Patent Description:
Insecticides are agents of chemical or biological origin which to control the growth of insect (arthropods) pests, primarily by either killing them or by inhibiting a critical step in the insect's lifecycle (e.g., a stage such as the egg, larva, nymph, pupa, and/or adult). Insecticides may either exert their effect acutely, such as after a single exposure, or after chronic exposure. There are a variety of ways in which insecticides can be disseminated, depending on the nature of the pest. Of the more than <NUM>-<NUM> million arthropod species known, including more than <NUM> million species of true insects known, many thousands are considered pests or parasites. Some are responsible for the destruction of crops, and this constitutes the bulk of commercial insecticide usage. Other insects cause disease in animals, including humans, companion animals (dogs, cats, horses, etc.), and livestock (e.g., poultry, cattle, pigs, sheep, goats).

Insecticides have a variety of modes of action, although primarily they affect the insect nervous system, usually as potent neurotoxins. For example, organochlorine insecticides (e.g., DDT) and pyrethroid insecticides promote sodium channel opening, organophosphate and carbamate insecticides inhibit the acetylcholinesterase enzyme, neonicotinoid insecticides inhibit the nicotinic acetylcholine receptor, and phenylpyrazole insecticides inhibit the GABA-gated chloride channel.

The phenylpyrazole class of insecticides are broad-spectrum insecticides and are among the most commonly used insecticides. Phenylpyrazole insecticides act on the GABA (gamma-aminobutyric acid) receptor on the neurons of insects, blocking chloride ion conduction. Because chloride ion influx into neurons is an inhibitory action, disruption of GABA receptor opening results in a hyperexcited state in the neurons. Fipronil was the first phenylpyrazole insecticide, introduced in <NUM>. These agents were developed specifically to address the widespread resistance of insects to older classes of insecticides. In particular fipronil has been used against insects which have become tolerant of pyrethroid, organophosphate, and carbamate insecticides, including cockroaches, beetles, ants, fleas, termites, ticks, weevils, mole crickets, flies, mosquitos, and moths. Other phenylpyrazole class insecticides include acetoprole, ethiprole, flufiprole, pyraclofos, pyrafluprole, pyriprole, pyrolan, and vanilliprole. While these compound share a N-phenylpyrazole ring, they are otherwise quite structurally diverse:
<CHM>
<CHM>
<CHM>.

After they were introduced, the phenylpyrazole insecticides played a crucial role in controlling pests in both crop, livestock, and companion animal uses. However, their extensive use has also already led to the development of resistance. In addition, while phenylpyrazoles are generally relatively safe to mammals (due to structural differences between insect and mammalian GABA receptors), fipronil has recently been banned in many countries for certain uses due to high toxicity to non-target beneficial organisms, including fish, aquatic invertebrates, and bees. See, e.g., <NPL>; <NPL>; <NPL>. Leemans et al. characterize fipronil as a widespread environmental contaminant, being found in soil, water, outdoor dust, and some food products. Fipronil is thought to be cytotoxic to epithelial cells, including intestinal epithelium. If ingested in sufficient quantities, fipronil can cause nausea, vomiting, abdominal pain, seizures, and toxic effects on the kidneys, liver and thyroid glands.

Fipronil was linked to mass bee die-offs in France in the <NUM>'s and <NUM>'s, resulting in it being banned as a crop insecticide by the European Commission in <NUM>, as well as banned for use on animals intended for human consumption. While environmental degradation of fipronil is slow, metabolic degradation in animals is rapid. Contributing to the risk of oral toxicity, the primary metabolites of fipronil-the sulfone, the sulfide, and the desulfinylated derivative, are considerably more toxic, persistent, bioaccumulative, and less selective than the parent compound. See Leemans et al.

Fipronil has long been used to treat ectoparasites infecting poultry, such as fleas, lice, and ticks (particularly the poultry mite or red mite, Dermanyssus gallinae). However, in the EU, fipronil was banned for use in poultry for human consumption from <NUM> to <NUM>. In <NUM>, a serious scandal rocked the continent when fipronil was detected in eggs on the European market. This was the result of pesticide vendors illegally selling fipronil-containing products to unknowing farmers. As a result, millions of eggs were pulled from the market, millions of chickens were slaughtered, and the proprietors of the pesticide vendors faced jail time. Nevertheless, in <NUM>, the judicial court of the EU reversed some aspects of the fipronil ban, on the ground of insufficient evidence, but there remain calls to completely ban fipronil from the EU marketplace.

Thus, there has been work done to try to replace fipronil. , observed that molecular modeling predicted an intramolecular hydrogen bond between the amino group and the sulfoxide oxygen in fipronil, and therefore devised a series of <NUM>,<NUM>,<NUM>,<NUM>-tetrahydropyrazolo[<NUM>,<NUM>-d]oxazine derivatives:
<CHM>.

prepared and tested <NUM> compounds, but only four of the compounds had measurable insecticidal activity, peaking at only about <NUM>% of fipronil's activity against Plutella xylostella none of which had insecticidal activity comparable to fipronil. The authors speculated that the disappointing result could have been due to differences in physicochemical properties,.

There exists a continuing need for effective methods for controlling pests in both crop, livestock, and companion animal uses, without the risk of accumulation of the agent in the animal, such as in food products derived from the animal.

The present disclosure provides new N-phenyl-<NUM>H-pyrazolo[<NUM>,<NUM>-b]pyridine-<NUM>-carbonitriles, effective as insecticidal agents for crop, livestock, human, companion animal, and other uses.

In a first aspect, the present disclosure provides a Compound <NUM>, wherein Compound <NUM> is a Compound of Formula Ib (Compound 1b) having the following general structure:
<CHM>
wherein:.

In a second aspect, the present disclosure a pharmaceutical or insecticidal composition (Composition <NUM>) comprising an effective amount of a Compound <NUM> of Formula I, wherein Compound <NUM> is a Compound of Formula Ib (Compound 1b), in free or salt form, in admixture with a pharmaceutically acceptable or insecticidally acceptable diluent or carrier. In certain embodiments, the compositions as described herein may be used to or is intended for use to treat, control, suppress, or eradicate, an infection or infestation of an insect (arthropod), such as adult insect, insect egg, insect larva, insect nymph, and/or insect pupa, such in a human, an animal, a product, or a location, in need thereof.

In another aspect, the present disclosure provides a method (Method <NUM>) for the treatment, control, suppression, or eradication, of an infection or infestation of an insect (arthropod), such as an adult insect, insect egg, insect larva, insect nymph, and/or insect pupa, comprising administering or applying an effective amount of a compound of Formula Ib, in free or salt form, or a composition comprising a compound of Formula Ib, in free or salt form, as described herein, to an animal or human patient in need thereof or to a product or location in need thereof.

The preceding general areas of utility are given by way of example only and are not intended to be limiting on the scope of the present disclosure and appended claims. Additional objects and advantages associated with the compositions, methods, and processes of the present disclosure will be appreciated by one of ordinary skill in the art in light of the instant claims, description, and examples. For example, the various aspects and embodiments of the invention may be utilized in numerous combinations, all of which are expressly contemplated by the present description. These additional advantages objects and embodiments are expressly included within the scope of the present disclosure. Where applicable or not specifically disclaimed, any one of the embodiments described herein are contemplated to be able to combine with any other one or more embodiments, even though the embodiments are described under different aspects of the disclosure.

The following is a detailed description provided to aid those skilled in the art in practicing the invention of the present disclosure.

In further embodiments of the first aspect, the present disclosure provides:.

In a second aspect, the present disclosure also provides a pharmaceutical or insecticidal composition (Composition <NUM>) comprising an effective amount of a Compound <NUM> of Formula I, wherein Compound <NUM> is a Compound of Formula Ib (Compound 1b, or any of Compounds <NUM>-<NUM>, in free or salt form, in admixture with a pharmaceutically acceptable or insecticidally acceptable diluent or carrier.

In further embodiments of the second aspect, the present disclosure provides:.

In a third aspect, the present disclosure provides a method (Method <NUM>) for the treatment, control, suppression, or eradication, of an infection or infestation of an insect (arthropod), such as an adult insect, insect egg, insect larva, insect nymph, and/or insect pupa, comprising administering or applying an effective amount of a Compound <NUM> of Formula I, wherein Compound <NUM> is a Compound of Formula Ib (Compound 1b), or any of Compounds <NUM>-<NUM>, in free or salt form, or a pharmaceutical Composition <NUM> or any of <NUM>-<NUM>, to an animal or human patient in need thereof, or comprising applying a Compound of Formula I, or any of <NUM>-<NUM>, or an insecticidal Composition <NUM> or any of <NUM>-<NUM>, to a product or location in need thereof.

In further embodiments of the third aspect the present disclosure provides:.

In a fourth aspect, the present disclosure provides a Compound <NUM> of Formula I, wherein Compound <NUM> is a Compound of Formula Ib (Compound 1b), or any of Compounds <NUM>-<NUM>, or a pharmaceutical or insecticidal Composition <NUM>, or any of <NUM>-<NUM>, for use in the treatment, control, suppression, or eradication, of an infection or infestation of an insect (arthropod), such as an adult insect, insect egg, insect larva, insect nymph, and/or insect pupa, for example, according to Method <NUM> or any of <NUM>-<NUM>.

In a fourth aspect, the present disclosure provides the use of a Compound <NUM> of Formula I, wherein Compound <NUM> is a Compound of Formula Ib (Compound 1b), or any of Compounds <NUM>-<NUM>, or a pharmaceutical or insecticidal Composition <NUM>, or any of <NUM>-<NUM>, for the treatment, control, suppression, or eradication, of an infection or infestation of an insect (arthropod), such as an adult insect, insect egg, insect larva, insect nymph, and/or insect pupa, for example, according to Method <NUM> or any of <NUM>-<NUM>.

The compounds of Formula I, and the Compositions comprising said compounds, are expected to be of broad-spectrum insecticidal activity. The compounds and compositions are generally effective to kill a wide variety of arthropods, including, but not limited to, Chelicerates, including arachnids, such as ticks, mites, and spiders, Myriapods, such as millipedes, centipedes, and symphylans, and Pancrustaceans including woodlice (Class Malacostraca, Order Isopoda), insects (Class Insecta) and springtails (Class Collembola). Thus, it is understood that the term "insect" as used herein is not limited to arthropods of the Class Insecta (i.e., true insects), but rather, includes a variety of arthropods that are not phylogenetically classified as insects. Likewise, the terms "insecticide" and "insecticidal" as used herein are indicative of the property of a compound or composition being effective to kill true insects, as well as other arthropods, including Chelicerates such as arachnids (ticks, mites, spiders), Myriapods and non-insect Pancrustaceans such as woodlice and springtails.

In particularly preferred embodiments of the present disclosure, the compounds and compositions disclosed herein are useful as, and used for, the control of ticks and mites, e.g., arthropods of the superorders Parasitiformes (mites and ticks) and Acariformes (mites), which belong to the Class Arachnida, of the subphylum Chelicerata. In particular, within the superorder Parasitiformes, this includes arthropods of the Orders Ixodida (ticks), and Mesostigmata (mites), and within the superorder Acariformes, this includes arthropods (mites) of the Orders Sarcoptiformes and Trombidiformes.

Compositions according to the present disclosure may generally be in the form of solids, liquids, or semisolids, including powders, granules, aggregates, blocks, solutions, suspensions, dispersions, emulsions, mists, aerosols, fogs, foams, gels, creams, pastes, ointments, and gums. Liquid compositions include water miscible concentrates, emulsifiable concentrates, flowable suspensions, wettable or soluble powders, which may be used to treat substrates or sites infested or liable to infestation by insects (arthropods), including premises, outdoor or indoor storage or processing areas, containers or equipment and standing or running water.

Compounds and compositions according to the present disclosure may generally be administered to animals by incorporation in feed or suitable orally-ingestible pharmaceutical formulations, edible baits, salt licks, dietary supplements, pour-on formulations, sprays, baths, dips, showers, jets, dusts, greases, shampoos, creams, or wax-smears.

Compounds and compositions according to the present disclosure may generally be applied to the environment, either in general or to specific locations where pests may lurk, including stored products, timber, household goods, and domestic and industrial premises, as sprays, fogs, dusts, smokes, wax-smears, lacquers, granules and baits, as tricklefeeds to waterways, wells, reservoirs and other running or standing water.

Compounds and compositions according to the present disclosure may generally be applied to growing crops as foliar sprays, dusts, granules, fogs and foams, or as suspensions of finely divided and encapsulated compounds; as soil and root treatments by liquid drenches, dusts, granules, smokes and foams; and as seed dressings by liquid slurries and dusts.

Solid compositions, such as dusts, granules or wettable powders, are generally prepared by impregnating solid diluents with solutions of the compound of Formula I in volatile solvents, evaporating the solvents and, if necessary, grinding the products so as to obtain powders and, if desired, granulating or compacting the products so as to obtain granules, pellets or briquettes or by encapsulating finely divided active ingredient in natural or synthetic polymers, e.g., gelatin, synthetic resins and polyamides. The wetting, dispersing and emulsifying agents which may be present, particularly in wettable powders, may be of the ionic or non-ionic types, for example sulphoricinoleates, quaternary ammonium derivatives or products based upon condensates of ethylene oxide with nonyl- and octylphenol, or carboxylic acid esters of anhydrous sorbitols which have been rendered soluble by etherification of the free hydroxy groups by condensation with ethylene oxide, or mixtures of these types of agents. Wettable powders may be treated with water immediately before use to give suspensions ready for application.

Liquid compositions may incorporate natural or synthetic polymers, wetting, dispersing or emulsifying agents. Liquid compositions may be prepared using aqueous, organic or aqueous-organic diluents, for example acetophenone, isophorone, toluene, xylene, mineral, animal or vegetable oils, and water-soluble polymers (and mixtures of these diluents), which may contain wetting, dispersing or emulsifying agents of the ionic or non-ionic types or mixtures thereof.

Compositions according to the present disclosure may also contain synergists (e.g., piperonyl butoxide or sesamex), stabilizing substances, other insecticides, acaricides, plant nematocides, anthelmintics or anticoccidials, fungicides (agricultural or veterinary as appropriate e.g., benomyl, iprodione), bactericides, arthropod or vertebrate attractants or repellents or pheromones, deodorants, flavoring agents, dyes, taste masking agents, and auxiliary therapeutic agents, e.g., trace elements. These may be designed to improve potency, persistence, safety, uptake where desired, spectrum of pests controlled or to enable the composition to perform other useful functions in the same animal or area treated.

The compositions of the present disclosure may typically comprise from <NUM>% to <NUM>% by weight, or more particularly from <NUM>% to <NUM>% by weight, of the Compound or Compounds of Formula I and/or of total active ingredients (Compounds of Formula I and any additional insecticidal agents, synergists, trace elements or stabilizers). The actual compositions employed, and their rate of application will be selected to achieve the desired effect(s) by the farmer, livestock producer, medical or veterinary practitioner, pest control operator or other person skilled in the art.

Solid and liquid compositions for application topically to animals, timber, stored products or household goods usually contain from <NUM>% to <NUM>%, more particularly from <NUM>% to <NUM>%, by weight of one or more Compounds of Formula I. For administration to animals orally or parenterally, including percutaneously solid and liquid compositions normally contain from <NUM>% to <NUM>% by weight of one or more Compound of Formula I. Medicated feedstuffs normally contain from <NUM>% to <NUM>% by weight of one or more Compounds of Formula I. Concentrates and supplements for mixing with feedstuffs normally contain from <NUM>% to <NUM>%, and preferably from <NUM>% to <NUM>%, by weight of one or more Compound of Formula I. mineral salt licks normally contain from <NUM>% to <NUM>% by weight of one or more Compound of Formula I.

Dusts and liquid compositions for application to livestock, persons, goods, premises or outdoor areas may contain <NUM>% to <NUM>%, and more especially <NUM>% to <NUM>%, by weight of one or more Compound of Formula I. Suitable concentrations in treated waters are between <NUM> ppm and <NUM> ppm, and more especially <NUM> ppm to <NUM> ppm of one or more Compound of Formula I and may also be used therapeutically in fish farming with appropriate exposure times. Edible baits may contain from <NUM>% to <NUM>% and preferably <NUM>% to <NUM>%, by weight of one or more Compound of Formula I.

When administered to vertebrates parenterally, orally or by percutaneous or other means, the dosage of Compound of Formula I will depend upon the species, age and health of the vertebrate and upon the nature and degree of its actual or potential infestation by the pest. A single dose of <NUM> to <NUM>, preferably <NUM> to <NUM>, per kg body weight of the animal or doses of <NUM> to <NUM>, preferably <NUM> to <NUM>, per kg body weight of the animal per day for sustained medication are generally suitable by oral or parenteral administration. By use of sustained release formulations or devices, the daily doses required over a period of months may be combined and administered to animals on a single occasion.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. The terminology used in the description is for describing particular embodiments only and is not intended to be limiting of the invention.

Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise (such as in the case of a group containing a number of carbon atoms in which case each carbon atom number falling within the range is provided), between the upper and lower limit of that range and any other stated or intervening value in that stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges is also encompassed within the invention, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either both of those included limits are also included in the invention.

The following terms are used to describe the present disclosure. In instances where a term is not specifically defined herein, that term is given an art-recognized meaning by those of ordinary skill applying that term in context to its use in describing the present invention.

The articles "a" and "an" as used herein and in the claims are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article unless the context clearly indicates otherwise.

The phrase "and/or" as used herein and in the claims should be understood to mean "either or both" of the elements so conjoined, i.e., elements that are conjunctively present in some cases and disjunctively present in other cases. Thus, as a non-limiting example, a reference to "A and/or B," when used in conjunction with open-ended language such as "comprising" can refer, in one embodiment, to A only (optionally including elements other than B); in another embodiment, to B only (optionally including elements other than A); in yet another embodiment, to both A and B (optionally including other elements); etc..

As used herein in the specification and in the claims, the term "or" should be understood to have the same meaning as "and/or" as defined above.

The term "about" and the like, as used herein, in association with numeric values or ranges, reflects the fact that there is a certain level of variation that is recognized and tolerated in the art due to practical and/or theoretical limitations. For example, minor variation is tolerated due to inherent variances in the manner in which certain devices operate and/or measurements are taken. In accordance with the above, the term "about" is normally used to encompass values within the standard deviation or standard error.

In the claims, as well as in the specification, all transitional phrases such as "comprising," "including," "carrying," "having," "containing," "involving," "holding," "composed of," and the like are to be understood to be open-ended, i.e., to mean "including without limitation. " Only the transitional phrases "consisting of" and "consisting essentially of" shall be closed or semi-closed transitional phrases, respectively.

It should also be understood, that although various compounds, compositions, and methods are described in "open" terms of "comprising," "including," or "having" various components or steps (interpreted as meaning "including without limitation"), the compounds, compositions, methods, and devices can also "consist essentially of" or "consist of" the various components and steps, and such terminology should be interpreted as defining essentially closed-member groups. This paragraph is not meant in any way to limit the meaning of "comprising," "having," or "including" (and other verb forms thereof) which are to be interpreted as open-ended phrases meaning "including without limitation" consistent with patent law and custom. The intent of this paragraph is merely to indicate that the closed-member groups defined by the "consisting of" or "consisting essentially of" language are lesser included groups within the open-ended descriptions and to provide support for claims employing the "consisting of" or "consisting essentially of" language.

As used herein in the specification and in the claims, the phrase "at least one," in reference to a list of one or more elements, should be understood to mean at least one element selected from anyone or more of the elements in the list of elements, but not necessarily including at least one of each and every element specifically listed within the list of elements and not excluding any combinations of elements in the list of elements.

It should also be understood that, in certain methods described herein that include more than one step or act, the order of the steps or acts of the method is not necessarily limited to the order in which the steps or acts of the method are recited unless the context indicates otherwise.

The terms "co-administration" and "co-administering" or "combination therapy" can refer to both concurrent administration (administration of two or more therapeutic agents at the same time) and time varied administration (administration of one or more therapeutic agents at a time different from that of the administration of an additional therapeutic agent or agents), as long as the therapeutic agents are present in the patient to some extent, preferably at effective amounts, at the same time. In some embodiments, one or more of the present compounds described herein, are co-administered in combination with at least one additional bioactive agent, such as another anticancer agent. In some embodiments, the co-administration of compounds results in synergistic activity and/or therapy, including anticancer activity.

The term "effective" can mean, but is in no way limited to, that amount/dose of the active pharmaceutical or insecticidal ingredient, which, when used in the context of its intended use, effectuates or is sufficient to provide the intended outcome, e.g., the treatment, control, suppression, or eradication, of an infection or infestation of an insect (arthropod), such as an adult insect, insect egg, insect larva, insect nymph, and/or insect pupa.

The effective amount depends on the type and severity of disease or infestation, the composition used, the route of administration or application, the type of animal or location being treated, concurrent treatment with or application of other insecticidal agents, and other factors which those skilled in the medical arts will recognize.

The term "pharmaceutically acceptable" can mean, but is in no way limited to, entities and compositions that do not produce an adverse, allergic or other untoward reaction when administered to a patient or subject.

The term "pharmaceutically acceptable carrier" can mean, but is in no way limited to, any and all solvents, excipients, coatings, and the like, compatible with pharmaceutical administration to a patient or subject. Such ingredients include diluents, binders, lubricants, dispersants, surfactants, tonicity agents, stabilizer, solubilizing agents, gelling agents, complexing agents, antioxidants, buffers, flavors, colors, and coatings. Suitable solvents and excipients include, but are not limited to water, ethanol, dimethylsulfoxide, polyethylene glycol, polypropylene glycol, poloxamers (ethylene oxide/propylene oxide block copolymers), mannitol, sorbitol, glycerin, carboxymethyl cellulose, sodium carboxymethyl cellulose, hydroxypropylmethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, methylcellulose, ethylcellulose, cellulose acetate, polyvinyl alcohol, polyvinyl acetate, polyvinyl acetate-polyvinyl alcohol copolymer, polyvinylpyrrolidone, polyacrylate, polymethyl acrylate, polymethyl methacrylate, xanthan gum, gum arabic, guar gum, karaya gum, agar, alginic acid, sodium alginate, carrageenan, gum tragacanth, locust bean gum, glucan, gellan gum, sodium lauryl sulfate, sodium laureth sulfate, cocamidopropyl betaine, polyoxyethylene sorbitan esters (e.g., polysorbate <NUM>, polysorbate <NUM>, polysorbate <NUM>, polysorbate <NUM>, polysorbate <NUM>), dimethicone, diethyl phthalate, stearic acid, cetyl alcohol, magnesium stearate, magnesium aluminum silicate, silicon dioxide, starch (e.g., pregelatinized starch), sodium starch glycolate, talc, triethyl citrate, glyceryl triacetate, dextrose, maltose, lactose, trehalose, calcium carbonate, dicalcium phosphate, sodium chloride, potassium chloride, sodium hydroxide, sodium carbonate, sodium bicarbonate, sodium citrate, sodium phosphate, sodium acetate, hydrochloric acid, acetic acid, citric acid, tartaric acid, phosphoric acid, propyl gallate, sodium metabisulfite, butylated hydroxyanisole, butylated hydroxytoluene, and alpha-tocopherol.

The term "insecticidally acceptable" can mean, but is in no way limited to, entities and compositions that do not produce an unacceptable adverse outcome when applied to a product or location, such as an unacceptably adverse hazardous or environmental outcome.

The term "insecticidally acceptable carrier" can mean, but is in no way limited to, any and all solvents, excipients, coatings, and the like, compatible with insecticidal administration to a product or location. Suitable carriers and excipients include all of the pharmaceutically acceptable and suitable carriers and excipients listed in the preceding paragraphs, and in addition: methanol, isopropanol, <NUM>-butoxyethanol, ethylene glycol, hexylene glycol, <NUM>,<NUM>-propanediol, <NUM>,<NUM>-propanediol, diethylene glycol, diethanolamine, aluminum oxide, aluminum oxide silicate, trisodium sulfosuccinate, cobalt propionate, <NUM>-ethylhexyl octadecenoate, ethylene oxide/oleic acid monoethanolamide, butyl <NUM>-propenoate polymers and copolymers, sodium dodecylnaphthalene sulfonate, decamethyl cyclopentasiloxane, cyclodextrins, sodium silicate, ammonium dodecanoate, sodium benzoate, boric acid, sodium borate, and mica.

The term "systemic administration" refers to a route of administration that is, e.g., enteral or parenteral, and results in the systemic distribution of an agent leading to systemic absorption or accumulation of drugs in the blood stream followed by distribution throughout the entire body. Suitable forms, in part, depend upon the use or the route of entry, for example oral (enteral) administration or parenteral (e.g., intravenous injection). Such forms should not prevent the composition or formulation from reaching a target cell (i.e., a cell to which the negatively charged polymer is desired to be delivered to). For example, pharmacological compositions injected into the blood stream should be soluble. Other factors are known in the art, and include considerations such as toxicity and forms which prevent the composition or formulation from exerting its effect.

The terms "patient" or "subject" may describe a cell, tissue, or animal, preferably a mammal, e.g., a human, livestock animal, or companion animal, to whom treatment, including prophylactic treatment, with the compositions according to the present disclosure is provided. For treatment of those infections, conditions or disease states which are specific for a specific animal such as a human patient, the term patient refers to that specific animal, including a domesticated animal such as a dog or cat or a farm animal such as a horse, cow, sheep, etc..

The term "compound," as used herein, unless otherwise indicated, refers to any specific chemical compound disclosed herein and includes tautomers, regioisomers, geometric isomers, and where applicable, stereoisomers, including optical isomers (enantiomers) and other stereoisomers (diastereomers) thereof, atropisomers, as well as salts, hydrates, solvates, polymorphs, and derivatives thereof where applicable, in context. Such isomers include, in the context of double and triple bonds and ring systems, both cis- and trans- isomers. Within its use in context, the term "compound" generally refers to a single compound, but also may include other compounds such as stereoisomers, regioisomers and/or optical isomers (including racemic mixtures) as well as specific enantiomers or enantiomerically enriched mixtures of disclosed compounds. The term also refers to any specific chemical compound in which one or more atoms have been replaced with one or more different isotopes of the same element. It is noted that in describing the present compounds, numerous substituents and variables associated with same, among others, are described.

"Tautomers" refer to compounds that are interchangeable forms of a particular compound structure, and that vary in the displacement of hydrogen atoms and electrons. Thus, two structures may be in equilibrium through the movement of pi-electrons and an atom (usually H). For example, enols and ketones are tautomers because they are rapidly interconverted by treatment with either acid or base. Another example of tautomerism is the aci- and nitro-forms of phenyl nitromethane, that are likewise formed by treatment with acid or base. Tautomeric forms may be relevant to the attainment of the optimal chemical reactivity and biological activity of a compound of interest.

It is understood by those of ordinary skill that molecules which are described herein are stable compounds as generally described hereunder. When the bond is shown, both a double bond and single bond are represented or understood within the context of the compound shown and well-known rules for valence interactions.

As used herein, "derivatives" can mean compositions formed from the native compounds either directly, by modification, or by partial substitution. As used herein, "analogs" can mean compositions that have a structure similar to, but not identical to, the native compound.

Amino refers to the group -NH<NUM>; hydroxy refers to the group "-OH"; cyano refers to the -CN group; nitro refers to the group -NO<NUM>.

"Halo" or "halogen" refers to fluoro (F), chloro (Cl), bromo (Br) or iodo (I).

"Alkyl" as used herein means an acyclic linear or branched fully saturated hydrocarbon moiety, preferably having one to six carbon atoms, or in some embodiments, one to four carbon atoms, or one to three carbon atoms. Exemplary alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, isobutyl, t-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, or any other acyclic hydrocarbon group having the general formula -CnH2n+<NUM>.

"Alkenyl" as used herein means an acyclic linear or branched unsaturated hydrocarbon moiety having at least one carbon-carbon double bond, preferably having two to six carbon atoms, or in some embodiments, two to four carbon atoms, or two to three carbon atoms. Preferably, said group is mono-unsaturated (having a single carbon-carbon double bond). Examples of alkenyl groups include, but are not limited to, vinyl and allyl, or any other hydrocarbon acyclic group having the general formula -CnH2n-<NUM>.

"Alkynyl" as used herein means an acyclic linear or branched unsaturated hydrocarbon moiety having at least one carbon-carbon triple bond, preferably having two to six carbon atoms, or in some embodiments, two to four carbon atoms, or two to three carbon atoms. Preferably, said group is mono-unsaturated (having a single carbon-carbon triple bond). Examples of alkynyl groups include, but are not limited to, ethynyl and propargyl, or any other hydrocarbon acyclic group having the general formula -CnH2n-<NUM>. As used herein, a hydrocarbon radical having both a double bond and a triple bond is considered an alkynyl radicals.

"Alkylene" as used herein means a -(CH<NUM>)n- group, e.g., wherein n is an integer generally from <NUM>-<NUM>, such as <NUM>-<NUM>.

"C<NUM>-<NUM> haloalkyl" as used herein means an alkyl group, as defined above, having from one to six carbon atoms, and substituted by at least on halogen atom (e.g., F, Cl, Br, or I), and up to the maximum number of halogen atoms permitted by the valence of the carbon atoms of the alkyl group. C<NUM>-<NUM> haloalkyl includes, for example, fluoromethyl, difluoromethyl, trifluoromethyl, <NUM>-fluoroethyl, <NUM>,<NUM>-difluoroethyl, <NUM>,<NUM>,<NUM>-trifluoroethyl.

"Alkoxy" as used herein means a group of the structure -O-(alkyl). Thus, for example, a C<NUM>-<NUM> alkoxy group is a group having the structure -O-(C<NUM>-<NUM> alkyl). An alkoxy radical attaches to the structure to which it is appended by its oxygen atom.

"Cycloalkyl" as used herein means a nonaromatic saturated or unsaturated free radical forming at least one ring, having, for example, <NUM> to <NUM> carbon atoms and a corresponding number of hydrogen atoms. The term "cycloalkyl" therefore includes carbocyclic rings having one or more double or triple bonds. Cycloalkyl groups can be monocyclic or polycyclic. Individual rings of such polycyclic cycloalkyl groups can have different connectivities, e.g., fused, bridged, spiro, etc., in addition to covalent bond substitution. Exemplary cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, norbornanyl, bicyclo[<NUM>. <NUM> ]octanyl, octahydro-pentalenyl, spiro[<NUM>]decanyl, cyclopropyl, adamantyl, etc. Of course, other cycloalkyl groups will be readily apparent to those of skill in the art given the benefit of the present disclosure.

"Cycloalkoxy" as used herein means a group of the structure -O-(cycloalkyl). Thus, for example, a C<NUM>-<NUM> cycloalkoxy group is a group having the structure -O-(C<NUM>-<NUM> cycloalkyl). A cycloalkoxy radical attaches to the structure to which it is appended by its oxygen atom.

"Halocycloalkyl" as used herein means a cycloalkyl group, as defined above, substituted by at least on halogen atom (e.g., F, Cl, Br, or I), and up to the maximum number of halogen atoms permitted by the valence of the carbon atoms of the alkyl group. Halocycloalkyl groups include, for example, <NUM>-fluorocyclopropyl, <NUM>-fluorocyclopropyl, <NUM>,<NUM>-difluorocyclopropyl, <NUM>,<NUM>-difluorocyclobutyl, <NUM>,<NUM>-difluorocyclopentyl, and the like.

"Heterocycloalkyl" as used herein means any cyclic nonaromatic ring system comprising at least one heteroatom (e.g., N, S, or O) ring atom. This includes <NUM>- to <NUM>-membered monocyclic and fused bicyclic ring systems, and any larger multi-ring fused ring systems, as long such ring systems do not comprise any aromatic carbocyclic or aromatic heterocyclic ring. Exemplary heterocycloalkyl groups include pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydropyranyl, pyranyl, thiopyranyl, azindinyl, azetidinyl, oxiranyl, methylenedioxyl, chromenyl, barbituryl, isoxazolidinyl, <NUM>,<NUM>-oxazolidin-<NUM>-yl, isothiazolidinyl, <NUM>,<NUM>-thiazolidin-<NUM>-yl, <NUM>,<NUM>-pyrazolidin-<NUM>-yl, <NUM>,<NUM>-pyrazolidin-<NUM>-yl, piperidinyl, thiomorpholinyl, <NUM>,<NUM>-tetrahydrothiazin-<NUM>-yl, <NUM>,<NUM>-tetrahydrothiazin-<NUM>-yl, tetrahydrothiadiazinyl, morpholinyl, <NUM>,<NUM>-tetrahydrodiazin-<NUM>-yl, <NUM>,<NUM>-tetrahydrodiazin-<NUM>-yl, tetrahydroazepinyl, piperazinyl, piperizin-<NUM>-onyl, piperizin-<NUM>-onyl, chromanyl, <NUM>-pyrrolinyl, <NUM>-pyrrolinyl, imidazolidinyl, <NUM>-imidazolidinyl, <NUM>,<NUM>-dioxanyl, <NUM>-azabicyclo[<NUM>. <NUM>]octanyl, <NUM>-azabicyclo[<NUM>. <NUM>]octanyl, <NUM>,<NUM>-diazabicyclo[<NUM>. <NUM>]octanyl, <NUM>,<NUM>-diazabicyclo[<NUM>. <NUM>]heptanyl, <NUM>,<NUM>-diazabicyclo[<NUM>. <NUM>]octanyl, octahydro-<NUM>-pyrido[<NUM> ,<NUM>-a]pyrazinyl, <NUM>-azabicyclo[<NUM>. <NUM>] heptanyl, <NUM>-azabicyclo[<NUM>. <NUM>]hexanyl, <NUM>-azaspiro[<NUM>]nonanyl, <NUM>-oxa-<NUM>-aza-spiro[<NUM>] nonanyl, <NUM>-azabicyclo[<NUM>. <NUM>]heptanyl, octahydro-<NUM>-indolyl, etc. In general, the heterocycloalkyl group typically is attached to the main structure via a carbon atom or a nitrogen atom. Of course, other heterocycloalkyl groups will be readily apparent to those of skill in the art given the benefit of the present disclosure.

"Aryl" as used herein means any carbocyclic aromatic ring system, i.e., any aromatic ring system comprising only carbon atoms as ring atoms. This includes <NUM>-membered monocyclic aryl ring systems and <NUM>-membered or <NUM>-membered fused bicyclic aryl ring systems, and larger fused ring systems, as long such ring systems comprise at least one <NUM>-membered aromatic carbocyclic ring (i.e., a benzene ring) within the fused ring system, and as long as no ring-atoms are heteroatoms. Aryl includes, but is not limited to, phenyl, naphthyl, phenanthryl, and anthracenyl.

"Heteroaryl" as used herein means any cyclic heteroaromatic ring system, i.e., any aromatic ring system comprising at least one heteroatom (e.g., N, S, or O) ring atom. This includes <NUM>-membered and <NUM>-membered monocyclic heteroaryl ring systems and <NUM>-membered or <NUM>-membered fused bicyclic heteroaryl ring systems, and larger fused ring systems, as long such ring systems comprise at least one aromatic carbocyclic or aromatic heterocyclic ring within the fused ring system and at least one heteroatom (e.g., N, S or O) ring-atom within the fused ring system (either in an aromatic ring or non-aromatic ring). Heteroaryl therefore includes, but is not limited to, bicyclic fused ring systems selected from aromatic-heteroaromatic, aromatic-heterocyclic, heteroaromatic-carbocyclic, heterocyclic-aromatic, and heteroaromatic-heteroaromatic, as well as larger fused ring systems comprising some combination of benzene, cycloalkane, heterocycloalkane and heteroaromatic rings. Exemplary heteroaryl groups include furyl, thienyl, thiazolyl, pyrazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyrrolyl, triazolyl, tetrazolyl, imidazolyl, <NUM>,<NUM>,<NUM>-oxadiazolyl, <NUM>,<NUM>,<NUM>-oxadiazolyl, <NUM>,<NUM>,<NUM>-oxadiazolyl, <NUM>,<NUM>,<NUM>-thiadiazolyl, <NUM>,<NUM>,<NUM>-thiadiazolyl, <NUM>,<NUM>,<NUM>-thiadiazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, pyridonyl, <NUM>,<NUM>,<NUM>-triazinyl, <NUM>,<NUM>,<NUM>-triazinyl, <NUM>,<NUM>,<NUM>-triazinyl, pyrazolo[<NUM>,<NUM>-b]pyridinyl, cinnolinyl, pteridinyl, purinyl, <NUM>,<NUM>-dihydro-SH-[<NUM>]pyridinyl, benzo[b]thiophenyl, <NUM>,<NUM>,<NUM>,<NUM>-tetrahydro-quinolin-<NUM>-yl, benzoxazolyl, benzofuranyl, isobenzofuranyl, benzothiazolyl, benzisothiazolyl, benzisoxazolyl, benzimidazolyl, benzothiadiazolyl, thianaphthenyl, isothianaphthenyl, benzofuranyl, isobenzofuranyl, isoindolyl, indolyl, indolizinyl, indazolyl, isoquinolinyl, quinolinyl, phthalazinyl, quinoxalinyl, quinazolinyl, naphthyridyl, phenanthridinyl, acridinyl, carbazolyl, carbazolinyl, permidinyl, phenanthrolinyl, phenacenyl, pyrrolopyrimidinyl, pyrrolopyridinyl, pyridopyrimidinyl, theinopyrimidinyl, furopyrimidinyl, furopyridyl, furopyrrolyl, pyrazoloxazolyl, thienofuranyl, imidazothiazolyl, imidazopyridyl, imidazotriazyl, imidazopyrimidinyl, pyrazinopyridazinyl, phenothiazinyl, furazanyl, phenoxazinyl, pyrazo benzoxazinyl, azaindolizinyl, dihydroquinolinyl, dihydroisoquinolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and pyridoxazinyl. It is understood that for heteroaryl systems in which the both ring carbon atoms and ring heteroatoms have open valencies, bonds can be formed to either such atom types (e.g., C-linked or N-linked). For example, where a pyrazolyl moiety is a substituent, and further substituted with other groups, either the core structure tow which the pyrazolyl is attached, and any one or more other substituents attached to the pyrazolyl, may be attached through a pyrazole ring nitrogen atom (N-linked) or a pyrazole ring carbon atom (C-linked).

As used herein, the term "unsubstituted" means that the indicated structure or group is further substituted only with hydrogen atoms.

It is understood that when describing the substituents attached in various positions to the core structure of Formula I, in some cases, the substituent may be referred to using the name of the corresponding chemical compound, especially in the case of rings, whereas in some cases the same substituent may be referred to using the name of the corresponding chemical radical (e.g., having an "-yl" suffix), but these terms are interchangeable. For example, when referring to the substituent Ar, or a heteroaryl ring R attached to the substituent Ar, the terms "pyridine" and "pyridyl" are equivalent, as are the terms "morpholine" and "morpholinyl. " The skilled artisan will recognize that such terms are used to denote attachment of, for example, pyridine or morpholine ring at the designated position, thus converting said ring to a pyridyl or morpholinyl substituent respectively. Absent an indication otherwise, such attachments may be made at any chemically permissible location of the attached ring.

As used herein, a range of carbon atoms which includes C<NUM> means that carbon is absent and is replaced with H (or deuterium). Thus, a range of carbon atoms which is C<NUM>-C<NUM> includes carbons atoms of <NUM>, <NUM>, <NUM>, <NUM>, <NUM> and <NUM> and for C<NUM>, H (or deuterium) stands in place of carbon.

The Compounds according to the first aspect may exist as tautomers, for example amide (NHC=O) ↔ hydroxyimine (N=COH) tautomers.

Exemplary non-limiting embodiments of the present disclosure are further described by reference to the following examples.

Unless otherwise noted, all starting materials and solvents were obtained from commercial sources (e.g., Acros Organics, Merck, Alfa Aesar, and TCI) and were used without further purification. All reactions were routinely monitored by thin-layer chromatography (TLC) performed on silica gel <NUM> F<NUM> (layer <NUM>) pre-coated aluminum foil (with fluorescent indicator UV254) (Sigma-Aldrich). Developed plates were air-dried and visualized under UV light (<NUM>/<NUM>) or by using KMnO<NUM> solutions. Automated flash column chromatography was performed on pre-packed silica (<NUM>Å, <NUM>) cartridges using Biotage® Selekt.

<NUM>H NMR and <NUM>C NMR spectra were recorded at <NUM> at <NUM> and <NUM>, respectively, using a Bruker Avance <NUM> spectrometer by using residual solvent peak as internal standard. Chemical shifts are reported in ppm (δ) and the coupling constants (J) are given in Hertz (Hz). Peak multiplicities are abbreviated as follow: s (singlet), bs (broad singlet), d (doublet), dd (double doublet), t (triplet), dt (double triplet), q (quartet), p (pentet), and m (multiplet). Data were acquired using Bruker TopSpin software and processed using MestreNova software.

High-Resolution Mass Spectroscopy (HRMS) spectra were registered on Agilent Technologies <NUM> UHD Accurate Mass Q-TOF LC-MS system or on Agilent <NUM> Infinity Series U-HPLC system (Agilent Technologies, Santa Clara, CA, USA) coupled with a Q TOF <NUM> high-resolution mass spectrometer and <NUM> Infinity Series DAD/UV-Vis detector (Agilent Technologies). The purity of the final compound was assessed as ><NUM>%, using UPLC-MS. The analyses were carried out according to the method listed below. The mobile phase was a mixture of water (solvent A) and acetonitrile (solvent B), both containing formic acid at <NUM>%. Method: Phenomenex Luna Omega <NUM> Polar (C18, <NUM> x <NUM>) column at <NUM>° C using a flow rate of <NUM>/min in a <NUM> gradient elution. Gradient elution was as follows: <NUM>:<NUM> (A/B) to <NUM>:<NUM> (A/B) over <NUM>, <NUM>:<NUM> (A/B) for <NUM>, and then reversion back to <NUM>:<NUM> (A/B) over <NUM>. The UV detection is an averaged signal from wavelength of <NUM> to <NUM> and mass spectra are recorded on a mass spectrometer using positive mode electro spray ionization.

Compounds described herein may be synthesized as described herein, using modified methods described herein or by methods known to a person of skill in the art.

Chemistry abbreviations: ACN, acetonitrile; Boc, tert-butoxycarbonyl; CD3OD, deuterated methanol; CDCl3, deuterated chloroform; DCE, dichloroethane; DCM, dichloromethane; DEE, diethyl ether; DIPEA, N,N'-diisopropylethylamine; DMA, dimethylacetamide; DMF, dimethylformamide; DMSO, dimethylsulfoxide; DMSO-d<NUM>, deuterated dimethylsulfoxide; EA, ethyl acetate; h, hour; EtOH, absolute ethanol; Et3N, triethylamine; HATU, <NUM>-(<NUM>-Aza-<NUM>-benzotriazole-<NUM>-yl)-<NUM>,<NUM>,<NUM>,<NUM>-tetramethyluronium hexafluorophosphate; min, minutes; HRMS, high-resolution mass spectroscopy; MeOH, methanol; NMR, nuclear magnetic resonance; tBu, tert-butyl; tBuOH, tert-butanol; TLC, thin-layer chromatography; PE, petroleum ether; rt, room temperature.

Compounds of general formula (I) may be prepared by the following general synthetic approaches described below, wherein according to the first aspect Ar is phenyl substituted by <NUM>-<NUM> R groups as described above:.

<CHM>
<CHM>
<CHM>
See, e.g., <NPL>); <NPL>); <NPL>); <NPL>); <NPL>); <NPL>); <NPL>); <NPL>);
<CHM>
See, e.g., <CIT>, <NPL>); <NPL>); <CIT>;
<CHM>
See, e.g., <NPL>); <NPL>); <NPL>); <NPL>).

<NUM>-amino-<NUM>-(<NUM>,<NUM>-dichloro-<NUM>-(trifluoromethyl)phenyl)-<NUM>H-pyrazole-<NUM>-carbonitrile (commercially available) (<NUM>, <NUM> mmol) and ethyl <NUM>,<NUM>,<NUM>-trifluoro-<NUM>-oxobutanoate (commercially available) (<NUM>, <NUM>µL, <NUM> mmol) are stirred in a pre-warmed pressure tube at <NUM> for <NUM>. After cooling, the crude product is purified by automated flash chromatography on silica gel (<NUM> cartridge, gradient petroleum ether/ethyl acetate <NUM>:<NUM> to <NUM>:<NUM>) to afford the title compound (<NUM>, <NUM>% yield) as white solid. <NUM>H NMR (<NUM>, DMSO-d<NUM>) δ <NUM> (bs, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>); <NUM>C NMR (<NUM>, DMSO-d<NUM>) δ <NUM>-<NUM> (m), <NUM>, <NUM> (2C), <NUM>, <NUM> (q, J = <NUM>), <NUM> (q, J = <NUM>), <NUM>-<NUM> (m), <NUM> (q, J = <NUM>), <NUM> (q, J = <NUM>), <NUM>, <NUM>, <NUM>, <NUM>; <NUM>F NMR (<NUM>, DMSO-d<NUM>) δ -<NUM>, -<NUM>. HRMS (ESI) m/z [M+Na]+ calcd for C<NUM>H<NUM>Cl<NUM>F<NUM>N<NUM>O <NUM>, found <NUM>.

To a solution of <NUM>-amino-<NUM>-(<NUM>,<NUM>-dichloro-<NUM>-(trifluoromethyl)phenyl)-<NUM>H-pyrazole-<NUM>-carbonitrile (commercially available) (<NUM>, <NUM> mmol) in acetic acid (<NUM>), is added ethyl <NUM>,<NUM>,<NUM>-trifluoro-<NUM>-oxobutanoate (commercially available) (<NUM>, <NUM>µL, <NUM> mmol) and the mixture is stirred at <NUM> for <NUM>. After cooling, the solvent is evaporated and the crude product is purified by automated flash chromatography on silica gel (<NUM> cartridge, gradient petroleum ether/ethyl acetate <NUM>:<NUM>) to afford the title compound (<NUM>, <NUM>% yield) as white solid. <NUM>H NMR (<NUM>, DMSO-d<NUM>) δ <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>); <NUM>C NMR (<NUM>, DMSO-d<NUM>) δ <NUM>, <NUM>, <NUM> (2C), <NUM>, <NUM> (q, J = <NUM>), <NUM> (q, J = <NUM>), <NUM>, <NUM> (q, J = <NUM>), <NUM>, <NUM>, <NUM>; <NUM>F NMR (<NUM>, DMSO-d<NUM>) δ -<NUM>. HRMS (ESI) m/z [M+Na]+ calcd for C<NUM>H<NUM>Cl<NUM>F<NUM>N<NUM>O <NUM>, found <NUM>.

To a solution of <NUM>-amino-<NUM>-(<NUM>,<NUM>-dichloro-<NUM>-(trifluoromethyl)phenyl)-<NUM>-pyrazole-<NUM>-carbonitrile (commercially available) (<NUM>, <NUM> mmol) in propionic acid (<NUM>), is added ethyl <NUM>,<NUM>,<NUM>-trifluoro-<NUM>-oxobutanoate (commercially available) (<NUM>, <NUM>µL, <NUM> mmol) and the mixture is stirred at <NUM> for <NUM>. After cooling, the solvent is evaporated and the crude product is purified by automated flash chromatography on silica gel (<NUM> cartridge, gradient petroleum ether/ethyl acetate <NUM>:<NUM>) to afford Example <NUM> (<NUM>, <NUM>% yield) as white solid. <NUM>H NMR (<NUM>, DMSO-d<NUM>) δ <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (q, J = <NUM>, <NUM>), <NUM> (t, J = <NUM>, <NUM>); <NUM>C NMR (<NUM>, DMSO-d<NUM>) δ <NUM>, <NUM>, <NUM>, <NUM> (2C), <NUM> (q, J = <NUM>), <NUM>-<NUM> (m), <NUM>, <NUM> (q, J = <NUM>), <NUM>, <NUM>, <NUM>, <NUM>; <NUM>F NMR (<NUM>, DMSO-d<NUM>) δ - <NUM>. HRMS (ESI) m/z [M+H]+ calcd for C14H9Cl2F3N4O <NUM>, found <NUM>.

To explore the therapeutic potential of the compounds of the present disclosure, live chickens with mite infestation (Dermanyssus gallinae) are treated topically with solutions of test compound, with Fipronil as a positive control. Test solutions are prepared by dissolving the test compound in a mixture of <NUM>-butoxyethanol, ethanol, and water (<NUM>:<NUM>:<NUM>) to reach a final concentration of <NUM>% (w/v).

The test solution is administered using a mechanical spray pump delivering <NUM> of solution per actuation. The test solution is sprayed over the entire surface of the animal's body from a distance of <NUM>-<NUM> centimeters, resulting in direct contact of the solution with the animal's skin. The dosage applied to each animal is <NUM>/kg, equivalent to <NUM>/kg (kg live weight). The dosage is achieved using six actuations of the spray per kg of body weight per day. The treatment is administered once daily for <NUM> days at <NUM> hours intervals. One animal is left untreated as a negative control.

Blood samples are taken from each animal before the first treatment (T=<NUM>), and at <NUM> hour intervals thereafter. Each egg produced after a treatment is collected for later analysis.

Plasma samples are derived from each blood sample using standard procedures immediately after blood collection. Plasma is stored frozen at - <NUM> until the time of analysis. Samples are then thawed at room temperature and extraction is performed by solid phase extraction (SPE) (Strata-X 33µ Polymeric Reversed Phase, <NUM>/<NUM>, Phenomenex) on an aliquot of sample (<NUM>µl) pretreated for protein precipitation with <NUM>µL of acetonitrile. After protein precipitation, the samples are well vortexed and then centrifuged (<NUM> rpm, <NUM>, RT). Supernatant is collected and transferred to clean Falcon tubes and diluted with <NUM> of water. Strata-X columns are conditioned with <NUM> of methanol and with <NUM> of water. Samples are loaded after conditioning, and the columns are washed with <NUM> of water, and analytes are eluted with <NUM> of methanol, dried under nitrogen and then resuspended in <NUM>µL of acetonitrile for mass spectral analysis. The recovery of the method is calculated by adding standards to not-treated plasma and it is found to be <NUM>%; calibration curves were made for both compounds (<NUM>-<NUM>).

A Thermo Q-exactive mass spectrometer (Thermo Fisher Scientific, Waltham, MA) is used. The LC system, governed by Chromeleon X-press software, consists of a binary pump, a thermostated autosampler, and a column compartment, all Dionex Ultimate <NUM> series modules (Thermo Fisher Scientific, Waltham, MA). A volume of <NUM>µL is injected for each sample. Chromatographic separation of analytes is conducted in reverse-phase chromatography. A Luna Omega <NUM> Polar (C18, <NUM> × <NUM>) is used, and the mobile phases are (A) water and (B) acetonitrile, both containing <NUM>% v/v formic acid. The flow rate is <NUM>/min over a <NUM> minute gradient elution as follows: <NUM>:<NUM> (A/B) to <NUM>:<NUM> (A/B) over <NUM> minutes, <NUM>:<NUM> (A/B) for <NUM> minutes, and then reversion back to <NUM>:<NUM> (A/B) over <NUM> minutes. The column is operated at a constant temperature of <NUM>. The LC effluents are introduced into the Q-Exactive mass spectrometer by an H-ESI source operating in the negative mode with a sheath gas flow rate of <NUM>; an auxiliary gas flow rate of <NUM>; a spray voltage of <NUM> kV; capillary temperature and auxiliary gas heater temperature, respectively, of <NUM> and <NUM>; and S-lens RF level <NUM>. The Q-Exactive mass spectrometer is operated with a resolution of <NUM> in FullMS mode. Data analysis is performed using Qual Browser Thermo Xalibur <NUM>. <NUM> (Thermo Fisher Scientific) and Lipostar <NUM> (Molecular Discovery ltd) software.

The following peaks are observed and quantified: Fipronil, RT <NUM> ([M-H]- ion <NUM>/z); Fipronil sulfone, RT <NUM> ([M-H]- ion <NUM>/z); Fipronil amide, RT <NUM> ([M-H]- ion <NUM>/z); Compound of Example <NUM>, RT <NUM> ([M-H]- ion <NUM>/z). No metabolites of the Compound of Example <NUM> are observed.

Plasma from four chickens is analyzed: two chickens treated with Fipronil and two treated with the Compound of Example <NUM>. Plasma samples from Day <NUM> and Day <NUM> are analyzed. The day <NUM> plasma samples from Chicken <NUM> were found to be unusable due to inhomogeneity. The results are shown in the following tables:.

The results demonstrate that the Compound of Example <NUM> undergoes substantially less systemic absorption and plasma accumulation compared to Fipronil.

A modified and optimized QuEChERS protocol was employed to extract the compounds from eggs. The shell is manually removed, and then each egg is transferred to a clean beaker and homogenized with an electric mixer for small volumes. <NUM> grams of homogenized egg is put into a <NUM> Falcon tube, and <NUM> of water is added and the samples are shaken for <NUM> minutes. Next, <NUM> of acetonitrile is added and the samples are shaken again for <NUM> minutes. Then the roQ salts (<NUM> MgSO4, <NUM> NaCl, <NUM> SCTD and <NUM> SCDS, Phenomenex P/N KS08909) are added to each sample, and the mixture is well shaken for <NUM> minutes. Samples are then centrifuged at <NUM> for <NUM> minutes at room temperature, and the supernatant is transferred to an HPLC vial for analysis. The same HPLC-MS method is used as described above.

One egg from two chickens is analyzed: one chicken treated with Fipronil and one chicken treated with the Compound of Example <NUM>. The eggs are both from Day <NUM>. The results are shown in the following tables expressed as relative LC-MS peak area:.

The results demonstrate that the Compound of Example <NUM> undergoes substantially less accumulation in the eggs of treated chickens compared to Fipronil.

To test the effectiveness of the chemical compounds, live chickens with natural mite infestation (Dermanyssus gallinae) are treated topically with solutions of test compound, with Fipronil as a positive control. Test solutions are prepared by dissolving the test compound in a mixture of <NUM>-butoxyethanol, ethanol, and water (<NUM>:<NUM>:<NUM>) to reach a final concentration of <NUM>% (w/v). The test solutions are sprayed on the animals from the bottom up, to facilitate their application and skin penetration, using a mechanical spray pump delivering <NUM> of solution per actuation. The dosage applied to each animal is <NUM>/kg, equivalent to <NUM>/kg (kg live weight). The dosage is achieved using six actuations of the spray per kg of body weight per day. The treatment is administered once daily for <NUM> days at <NUM> hours intervals.

Two animals are treated with the compound of Example <NUM>, and two animals are treated with Fipronil, while one animal is left untreated as a negative control. The infestation by D. gallinae is monitored by checking the density of mites in particular areas of the animal's bodies (feet, legs, and wings), comparing pre-treatment levels of infestation to post-treatment levels.

When all pests have been eliminated, a <NUM>% pest control capability was achieved. In the case of some chemical products, when the pest were not fully eliminated, the disinfestation capacity was measured by averaging the number of infesting organisms located in three distinct areas of the treated animals, at the beginning of the treatment, and at the end of the treatment.

By day <NUM> of treatment, all chickens treated with either the Compound of Example <NUM> or Fipronil were <NUM>% free of mites. Untreated chickens (negative control) retained substantial amounts of mites similar to the initial amounts.

The experiments are repeated using the compounds of Example <NUM> and <NUM> as comparative examples. After five days of treatment, levels of mite infestations are reduced by only about <NUM>-<NUM>%, which could be explained by non-specific solvent effects on the mites.

Claim 1:
A compound of the following general structure,
<CHM>
wherein:
R<NUM> and R<NUM> are each independently selected from H, halo, cyano, nitro, hydroxy, amino, C<NUM>-<NUM> alkyl, C<NUM>-<NUM> cycloalkyl, C<NUM>-<NUM> haloalkyl, C<NUM>-<NUM> alkoxy, C<NUM>-<NUM> haloalkoxy,
C<NUM>-<NUM> cycloalkoxy, C<NUM>-<NUM> halocycloalkyl, NH(C<NUM>-<NUM> alkyl), N(C<NUM>-<NUM> alkyl)(C<NUM>-<NUM> alkyl), and <NUM>-<NUM> membered heterocycloalkyl; and
each R is independently selected from halo, cyano, nitro, hydroxy, amino, C<NUM>-<NUM> alkyl, C<NUM>-<NUM> cycloalkyl, C<NUM>-<NUM> haloalkyl, C<NUM>-<NUM> alkoxy, C<NUM>-<NUM> haloalkoxy, and C<NUM>-<NUM> halocycloalkyl;
in free or salt form.