

[0134] LC-MS-2 (Method 2A): Rt 1.73 min, MS m / z 402.0 = [M+H]+(93 % @ 254 nm).

[0135] 1H NMR (400 MHz, CDCh) 6 7.67 - 7.63 (m, 1 H), 7.54 - 7.47 (m, 2H), 7.44 - 7.35 (m, 5H), 5.81 (d, J = 2.6 Hz, 1 H), 2.60 - 2.54 (m, 3H).Example 5: 1-(2-ff 1-(o-Chlorophenyl)-3-pyrazolyloxyl(2H2}methyl}-3- (2H3)methylltolyl)-4- f I2!- / 3) methyl]- 1 ,4-dihydro-5-tetraazolone

[0136] Methylboronic acid-d3 (6 mg, 0.10 mmol), tripotassium phosphate (41 mg, 0.19 mmol), palladium (II) bis(diphenylphosphino)ferrocene dichloride (5 mg, 0.006 mmol) and water (30 pL) were added to a solution of 1-(3-bromo-2-{[1-(p-chlorophenyl)-3- pyrazolyloxy](2H2)methyl}phenyl)-4-[(2H3)methyl]-1 ,4-dihydro-5-tetraazolone (intermediate X15) (30 mg, 0.06 mmol) in dioxane (0.5 mL). The reaction mixture was heated at 80 °Cfor 3 h. Upon cooling the reaction mixture was diluted with water (5 mL) and the crude product extracted into ethyl acetate (4 x 5 mL). The combined organics were washed with brine (5 mL), dried over MgSCU and concentrated in vacuo. The crude product was purified by flash column chromatography on the Combiflash® Nextgen 300+ (4 g MODUS silica cartridge, eluting 2 to 30% ethyl acetate in petroleum ether) followed by further purification by flash column chromatography on the Interchim Puriflash® 450 (5.4 g MODUS C cartridge, 5 to 70% acetonitrile in water (0.1% formic acid) followed by lyophilisation to give the title compound as a white solid (7 mg).

[0137] LC-MS-1 (Method 2A): Rt 1.61 mins; MS m / z 403.3 = [M-H]’ (95% @ 254nm).

[0138] 1H NMR (400 MHz, CDCh) 6 7.67 - 7.62 (m, 1H), 7.53 - 7.47 (m, 2H), 7.44 - 7.33 (m, 5H), 5.82 (d, J = 2.6 Hz, 1 H).Example 6: 4-(2-ff 1 -(p-Chlorophenyl)(5-2H)-1 H-pyrazol-3-yloxylmethyl}-3-tolyl)-1 -methyl- 1 , 4-dihydro- 1,2,3, 4-tetraazol-5-one

[0139] Methylboronic acid (8.5 mg, 0.14 mmol), tripotassium phosphate (61 mg, 0.29 mmol), palladium (II) bis(diphenylphosphino)ferrocene dichloride (7 mg, 0.009 mmol) and water (48 pL) were added to a solution of 4-(3-bromo-2-{[1-(p-chlorophenyl)(5-2H)-1 / 7- pyrazol-3-yloxy]methyl}phenyl)-1-methyl-1,4-dihydro-1 ,2,3,4-tetraazol-5-one (intermediate X17) (44 mg, 0.095 mmol) in dioxane (0.76 mL). The reaction mixture was heated at 80 °C for 5.5 h. Then further methylboronic acid (3 mg, 0.05 mmol) and palladium (II) bis(diphenylphosphino)ferrocene dichloride (4 mg, 0.005 mmol) were added and the reaction mixture was stirred at 80 °C for 5 h. Upon cooling the reaction mixture was diluted with water (10 mL) and the crude product extracted into ethyl acetate (4 x 10 mL). The combined organics were washed with brine (10 mL), dried over MgSCU and concentrated in vacuo. The crude product was purified by flash column chromatography on the Combiflash® Nextgen 300+ (25 g MODUS silica cartridge, eluting 0 to 46% ethyl acetate in petroleum ether) followed by lyophilisation to give the title compound as a white solid (18 mg).

[0140] LC-MS-1 (Method 2B): Rt 1.63 min, MS m / z 399.2 = [M+H]+(100 % @ 254 nm).

[0141] 1H NMR (400 MHz, CDCh) 6 7.52 - 7.49 (m, 2H), 7.44 - 7.34 (m, 5H), 5.82 (s, 1 H), 5.38 (d, J = 41.6 Hz, 2H), 3.64 (d, J = 8.2 Hz, 3H), 2.57 (d, J = 11.7 Hz, 3H).Example 7: 1-(p-Chlorophenyl)-3-pyrazolyloxy]methyl}-3-tolyl)-4-[(2H3)methyl]-1,4- dihydro-5-tetraazolone

[0142] Methylboronic acid (11 mg, 0.18 mmol), tripotassium phosphate (75 mg, 0.36 mmol), palladium (II) bis(diphenylphosphino)ferrocene dichloride (8.5 mg, 0.01 mmol) and water (6 pL) were added to a solution of 1-(3-bromo-2-{[1-(p-chlorophenyl)-3- pyrazolyloxy]methyl}phenyl)-4-[(2H3)methyl]-1 ,4-dihydro-5-tetraazolone (intermediate X20) (55 mg, 0.12 mmol) in dioxane (0.9 mL). The reaction mixture was heated at 80 °C for 2 h then, upon cooling, was diluted with water (10 mL) and the crude product extracted into ethyl acetate (3 * 10 mL). The combined organics were washed with brine (10 mL), dried over MgSCU and concentrated in vacuo. The crude product was purified by flash column chromatography on the Combiflash® Nextgen 300+ (12 g MODUS silica cartridge, eluting 0 to 40% ethyl acetate in petroleum ether) to give the title compound as a yellow oil (42 mg).

[0143] LC-MS-1 (Method 3A): Rt 2.18 mins; MS m / z = 400.2 [M+H]+(87% @ 254nm).

[0144] 1H NMR (500 MHz, Chloroform-d) 5 7.64 (d, J = 2.6 Hz, 1 H), 7.52 - 7.49 (m, 2H), 7.41 - 7.34 (m, 5H), 5.82 (d, J = 2.6 Hz, 1 H), 5.33 (s, 2H), 2.56 (s, 3H).Fungicidal ActivityExperiment 1: Broth Assay

[0145] Metyltetraprole and compounds of the invention were screened in 96 well plates with one compound screened against six pathogens per plate, with each test plate replicated twice. Each test compound was screened at 100, 10, 2, 0.04, 0.008 and 0.0016 ppm and tested against Zymoseptoria tritici (Az reduced sensitivity), Zymoseptoria tritici (SDHI resistant), Alternaria alternata, Phytophthora cactorum, Fusarium graminearum and Microdochium nivale.

[0146] A x100 stock solution in DMSO was produced for each dose and 2 pl of these added to the appropriate wells on the 96 well plate, and an equivalent amount of DMSO added to control wells.

[0147] For each pathogen a spore suspension (containing 1 ,000 spores / ml) was produced in GPM broth and 198 pl of this added to the appropriate wells on the 96 well plate to give the required final well concentrations.

[0148] Plate absorbance was read at 405 nm immediately after plate set up and then again after plates were incubated for 4 days at 18°C.

[0149] EC50 values for each compound and pathogen were calculated based on a comparison of growth in the control and test wells. Results are shown in Table 1 .Table 1

[0150] As can be seen from Table 1 , certain compounds of the invention, e.g. Examples 2, 3 and 5, display superior activity against certain pathogenic strains, e.g. SDHI resistant Zymoseptoria tritici, compared to metyltetraprole. Notably, Example 3 shows an order of magnitude greater control over Microdochium nivale than metyltetraprole.Experiment 2: In planta test

[0151] Metyltetraprole and Examples 2, 3 and 5 were screened at five concentrations (150, 37.5, 9, 2.3 and 1 g / ha) for efficacy against an SDHI resistant isolate of Zymoseptoria tritici.

[0152] Treatments were applied to 2-week-old winter wheat (var. JB Diego) grown in 9 cm diameter pots (10 plants per pot). Compounds were formulated in a 25% acetone and 75% 0.1% Tween 20 solution and sprayed using a track sprayer in a spray volume of 200 L / ha.

[0153] Each treatment was replicated three times with treatments applied 24 h prior to inoculation with spores of Z. tritici (SDHI resistant). Treated and the appropriate control plants were inoculated with 2 ml of a spore suspension at 1x106spores / ml. Inoculated plants were kept under high humidity conditions for 72 hours and then grown at 21 °C until they were ready to be assessed.

[0154] Plants were assessed once disease symptoms had fully developed (21 days after inoculation). The leaf area covered by pycnidia was carried out on the second leaf to emerge for 5 plants per replicate pot.

[0155] The assessment was based on the % control compared to the untreated controls. Results are shown in Figure 1. As can be seen from Figure 1 each tested example displayed superior activity in planta compared to metyltetraprole at each tested dose.

Claims

CLAIMS1 . A compound of Formula (I):agronomically acceptable salt or / V-oxide thereof, wherein:R1is selected from D and H;R2is independently at each occurrence selected from D and H;R3is selected from CH3, CH2D, CHD2 and CD3; andR4is selected from CH3, CH2D, CHD2 and CD3, wherein R1, R2, R3and R4are selected so that at least one of the groups selected from R1, the two R2groups, R3and R4is D or comprises D.

2. A compound of claim 1 , wherein R1is H.

3. A compound of claim 1 , wherein R1is D.

4. A compound of any preceding claim, wherein at least one R2is D.

5. A compound of any preceding claim, wherein R2is at each occurrence D.

6. A compound of any one of claims 1 to 3, wherein R2is at each occurrence H.

7. A compound of any preceding claim, wherein R3is CH3.

8. A compound of any one of claims 1 to 6, wherein R3is CD3.

9. A compound of any preceding claim, wherein R4is CH3.

10. A compound of any one of claims 1 to 8, wherein R4is CD3.

11. A compound of claim 1 , wherein the compound of Formula (I) is selected from:

12. A fungicidal formulation comprising a compound of any one of claims 1 to 11.

13. A method for preventing, controlling, or treating a fungal disease, the method comprising applying an agronomically effective and substantially non-phytotoxic quantity of a compound of any one of claims 1 to 11 , or a fungicidal formulation of claim 12, to seeds of plants, to plants themselves or to an area where it is intended that plants will grow or are growing.

14. The method of claim 13, wherein the fungal disease is caused by Zymoseptoria tritici.

15. The method of claim 14, wherein the Zymoseptoria tritici has a G143A mutation.

16. The method of any one of claims 13 to 15, wherein the plants are cereal crops.

17. The method of claim 16, wherein the cereal crops are wheat crops.

18. A use of a compound of any one of claims 1 to 11 , or a fungicidal formulation of claim 12, as a fungicide.