RU2811411C2 - Compositions of grapiprante and methods of their use - Google Patents

Abstract

FIELD: veterinary medicine.

SUBSTANCE: invention discloses a method of treating pain or inflammation in a dog or cat in need thereof, comprising administering to the dog or a cat a pharmaceutical composition containing a therapeutically effective amount of grapiprant, a surfactant and at least one excipient, wherein the therapeutically effective amount of grapiprant administered orally at a dose rate of 2 to 4 mg per kilogram of a dog or cat body weight at least once daily; wherein the surfactant is selected from poloxamer or sodium lauryl sulfate and is present in an amount of from 0.1% to 5% by weight of the total weight of the composition, wherein at least one excipient includes one or more selected from the group consisting of lactose, sodium starch glycolate, microcrystalline cellulose, colloidal silicon dioxide, magnesium stearate and copovidone.

EFFECT: claimed invention provides high efficiency in the treatment of pain or inflammation in a dog or cat when using low doses of grapiprant.

14 cl, 20 dwg, 19 tbl, 15 ex

Description

CROSS REFERENCE TO RELATED APPLICATIONS

Pursuant to 35 U.S.C. 119(e), this application claims priority to U.S. Provisional Patent Application Serial No. 61/948,957, filed March 6, 2014, entitled “Grapiprant Compositions and Methods for Using Them,” and U.S. Provisional Patent Application Serial No. 62/089713, filed December 9, 2014, entitled “Grapiprant Compositions and Methods for Use Thereof,” the entire contents of which are incorporated herein by reference.

FIELD OF TECHNOLOGY TO WHICH THE INVENTION RELATES

The present invention generally relates to grapiprant compositions and methods of using them.

PREREQUISITES FOR CREATION OF THE INVENTION

Grapiprant is a prostaglandin E2 receptor subtype 4 (EP ₄ ) antagonist useful for the treatment of pain and inflammation. Grapiprant formulations are valuable because they alter the pharmacokinetic properties of the drug in the body, such as peak plasma concentration, time reached, half-life, and area under the plasma concentration-time curve ( _Cmax , _Tmax , _t½ , and AUC , respectively). Another consideration when formulating a pharmaceutical composition is the palatability of the dosage form, which makes the animal more willing to accept the drug.

SUMMARY OF THE INVENTION

Thus, briefly, one aspect of the present invention relates to a method of treating pain or inflammation in a non-human animal in need thereof. The method includes administering to a non-human animal a pharmaceutical composition containing a therapeutically effective amount of grapiprant. The pharmaceutical composition can be administered orally. Upon administration, a Grapiprant C _max of about 375 ng/mL to about 10,000 ng/mL can be achieved with a T _max of about 0.4 to about 3.4 hours, for example, a Grapiprant C _Max of about 750 ng/mL to about 4000 ng/ml or about 1300 ng/ml to about 4000 ng/ml. Grapiprant's _Cmax can also be achieved at a _Tmax of about 0.7 to about 1.7 hours, for example, a _Tmax of about 0.5 to about 1.0 hours. Grapiprant is administered at a dose rate of about 1 to about 10 mg per kilogram of non-human animal body weight per day (mg/kg/day), more specifically at a dose rate of about 2 to about 4 mg/kg/day. The pharmaceutical composition can be administered at least once a day or at least twice a day, for example at least three times a day. The pharmaceutical composition can be administered until the cause of the pain disappears, for example, for a period of time from about 6 days to about 9 months. The non-human animal may be a companion animal such as a dog, cat or horse. In illustrative embodiments, the pharmaceutical drug can be administered twice daily at a dose of about 2 to about 4 mg/kg/day for a period of time from about 9 to about 21 days, and upon administration, a grapiprant _Cmax of about 750 can be achieved. ng/ml to about 2200 ng/ml with a _Tmax of about 0.7 to about 1.7 hours. In other embodiments, the pharmaceutical preparation may be administered to a non-human animal approximately 10 to 18 hours prior to surgery.

Another aspect of the invention relates to a pharmaceutical composition for treating pain or inflammation in a non-human animal in need thereof. The pharmaceutical composition contains a therapeutically effective amount of grapiprant and an excipient. The excipient may include one or more selected from the group consisting of lactose, sodium starch glycolate, microcrystalline cellulose, colloidal silicon dioxide, magnesium stearate, copovidone and poloxamer. In specific embodiments, the pharmaceutical composition may contain from about 5% to about 15% grapiprant (by weight of the total weight of the composition), from about 20% to about 80% lactose (by weight of the total weight of the composition), from about 15% to about 80% microcrystalline cellulose (by weight of total composition), about 1% to about 10% sodium starch glycolate (by weight of total composition), about 1% to about 10% copovidone (by weight of total composition ), about 0.5% to about 3% magnesium stearate (by weight, based on the total weight of the composition), from about 0.5% to about 4% poloxamer (by weight, based on the total weight of the composition), and from about 0.1% to approximately 1% colloidal silica (by weight based on the total weight of the composition). The pharmaceutical preparation may further contain from about 1% to about 30% flavoring agent (by weight based on the total weight of the composition), such as from about 5% to about 15% flavoring agent (by weight based on the total weight of the composition).

The invention also relates to a method of treating pain or inflammation in a non-human animal in need thereof, comprising orally administering to the non-human animal a pharmaceutical composition containing a therapeutically effective amount of grapiprant; Moreover, upon administration, the C _max of grapiprant is achieved from 675 ng/ml to 5000 ng/ml within 4 hours after administration and the half-life is less than 14 hours. _Cmax can be achieved within 1 hour after administration. The pharmaceutical drug can be administered once daily at a dose rate of 3 mg/kg/day to 15 mg/kg/day for 28 days, for example, at a dose rate of 3 mg/kg/day, 9 mg/kg/day, or 15 mg/kg/day.

Additional embodiments and features of the invention are set forth in part in the description of the invention that follows and in part will become apparent to those skilled in the art upon examination of the specification or may be learned by practice of the embodiments described herein. A more complete understanding of the nature and advantages of specific embodiments can be achieved by studying the remainder of the specification, drawings, chemical structures and descriptions that form part of this invention.

BRIEF DESCRIPTION OF THE DRAWINGS

In addition to the illustrative aspects and embodiments described above, additional aspects and embodiments will become apparent from reading the drawings and the following descriptions.

In fig. 1 shows individual serum concentrations of grapiprant over time for group 1 dogs in the experiment described in example 3.

In fig. Figure 2 shows individual serum concentrations of grapiprant over time for group 2 dogs in the experiment described in Example 3.

In fig. Figure 3 shows individual serum concentrations of grapiprant over time for group 3 dogs in the experiment described in Example 3.

In fig. Figure 4 shows individual serum concentrations of grapiprant over time for a group of 4 dogs in the experiment described in Example 3.

In fig. Figure 5 shows individual serum concentrations of grapiprant over time for a group of 5 dogs in the experiment described in Example 3.

In fig. Figure 6 shows individual serum concentrations of grapiprant over time for a group of 6 dogs in the experiment described in Example 3.

In fig. 7 presents pooled individual serum concentrations of grapiprant over time for groups 1-6 dogs in the experiment described in Example 3, which are separately presented in FIG. 3-8.

In fig. Figure 8 shows individual serum concentrations of grapiprant over time for Group 1 male dogs treated with A34 in the experiment described in Example 4.

In fig. Figure 9 shows individual serum concentrations of grapiprant over time for Group 1 female dogs treated with A34 in the experiment described in Example 4.

In fig. 10 shows individual serum concentrations of grapiprant over time for a group of 2 male dogs treated with A27 in the experiment described in Example 4.

In fig. 11 shows individual serum concentrations of grapiprant over time for group 2 female dogs treated with A27 in the experiment described in Example 4.

In fig. 12 shows individual serum concentrations of grapiprant over time for a group of 3 male dogs treated with A29 in the experiment described in Example 4.

In fig. 13 shows individual serum concentrations of grapiprant over time for a group of 3 female dogs treated with A29 in the experiment described in Example 4.

In fig. 14 shows individual serum concentrations of grapiprant over time for a group of 4 male dogs treated with A31 in the experiment described in Example 4.

In fig. 15 shows individual serum concentrations of grapiprant over time for a group of 4 female dogs treated with A31 in the experiment described in Example 4.

In fig. 16 presents mean serum concentrations of grapiprant over time for group 1 dogs, divided into males and females, from the experiment described in Example 4, based on the concentrations individually presented in FIG. 8 and 9.

In fig. 17 presents mean serum concentrations of grapiprant over time for Group 2 of male and female dogs from the experiment described in Example 4, based on the concentrations individually presented in FIG. 10 and 11.

In fig. 18 presents mean serum concentrations of grapiprant over time for group 3 of dogs, divided into males and females, from the experiment described in Example 4, based on the concentrations individually presented in FIG. 12 and 13.

In fig. 19 presents mean serum concentrations of grapiprant over time for a group of 4 dogs, divided into males and females, from the experiment described in Example 4, based on the concentrations individually presented in FIG. 14 and 15.

In fig. 20 presents mean serum concentrations of grapiprant over time for groups 1-4 dogs from the experiment described in Example 4, based on the concentrations individually presented in FIG. 8-19.

DETAILED DESCRIPTION OF THE INVENTION

Grapiprant is a prostaglandin E2 receptor subtype 4 (EP ₄ ) antagonist. Grapiprant has CAS registration number 415903-37-6 and also has other names: AT-001, CJ-023,423, RQ-7, RQ-00000007, MR10A7, AAT-007, N -{2-[4-(2-ethyl -4,6-dimethyl-1H-imidazo[4,5- c ]pyridin-1-yl)phenyl]ethyl}- N' -[(4-methylphenyl)sulfonyl]urea, N -[[[2-[4 -(2-ethyl-4,6-dimethyl-1H-imidazo[4,5- c ]pyridin-1-yl)phenyl]ethyl]amino]carbonyl]-4-methylbenzenesulfonamide or 2-ethyl-4,6-dimethyl -3-(4(2-(((((4-methylphenyl)sulfonyl)amino)carbonyl)amino)ethyl)phenyl)-3H-imidazo[4,5- c ]pyridine. The chemical structure and synthesis of grapiprant are described in WO 2002/032900 and US Patent Nos. 6,710,054, 7,141,580 and 7,479,564, the entire contents of which are incorporated herein by reference. Grapiprant has the following chemical structure:

Without being bound by any particular theory, prostaglandin E2 (PGE2) is a potent modulator involved in the pathogenesis of various diseases such as inflammation, pain, arthritis and cancer. PGE2 binds to at least four types of PGE receptors, which are designated EP ₁ , EP ₂ , EP ₃ and EP ₄ . Molecular pharmacological studies have shown that all subtypes are 7-transmembrane domain receptors belonging to the G protein-coupled receptor superfamily. Activation of EP ₁ stimulates the release of intracellular calcium; stimulation of both EP ₂ and EP ₄ activates adenylate cyclase, however, there are differences in their response to some ligands; and stimulation of EP ₃ inhibits adenylate cyclase through inhibitory G proteins.

In vivo, grapiprant inhibits the binding of [ ³ H]PGE to human, rat and dog EP ₄ receptors with _Ki values of 13 ± 4 nM, 20 ± 1 nM and 24.1 ± 2.7 nM, respectively. Grapiprant is highly selective for the EP ₄ receptor compared to other human prostanoid receptor subtypes and inhibits the PGE ₂ -induced increase in intracellular cAMP at human and rat EP ₄ receptors with p A ₂ values of 8.3 ± 0.03 and 8.2 ± 0.2 nM, respectively. Oral administration of grapiprant leads to a significant reduction in thermal hyperalgesia caused by intraplantar injection of PGE ₂ (ED ₅₀ =12.8 mg/kg). Grapiprant is effective in models of acute and chronic inflammatory pain. Grapiprant significantly reduces the model of mechanical hyperalgesia induced by carrageenan and leads to the resolution of the chronic inflammatory pain response induced by Freund's complete adjuvant. In summary, grapiprant is a potent and selective antagonist of human and rat EP ₄ receptors, producing anti-hyperalgesic effects in animal models of inflammatory pain.

Grapiprant can exist in any of several polymorphic forms. Polymorphic forms differ from each other in their physical properties, spectral characteristics, stability and methods of preparation. Some crystalline forms have already been described, for example, Form A, Form B, Form C, Form D and Form G described in US Pat. No. 7,960,407, ethyl acetate solvate Form I and Form II described in WO 2012/157288, and Form X. Form X2, Form X3 and Form B4 described in the co-pending application entitled "Crystal Forms of Grapiprant", the entire contents of which are incorporated herein by reference.

(I) Pharmaceutical compositions

One aspect of the invention relates to a pharmaceutical composition for treating pain or inflammation in a non-human animal in need thereof, comprising a therapeutically effective amount of grapiprant and an excipient.

Grapiprant can be included in the composition in one or more concentrations. Typically, the concentration of grapiprant may range from about 1% to about 30% (by weight) of the total weight of the composition; that is, the amount of grapiprant may be from about 1% to about 30% by weight, based on all components in the pharmaceutical composition, including grapiprant and excipients. In various embodiments, the concentration of grapiprant can be from about 1% to about 2%, from about 2% to about 3%, from about 3% to about 4%, from about 4% to about 5%, from about 5% to about 6%, from about 6% to about 7%, from about 7% to about 8%, from about 8% to about 9%, from about 9% to about 10%, from about 10% to about 11%, from about 11% to about 12%, about 12% to about 13%, about 13% to about 14%, about 14% to about 15%, about 15% to about 16%, about 16% to about 17 %, from about 17% to about 18%, from about 18% to about 19%, from about 19% to about 20%, from about 20% to about 21%, from about 21% to about 22%, from about 22 % to about 23%, from about 23% to about 24%, from about 24% to about 25%, from about 25% to about 26%, from about 26% to about 27%, from about 27% to about 28% , from about 28% to about 29%, or from about 29% to about 30% (by weight) of the total weight of the composition.

In illustrative embodiments, the concentration of grapiprant may range from about 5% to about 15% (by weight) of the total weight of the composition. In one embodiment, the concentration of grapiprant may be less than about 30% (by weight) of the total weight of the composition. In another embodiment, the concentration of grapiprant may be greater than about 1% (by weight) of the total weight of the composition.

In some embodiments, grapiprant may be provided in a pharmaceutical composition at a concentration suitable for relieving pain or inflammation in a non-human animal. Grapiprant may be administered at a concentration appropriate to relieve pain or inflammation in a non-human animal in need for about 6 days, for about 7 days, for about 8 days, for about 9 days, for for about 10 days, for about 11 days, for about 12 days, for about 13 days, for about 14 days, for about 15 days, for about 16 days, for about 17 days, for about 18 days, for about 19 days, for about 20 days, for about 21 days, for about 22 days, for about 23 days, for about 24 days, for about 25 days, for about 26 days, for about 27 days or for about 28 days.

In illustrative embodiments, grapiprant can be administered at a concentration suitable to relieve pain or inflammation in a non-human animal in need thereof for a period of from about 9 days to about 21 days. In other illustrative embodiments, grapiprant can be administered at a concentration suitable to relieve pain or inflammation in a non-human animal in need thereof for a period of from about 12 days to about 14 days. In some embodiments, grapiprant may be administered at a concentration suitable to relieve pain or inflammation in a non-human animal in need thereof for about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, approximately 6 months, approximately 7 months, approximately 8 months, or approximately 9 months, or longer. In one embodiment, grapiprant can be administered at a concentration suitable to relieve pain or inflammation in a non-human animal in need thereof for a period of less than about 28 days. In another embodiment, grapiprant can be administered at a concentration suitable to relieve pain or inflammation in a non-human animal in need thereof for at least about 9 days.

In other embodiments, the grapiprant may be at least partially dissolved in an aqueous solvent (eg, deionized and/or purified water). In some other embodiments, the grapiprant may be formulated as a suspension. The concentration of grapiprant in the composition may, at least in part, depend on the route of administration and/or the number of administrations over a predetermined period of time when the composition is administered to a non-human animal. For example, one or more compositions may be formulated for administration by injection. As a result, grapiprant in the composition can be delivered directly into the circulatory system (for example, by intravenous administration), thereby eliminating the need for absorption in the gastrointestinal tract. Accordingly, larger quantities of grapiprant can reach intended targets compared to oral formulations, resulting in lower required concentrations of grapiprant in a sterile injectable formulation.

In other embodiments, the pharmaceutical composition can be orally administered one or more times per day, for example, at least twice a day or at least three times a day. For example, the pharmaceutical composition may be administered as a solution, suspension, solid, or viscous liquid preparation. Accordingly, the more times per day the composition is administered to a non-human animal, the less amount of grapiprant can produce the intended result. In illustrative embodiments, the pharmaceutical composition may be formulated for oral administration, for example, as an oral solution or oral suspension, or an oral gel.

Various excipients commonly used in pharmaceutical preparations can be selected based on several criteria, such as, for example, the desired dosage form and the release profile of the dosage form. Non-limiting examples of suitable excipients include those selected from the group consisting of a binder, a bulking agent, a non-effervescent disintegrant, an effervescent disintegrant, a preservative, a diluent, a flavoring agent, a sweetener, a lubricant, an oral dispersant, a coloring agent, a flavoring agent, a pH modifier, stabilizer, sealant, or combinations of any of these agents.

In one embodiment, the excipient may be a binder that holds the pharmaceutical composition together prior to administration. Suitable binders include starches, pregelatinized starches, gelatin, polyvinylpyrrolidone, cellulose, methylcellulose, sodium carboxymethylcellulose, ethylcellulose, polyacrylamides, polyvinyloxoazolidone, polyvinyl alcohols, C ₁₂ -C ₁₈ fatty acid alcohols, polyethylene glycol, polyols, saccharides, oligosaccharides, polypeptides, peptides , as well as their combinations.

In another embodiment, the excipient may be an excipient that adds volume to the pharmaceutical composition for ease of handling and more accurate dosing. Suitable excipients include carbohydrates, inorganic compounds and polyvinylpyrrolidone. By way of non-limiting example, the filler may be calcium sulfate, such as either dibasic or tribasic calcium sulfate; starch, calcium carbonate, magnesium carbonate, microcrystalline cellulose, dibasic calcium phosphate, magnesium carbonate, magnesium oxide, calcium silicate, talc, modified starches, lactose, sucrose, mannitol and sorbitol.

The excipient may be a non-effervescent disintegrant that allows the pharmaceutical composition to dissolve more easily after administration without evolution of gas. Suitable examples of non-effervescent disintegrants include starches (such as corn starch, potato starch and the like), pregelatinized and modified starches derived therefrom, sweeteners, clays (such as bentonite), microcrystalline cellulose, alginates, sodium starch glycolate and gums (such as agar, guar gum, locust bean gum, karaya gum, pectin and gum tragacanth).

In another embodiment, the excipient may be an effervescent disintegrant that allows the pharmaceutical composition to more readily dissolve during administration to release gas. By way of non-limiting example, suitable effervescent disintegrants include sodium bicarbonate in combination with citric acid and sodium bicarbonate in combination with tartaric acid.

The excipient may include a preservative that enhances the stability and shelf life of the pharmaceutical composition, in particular by delaying undesirable degradation of the active ingredient. Suitable examples of preservatives include antioxidants (such as alpha-tocopherol or ascorbate) and antimicrobial agents (such as parabens, chlorobutanol or phenol). In other embodiments, an antioxidant such as butylated hydroxytoluene (BHT) or butylated hydroxyanisole (BHA) may be used.

The excipient may include a diluent that reduces the relative concentrations of other components in the pharmaceutical composition. Diluents suitable for use include pharmaceutically acceptable saccharides such as sucrose, dextrose, lactose, microcrystalline cellulose, fructose, xylitol and sorbitol; polyhydric alcohols; starches; pre-formulated diluents for direct compression; as well as mixtures of any of the above.

The excipient may include a surfactant that alters the solubility parameters of other components in the pharmaceutical composition. In various embodiments, the surfactant may be an alkyl aryl polyether alcohol such as Triton™ X-100, Surfonic™ N-100 (nonoxynol-10) or Witconol™ NP-100; or a poloxamer such as Pluronic™, Synperonic™ or Kolliphor™. Other suitable examples of surfactants include, for example, 2-acrylamido-2-methylpropanesulfonic acid, alkyl polyglycoside, ammonium perfluorononanoate, benzalkonium chloride (BAC), benzethonium chloride (BZT), 5-bromo-5-nitro-1,3-dioxane, cetyltrimethylammonium bromide (CTAB, hexadecyltrimethylammonium bromide, cetyltrimethylammonium chloride), cetylpyridinium chloride (CPC), cyclohexyl-1-hexyl-maltopyranoside, decyl-maltopyranoside, decyl-polyglucose, dimethyldioctadecylammonium chloride, dioctadecyldimethylammonium bromide (DODAB), dipalmitoylphosphatidylcho lin, lauryldimethylamine oxide, dodecyl maltopyranoside, magnesium laureth sulfate, polyethoxylated thalamine (POEA), octenidine dihydrochloride, octylphenoxypolyethoxyethanol (Igepal™ CA-630), octylthioglucopyranoside (OTG), ox bile, sodium nonanoyloxybenzene sulfonate, sorbitan monolaurate, surfactin and tonsonia bromide. In illustrative embodiments, the surfactant may be a poloxamer or sodium lauryl sulfate.

The excipient may be a lubricant that allows the pharmaceutical composition to be more easily removed from the molds during manufacture and may facilitate administration of the pharmaceutical composition. Suitable non-limiting examples of lubricants include magnesium stearate, calcium stearate, zinc stearate, hydrogenated vegetable oils, Sterotex, polyoxyethylene monostearate, talc, polyethylene glycol, sodium benzoate, sodium lauryl sulfate, magnesium lauryl sulfate and light mineral oil.

The excipient may be a dispersant that promotes the dispersion of the components of the pharmaceutical composition in the body of the subject after administration. Suitable dispersants may include starch, alginic acid, polyvinylpyrrolidones, guar gum, kaolin, bentonite, purified wood cellulose, sodium starch glycolate, isomorphic silicate and microcrystalline cellulose.

Depending on the embodiment, it may be desirable to include a colorant that improves the appearance and facilitates identification of the pharmaceutical composition. Suitable colorants include food, drug and cosmetic colors (FD&C), drug and cosmetic colors (D&C), or topical drug and cosmetic colors (Ext. D&C). These color additives or dyes, along with their corresponding varnishes, as well as certain natural and artificial dyes, may be suitable for use in the present invention depending on the embodiment.

In various embodiments, the excipient may include a pH modifier that can alter the solubility profile and bioavailability parameters of the components in the pharmaceutical composition. In specific embodiments, the pH modifier may include sodium carbonate or sodium bicarbonate.

The weight fraction of the excipient or combination of excipients in the pharmaceutical composition may be about 98% or less, about 95% or less, about 90% or less, about 85% or less, about 80% or less, about 75% or less, about 70% or less, about 65% or less, about 60% or less, about 55% or less, about 50% or less, about 45% or less, about 40% or less, about 35% or less, about 30% or less , about 25% or less, about 20% or less, about 15% or less, about 10% or less, about 5% or less, about 2%, or about 1% or less, by weight of the total pharmaceutical composition.

In specific embodiments, the excipient may include one or more of the group consisting of lactose, sodium starch glycolate, microcrystalline cellulose, colloidal silica, magnesium stearate, copovidone, surfactant, poloxamer, and sodium lauryl sulfate.

The concentration of lactose in the pharmaceutical composition may vary. Typically, the lactose concentration may range from about 10% to about 95% (by weight) of the total weight of the composition. In various embodiments, the lactose concentration may be from about 10% to about 15%, from about 15% to about 20%, from about 20% to about 25%, from about 25% to about 30%, from about 30% to about 35%, from about 35% to about 40%, from about 40% to about 45%, from about 45% to about 50%, from about 50% to about 55%, from about 55% to about 60%, from about 60% to about 65%, from about 65% to about 70%, from about 70% to about 75%, from about 75% to about 80%, from about 80% to about 85%, from about 85% to about 90 % or from about 90% to about 95% (by weight) of the total weight of the composition. In illustrative embodiments, the lactose concentration may range from about 20% to about 80% (by weight) of the total weight of the composition.

The concentration of microcrystalline cellulose in the pharmaceutical composition may vary. Typically, the concentration of microcrystalline cellulose may range from about 5% to about 95% (by weight) of the total weight of the composition. In various embodiments, the concentration of microcrystalline cellulose can be from about 5% to about 10%, from about 10% to about 15%, from about 15% to about 20%, from about 20% to about 25%, from about 25% to about 30%, from about 30% to about 35%, from about 35% to about 40%, from about 40% to about 45%, from about 45% to about 50%, from about 50% to about 55%, from about 55% to about 60%, about 60% to about 65%, about 65% to about 70%, about 70% to about 75%, about 75% to about 80%, about 80% to about 85%, about 85% to about 90%, or about 90% to about 95% (by weight) of the total weight of the composition. In illustrative embodiments, the concentration of microcrystalline cellulose may range from about 15% to about 80% (by weight) of the total weight of the composition.

The concentration of sodium starch glycolate in the pharmaceutical composition may vary. Typically, the concentration of sodium starch glycolate may range from about 1% to about 20% (by weight) of the total weight of the composition. In various embodiments, the sodium starch glycolate concentration may be from about 1% to about 2%, from about 2% to about 3%, from about 3% to about 4%, from about 4% to about 5%, from about 5% % to about 6%, from about 6% to about 7%, from about 7% to about 8%, from about 8% to about 9%, from about 9% to about 10%, from about 10% to about 11% , from about 11% to about 12%, from about 12% to about 13%, from about 13% to about 14%, from about 14% to about 15%, from about 15% to about 16%, from about 16% to about 17%, from about 17% to about 18%, from about 18% to about 19%, or from about 19% to about 20% (by weight) of the total weight of the composition. In illustrative embodiments, the concentration of sodium starch glycolate may be from about 1% to about 10% (by weight) of the total weight of the composition.

The concentration of copovidone in the pharmaceutical composition may vary. Typically, the concentration of copovidone may range from about 1% to about 20% (by weight) of the total weight of the composition. In various embodiments, the concentration of copovidone may be from about 1% to about 2%, from about 2% to about 3%, from about 3% to about 4%, from about 4% to about 5%, from about 5% to about 6%, from about 6% to about 7%, from about 7% to about 8%, from about 8% to about 9%, from about 9% to about 10%, from about 10% to about 11%, from about 11% to about 12%, about 12% to about 13%, about 13% to about 14%, about 14% to about 15%, about 15% to about 16%, about 16% to about 17 %, from about 17% to about 18%, from about 18% to about 19%, or from about 19% to about 20% (by weight) of the total weight of the composition. In illustrative embodiments, the concentration of copovidone may range from about 1% to about 10% (by weight) of the total weight of the composition.

The concentration of magnesium stearate in the pharmaceutical composition may vary. Typically, the concentration of magnesium stearate may range from about 0.1% to about 5% (by weight) of the total weight of the composition. In various embodiments, the concentration of magnesium stearate can be from about 0.1% to about 0.5%, from about 0.5% to about 1%, from about 1% to about 1.5%, from about 1.5% to about 2%, from about 2% to about 2.5%, from about 2.5% to about 3%, from about 3% to about 3.5%, from about 3.5% to about 4%, from about 4% to about 4.5%, or about 4.5% to about 5% (by weight) of the total weight of the composition. In illustrative embodiments, the concentration of magnesium stearate may range from about 0.5% to about 3% (by weight) of the total weight of the composition.

The concentration of surfactant in the pharmaceutical composition may vary. Typically, the surfactant concentration may range from about 0.1% to about 5% (by weight) of the total weight of the composition. In various embodiments, the surfactant concentration may be from about 0.1% to about 0.5%, from about 0.5% to about 1%, from about 1% to about 1.5%, from about 1.5% to about 2%, from about 2% to about 2.5%, from about 2.5% to about 3%, from about 3% to about 3.5%, from about 3.5% to about 4%, from about 4% to about 4.5%, or from about 4.5% to about 5% (by weight) of the total weight of the composition. In illustrative embodiments, the surfactant concentration may range from about 0.5% to about 4% (by weight) of the total weight of the composition.

The concentration of colloidal silica in the pharmaceutical composition may vary. Typically, the concentration of colloidal silica may range from about 0.1% to about 2% (by weight) of the total weight of the composition. In various embodiments, the concentration of fumed silica can be from about 0.1% to about 0.2%, from about 0.2% to about 0.3%, from about 0.3% to about 0.4%, from about 0.4% to about 0.5%, about 0.5% to about 0.6%, about 0.6% to about 0.7%, about 0.7% to about 0.8% , from about 0.8% to about 0.9%, from about 0.9% to about 1.0%, from about 1.0% to about 1.1%, from about 1.1% to about 1, 2%, about 1.2% to about 1.3%, about 1.3% to about 1.4%, about 1.4% to about 1.5%, about 1.5% to about 1.6%, from about 1.6% to about 1.7%, from about 1.7% to about 1.8%, from about 1.8% to about 1.9%, or from about 1.9% to about 2.0% (by weight) of the total weight of the composition. In illustrative embodiments, the concentration of colloidal silica may be from about 0.5% to about 1% (by weight) of the total weight of the composition.

In an illustrative embodiment, the pharmaceutical composition may contain from about 5% to about 15% grapiprant (by weight), from about 20% to about 80% lactose (by weight), from about 15% to about 80% microcrystalline cellulose (by weight) , from about 1% to about 10% sodium starch glycolate (by weight), from about 1% to about 10% copovidone (by weight), from about 0.5% to about 3% magnesium stearate (by weight), from about 0.5% to about 4% surfactant (by weight) and from about 0.1% to about 1% colloidal silica (by weight).

(i) Flavors

It may be necessary to add one or more additional compounds to the pharmaceutical composition to enhance palatability. The excipient may include a flavoring agent, which may act as a flavoring agent and/or a flavor-modifying agent. In some embodiments, a flavoring agent may include one or more of a sweetener, a savory agent (ie, a substance that imparts a salty taste to the pharmaceutical composition), a bittering agent, and an acidifying agent. Flavoring agents may be selected from synthetic flavor oils and flavoring agents and/or natural oils, extracts from plants, leaves, flowers, fruits, and combinations thereof. By way of example, these may include cinnamon oil, wintergreen oil, peppermint oil, clove oil, hay oil, anise oil, eucalyptus oil, vanilla oil, citrus oil (such as lemon oil, orange oil, grapeseed oil and grapefruit oil), and also fruit essences (for example, from apple, pear, peach, strawberry, raspberry, cherry, plum, pineapple and apricot). In some embodiments, flavorings and/or flavoring agents may include a vanilla-containing composition, e.g., ethylvanillin, vanillin-RHD, vanillin-Merck, vanilla-TG-old, as well as appropriate solvents (e.g., ethanol and/or water). In other embodiments, other flavorings may be added that impart a different flavor to the composition, such as banana, pork liver, beef, and so on.

In some embodiments, flavorings and/or flavoring agents may include a vanilla-containing composition such as, but not limited to, ethylvanillin, vanillin (vanillin-RHD), natural vanilla flavor (vanillin-Merck), vanilla flavor , identical to natural (vanilla-TG-old), as well as appropriate solvents (for example, ethanol and/or water).

In other embodiments, the flavorings and/or flavoring agents may include flavors selected from chicken, bacon, beef, pork, liver, fish, honey, caramel, and banana.

In another embodiment, the excipient may include a sweetener. By way of non-limiting example, the sweetener may be selected from glucose (corn syrup), dextrose, invert sugar, fructose, and mixtures thereof (if not used as carriers); saccharin and various salts thereof, such as sodium salt; dipeptide sweeteners such as aspartame; dihydrochalcone compounds, glycyrrhizin; stevia-based sweeteners; chlorinated derivatives of sucrose such as sucralose; sugar alcohols such as sorbitol, mannitol, xylitol and the like. The use of hydrogenated starch hydrolysates and the synthetic sweetener 3,6-dihydro-6-methyl-1,2,3-oxathiazin-4-one-2,2-dioxide, in particular its potassium salt (acesulfame-K), and also sodium and calcium salts. In specific embodiments, the pharmaceutical composition may be formulated for oral administration and contain one or more of the following flavoring agents (e.g., sweeteners): sucralose, ^MagnaSweet® , Di- ^Pac® compressed sugar (i.e., a 97:3 mixture of sucrose and maltodextrin), Thaumatin T200X, Talin-Pure, OptisweetSD, Stevia Rebaudioside A Extract and/or Neotame.

In some embodiments, the pharmaceutical composition, which may be formulated for oral administration, may contain one or more of the following flavoring agents and/or flavoring agents (eg, sweeteners): sucralose; licorice dispersion, licorice derivatives and licorice extract (glycyrrhizic acid/monoammonium glycyrrhizinate); ^MagnaSweet® ; mixture of sodium saccharin and neohesperidin dihydrochalcone (Optisweet™ SD), 97:3 (by weight) mixture of sucrose and maltodextrin (Di-Pac ^® ), thaumatin 7% (sweetener) mixed with inactive maltodextrin (Thaumatin T200X), pure thaumatin (Talin -Pure), rebaudioside A from stevia extract (steviol glycosides), neotame and/or polyols (sugar alcohols) such as sorbitol, maltitol, isomalt, xylitol and glycerin.

As used herein, " ^MagnaSweet® " means a composition consisting primarily of one or more sweeteners selected from the group consisting of glycyrrhizic acid (GA), monoammonium glycyrrhizinate (MAG), rebaudioside A and glycerol. In some embodiments ^{, MagnaSweet®} consists primarily of glycyrrhizic acid (GA), monoammonium glycyrrhizinate (MAG), rebaudioside A, and glycerol. In other embodiments ^{, MagnaSweet®} consists primarily of glycyrrhizic acid (GA), monoammonium glycyrrhizinate (MAG), and glycerol. In some embodiments ^{, MagnaSweet®} contains from about 0.5% to about 25% GA/MAG, from about 0% to about 15% rebaudioside A, and from about 75% to about 99.5% glycerol. In other embodiments ^{, MagnaSweet®} contains from about 1.5% to about 17% GA/MAG, from about 0% to about 7.5% rebaudioside A, and from about 83% to about 91% glycerol. In illustrative embodiments, ^MagnaSweet® contains about 1.5% GA/MAG, about 7.5% rebaudioside A, and about 91% glycerol. In other illustrative embodiments ^{, MagnaSweet®} contains about 9% GA/MAG and about 91% glycerol. In another illustrative embodiment ^{, MagnaSweet®} contains about 17% GA/MAG and about 83% glycerol.

In particular, certain sugar-containing sweeteners, such as sucrose-containing materials, sucrose, glucose, fructose and maltodextrin, may at least partially degrade the capromorelin in the composition. Accordingly, high concentrations of some sugar sweeteners should be avoided.

In illustrative embodiments, flavoring or flavoring agents may include at least one of thaumatin, sucralose, neotame, sodium saccharin, neohesperidin dihydrochalcone, rebaudioside A, steviol glycoside, licorice, glycyrrhizic acid, monoammonium glycyrrhizinate, sucrose, glucose, fructose , maltodextrin, sorbitol, maltitol, isomalt, glycerin and a vanilla-containing composition.

The excipient may include a substance that corrects the taste of the drug. Taste-correcting materials include hydroxypropyl cellulose ethers (HPC); low-substituted hydroxypropyl cellulose ethers (L-HPC); hydroxypropyl methyl cellulose ethers (HPMC); methylcellulose polymers and mixtures thereof; polyvinyl alcohol (PVA); hydroxyethylcellulose; carboxymethylcelluloses and their salts; copolymers of polyvinyl alcohol and polyethylene glycol; monoglycerides or triglycerides; polyethylene glycols; acrylic polymers; mixtures of acrylic polymers with cellulose ethers; cellulose acetate phthalate; as well as their combinations.

In some embodiments, the flavoring agent may comprise, as a percentage by weight of the final volume of the pharmaceutical composition, from about 50% to about 0.001%, depending on the agent selected, for example, from about 40% to about 0.01%, from about 30% to about 0.01%, from about 1% to about 30%, or from about 5% to about 15%. As mentioned previously, a pharmaceutical composition may contain more than one flavoring agent.

(ii) Dosage form

The pharmaceutical compositions described herein may be formulated in one or more dosage forms. Suitable dosage forms include tablets, including suspension tablets, chewable tablets, effervescent tablets or caplets; pills; powders, for example, sterile packaged powder, dispersible powder, and effervescent powder; capsules, including soft or hard gelatin capsules such as HPMC capsules; lozenges; sachet; granule-filled capsules; reconstituted powder or cocktail; lozenges; drops such as sublingual or cheek drops; granules; liquids for oral or parenteral administration; suspensions; emulsions; semi-solid formulations or gels. Other suitable dosage forms include transdermal systems or patches. The transdermal system may be a matrix system, a reservoir system, or a system without rate-controlling membranes.

Dosage forms can be prepared using generally accepted pharmacological methods. Conventional pharmacological methods include, for example, one or a combination of: (1) dry mixing, (2) direct compression, (3) milling, (4) dry or non-aqueous granulation, (5) wet granulation, or (6) fusion. See, for example, Lachman et al ., The Theory and Practice of Industrial Pharmacy (1986). Other methods include, for example, prilling, spray drying, panning, melt granulation, granulation, Wurster coating, tangential coating, surface spraying, extrusion, coacervation and the like.

The amount of active ingredient that is administered to a subject can and will vary depending on various factors, such as the age and general health of the subject, as well as the specific route of administration. Those skilled in the art will appreciate that dosages can also be determined using the guidance provided in Goodman &Gilman's The Pharmacological Basis of Therapeutics , tenth edition (2001), Appendix II, pp. 475-493, and Physicians' Desk Reference (PDR).

(II) Methods of treatment using the grapiprant composition

Another aspect of the present invention relates to a method of treating pain or inflammation in a non-human animal in need thereof. The method includes administering to a non-human animal a pharmaceutical composition containing a therapeutically effective amount of grapiprant.

The dose of grapiprant may range from about 0.01 milligrams of grapiprant per kilogram of non-human animal body weight (“mg/kg”) to about 75 mg/kg, such as from about 0.1 mg/kg to about 7 .5 mg/kg. In some embodiments, the dose of grapiprant may range from about 0.75 mg/kg to about 6 mg/kg. In some embodiments, the dose of grapiprant may range from about 6 mg/kg to about 50 mg/kg. In other embodiments, the dose of grapiprant may be at least about 0.1 mg/kg, about 0.2 mg/kg, about 0.3 mg/kg, about 0.33 mg/kg, about 0.5 mg/kg , approximately 0.75 mg/kg, approximately 1.0 mg/kg, approximately 2.0 mg/kg, approximately 3.0 mg/kg, approximately 4.0 mg/kg, approximately 5.0 mg/kg, approximately 6.0 mg/kg, approximately 9.0 mg/kg, or approximately 15 mg/kg.

In illustrative embodiments, a non-human animal may receive a dose of about 2 mg/kg of grapiprant. In another illustrative embodiment, a non-human animal may receive a dose of about 3 mg/kg grapiprant. In one illustrative embodiment, a non-human animal may receive a dose of about 4 mg/kg of grapiprant. In another illustrative embodiment, a non-human animal may receive a dose of greater than about 2 mg/kg of grapiprant. In another illustrative embodiment, the non-human animal may receive a dose of less than about 50 mg/kg of grapiprant. In one illustrative embodiment, the non-human animal may receive a dose of less than about 6 mg/kg of grapiprant.

In one embodiment, a non-human animal may receive a dose of about 3 mg/kg grapiprant. In one embodiment, a non-human animal may receive a dose of about 6 mg/kg grapiprant. In one embodiment, a non-human animal may receive a dose of about 9 mg/kg of grapiprant. In one embodiment, a non-human animal may receive a dose of about 15 mg/kg of grapiprant. In one embodiment, a non-human animal may receive a dose of about 50 mg/kg of grapiprant.

Dosing can be accomplished using a variety of dosing regimens depending on the severity of symptoms in the non-human animal. For example, in some embodiments, the pharmaceutical composition may be administered to a non-human animal in need at least once per day, such as at least twice per day, at least three times per day, or several times per day. day. In other embodiments, the pharmaceutical composition can be administered to a non-human animal in need about 10 to 18 hours before surgery, e.g., about 11 hours, about 12 hours, about 13 hours, about 14 hours , approximately 15 hours, approximately 16 hours, approximately 17 hours or approximately 18 hours before surgery.

The dose of grapiprant can also be expressed in terms of dose intensity; that is, the total amount of grapiprant administered to a non-human animal per kilogram of body weight over a 24-hour period (mg/kg/day). In some embodiments, the dose intensity of grapiprant may range from about 1.5 mg/kg/day to about 12 mg/kg/day. In other embodiments, the dose of grapiprant may be at least about 0.2 mg/kg/day, about 0.4 mg/kg/day, about 0.6 mg/kg/day, about 0.66 mg/kg/day , approximately 1.0 mg/kg/day, approximately 1.5 mg/kg/day, approximately 2.0 mg/kg/day, approximately 4.0 mg/kg/day, approximately 6.0 mg/kg/day , about 8.0 mg/kg/day, about 10.0 mg/kg/day, about 12.0 mg/kg/day, or about 15.0 mg/kg/day. In illustrative embodiments, a non-human animal may receive grapiprant at a dose intensity of from about 1 to about 8 mg/kg/day. In other illustrative embodiments, a non-human animal may receive grapiprant at a dose intensity of from about 2 to about 4 mg/kg/day. In one embodiment, a non-human animal may receive grapiprant at a dose intensity of from about 3 to about 15 mg/kg/day.

The doses administered may be in the form of solid or liquid preparations. For example, certain non-human animals, such as dogs, may receive one or more solid oral preparations, for example, a pharmaceutical composition formulated for administration in capsules, gelatin capsules, gelled liquids (i.e., viscous liquids), solutions, suspensions, pills, caplets, tablets or other solid, liquid or spray forms. For example, capsules or other forms may contain grapiprant in different concentrations so that doses can be administered to non-human animals of different body weights. By way of example only, capsules may be formulated with a grapiprant concentration of 20 mg per capsule, 35 mg per capsule, or 75 mg per capsule. As a result, different combinations of capsules can be administered to non-human animals in need of treatment. By way of example only, a non-human animal weighing approximately 15 kg and prescribed a 3 mg/kg treatment regimen may receive approximately 45 mg of grapiprant per dose. Accordingly, a non-human animal may receive two 20 mg capsules to achieve a dose of grapiprant that is close to 45 mg (i.e., within 5 and 10 milligrams of the desired dose based on body weight, or within the dosing range) . Animals other than humans, of different sizes, and with different prescribed treatment regimens may receive similarly calculated doses to obtain an effective amount of grapiprant.

Upon administration, a Grapiprant C _max of about 375 ng/mL to about 10,000 ng/mL can be achieved with a T _max of about 0.4 to about 3.4 hours, for example, a Grapiprant C _Max of about 750 ng/mL to about 8000 ng/ml or from about 1300 ng/ml to about 4000 ng/ml. In other embodiments, the _Cmax of grapiprant can also be achieved at a _Tmax of about 0.7 to about 1.7 hours, for example, a _Tmax of about 0.5 to about 1.0 hours.

In some embodiments, the _Cmax may vary between non-human animals, such that the _Cmax may be even higher. For example, _Cmax may be about 375 ng/ml, about 400 ng/ml, about 500 ng/ml, about 600 ng/ml, about 700 ng/ml, about 800 ng/ml, about 900 ng/ml, about 1000 ng/ml, approximately 1500 ng/ml, approximately 2000 ng/ml, approximately 2500 ng/ml, approximately 3000 ng/ml, approximately 3500 ng/ml, approximately 4000 ng/ml, approximately 4500 ng/ml, approximately 5000 ng/ml ml, approximately 5500 ng/ml, approximately 6000 ng/ml, approximately 6500 ng/ml, approximately 7000 ng/ml, approximately 7500 ng/ml, approximately 8000 ng/ml, approximately 8500 ng/ml, approximately 9000 ng/ml, about 9500 ng/ml or about 10,000 ng/ml. In illustrative embodiments, the _Cmax of grapiprant may range from 375 ng/ml to 10,000 ng/ml. In other illustrative embodiments, the C _max of grapiprant may be from 750 ng/ml to 400 ng/ml. In other illustrative embodiments, the C _max of grapiprant may be from 1300 ng/ml to 400 ng/ml. In some embodiments, the _Cmax of grapiprant may be greater than 375 ng/ml. In other embodiments, the _Cmax of grapiprant may be less than 10,000 ng/ml.

In other embodiments, the _Tmax may be achieved in an individual non-human animal over a 30-minute, 1-hour, or 2-hour time interval, with a range of about 30 minutes to 2 hours to achieve the _Tmax . T _max may be about 0.4 hour, about 0.5 hour, about 0.6 hour, about 0.7 hour, about 0.8 hour, about 0.9 hour, about 1.0 hour, about 1.1 hour, approximately 1.2 hour, approximately 1.3 hour, approximately 1.4 hour, approximately 1.5 hour, approximately 1.6 hour, approximately 1.7 hour, approximately 1.8 hour, approximately 1.9 hour, about 2.0 hour, about 2.2 hour, about 2.4 hour, about 2.6 hour, about 2.8 hour, about 3.0 hour, about 3.2 hour or about 3.4 hour. In one embodiment, T _max may be from 0.4 to 3.4 hours. In illustrative embodiments, the T _max may be from 0.7 to 1.7 hours. In other illustrative embodiments, the T _max may be from 0.5 to 1.0 hour. In some embodiments, the _Tmax may be greater than 0.4 hours. In other embodiments, the _Tmax may be less than 3.4 hours.

The AUC value may range from about 1000 h*ng/ml to about 11,000 h*ng/ml. In illustrative embodiments, the AUC may range from about 1000 h*ng/mL to about 1500 h*ng/mL, from about 1500 h*ng/mL to about 2000 h*ng/mL, from about 2000 h*ng/mL. ml to about 2500 h*ng/ml, from about 2500 h*ng/ml to about 3000 h*ng/ml, from about 3000 h*ng/ml to about 3500 h*ng/ml, from about 3500 h*ng /ml to about 4000 h*ng/ml, from about 4000 h*ng/ml to about 4500 h*ng/ml, from about 4500 h*ng/ml to about 5000 h*ng/ml, from about 5000 h* ng/ml to about 5500 h*ng/ml, from about 5500 h*ng/ml to about 6000 h*ng/ml, from about 6000 h*ng/ml to about 6500 h*ng/ml, from about 6500 h *ng/ml to about 7000 h*ng/ml, from about 7000 h*ng/ml to about 7500 h*ng/ml, from about 7500 h*ng/ml to about 8000 h*ng/ml, from about 8000 h*ng/ml to about 8500 h*ng/ml, from about 8500 h*ng/ml to about 9000 h*ng/ml, from about 9000 h*ng/ml to about 9500 h*ng/ml, from about 9500 h*ng/ml to about 10,000 h*ng/ml, from about 10,000 h*ng/ml to about 10,500 h*ng/ml, or from about 10,500 h*ng/ml to about 11,000 h*ng/ml.

The half-life (t _1/2 ) may range from about 1.5 hours to about 9 hours. In some embodiments, t _1/2 may range from about 3 hours to about 14 hours. In illustrative embodiments, t _1/2 may range from about 1.5 hour to about 2 hour, from about 2 hour to about 2.5 hour, from about 2.5 hour to about 3 hour, from about 3 hour to about 3.5 hours, from about 3.5 hours to about 4 hours, from about 4 hours to about 4.5 hours, from about 4.5 hours to about 5 hours, from about 5 hours to about 5.5 hours, from about 5.5 hours to about 6 hours, from about 6 hours to about 6.5 hours, from about 6.5 hours to about 7 hours, from about 7 hours to about 7.5 hours, from about 7.5 hours to about 8 hours, from about 8 hours to about 8.5 hours, from about 8.5 hours to about 9 hours, from about 9 hours to about 9.5 hours, from about 9.5 hours to about 10 hours, from from about 10 hour to about 10.5 hour, from about 10.5 hour to about 11 hour, from about 11 hour to about 11.5 hour, from about 11.5 hour to about 12 hour, from about 12 hour to about 12 .5 hour, from about 12.5 hour to about 13 hour, from about 13 hour to about 13.5 hour, or from about 13.5 hour to about 14 hour.

The duration of administration can and will vary. Typically, the pharmaceutical composition can be administered to a non-human animal in need thereof for a period of time ranging from about 6 days to about 9 months. In specific embodiments, the pharmaceutical composition can be administered to a non-human animal in need for about 6 days, for about 7 days, for about 8 days, for about 9 days, for about 10 days, in for about 11 days, for about 12 days, for about 13 days, for about 14 days, for about 15 days, for about 16 days, for about 17 days, for about 18 days, for about 19 days, for about 20 days, for about 21 days, for about 22 days, for about 23 days, for about 24 days, for about 25 days, for about 26 days, for about 27 days or within approximately 28 days.

In illustrative embodiments, the pharmaceutical composition can be administered to a non-human animal in need thereof for a period of time from about 9 days to about 21 days. In other illustrative embodiments, the pharmaceutical composition can be administered to a non-human animal in need thereof for a period of time from about 12 days to about 14 days. In some embodiments, the pharmaceutical composition can be administered to a non-human animal in need for about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months or approximately 9 months. In one embodiment, the pharmaceutical composition can be administered to a non-human animal in need thereof for about 28 days.

In some illustrative embodiments, the pharmaceutical drug can be administered twice daily at a dose of about 2 mg/kg/day to about 4 mg/kg/day for a period of time from about 9 days to about 21 days, and upon administration can achieve Grapiprant C _max is from about 750 ng/ml to about 2200 ng/ml with a T _max of about 0.7 to about 1.7 hours.

In some embodiments, the pharmaceutical composition may be a liquid oral preparation that can be used in the same manner as a solid oral preparation described above. Additionally, liquid medications can be administered using a syringe or sprinkled onto the pet's food, treats, or chews. If the non-human animal is livestock, the pharmaceutical composition can be sprayed onto or included in the feed. For example, a liquid formulation may be formulated containing about 20 mg/mL, about 30 mg/mL, about 40 mg/mL, or about 60 mg/mL of grapiprant in the liquid formulation.

As with the solid formulations described above, different concentrations of the liquid formulation can be used to dose non-human animals of varying body weights. As a result, different volumes of different solutions can be administered to non-human animals to achieve a dose of grapiprant. By way of example only, a non-human animal weighing approximately 15 kg and prescribed a 3 mg/kg treatment regimen may receive approximately 45 mg of grapiprant per dose. Accordingly, a non-human animal can receive approximately 2.3 ml of a 20 mg/ml solution or 1.1 ml of a 40 mg/ml solution to achieve a dose of grapiprant approaching 45 mg. Similarly, if a 4.5 mg/kg treatment regimen was prescribed for the same non-human animal, the animal could receive 2.3 mL of a 30 mg/mL solution or 1.1 mL of a 60 mg/mL solution. so that a dose of grapiprant close to 67.5 mg is achieved (that is, the dose that a 15 kg animal should receive with this treatment regimen). Animals other than humans, of different sizes, and with different prescribed treatment regimens may receive similarly calculated doses to obtain an effective amount of grapiprant.

In some embodiments, the pharmaceutical composition can be administered using any of a variety of routes of administration. The pharmaceutical composition can be administered orally, parenterally and/or topically. For example, the pharmaceutical composition may be formulated for oral administration in a liquid and/or solid formulation such that the composition can be administered using at least one of an aerosol, syringe, pill, tablet, caplet, gelatin capsule, or other liquid formulation administration options. .

In other embodiments, the pharmaceutical composition may be formulated for administration by subcutaneous, intradermal, intravenous, intramuscular, intracranial, intraperitoneal, or intrathecal routes of administration (eg, by injection or via a composition-metering pump). The pharmaceutical composition can be formulated as a parenterally administered depot drug that can be configured for sustained release of grapiprant (eg, release over a period of several days to several months). In addition, the pharmaceutical composition can be administered in the form of a gel, which contacts and is accordingly absorbed through the skin or other tissues of the animal. Alternatively, the pharmaceutical composition can be administered using an electrophoretic system to direct the composition into the circulatory system of a non-human animal. In other embodiments, the pharmaceutical composition may be formulated for transdermal and/or transmucosal administration (eg, via a buccal film or patch that is applied to the inner cheek of a non-human animal). Additionally, in some embodiments, the pharmaceutical composition may be administered intranasally or in the form of one or more suppositories. In some embodiments, the pharmaceutical composition can be formulated as an implant that can be placed in the soft tissue of non-human animals. For example, an implant containing the composition can be implanted into the skin, subcutaneous and/or muscle tissue of a non-human animal for sustained release of the drug. In addition, the pharmaceutical composition can also be formulated for application to the skin of a non-human animal directly at the point of need. In other embodiments, the pharmaceutical composition can be formulated for any other suitable route of administration known in the art. In illustrative embodiments, the pharmaceutical preparation may be administered orally and may be selected from any of the pharmaceutical preparations described in section (I) above.

In some embodiments, the pharmaceutical composition can be administered to a non-human animal as part of a daily feeding regimen. For example, a pharmaceutical composition may be formulated for mixing with a food or other food product intended for a non-human animal such that when the non-human animal consumes its daily food (e.g., dry food or soft food), the animal, non-human also consumes the pharmaceutical composition. In particular, the pharmaceutical composition can be formulated as a liquid or powder such that, before feeding to a non-human animal, the pharmaceutical composition can be applied (eg, sprayed) to the food. Additionally, in some embodiments, the food provided to the non-human animal may be provided with the pharmaceutical composition already added so that the person caring for the non-human animal only needs to provide the drug-containing food to the non-human animal .

Other food products intended for non-human animals may also contain the grapiprant composition as an additive. For example, soft or hard treats or chewable treats (e.g., rawhide or other animal products given to non-human animals for pleasure and/or dietary supplementation) may contain a grapiprant composition as an additive, wherein the grapiprant composition may be either included in a treat or chew, or sprinkled onto a treat or chew. In some aspects, treats or chews may be purchased in a form that already contains the grapiprant composition. In other aspects, the grapiprant composition can be added later to treats or chews when individually feeding the animal.

Additionally, in some embodiments, dry food, treats, and/or chews may be mixed with a maintenance dose of the grapiprant-containing composition. Preferably, the animal receiving the maintenance dose is able to maintain a certain level of food intake. For example, as described above, a maintenance dose (eg, 0.2 mg/kg) can be given to the animal regularly or irregularly to introduce lower doses of the active ingredient. If these maintenance doses are provided along with food (eg, dry food), treats, and/or chews, the animal can experience the maintenance doses without discomfort so that there is little or no loss of active ingredient during administration.

DEFINITIONS

The compounds described herein have asymmetric centers. The compounds of the present invention containing an asymmetrically substituted atom can be isolated in optically active or racemic form. All chiral, diastereomeric, racemic forms and all geometric isomeric forms of the structure are contemplated by the invention unless specifically stated otherwise for any particular stereochemical or isomeric form.

The term "animal" refers to non-human animals such as "livestock" and "companion animals". “Livestock” includes cows, sheep, pigs, poultry (such as chickens, turkeys, quail, etc.), goats, llamas, and other similar animals. The term "companion animal" includes, but is not limited to, dogs, cats, rabbits, ferrets, horses and hamsters. In illustrative embodiments, the companion animal is a dog, cat, or horse.

The term "composition" refers to any solid, semi-solid or liquid composition designed to contain a specific predetermined amount (dose) of a particular ingredient, for example, an active pharmaceutical ingredient, as mentioned previously and as discussed below. Suitable compositions may be pharmaceutical drug delivery systems, including those intended for oral administration, buccal administration, rectal administration, topical or transmucosal administration, or subcutaneous implants or other implantable drug delivery systems; or compositions for the delivery of minerals, vitamins and other biologically active additives, oral care products, flavorings, substances that correct the taste of a medicinal product, and the like. In one embodiment, the compositions are typically liquid, however they may contain solid or semi-solid components. Typically, the dosage form is an orally administered system for delivering a pharmaceutically active ingredient to the gastrointestinal tract of a companion animal.

The expression "therapeutically effective amount" means an amount effective, in the required dosages and for the required period of time, to achieve the desired therapeutic result. The therapeutically effective amount of grapiprant can be determined by one skilled in the art (e.g., a veterinarian) and may vary depending on factors such as the clinical condition, age, sex and body weight of the companion animal, the bioavailability of grapiprant, and the ability of the active agent(s) to evoke a desired response in a companion animal. A therapeutically effective amount is also an amount in which the therapeutically beneficial effects outweigh any toxic or harmful effects of the active agent(s). A therapeutically effective amount also includes an amount that is effective, in the required doses and for the required period of time, to achieve the desired result (eg, weight gain by adding muscle mass).

The term "q.s." means adding a quantity (eg volume or mass) of an ingredient until a final quantity (eg volume or mass) is reached.

The term "w/v" means the concentration of a substance, measured as the mass of the substance per unit volume of a solution or composition.

When describing elements of the present invention or embodiment(s) thereof, the singular number used, as well as the qualifier "specified" should mean that one or more elements are being referred to. The terms “comprising,” “including,” and “having” are inclusive and mean that additional items other than those listed may be included.

Having described the invention in detail, it will be understood that modifications and variations thereof are possible without deviating from the scope of the invention as defined in the appended claims.

EXAMPLES

The following examples are included to demonstrate specific embodiments of the invention. Those skilled in the art will appreciate that the methods described in the examples are methods that the inventors have found to be well suited for practicing the present invention. However, those skilled in the art in light of the present disclosure will recognize that many changes can be made to the specific embodiments that are described and still obtain a similar or similar result without departing from the spirit and scope of the invention, therefore, all material set forth herein should be interpreted as illustrative but not limiting.

Example 1 - Study of the palatability of grapiprant on dogs

Fifty privately owned dogs were used in the study. The dogs were of different breeds, different sexes (male and female, all neutered), ranging in age from 1.5 years to 13 years, and weighing from 14.5 to 113 pounds. The drug was a flavored tablet containing 12.5 mg of grapiprant. Four different flavored tablets (A, B, C and D) were used for each dog, regardless of body weight. The flavors used were beef flavor, pork liver flavor, and two other flavor combinations. All drugs tested were manufactured by Argenta Manufacturing, Manurewa, Auckland, New Zealand. The composition of the tested drugs is given in Table 1.

Table 1: Grapiprant preparations used in palatability studies.

Material name Formula (% by mass) Tablet A Tablet B Tablet C Tablet D Grapiprant 2.4 2.4 2.4 2.4 Microcrystalline cellulose 40.0 35.0 35.0 35.0 Lactose Supertab, spray dried 45.6 40.6 40.6 40.6 Sodium starch glycolate 5.0 5.0 5.0 5.0 Flavor Flavoring powder
Protex 3496 "A"
5.0 Flavoring powder
Spray Dried Pork Liver
15.0 Artificial beef flavoring powder PC-0125
15.0 Artificial flavor powder PC-0335
15.0 Colloidal silicon dioxide 0.5 0.5 0.5 0.5 Magnesium stearate 1.5 1.5 1.5 1.5

Table 2: Evaluation of palatability, results.

Reaction Tablet A
N=12 Tablet B
N=13 Tablet C
N=12 Tablet D
N=13 The dog ate everything at once 1 (8.3%) 11 (84.6%) 8 (66.7%) 7 (53.8%) The dog played with the tablet but eventually ate it 2 (16.7%) 1 (7.7%) 2 (16.7%) 3 (23.1%) The dog took it in his mouth but spat it out 3 (25.0%) 0 (0.0%) 1 (8.3%) 1 (7.7%) The dog resolutely refused to eat 6 (50.0%) 1 (7.7%) 10 (83.3%) 10 (76.9%) Acceptability/palatability 3 (25.0%) 12 (92.3%) 10 (83.3%) 10 (76.9%) Tastes attractive to dogs, according to the owner 3 (25.0%) 11 (84.6%) 10 (83.3%) 9 (69.2%) Tastes unappealing to the dog, according to the owner 9 (75.0%) 2 (15.4%) 2 (16.7%) 4 (30.8%)

This study was a randomized, blinded, non-GCP clinical trial. Based on these data, the animals in the experiment preferred tablet B, followed by tablets C and D. Tablet A was rejected by the majority (75.0%) of the dogs participating in the experiment. In Experimental Groups A and C, owners' ratings of dog palatability were consistent with the dogs' pill-eating results. In these two cases, the dogs ended up eating the tablet; however, owners reported that their dogs probably did not like the taste of the tablets. Overall, Tablet B was found to have the greatest palatability across both assessments. These data were not subjected to any statistical processing.

Example 2 - Another study on the palatability of grapiprant in dogs.

A total of 40 privately owned dogs at two veterinary clinics took part in the study. The dogs were of different breeds, different sexes (male and female, all neutered except two females), ranging in age from 15 months to 13 years 10 months, and weighing from 19.6 to 109 pounds. There were a total of four protocol deviations related to the selection criteria.

The drug was a flavored tablet containing 20 mg of grapiprant. Four different flavored tablets (A27, A29, A31 and A34) were used in all dogs, regardless of body weight. The flavoring used was derived from pork liver. The tablets varied in the amount (5%, 10%, or 15%) of flavoring added. Two of the products evaluated (labeled A27 and A34) contained 15% pork liver flavoring; the difference between the two drugs was the surfactant component: drug A contained poloxamer, while drug D contained sodium lauryl sulfate. All drugs tested were manufactured by Argenta Manufacturing (Manurewa, Auckland, New Zealand) and obtained by Ricerca Laboratories (Concord, OH). The composition of the tested drugs is given in Table 3.

Table 3: Grapiprant preparations.

Material* Preparation A27 Preparation A29 Preparation A31 Preparation A34 Grapiprant 8.0 8.0 8.0 8.0 Lactose
(Super-Tab) 36.0 36.0 41.0 35.0 Sodium starch glycolate 5.0 5.0 5.0 5.0 Microcrystalline cellulose 28.0 38.0 28.0 28.0 Colloidal silicon dioxide 0.5 0.5 0.5 0.5 Magnesium stearate 1.5 1.5 1.5 1.5 Flavoring powder -
pork liver 15.0 5.0 10.0 15.0 Copovidon
(Kollidon VA64) 5.0 5.0 5.0 5.0 Surfactant 1.0 1.0 1.0 2.0

*The amounts of each listed component are given as a mass percentage of the total mass of the drug.

After dogs ingested the tablets (if applicable), owners' opinions on whether the tablet tasted attractive to their dogs were recorded. Dogs were observed for 5 minutes for regurgitation/vomiting or adverse reactions to administration (eg, foaming/foaming at the mouth, pawing at the mouth, other abnormal behavior). The dogs' acceptance or rejection of the test product was recorded.

The relationship between each adverse effect and the test drug was classified in terms of relevance to the study as follows:

1=Unrelated: Undoubtedly pre-existing or caused by a specific extraneous factor.

2=Possibly related: Possibly drug related, as indicated by the adverse effect being related to the study drug and confounding factors.

3=Related: Strong suspicion of drug-drug-related, given timing, and the relationship of the side effect to the study drug and extraneous factors.

Data analysis and results. After entering the results into the database, categorical statistical processing of the summary data was performed to assess the palatability of the four drugs. The summary results are shown in Table 4 below. “Acceptability/palatability” was determined based on the percentage of dogs offered each drug that ate the tablet within a 5-minute time period, regardless of whether they ate it willingly or played with the tablet before eating it.

Table 4: Results of palatability assessment.

Dog behavior A drug
A27
N=9 Preparation A29
N=11 Preparation A31
N=10 Preparation A34
N=10 The dog ate everything at once 4 3 4 2 The dog played with the tablet but eventually ate it 1 2 1 3 The dog took it in his mouth but spat it out 1 1 1 1 The dog resolutely refused to eat 3 5 4 4 Acceptability/palatability 55.6% 45.5% 50.0% 50.5% Tastes attractive to dogs, according to the owner 55.6% 36.4% 40.0% 50.0% Tastes unappealing to the dog, according to the owner 44.4% 63.6% 60.0% 50.0%

Based on these data, A27 was the most acceptable drug for the animals in the experiment, with tablet consumption without reward corresponding to 55.6%. Drugs A31 and A34 were accepted and rejected by an equal number (50%) of the dogs in the experiment. Drug A29 was rejected by more than half (55.4%) of the dogs in the experiment. Although palatability ratings did not differ markedly for the four drugs, there may have been a trend for a dose response effect related to the level of flavor compound: 5% pork liver powder < [10% pork liver powder=15% pork liver powder (sodium lauryl sulfate in as a surfactant] < 15% pork liver powder (poloxamer). In all but two cases (A31 and A29), owner ratings of palatability for dogs were consistent with the results of dogs eating the tablets.

In these cases, the dog eventually ate the tablet; however, owners reported that the dogs probably did not like the taste of the tablets. These data were not subjected to any statistical processing. In this single-dose study, no adverse effects were noted and there were no unusual behaviors (foaming/foaming at the mouth, pawing at the mouth, other abnormal behavior).

Example 3 - Pharmacokinetic studies of grapiprant in dogs.

The purpose of this study was to compare the pharmacokinetic profile of several grapiprant tablet formulations with 20 mg doses of grapiprant capsule and aqueous methylcellulose suspension. Appropriate quantities of the test drug/excipient mixture were placed in gelatin capsules to administer 20 mg of grapiprant to each animal. Since the content of grapiprant in the mixture with the excipient was 45% of the mass of the mixture, the total mass of the mixture in each capsule was 0.0440 g. The permissible mass deviation was within ± 0.005 g of the nominal mass of the capsule contents.

For the aqueous suspension (group 2), the medium was 0.5% methylcellulose (400 centipoise) in water. A suspension of grapiprant at a concentration of 0.3 mg/ml was prepared by adding grapiprant to a calibrated beaker and adding approximately 60-90% of a 0.5% aqueous suspension of methylcellulose to the beaker. A homogenizer was used to mix the material. A magnetic stirrer arm was placed in the beaker and the suspension was stirred for approximately 5 minutes. The magnetic stirrer arm was temporarily removed to adjust the volume by adding medium, and the suspension was then stirred for an additional 5 minutes before being transferred to a labeled dispensing container. The drug was constantly stirred during dosing.

Dogs received one oral dose as a capsule, suspension, or tablet as shown in Table 5.

Table 5: Drugs used in this study.

Group Number of animals A drug Nominal dose level
(mg/kg)* 1 5 20 mg API per capsule 2 2 5 0.3 mg/ml suspension in methylcellulose
Dose volume: 6.67 ml/kg 2 3 5 20 mg tablet, drug A29 2 4 5 20 mg tablet, drug A33 2 5 5 20 mg tablet, drug A31 2 6 5 20 mg tablet, drug 120314-1 2

*based on a 10 kg animal. Only whole tablets were used in groups 3-6.

Statistical analyzes were performed to determine differences in _Cmax , _Tmax and AUC. Values were normalized to mg/kg and group means and standard deviations were calculated for numerical data. Actual doses administered ranged from 1.81 to 2.82 mg/kg, with a mean of 2.2 mg/kg. No animals died or were near death during the study. All animals were in normal health at the time of scheduled examination 2 hours after dosing. Average pharmacokinetic parameters, in addition to dose-normalized _Cmax and AUC _0-∞ values, are given in Table 6.

Table 6: Mean pharmacokinetic parameters of grapiprant in male dogs on day 0.

Group Dose (mg/kg) C _max
(ng/ml) T _max
(hour) AUC _last
(hour*ng/ml) AUC _{0- ∞}
(hour*ng/ml) t _1/2
(hour) C _max /Dose
ng/ml)/
(mg/kg) AUC _{0- ∞} /
Dose
(hour*ng/ml)/
(mg/kg) 1 2.16 804 1.08 1580 1640 5.5 359 724 2 2.00 751 0.786 1650 1870 4.1 376 937 3 2.2 1200 0.891 2330 2340 2.17 546 1070 4 2.2 887 0.995 2050 2320 8.58 409 1160 5 2.29 697 1.4 1690 1860 5.87 311 757 6 2.14 816 1.09 2020 2560 7.52 377 1150

Following administration of a single oral dose of 20 mg grapiprant capsule to dogs in Group 1, the mean _Tmax was 1.08 hours, the mean terminal half-life was 5.5 hours, the mean _Cmax was 804 ng/mL, and the mean AUC _0-∞ was 1640 hours. *ng/ml. Following administration of a single oral dose of a 2 mg/kg grapiprant suspension to dogs in Group 2, the mean _Tmax was 0.786 hours, the mean terminal half-life was 4.1 hours, the mean _Cmax was 751 ng/ml, and the mean AUC _0-∞ was 1870 hours* ng/ml.

Following administration of a single oral dose of 20 mg grapiprant tablet, Drug A29, to dogs in Group 3, the mean _Tmax was 0.891 hours, the mean terminal half-life was 2.17 hours, the mean _Cmax was 1200 ng/mL, and the mean AUC _0-∞ was 2340 hours*ng/ml. Following administration of a single oral dose of 20 mg grapiprant tablet, Drug A33, to dogs in Group 4, the mean _Tmax was 0.995 hours, the mean terminal half-life was 8.58 hours, the mean _Cmax was 887 ng/mL, and the mean AUC _0-∞ was 2320 hours*ng/ml. Following administration of a single oral dose of 20 mg grapiprant tablet, Drug A31, to dogs in Group 5, the mean _Tmax was 1.40 hours, the mean terminal half-life was 5.87 hours, the mean _Cmax was 697 ng/mL, and the mean AUC _{was 0- ∞} was 1860 h*ng/ml. Following administration of a single oral dose of 20 mg grapiprant tablet, Lot 120322-1, to dogs in Group 6, the mean _Tmax was 1.09 hours, the mean terminal half-life was 7.52 hours, the mean _Cmax was 816 ng/mL, and the mean AUC _0-∞ was 2560 h*ng/ml.

The mean _Tmax value was lowest after administration of the grapiprant suspension to dogs, and the AUC _0-∞ value was lowest after administration of the grapiprant capsule. The average t _1/2 value was the smallest, but the C _max value was highest after administration of the tablet (drug A29). The average t _1/2 value was greatest after administration of the tablet (drug A33). The average T _max value was the highest, but the C _max value was the smallest after administration of the tablet (drug A31).

When normalized to the dose received, _Cmax and AUC _0-∞ values were similar among groups, with only slightly lower absorption ( _Cmax and AUC _0-∞ ) in the capsule (group 1), suspension (group 2) and in group 5, taking the drug in tablet form; however, analysis of variance (ANOVA) showed that there was no significant difference between groups in terms of _Cmax , _Tmax and AUC _0-∞ . Figures 1-6 show individual serum concentrations of grapiprant over time for groups 1-6 dogs, respectively. Figure 7 shows the pooled individual serum concentrations of grapiprant over time for groups 1-6 dogs.

Following administration of a single nominal oral dose of 2 mg/kg grapiprant using either a capsule, a methylcellulose suspension, or one of four different formulation tablets, pharmacokinetic parameters varied depending on the dosage form. When normalized to the dose received, _Cmax and AUC _0-∞ values were similar among groups, with only slightly lower absorption ( _Cmax and AUC _0-∞ ) in the capsule (group 1), suspension (group 2) and in group 5, taking the drug in tablet form; however, ANOVA analysis showed that there was no significant difference between groups in terms of _Cmax , _Tmax and AUC _0-∞ .

Example 4 - Additional pharmacokinetic studies of grapiprant in dogs.

The purpose of this study was to determine the pharmacokinetic profile of several formulations of grapiprant tablets administered to male and female beagle dogs. Four drugs were tested: group 1 (drug A34), group 2 (drug A27), group 3 (drug A29) and group 4 (drug A31), as described above in table 4 of example 2. Each tablet of the test drug contained 20 mg of active pharmaceutical ingredient in a specific mixture of excipients. The test drug was used without adjustment for purity or salt content. Dogs received one oral dose in one whole tablet as shown in Table 7 below.

Table 7: Drugs used in this study.

Group Number of animals A drug Nominal dose level
(mg/kg)* 1 4/4 20 mg tablet, drug A34 2 2 4/4 20 mg tablet, drug A27 2 3 4/4 20 mg tablet, drug A29 2 4 4/4 20 mg tablet, drug A31 2

*based on animal body weight 10 kg.

Animals underwent a detailed clinical examination prior to study entry/randomization on day -6. The animals' body weight was within the normal range for beagles of this age and gender. No signs of belching or vomiting were observed after dosing. Dogs received a nominal oral dose of 2 mg/kg grapiprant using four different drug tablets.

Actual dose levels for Group 1 ranged from 2.05 to 3.23 mg/kg, for Group 2 ranged from 1.82 to 3.15 mg/kg, and for Group 3 ranged from 1.69 to 3.23 mg/kg and for group 4 ranged from 2.05 to 3.15 mg/kg. Following administration of a single oral dose of 20 mg grapiprant tablet, Formulation A34, to dogs in Group 1, the mean _Tmax ranged from 0.975 to 0.992 hours. The mean terminal half-life ranged from 5.71 to 6.88 hours. The mean _Cmax ranged from 1750 to 2180 ng/ml and the mean AUC _0-∞ ranged from 4420 to 4650 h*ng/ml. Following administration of a single oral dose of 20 mg of Grapiprant tablet, Formulation A27, to dogs in Group 2, the mean T _max ranged from 0.704 to 0.709 hours. The mean terminal half-life ranged from 6.41 to 9.22 hours. The mean _Cmax ranged from 1430 to 2460 ng/ml and the mean AUC _0-∞ ranged from 3190 to 5160 h*ng/ml. Following administration of a single oral dose of 20 mg grapiprant tablet, Formulation A29, to dogs in Group 3, the mean _Tmax ranged from 0.704 to 0.933 hours. The mean terminal half-life ranged from 3.11 to 6.83 hours. The mean _Cmax ranged from 1270 to 1900 ng/ml and the mean AUC _0-∞ ranged from 2840 to 3530 h*ng/ml. Following administration of a single oral dose of 20 mg of Grapiprant tablet, Formulation A31, to dogs in Group 4, the mean _Tmax ranged from 0.817 to 0.933 hours. The mean terminal half-life ranged from 5.15 to 7.42 hours. The mean _Cmax ranged from 1430 to 2160 ng/ml and the mean AUC _0-∞ ranged from 2860 to 3410 h*ng/ml.

Figures 8 and 9 show individual serum concentrations of grapiprant over time for Group 1 male and female dogs, respectively, treated with A34. Figures 10 and 11 show individual serum concentrations of grapiprant over time for a group of 2 male and female dogs, respectively, treated with A27. Figures 12 and 13 show individual serum concentrations of grapiprant over time for a group of 3 male and female dogs, respectively, treated with A29. Figures 14 and 15 show individual serum concentrations of grapiprant over time for a group of 4 male and female dogs, respectively, treated with A31.

Figure 18 presents mean serum concentrations of grapiprant over time for Group 1 dogs, divided into males and females, based on the concentrations individually presented in Figures 8 and 9. Figure 12 presents mean serum concentrations of Grapiprant over time for Group 2 dogs. divided into males and females, based on the concentrations individually presented in Figures 10 and 11. Figure 18 presents the mean serum concentrations of grapiprant over time for group 3 dogs, divided into males and females, based on the concentrations individually presented in Figures 12 and 13. Figure 19 presents mean serum concentrations of grapiprant over time for a group of 4 dogs, divided into males and females, based on the concentrations individually presented in Figures 16 and 17 . Figure 20 presents mean serum concentrations of grapiprant over time for groups 1-4 dogs based on the concentrations individually presented in Figures 8-19.

No gender differences were observed for any of the drug tablets in this study. The mean T _max was greatest and variability for AUC was smallest following administration of a single oral dose of 20 mg grapiprant tablet with A34. The mean _Tmax value was the smallest, the t _1/2 value was the largest, and the variability for _Cmax was the smallest after taking the A27 tablet. The A29 tablet had the lowest mean _Cmax and t1 _/2 values, but had the greatest variability for _Cmax and AUC. Overall, all four drugs were very similar.

After administration of one nominal oral dose of 2 mg/kg grapiprant using four tablets of different formulations, _Cmax values among the preparations were not significantly different (p>0.05), but _Tmax values were significantly different (p<0.05). The _Tmax value for the A34 tablet was significantly greater than for the A27, A29 and A31 tablets (p < 0.05), and no other pairwise comparisons showed significant differences (p > 0.05). The AUC _{of last} tablet of either drug was not significantly different from the others in pairwise comparisons (p >0.05).

Example 5 - Study of the effectiveness of grapiprant on cats.

The purpose of this pilot study was to confirm that this pain scoring model/score system can distinguish a positive outcome from a negative outcome of control animals in the management of pain and inflammation associated with onychectomy, and to test the effectiveness of a single dose of grapiprant for the control of pain and inflammation associated with onychectomy. in cats.

After an acclimation period of 7, 8 or 9 days, a total of 30 adult cats were randomly assigned to three treatment groups, with ten animals of either sex in each group. All animals were randomly assigned to a specific day of surgery such that the same number of cats from each treatment group underwent surgery each day. Each day, the order of operations was also random. On day 0, all experimental animals were administered butorphanol by subcutaneous injection at a target dose of 0.4 mg/kg body weight immediately before administration of the drug prior to surgery.

All animals in Group 1 were orally administered a 6 mg positive control tablet (Onsior™, robenacoxib), animals in Group 2 were administered an empty gelatin capsule as a negative control, and animals in Group 3 were administered a 20 mg Grapiprant tablet (Drug A34, see Table 4 in Example 2 above), all approximately 30 minutes before onychectomy. After administration, the drugs were not allowed to be washed down with water. On days 1 and 2, animals in group 1 were administered a 6 mg positive control tablet, while animals in groups 2 and 3 were administered an empty gelatin capsule. The detailed experimental design is shown in Table 8.

Table 8: Description of study design.

Group Number of animals Dose Route of administration Dose 1 10 Positive control (onsior™) By Days 0, 1 and 2:
6 mg/animal
(whole 6 mg tablet) 2 10 Negative control
(gelatin capsule) By Days 0, 1 and 2:
0 mg (empty capsule) 3 10 Grapiprant
(20 mg tablet)
Preparation A34 By Day 0: ~20 mg/animal
(whole 20 mg tablet)
Days 1 and 2:
0 mg (empty capsule)

The variables assessed and the intervals at which they were determined are shown in Table 9.

Table 9: Variables assessed and description of study design.

Research day Activity/Parameter Throughout the study Assess general health (at least once daily) Until days -9 to -1 Acclimatization Day -4 Physical examination Day 1 Body weight measurement Day 1 Randomization Day 0 (0.5 hour, 1 hour and every hour until the animal receives two consecutive “one” ratings) Sedation score Days 0 (0.5, 1, 3, 5 and 8 hours after extubation),
1 (24 hours after dosing on day 0, 12.00-14.00 and 17.00-19.00),
2 (48 hours after dosing on day 0, 12.00-14.00 and 17.00-19.00) and
3 (72 hours after dosing on day 0) Analgesia Score

On day 0, cats were dosed orally with positive control (group 1), negative control (group 2), or grapiprant (group 3) approximately 30 minutes before onychectomy. On days 1 and 2, animals in group 1 were dosed orally with the positive control, while animals in groups 2 and 3 were dosed with the negative control.

Surgical procedure .Surgical operations for the study were performed on a staggered schedule on different days. Experimental animals were divided into groups of 9, 9 and 12 cats to perform surgical procedures over three days. The day of surgery was considered day 0 for this group. All animals were administered butorphanol by subcutaneous injection at a target dose of 0.4 mg/kg body weight immediately before administration of the drug prior to surgery. Cats were premedicated with acepromazine (target dose 0.05 mg/kg, subcutaneous) and anesthetized with propofol (maximum 8 mg/kg intravenously, slowly until effective). Anesthesia was maintained with isoflurane (0.5-5% in 100% oxygen). Animals were intubated at the discretion of veterinary staff. During surgical procedures, each animal was monitored for at least heart rate and respiratory rate. Onychectomy of the forelegs of all animals was performed with a surgical scalpel, and after completion of the operation, the soft tissue and skin were closed with GLUture™ external tissue adhesive. After surgery, each animal was monitored periodically until complete recovery. Each cat was extubated when recovery of the swallowing reflex was observed. Extubation time (zero time=T₀) were recorded after the end of the operation.

The sedative and analgesic effect in all experimental animals was assessed in points in accordance with the scheme given in Table 10.

Table 10: Scheme for scoring the sedative and analgesic effect.

Day Time Grade Comments 0 0 Extubation 30 ± 10 min Sedation, in points
Analgesia, in points Performed by an expert unaware of treatment groups 1 hour ± 10 min Sedation, in points
Analgesia, in points As mentioned above 3 hours ± 30 min Sedation, in points (or until the animal receives two consecutive scores of 1)
Analgesia, in points As mentioned above 5 hours ± 30 min Sedation, in points (or until the animal receives two consecutive scores of 1) As mentioned above 8 hours ± 30 min Sedation, in points (or until the animal receives two consecutive scores of 1) As mentioned above 12 hours ± 30 min Analgesia, in points As mentioned above 16 hours ± 30 min Analgesia, in points As mentioned above 20 hours ± 30 minutes Analgesia, in points As mentioned above 24 hours ± 30 min Analgesia, in points As mentioned above 1 24 hours ± 1 hour
(after day 0) Analgesia, in points As mentioned above Middle of a day
12.00-14.00 hour Analgesia, in points As mentioned above Early evening
17.00-19.00 hour Analgesia, in points As mentioned above 2 4 hours ± 1 hour
(after day 0) Analgesia, in points As mentioned above Middle of a day
12.00-14.00 hour Analgesia, in points As mentioned above Early evening
17.00-19.00 hour Analgesia, in points As mentioned above 3 72 hours ± 1 hour
(after day 0) Analgesia, in points As mentioned above

Each experimental animal was observed and sedation scored at specified time points throughout the study or until the animal received a score of 1 on two consecutive occasions. A five-point system was used to assess postoperative sedation.

Analgesic effect scores were determined at specified time points throughout the study. An examiner unaware of the treatment groups observed the cat in its cage and then gently palpated the surgical site to assess pain. The analgesic effect was scored by the same expert during the first 8 hours of the study. Observations after 8 hours were performed by other experts as necessary.

Results. The cat was evaluated as an experimental unit. Differences between treatment groups were assessed at an alpha value of 0.05. Sedative and analgesic effects were scored using methods suitable for assessing ordinal data measured repeatedly over time, such as the GLIMMIX method (SAS Institute, Cary, NC, version 9.3). A multinomial distribution was assumed and a cumulative logit link was used. The model included treatment group, time, and group-time interaction effects as fixed effects. Considering the uneven spacing of time intervals, a complex symmetric (CS) or heterogeneous CS structure was assumed for the covariance matrix, depending on the Akaike information criterion. If the group-time interaction effect was significant, within-time comparisons were made and groups were compared in pairs. If the interaction effect was not significant, the average effect for the group was estimated. If the main effect for group was significant, pairwise comparisons were performed among groups. If the algorithm used in the model fitting process did not converge, alternative models were evaluated. Average values were used to summarize results. In addition, each cat was categorized as success or failure based on the need for pain intervention. Success rates across treatment groups were estimated using methods appropriate for estimating binary outcomes, assuming a binomial distribution and a logit link. The model included treatment group as a fixed effect. If the treatment effect was significant, pairwise comparisons between groups were performed. Percentages of failures within groups and 95% confidence intervals are presented. Intervention outcomes were also assessed using methods appropriate for assessing binary data measured repeatedly over time. A binomial distribution was assumed and a logit link was used.

All animals were successfully dosed with the specific test drug on day 0 using the positive control, negative control and grapiprant drug. From the one-hour time point after surgery through the entire observation period (72 hours), animals in the negative control group consistently had higher analgesia scores than animals in the positive control and grapiprant groups. However, this trend only reached statistical significance at the midday time point on Day 1 (P < 0.05). These results were also obtained when three animals with limb problems unrelated to treatment (swollen paws and paw injury) were excluded from the analysis, but the differences did not reach statistical significance at any time point. Animals in the placebo group required "emergency" analgesia (butorphanol) more frequently and in greater quantities than animals in either of the other two groups, although these differences were not statistically significant.

In summary, administration of the test drug, grapiprant, was associated with a decrease in analgesia scores (reduction in pain) with no apparent effect on sedation scores when compared with a placebo control group. These relationships were similar to those found for the positive control group.

Example 6 - Oral grapiprant tablet for the treatment of osteoarthritis in dogs.

In a blinded, multicenter, dose-ranging field study, dogs with naturally occurring osteoarthritis were treated with grapiprant 2 mg/kg once daily (SID), 5 mg/kg once daily, 4 mg/kg twice daily (BID), or placebo twice daily to assess pain and inflammation control. Dogs were randomly assigned to one of four treatment groups and received oral A34 tablets containing 20, 60 and 100 mg or placebo tablets (corresponding to the 4 mg/kg dose group). (See example 2 above, table 4). Pain and inflammation control were assessed by the owner using the validated Canine Brief Pain Inventory (CBPI), which consists of a Pain Severity Score (PSS), a Pain Interference Score (PIS), and the owner's assessment of the dog's quality of life as an assessment tool. CBPI assessments, as well as veterinarian assessments of osteoarthritis, were performed on study day 0 (baseline) and days 7, 14, 21, and 28.

A total of 476 dogs in the study were screened and 108 were not screened, resulting in a per protocol population of 336 dogs. Table 11 shows the number of animals from each group in the “protocol” population.

Table 11: Number of animals in the study population for each treatment group.

Treatment group Population “according to the protocol” Placebo 83 2 mg/kg SID 86 5 mg/kg SID 82 4 mg/kg BID 85

There were no differences in mean PSS or PIS scores among groups at baseline. Positive responses (decrease in PSS and PIS scores) were observed in all treatment groups, with statistically significant differences observed in the 2 and 5 mg/kg SID groups compared to placebo. On days 14, 21, and 28, PSS scores differed significantly (p ≤0.05) among groups, with statistically significant major improvements in the 2 mg/kg SID group in pairwise comparisons with the placebo group on days 14 and 28, and on days 14, 21 and 28 in the 5 mg/kg group. For PIS scores, the groups differed significantly (p ≤0.05) at day 28, with statistically significant major improvements in the 2 and 5 mg/kg SID groups in pairwise comparisons with the placebo group. Descriptive statistics for the PSS and PIS scores from the CBPI questionnaire are shown in Tables 12 and 13.

Similar efficacy was observed across dose groups, with no increase in benefit with increasing dose, as demonstrated by descriptive statistics for the CBPI PSS and PIS scores (Tables 12 and 13). The mean reductions from baseline in PSS were 48, 48.53, and 44.44 for the 2 mg/kg SID, 5 mg/kg SID, and 4 mg/kg BID groups, respectively, compared with only 28 for the placebo group. For PIS scores, the mean reductions from baseline were 53.25, 55.27, and 52.27 for the 2 mg/kg SID, 5 mg/kg SID, and 4 mg/kg BID dose groups, respectively, compared with only 39.13 for the placebo group. These data suggest that both doses of 2 and 5 mg/kg SID were effective and that twice daily dosing did not increase the beneficial effect.

Table 12: Descriptive statistics of pain severity scores (PSS).

Research day Placebo 2 mg/kg SID 4 mg/kg BID 5 mg/kg SID p-value* Day 0 N 83 86 85 82 0.5217(r) Average 5.00 5.25 5.25 5.00 p-value+ 0.9902 0.9999 0.5325 % Changes**:
Day 7 N 79 84 83 81 0.1024(r) Average -14.29 -24.26 -17.39 -22.22 p-value+ 0.0719 0.9452 0.2785 % Changes**:
Day 14 N 81 85 80 81 0.0441(r) Average -22.22 -32.00 -28.29 -30.00 p-value+ 0.0380 0.1739 0.0370 % Changes**:
Day 21 N 75 81 77 77 0.0478(r) Average -25.93 -35.00 -38.89 -44.44 p-value+ 0.1260 0.1102 0.0207 % Changes**:
Day 28 N 77 84 75 78 0.0109(r) Average -28.00 -48.00 -44.44 -48.53 p-value+ 0.0250 0.1028 0.0048

* Overall ANOVA p-values for treatment, center, and treatment-center interaction effect; + pairwise comparison with placebo; ** percentage change relative to day 0.

Table 13: Descriptive statistics of pain interference scores (PIS).

Research day Placebo 2 mg/kg SID 4 mg/kg BID 5 mg/kg SID p-value* Day 0 N 83 86 85 82 0.5434(r) Average 5.67 6.00 5.83 5.50 p-value+ 0.7963 0.9902 0.3876 % Changes**:
Day 7 N 79 84 83 81 0.3295(r) Average -15.91 -27.97 -28.57 -26.92 p-value+ 0.3039 0.4652 0.1953 % Changes**:
Day 14 N 81 85 80 81 0.2033(r) Average -27.45 -33.33 -35.24 -40.00 p-value+ 0.2817 0.3693 0.0836 % Changes**:
Day 21 N 75 81 77 77 0.1166(r) Average -35.71 -47.83 -42.11 -53.85 p-value+ 0.1777 0.2755 0.0541 % Changes**:
Day 28 N 77 84 75 78 0.0321(r) Average -39.13 -53.25 -52.27 -55.27 p-value+ 0.0416 0.1146 0.0195

* Overall ANOVA p-values for treatment, center, and treatment-center interaction effect; + pairwise comparison with placebo; ** percentage change relative to day 0.

Based on the results for PSS and PIS scores from the CBPI questionnaire, doses of 2 mg/kg SID and 5 mg/kg SID were equally effective in controlling pain and inflammation in osteoarthritis in dogs. Preliminary estimated mean effective dose for dogs 2 mg/kg SID, obtained by preliminary extrapolation of calculations from pharmacokinetics/pharmacodynamics studies in phase 1 and 2 trials in humans, protein binding studies, and based on the comparative receptor affinities of grapiprant in humans and dogs, was confirmed in this field study with a range of doses. The dose was effective under field conditions without food restrictions. In summary, these results support the use of a dose of 2 mg/kg SID, given with or without food, to control pain and inflammation associated with osteoarthritis in dogs.

Example 7-9 month oral toxicity study of grapiprant in dogs with 4 week recovery period (06NG032).

Grapiprant was administered orally once daily for nine consecutive months to Beagle dogs at doses of 0 (0.5% methylcellulose), 1, 6, and 50 mg/kg/day in a dose volume of 5 ml/kg. Four animals of each sex were used in each dose group and two additional animals of each sex were used in the 50 mg/kg dose group for the recovery study. Clinical signs and food intake were assessed daily. Body weight was measured weekly. Ophthalmologic examination was performed at weeks 20 and 38 of the dosing phase and at week 4 of the recovery phase. Electrocardiograms were taken at weeks 13, 26, and 38 of the dosing phase and at week 4 of the recovery phase. Hematologic parameters, blood coagulation, and serum chemistries were monitored at weeks 13, 26, and 39 of the dosing phase and at week 4 or 5 of the recovery phase. Urinalysis was performed at week 37 of the dosing phase and at week 3 or 4 of the recovery phase. Serum concentrations of grapiprant were measured at 0.5, 1, 2, 4, 8, and 24 hours post-dose on Day 1 (50 mg/kg only) and Week 38. At the end of the dosing or recovery period, dogs were euthanized and necropsied. . After macroscopic examination, individual organs were weighed, and a complete set of tissues was collected and processed for microscopic examination.

During the 9-month dosing period, there were no deaths or drug-related effects on body weight, food intake, ophthalmic parameters, electrocardiograms, hematological parameters, blood clotting, organ weights, or serious cases of pathology at doses up to 50 mg/day. kg. Gastrointestinal effects, such as soft or mucous stools that sometimes contained slightly bloody or red material, were observed in all groups, including controls. The incidence of these events was higher in some animals receiving doses of 1-50 mg/kg compared to the incidence in control animals. Significant reductions in mean serum albumin levels were observed at weeks 26 and 39 (up to -14% versus control) at the 50 mg/kg dose, as well as mean albumin/globulin (A/G) ratio at week 39 at the 6 mg dose. /kg (-16%). Individually, a dose-dependent trend toward increased incidence and decreased time to event onset was observed, with decreases in albumin levels (up to -41% versus pre-study values), total protein (up to -30%), and/or calcium (down to -18 %). These values returned to the normal range one month after discontinuation of the drug. The changed serum parameters were restored by the end of the recovery period. There were no noteworthy abnormalities during or at the end of the 4-week recovery period.

Following single or intermittent oral administration, there were no sex-related differences in systemic exposure of grapiprant, and no accumulation of grapiprant was observed following dosing regimens of 1, 6, and 50 mg/kg/day. The combined mean systemic exposure of grapiprant increased proportionally with dose in the dose range of 1-6 mg/kg/day, more than proportionally in the dose range of 6-50 mg/kg/day. Due to moderate regeneration of the ileal mucosal epithelium, one male in the 50 mg/kg dose group had a dose level without observed adverse effects of 6 mg/kg. The combined mean _Cmax was 3480 ng/mL and AUC _0-24 was 10600 at the 6 mg/kg dose. The combined mean C _max was 49,283 ng/mL and AUC _0-24 was 138,667 ng⋅h/mL.

During the 9-month dosing period, there were no drug-related effects on mortality, body weight, food intake, ophthalmologic parameters, electrocardiograms, hematologic parameters, blood clotting, organ weights, or serious morbidity at doses up to 50 mg/kg. . Although functional effects such as soft or mucoid stools and decreased levels of total protein, albumin, and calcium in serum chemistries were observed at doses above 1 mg/kg, these were secondary gastrointestinal effects caused by EP ₄ antagonism and therefore not were adverse effects of the drug. At the same time, histological changes in the mucosal epithelium in the ileum observed in the case of a dose of 50 mg/kg were considered an adverse effect of the drug.

Example 8 - Effect of Grapiprant on Lameness and Pain in Dogs (CL-001).

Twenty drug-naïve adult female beagles underwent surgery to remove the meniscus of the right stifle and were allowed to develop osteoarthritis for at least 8 weeks. Signs of osteoarthritis were confirmed radiographically in all dogs approximately 8 weeks after surgery. Animals were randomly assigned to three groups: a negative control group (n=6), a positive control group (n=7) receiving 5 mg/kg firocoxib per day, and a group receiving grapiprant (n=7) administered once daily. at a dose of 30 mg/kg. Dosing was based on body weight at baseline.

Baseline physical examination, body weight, hematologic parameters, serum chemistries, and urinalysis were performed on study day -1 and repeated on day 13. Orthopedic evaluation was performed on day -1, day 2, day 6, and day 13, including assessment of kinetics with using the GAITRite system to determine the left hindlimb:right hindlimb ratio for peak pressure, distance between two successive steps of the same leg, stride length, and stance phase time of walking, 5-point lameness rating, VAS ratings right hind limb function, pain and joint effusion in the right knee joint, and measurements of comfortable range of motion for both knee joints using a goniometer.

The groups did not differ significantly on any orthopedic parameters, with the exception of a lower lameness score in the grapiprant group on day 2 compared with the negative and positive control groups, and a higher function score (less lameness) in the negative control group at the beginning of the study compared with the positive control and grapiprant groups. Both the grapiprant and negative control groups had statistically significant lower pain scores on days 2 and 6 relative to baseline. In the positive control group, there was a statistically significant improvement in mean peak pressure ratios on day 13, as well as mean lameness scores and mean function scores on days 6 and 13 relative to baseline. In animals receiving grapiprant, a decrease in the average level of total protein, albumin and globulin was observed.

Grapiprant administered at a dose of 30 mg/kg for 14 days was effective on day 2 in reducing claudication and pain in a model of osteoarthritis induced by meniscus ablation. Grapiprant administered at a dose of 30 mg/kg was found to be safe for at least a 14-day treatment period; however, there was a slight decrease in serum protein levels.

Example 9 - Efficacy of Grapiprant for Control of Pain and Inflammation in Feline Onychectomy (FCL-12-002).

In this controlled, non-GLP laboratory study, three treatment groups were used: animals in group 1 received 20 mg grapiprant A34 tablets the night before (approximately 12 hours) and 30 minutes before surgery; group 2 was a negative control group in which animals received empty gelatin capsules, and animals in group 3 received two 20 mg grapiprant A34 tablets 30 minutes before surgery.

There were ten adult cats in each group. Acclimatization of cats continued for 7 days. The cats were divided into two groups of 15 cats each so that the operations were carried out over two days. The day of surgery was considered day 0 for each group. One cat from each grapiprant-treated group was removed prior to surgical procedures. On day -1/0 for a given group, all animals were administered the appropriate test drug before onychectomy. All animals received a dose of butorphanol before surgery. After day 2 procedures, all animals were transferred to a free cat colony. Parameters of interest in the study included statistical analysis of recovery outcomes, sedation scores, analgesia scores, and clinical pathology events.

In summary, administration of the test drug, grapiprant, was associated with a decrease in analgesia scores (reduction in pain) with no apparent effect on sedation scores when compared with a placebo control group. Animals in both grapiprant groups required fewer interventions for pain than those in the placebo control group. The treatment regimen in group 1 was associated with fewer interventions for pain compared with group 3. Thus, administering a dose of grapiprant to cats the night before surgery was more effective than administering the same total amount of grapiprant to cats just 30 minutes before surgery .

Example 10 - Evaluation of dose linearity and feeding effect in an experiment on cats (FPK-11-001).

Grapiprant was mixed with excipients as indicated in Table 14 and administered orally in capsules to male and female cats. Animals were distributed into groups as indicated in Table 15.

Table 14: Grapiprant drug used in this study.

Ingredient Component, % (by mass) Grapiprant 45 Lactose 200 mesh 23 Dicalcium phosphate dihydrate 15 Pregelatinized starch 5 Microcrystalline cellulose 6 Povidone 5 Polaxamer 188 1

Table 15: Groups in the study.

Group Number of animals (male/female) Dose level (mg/kg) Feeding/Fasting 1 3/3 2 feeding 2 3/3 2 starvation 3 3/3 6 starvation 4 3/3 10 starvation

The day of dosing was considered day 0. After dosing, animals were assessed for clinical signs. Blood samples were collected before dosing and at 0.25, 0.5, 1, 2, 4, 8, 12, and 24 hours postdose. Serum was obtained from blood samples and analyzed for grapiprant concentration. There were no clinical signs associated with the test drug. The average pharmacokinetics of grapiprant are given in Table 16.

Table 16.

Group Dose (mg/kg) Food Floor C _max
(ng/ml) T _max (hour) AUC _last (hour*ng/ml) AUC _{0- ∞}
(hour*ng/ml) t _1/2
(hour) 1 2 feeding males 407 1.65 1030 1220 3.12 1 2 feeding females 327 1.33 990 1070 2.4 2 2 starvation males 1100 1.01 2290 1970 2.16 2 2 starvation females 1630 1.01 2780 2850 1.47 3 6 starvation males 2970 0.99 3650 3750 2.79 3 6 starvation females 3110 1.17 6130 6350 4.98 4 10 starvation males 4280 1.33 7430 7580 4.03 4 10 starvation females 7130 0.73 10700 10900 4.19

There were no sex-related differences, although _Cmax and AUC were higher in females than males in fasted cats and _Cmax and AUC were lower in females in fed cats. Following a single oral dose of 2 mg/kg grapiprant capsule in Group 1 fed cats, the mean _Tmax ranged from 1.33 to 1.65 hours. The mean terminal half-life ranged from 2.40 to 3.12 hours. The mean _Cmax ranged from 327 to 407 ng/ml and the mean AUC _0-∞ ranged from 1070 to 1220 h*ng/ml.

Following a single oral dose of 2 mg/kg grapiprant capsule in a group of 2 fasted cats, the mean _Tmax was 1.01 hours. The mean terminal half-life ranged from 1.47 to 2.16 hours. The mean _Cmax ranged from 1100 to 1630 ng/ml and the mean AUC _0-∞ ranged from 1970 to 2850 h*ng/ml. Following an oral dose of 2 mg/kg grapiprant capsule, _Cmax and AUC values were significantly higher (p < 0.05) in fasted cats than in fed cats in both males and females. However, the values of T _max and t _1/2 did not differ significantly.

Following a single oral dose of 6 mg/kg grapiprant capsule in a group of 3 fasted cats, the mean _Tmax ranged from 0.99 to 1.17 hours. The mean terminal half-life ranged from 2.79 to 4.98 hours. The mean _Cmax ranged from 2970 to 3110 ng/ml and the mean AUC _0-∞ ranged from 3750 to 6350 h*ng/ml.

Following a single oral dose of 10 mg/kg grapiprant capsule in a group of 4 fasted cats, the mean _Tmax ranged from 0.728 to 1.33 hours. The mean terminal half-life ranged from 4.03 to 4.19 hours. The average _Cmax ranged from 4280 to 7130 ng/ml and the average AUC _0-∞ ranged from 7580 to 10900 h*ng/ml. Greater exposure occurred when animals were dosed in the fasted state than in the fed state, as evidenced by larger AUC and _Cmax values in Group 2 (fasted) than in Group 1 (fed) animals.

Exposure (AUC _0-∞ ) in groups of fasted animals (groups 2-4) was approximately linearly dependent on dose, but was less than proportional to dose. Single administration of grapiprant (oral, capsule) to male and female cats at dose levels of 2 (fed and fasted), 6 and 10 mg/kg was well tolerated without any test drug-related clinical signs. Significantly greater (p < 0.05) exposure occurred when doses were administered to animals in the fasted state than in the fed state, as evidenced by larger AUC and _Cmax values in group 2 (fasted) than in group 1 (fed). animals. Exposure (AUC _0-∞ ) in groups of fasted animals (groups 2-4) was approximately linearly dependent on dose, but was less than proportional to dose.

Example 11 - Evaluation of the binding of grapiprant to cat serum proteins by equilibrium dialysis (FPK-12-002).

Grapiprant at concentrations of 200 and 1000 ng/ml in cat serum was subjected to equilibrium dialysis using a rapid equilibrium dialysis device (Rapid Equilibrium Dialysis (RED)). Following dialysis, protein binding to grapiprant was high in cat serum. The percentage binding of grapiprant in cat serum at concentrations of 200 and 1000 ng/ml was 95% and 92%, respectively. Under these conditions, changing the concentration of grapiprant did not have a significant effect on the binding of cat serum proteins. The positive control compound warfarin (10 μM) was 99.2% bound to human plasma proteins.

Example 12 - Pharmacokinetics of grapiprant in cats (FPK-12-003).

This controlled, non-GLP laboratory study used two treatment groups of six adult cats (three of each sex), for a total of 12 experimental animals. All cats underwent an 8-day acclimatization phase. On day 0, fasted animals in Group 1 were orally administered a 20 mg grapiprant tablet (Formulation A34), while fasted animals in Group 2 were orally administered a 20 mg Grapiprant tablet (Formulation A29). Blood samples were collected and serum was obtained for pharmacokinetic (PK) analysis. Four hours after dosing, all cats were offered food. After collecting samples for PK analysis on day 1, all animals were released into a free cat colony. The variable of interest in this study was the serum PK profile. All animals were successfully dosed with the prescribed test drug, Grapiprant. No abnormalities in general health were noted during the study. Serum concentrations of grapiprant were successfully measured to determine the PK profile presented in Table 17.

Table 17.

A drug Floor Statistics C _max (ng/ml) T _max (hour) AUC _last (hour*ng/ml) AUC _last /Dose
(hour*mg/ml) t _1/2
(hour) A29 males average 1029 1.3 4760 1182 1.7 Art. off 818 0.3 3564 832 0.4 CV% 79.5 43.3 74.9 - 25.3 females average 4413 1.2 17089 2905 1.7 Art. off 1535 0.8 4189 1253 0.3 CV% 34.8 65.5 24.5 - 19.3 A34 males average 1664 0.8 5785 1419 5.0 Art. off 1090 0.3 2534 638 2.9 CV% 65.5 34.6 43.8 - 57.9 females average 9630 1.2 29691 4606 2.5 Art. off 4901 0.8 8780 968 0.7 CV% 50.9 65.5 29.6 - 27.7

Generally, no abnormalities in general health were noted during the study. Greater systemic exposure was observed in female cats than in male cats for both A29 and A34. In male cats, similar systemic exposure was observed for A29 and A34. Female cats treated with grapiprant in A34 had greater systemic exposure to grapiprant than female cats treated with A29. All female cats received a higher dose (mg/kg) than the male cat dose; however, after adjustment for dose administered (mg/kg), female cats still had greater grapiprant exposure than male cats.

Example 13 - Pharmacokinetics of grapiprant in cats (FPK-13-004).

In this controlled, non-GLP laboratory study, three treatment groups were used: animals in group 1 received grapiprant at a dose of 2.5 mg/animal; animals in group 2 were administered grapiprant at a dose of 5 mg/animal, and animals in group 3 were administered grapiprant at a dose of 10 mg/animal. Each dose was administered once on day 0. Blood samples were collected and serum was obtained for pharmacokinetic (PK) analysis. Four hours after dosing, all cats were offered food. After collecting samples for PK analysis on day 1, all animals were released into a free cat colony.

The mean pharmacokinetic parameters of grapiprant are presented in Table 18 below. Pharmacokinetic parameters were similar in males and females within each dosing group and, therefore, mean values were determined by pooling data obtained from all animals.

Table 18 - Pharmacokinetic data.

Dose (mg) Dose (mg/kg) T _max (h) C _max (ng/ml) C _max /Dose (kg*ng/ml/mg) AUC _last (h*ng/ml) AUC _last /Dose (h*ng/ml) t _1/2 (h) 2.5 0.5 1.50 1060 2120 4190 8380 1.58 5 1 1.13 2690 2500 10300 9450 1.13 10 2 1.25 4430 2150 21200 10300 3.44

Mean _Tmax values were similar between groups, ranging from 1.13 to 1.50 hours. Half-life values were similar for the 0.5 and 1 mg/kg doses (1.58 and 1.13 hours, respectively) and increased slightly for the 2 mg/kg dose (3.44 hours). The increase in C _max was approximately dose proportional, and the increase in AUC _last values was slightly greater than dose proportional. With a 4-fold increase in dose from 0.5 to 2 mg/kg, C _max increased by 4.18 times and AUC _last values increased by 5.06 times. Overall, the data suggest that the 2 mg/kg dose of grapiprant was most effective when administered to cats.

Example 14 - Safety and toxicokinetic profiles of grapiprant in cats.

The purpose of this study was to evaluate the safety and toxicokinetic profiles of grapiprant in cats. Twenty-four domestic shorthair cats were randomly assigned to receive either placebo or 3, 9, or 15 mg/kg doses of grapiprant capsule (3 males and 3 females per group) once daily for 28 days. All cats received their prescribed medications according to protocol. Food intake and behavior were monitored daily, body weight was determined weekly, CBCs were performed on days -7, 14 and 25, and urinalysis was performed on days -7 and 25. Serial blood samples for toxicokinetic analyzes were collected after dosing on day 0 and day 27. A complete autopsy and histopathological analysis were performed after humane euthanasia on day 28.

Grapiprant was well tolerated and no adverse effects were observed at daily doses up to 15 mg/kg for 28 days. All animals had normal appearance throughout the study. Grapiprant did not cause death or drug-related effects on body weight, food intake, clinical parameters, organ weights, gross pathology, or histopathology.

Grapiprant reached peak serum concentrations rapidly and remained at significant levels throughout the study. Grapiprant exposure varied depending on dose, but similar mean AUC _last values were observed in the 9 mg/kg and 15 mg/kg groups. By day 27, maximum serum concentrations of grapiprant ranged from 683 ng/mL to 4950 ng/mL and were achieved 1 to 4 hours after administration. The mean elimination half-lives on day 27 ranged from about 3 to about 14 hours, with a mean of about 5 to about 6 hours.

Grapiprant capsule formulation was well tolerated when administered for 28 days with a serum exposure of 4950 ng/mL.

Example 15 - Additional pharmacokinetic study of grapiprant in dogs.

The purpose of this study was to determine and compare the pharmacokinetic parameters of grapiprant in Beagle dogs after a single oral administration of the drug in the form of a flavored tablet and a drug in the form of a suspension in methylcellulose at nominal doses of 6 mg/kg and 50 mg/kg. For this study, sixteen beagles, two of each sex at each dose, were divided into two groups based on nominal doses of grapiprant, either 6 mg/kg in groups 3A/B or 50 mg/kg in groups 4A/B. Animals in each group were administered the appropriate oral dose of methylcellulose tablet or formulation using a crossover study as shown in Table 19 below on days 0 and 15 of the study. Blood samples were collected, serum was obtained, frozen and analyzed for grapiprant concentration.

The medium was 0.5% methylcellulose (400 centipoise) in water. Approximately half the volume of distilled water was heated to a temperature of approximately 40-50°C. Methylcellulose was weighed to obtain a final concentration of 0.5% (w/v) and dissolved in heated distilled water with stirring. The flask with the methylcellulose solution was immediately cooled. The remaining distilled water was added to the flask to adjust the volume. The final solution was stirred for at least 30 minutes.

To prepare 1.2 mg/mL and 10 mg/mL grapiprant suspensions in media, the appropriate amount of grapiprant was added to a calibrated beaker containing 0.5% aqueous methylcellulose and mixed using a vortex mixer. The suspension was homogenized using a Polytron™ mixer equipped with an anti-foaming attachment. A magnetic stirrer arm was placed in the beaker and the suspension was stirred for approximately 5 minutes. The magnetic stirrer arm was temporarily removed to adjust the volume, and the suspension was then stirred for an additional 5 minutes before transferring to a labeled amber dispensing container or foil-wrapped glass beaker.

The dosing suspension was re-prepared on each dosing day. On both dosing days, the depth of the mixing funnel ranged from 75% to 100% of the depth of the container containing 500 mg of suspension. Stirring sufficiently vigorous to achieve a funnel of this depth results in a homogeneous mixture, as has been demonstrated for all drug samples ranging from 93% to 100% of the nominal concentration. Samples of drug doses were transferred to the animal room prior to collection and collected (while mixing the drugs) immediately before dosing the first animal of each group and immediately before dosing the last animal of each group. These measures prevented sedimentation of the 500 mg suspension during transport from the laboratory where the drugs were prepared to the animal room.

Table 19 - Experimental groups and dose levels.

Group Number of animals Type of dosing Day 0 Day 15 3A 2 males/2 females Tablet/Suspension 60 mg tablet 60 mg suspension 3B 2 males/2 females Suspension/Tablet 60 mg suspension 60 mg tablet 4A 2 males/2 females Tablet/Suspension Five 100 mg tablets 500 mg suspension 4B 2 males/2 females Suspension/Tablet 500 mg suspension Five 100 mg tablets

The gastric tube was carefully inspected after dosing in all animals on both days. No appreciable amounts of the test drug were observed in any of the gastric tubes. There were no cases of animal death. A slight increase in the incidence of fecal abnormalities was noted and was considered related to grapiprant exposure.

The _Tmax value on day 0 at a dose of 60 mg was 1.0 or 2.0 hours in the case of tablets and 1.0 hours in the case of suspension. The average _Cmax for dogs of both sexes combined was 5330 ng/ml for 60 mg tablets and 4050 ng/ml for 60 mg suspension. The average AUC _last value for dogs of both sexes in total was 19,600 hours * ng/ml in the case of 60 mg tablets and 14,500 hours * ng/ml in the case of 60 mg suspension.

The _Tmax on day 0 at the 500 mg dose was 1.0, 2.0 or 4.0 hours for tablets and 0.5, 1.0 or 2.0 hours for suspension. The mean _Cmax for dogs of both sexes combined was 96,100 ng/ml for 500 mg tablets and 76,800 ng/ml for 500 mg suspension. The mean AUC _last value for dogs of both sexes combined was 455,000 h*ng/ml for 500 mg tablets and 293,000 h*ng/ml for 500 mg suspension.

The _Tmax value on day 15 at a dose of 60 mg was 1.0 or 2.0 hour in the case of tablets and 1.0 hour in the case of suspension. The mean _Cmax for dogs of both sexes combined was 6110 ng/ml for 60 mg tablets and 3810 ng/ml for 60 mg suspension. The average AUC _last value for dogs of both sexes combined was 17,400 h*ng/ml for 60 mg tablets and 12,500 h*ng/ml for 60 mg suspension.

The _Tmax value on day 15 at a dose of 500 mg was 1.0 or 2.0 hour in the case of tablets and 1.0 hour in the case of suspension. The mean _Cmax for dogs of both sexes combined was 101,000 ng/ml for 500 mg tablets and 74,200 ng/ml for 500 mg suspension. The average AUC _last value for dogs of both sexes combined was 430,000 h*ng/ml for 500 mg tablets and 209,000 h*ng/ml for 500 mg suspension.

_Tmax , _Cmax and AUC _last values were higher at the 500 mg dose than at the 60 mg dose. _Cmax and AUC _last values were similar for a given dose on days 0 and 15. There were differences in pharmacokinetic parameters between tablets and suspension at both doses, with exposure (AUC _last ) being greater for the tablet formulation compared to the tablet formulation. suspension form. ANOVA results indicate that when tablet and suspension formulations were compared, there was a significant difference (p ≤0.05) in _Cmax and AUC _last at the 60 mg dose and a significant difference (p ≤0.05 or p ≤ 0.01) in _Tmax and AUC _last values at a dose of 500 mg. Statistical differences occurred with and without data from the animal that vomited within the first 11 minutes of dosing. Overall, coefficients of variation for AUC _last for the 500 mg suspension were 53.7% (pharmacokinetic data from the vomiting animal excluded) on day 0 and 28.7% on day 15.

Thus, the results showed a higher exposure (AUC) in the case of the drug in the form of tablets compared to the suspension. The difference between the AUC values for the tablets and the suspension was statistically significant for both doses, 60 mg and 500 mg, both when data from the vomiting animal were included in the analysis and when they were excluded. The relative bioavailability ratios (tablet/suspension) for both dose levels in both trials were consistently greater than 100, indicating that the bioavailability of the tablets was greater than that of the suspension. Relative bioavailability ratios greater than 100 were maintained regardless of whether data from a vomiting animal were included in the analysis.

Claims (17)

1. A method of treating pain or inflammation in a dog or cat in need thereof, comprising administering to the dog or cat a pharmaceutical composition containing a therapeutically effective amount of grapiprant, a surfactant and at least one excipient, wherein a therapeutically effective amount of grapiprant is administered orally at a dose rate of 2 to 4 mg per kilogram of body weight of the dog or cat at least once daily; wherein the surfactant is selected from poloxamer or sodium lauryl sulfate and is present in an amount of from 0.1% to 5% by weight of the total weight of the composition, wherein the at least one excipient includes one or more selected from the group consisting of lactose, sodium starch glycolate, microcrystalline cellulose, colloidal silicon dioxide, magnesium stearate and copovidone. 2. The method according to claim 1, characterized in that upon administration the C _max of grapiprant is achieved from 750 ng/ml to 4000 ng/ml. 3. The method according to claim 2, characterized in that upon administration the C _max of grapiprant is achieved from 1300 ng/ml to 4000 ng/ml. 4. The method according to claim 2, characterized in that upon administration, the _Cmax of grapiprant is achieved at a _Tmax of 0.7 to 1.7 hours. 5. The method according to claim 4, characterized in that upon administration the _Cmax of grapiprant is achieved at a _Tmax of 0.5 to 1.0 hour. 6. The method according to claim 1, characterized in that the pharmaceutical composition additionally contains from 1% to 30% flavoring (by weight of the total weight of the composition). 7. The method according to claim 6, characterized in that the pharmaceutical composition contains from 5% to 15% flavoring (by weight of the total weight of the composition). 8. The method according to claim 1, characterized in that the pharmaceutical composition is administered at least twice a day. 9. The method according to claim 1, characterized in that the pharmaceutical composition is administered over a period of time from 6 days to 9 months. 10. The method according to claim 9, characterized in that the pharmaceutical composition is administered over a period of time from 9 to 21 days. 11. The method according to claim 10, characterized in that the pharmaceutical composition is administered over a period of time from 12 to 14 days. 12. The method according to claim 10, characterized in that the pharmaceutical composition is administered once a day and upon administration, the _Cmax of grapiprant is achieved from 750 ng/ml to 4000 ng/ml with a _Tmax from 0.7 to 1.7 hours. 13. The method according to claim 10, characterized in that the pharmaceutical composition is administered twice a day and upon administration, the _Cmax of grapiprant is achieved from 750 ng/ml to 4000 ng/ml with a _Tmax from 0.7 to 1.7 hours. 14. The method according to claim 1, characterized in that the pharmaceutical composition is administered twice a day and the administration is performed approximately 10-18 hours before surgery on the dog or cat.