HK1144939B - Particulates of a crth2 antagonist - Google Patents

Description

Microparticles of CRTH2 antagonist

Technical Field

The present invention provides microparticles of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid in amorphous or crystalline form, and methods of preparation and pharmaceutical compositions thereof. The invention also provides methods of using them to treat, prevent or ameliorate one or more symptoms of a CRTH 2-mediated disorder or disease.

Background

CRTH2 is a G protein-coupled chemoattractant receptor expressed on Th2 cells, eosinophils and basophils (Nagata et al, J.Immunol.1999, 162, 1278-minus 1286; Hirai et al, J.Exp.Med.2001, 193, 255-minus 261). Prostaglandin D, the main inflammatory mediator produced by mast cells₂(PGD₂) Is a natural ligand of CRTH 2. Recently, it has been discovered that the PGD₂The induced activation of CRTH2 induced migration and activation of Th2 cells and eosinophils, suggesting CRTH2May have a pro-inflammatory effect in allergic diseases (Hirai et al, J.Exp.Med.2001, 193, 255-. It was also found that circulating T cells expressing CRTH2 appeared to be increased in atopic dermatitis patients, and that CRTH2 had an association with the severity of the disease (Cosmi et al, Eur. J. Immunol.2000, 30, 2972-2979; Iwazaki et al, J. investigative Dermatology 2002, 119, 609-616). PGD₂The role in the initiation and maintenance of allergic inflammation is further demonstrated in the mouse asthma model, which shows passage through PGD₂Overproduction of PGD by synthetase in vivo₂Airway inflammation can be exacerbated (Fujitani et al, J.Immunol.2002, 168, 443-449). Thus, CRTH2 antagonists have potential use in the treatment of CRTH 2-mediated conditions or diseases, such as allergic rhinitis, allergic asthma, bronchoconstriction, atopic dermatitis or systemic inflammatory diseases.

Brief description of the invention

The present invention provides microparticles of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid having formula I, in amorphous form or in crystalline form I or II:

the compound of formula I was identified as a CRTH2 antagonist (WO 2004/0022218).

In one embodiment, the particles of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid are form I having an X-ray powder diffraction pattern substantially as shown in figure 1 or a differential scanning calorimetry thermogram substantially as shown in figure 2.

In another embodiment, the microparticles of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid are form II having an X-ray powder diffraction pattern substantially as shown in figure 6.

In another embodiment, the microparticles of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid are amorphous.

Also provided are pharmaceutical compositions comprising microparticles of 4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid in amorphous, form I or form II crystalline form or mixtures thereof, and one or more pharmaceutically acceptable carriers or excipients.

Further provided are methods of treating, preventing, or ameliorating one or more symptoms of a CRTH 2-mediated disorder or disease, comprising administering microparticles of 4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid or mixtures thereof in amorphous, form I or form II crystalline forms.

In addition, methods of preparing microparticles of 4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid in amorphous form or in crystalline form I or form II are provided.

Drawings

Figure 1 shows an X-ray powder (XRP) diffraction pattern of particles of {4, 6-bis (dimethyl-amino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid as form I crystals.

Figure 2 shows a Differential Scanning Calorimetry (DSC) thermogram for microparticles of the acid of formula I, described as form I.

FIG. 3 shows a thermogram of Thermogravimetry (TG) of microgranules of an acid of said formula I, form I.

FIG. 4 shows Scanning Electron Microscope (SEM) photographs of microparticles of the acid of formula I in form I at magnifications of 2,500(A), 5,000(B), and 10,000 (C).

Figure 5 shows the particle size distribution diagram for microparticles of the acid of formula I described in form I.

Figure 6 shows the XRP diffractogram of the particles of said acid of formula I in form II.

Figure 7 shows SEM photographs of microparticles of the acid of formula I in form II at magnifications of 370(a), 2,000(B), 5,500(C) and 10,000 (D).

Figure 8 shows the XRP diffraction pattern of the particles of the acid form I obtained by phase equilibrium between form I and form II particles.

Figure 9 shows the XRP diffractogram of the amorphous form of the acid of formula I.

FIG. 10 shows a dose response curve for {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid of formula I in a competitive radioligand binding assay using CRTH 2-transfected cells.

FIG. 11 shows dose response curves for {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid of formula I (. tangle-solidup.) and DPI-selective antagonist BWA 868C (■) in a competitive radioligand binding assay using DP 1-transfected cells.

Detailed Description

To facilitate an understanding of the disclosure of the present invention, a number of terms are defined below.

As used herein, the singular forms "a," "an," and "the" may refer to a plurality of items unless otherwise specified. Generally, the nomenclature used herein and the laboratory procedures in organic chemistry, medicinal chemistry, and pharmacology described herein are those well known and commonly employed in the art. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Where there are multiple definitions of a term herein, the definitions used in this section should be used unless otherwise indicated.

The term "anti-solvent" refers to a liquid that is added to a solvent to reduce the solubility of a compound in the solvent, thereby causing the compound to precipitate.

The term "subject" refers to an animal, including, but not limited to, a primate (e.g., human), cow, sheep, goat, horse, dog, cat, rabbit, rat, or mouse. Typically, the terms "subject" and "patient" are used interchangeably herein to refer to, for example, a mammalian subject, particularly a human subject.

The term "treating" is intended to include alleviating or eliminating a condition or disease or one or more symptoms associated with the condition or disease; or to alleviate or eliminate the cause of the condition or disease itself.

The term "preventing" refers to a method of delaying or preventing the onset of a disease and/or its attendant symptoms, discouraging an individual from becoming ill, or reducing the risk of becoming ill in an individual.

The term "therapeutically effective amount" refers to an amount of a compound that, when administered, is sufficient to prevent or to reduce to some extent the progression of one or more symptoms of the condition or disease to be treated. The term "therapeutically effective amount" also refers to the amount of a compound that will elicit the biological or medical response of a tissue, system, animal or human that is desired by the researcher, veterinarian, medical doctor or clinician.

The term "pharmaceutically acceptable carrier", "pharmaceutically acceptable excipient", "physiologically acceptable carrier" or "physiologically acceptable excipient" refers to a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material. Each component must be "pharmaceutically acceptable" in the sense of being compatible with the other ingredients of the pharmaceutical formulation. It must also be suitable for use in contact with the tissues or organs of humans and animals without excessive toxicity, irritation, allergic response, immunogenicity, or other problems or complications, commensurate with a reasonable benefit/risk ratio.

The term "naturally-occurring" or "native" as used in reference to biological material (e.g., nucleic acid molecules, polypeptides, host cells, etc.) refers to material that is found in nature and has not been manipulated by man. Similarly, "non-naturally occurring" or "non-natural" refers to a material that is not found in nature or that has been structurally modified or synthesized by man.

The term "CRTH 2 refers to a CRTH2 receptor protein or variant thereof, which is capable of mediating PGD in vitro or in vivo₂The cellular response of (a). CRTH2 variants include proteins substantially homologous to native CRTH2, i.e., proteins having one or more naturally or non-naturally occurring amino acid deletions, insertions, or substitutions as compared to the amino acid sequence of native CRTH2 (e.g., CRTH2 derivatives, homologs, and fragments). The amino acid sequence of the CRTH2 variant has at least about 80%, at least about 90%, or at least about 95% identity to native CRTH 2.

The term "other PGD₂Receptor "refers to a prostanoid receptor protein other than CRTH2 or a variant thereof, which is capable of mediating PGD in vitro or in vivo₂The cellular response of (a). "other PGD₂Receptor for PGD₂(e.g., DP) or one or more other prostanoids. "other PGD₂Receptor "variants" include proteins that are substantially homologous to the corresponding native prostanoid receptor other than CRTH2, i.e., proteins having one or more deletions, insertions, or substitutions of naturally or non-naturally occurring amino acids (e.g., derivatives, homologs, and fragments of a native prostanoid receptor other than CRTH 2). Natural "other PGD₂Receptor variants having amino acid sequences corresponding to the natural PGD' s₂The receptor "has at least about 80%, at least about 90%, or at least about 95% identity.

The term "CRTH 2 antagonist" refers to, for example, a partial or complete block, reduction, prevention, inhibition, or downregulation of CRTH2 activity and/or one or more other PGDs₂A receptor active compound. The term "CRTH 2 antagonist" also refers to a compound that binds to CRTH2 or one or more other PGDs₂Compounds that bind to, delay activation of, inactivate or desensitize the receptor. CRTH2 antagonists may interfere with PGD₂With CRTH2 or one or more other PGDs₂The receptor interacts to produce an effect.

The terms "CRTH 2-mediated disorder, condition, or disease" and "CRTH 2-mediated disorder, condition, or disease" refer to a disorder, condition, or disease characterized by inappropriate (e.g., less than or greater than normal) CRTH2 activity. Inappropriate CRTH2 functional activity may cause cells that do not normally express CRTH2 to express CRTH2, increasing CRTH2 expression or intracellular activation, leading to, for example, inflammatory and immune-related disorders or diseases; or may cause a decrease in expression of CRTH 2. The disorder, condition or disease mediated by CRTH2 may be mediated, in whole or in part, by inappropriate CRTH2 activity. In particular, in a CRTH 2-mediated condition, disorder or disease, the disease is mediated by CRTH2 or one or more other PGDs₂Modulation of the receptor may have some effect on the underlying disorder or condition, e.g., a CRTH2 antagonist or agonist may result in some improvement in at least a portion of the patient being treated.

The term "tapped bulk density" refers to the bulk density in a densely packed state obtained with the aid of tapping, for example by repeatedly dropping a specimen-filled container from a predetermined height to provide a slight impact to the bottom of the container. Tapping bulk density can be measured by, for example, Powder Tester PT-D (hosokawa micron Corporation, osaka, japan).

The term "uniformity coefficient" refers to a parameter representing the uniformity of the particle size distribution of microparticles, and is defined as a value obtained by dividing the particle size of 60% by the particle size of 10% obtained from a particle distribution curve. The more uniform the particle size distribution, the closer the uniformity coefficient is to 1.

Microparticles

The present invention provides microparticles of 4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid in amorphous form or in crystalline form I or II. The particles of the acid of formula I can be characterized by a variety of methods known to those skilled in the art, including single crystal X-ray diffraction, X-ray powder diffraction (XRPD), microscopy (e.g., Scanning Electron Microscopy (SEM)), thermal analysis (e.g., Differential Scanning Calorimetry (DSC), thermogravimetric analysis (TGA), and hot stage microscopy), and spectroscopy (e.g., infrared, raman, solid state nuclear magnetic resonance). Particle size and size distribution can be determined by conventional methods, such as laser light scattering techniques. The purity of the microparticles of the acid of formula I can be determined by standard analytical methods, such as Thin Layer Chromatography (TLC), gel electrophoresis, gas chromatography, High Performance Liquid Chromatography (HPLC), and Mass Spectrometry (MS).

In one embodiment, the microparticles of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid are form I crystals. In one embodiment, the type I particles have an X-ray powder diffraction pattern substantially as shown in figure 1. In another embodiment, the particles of form I have characteristic XRP diffraction peaks at 2 θ angles of about 9.8, 13.1, 22.0, and 26.4 °. In another embodiment, the particles of form I have a characteristic XRP diffraction peak at a 2 θ angle of about 9.8, 13.1, 22.0, or 26.4 °. In another embodiment, the particles of form I have a characteristic XRP diffraction peak at a 2 Θ angle of about 22.0 °. In another embodiment, the microparticles of formula I have a differential scanning calorimetry thermogram substantially as shown in figure 2. In another embodiment, the particulates of form I have a DSC thermogram with an endotherm with a peak temperature of about 224 ℃ and an onset temperature of about 220 ℃. In another embodiment, the crystalline morphology of the form I particles is substantially needle-like (fig. 4). In another embodiment, the microparticles of form I have a water solubility of about 3 μ g/mL at room temperature.

In one embodiment, the microparticles of form I have an average particle size in the range of about 0.1 to about 150 μm, about 0.5 to about 100 μm, about 1 to about 50 μm, about 1 to about 25 μm, about 1 to about 20 μm, about 1 to about 10 μm, about 2 to about 10 μm, or about 2 to about 7.5 μm. In another embodiment, the microparticles of form I have a uniform particle size distribution, as determined by the uniformity coefficient, in the range of from about 1 to about 20, from about 1 to about 10, from about 2 to about 5, or from about 3 to about 4. In another embodiment, the microparticles of form I have a tapped bulk density in the range of about 0.1 to about 1.0, about 0.15 to 0.8, about 0.2 to about 0.6, about 0.25 to about 0.5, or about 0.3 to about 0.4 g/mL.

Microparticles having such particle characteristics can be directly prepared by the preparation method of the present invention. Alternatively, microparticles of this size may also be prepared using any conventional particle processing method (e.g., milling, micronization, or granulation). The microparticles of form I provided herein are suitable for the direct preparation of the acid of formula I as a therapeutic product, thereby simplifying the manufacturing process and eliminating the risks associated with conventional granulation processes, such as undesired polymorph conversion.

In certain embodiments, the particulates of the acid of formula I in form I may comprise no less than about 95%, no less than about 97%, no less than about 98%, no less than about 99%, or no less than about 99.5% by weight of the acid of formula I. The microparticles may also comprise no less than about 90%, no less than about 95%, no less than about 98%, no less than about 99%, or no less than about 99.5% by weight of said form I microparticles.

In certain embodiments, the particulates of the acid of formula I in form I have a residual organic solvent content of no greater than about 5%, no greater than about 2%, no greater than about 1%, no greater than about 0.9%, no greater than about 0.8%, no greater than about 0.7%, no greater than about 0.6%, no greater than about 0.5%, no greater than about 0.4%, no greater than about 0.3%, no greater than about 0.2%, no greater than about 0.1%, no greater than about 0.05%, or no greater than about 0.01% by weight.

In certain embodiments, the particulates of the acid of formula I in form I have a residual methanol content of no greater than about 100,000ppm, no greater than about 10,000ppm, no greater than about 5,000ppm, no greater than about 4,000ppm, no greater than about 3,000ppm, no greater than 2,000ppm, no greater than about 1,000ppm, no greater than about 500ppm, or no greater than about 100 ppm.

In another embodiment, the microparticles of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid are form II crystals. The form II particles have an X-ray powder diffraction pattern substantially as shown in figure 6. The particles of form II have a characteristic XRP diffraction peak at an angle 2 theta of about 31.7 deg..

In one embodiment, the microparticles of form II have an average particle size in the range of about 0.1 to about 150 μm, about 0.5 to about 100 μm, about 1 to about 50 μm, about 1 to about 25 μm, about 1 to about 20 μm, about 1 to about 10 μm, about 1 to about 5 μm, or about 2 to about 5 μm. In another embodiment, the microparticles of form II have a uniform particle size distribution, as determined by the uniformity coefficient, in the range of from about 1 to about 20, from about 1 to about 10, from about 2 to about 5, or from about 3 to about 4. In another embodiment, the microparticles of form II have a tapped bulk density in the range of about 0.1 to about 1.0, about 0.15 to 0.8, about 0.2 to about 0.6, about 0.25 to about 0.5, or about 0.3 to about 0.4 g/mL.

Microparticles having such particle characteristics can be directly prepared by the preparation method of the present invention. Alternatively, microparticles of this size may also be prepared by any conventional method (e.g., milling, micronization or granulation).

In certain embodiments, the particulates of the form II acid of formula I may comprise no less than about 95%, no less than about 97%, no less than about 98%, no less than about 99%, or no less than about 99.5% by weight of the acid of formula I. The microparticles may also comprise not less than about 90%, not less than about 95%, not less than about 98%, not less than about 99%, or not less than about 99.5% by weight of said type II microparticles.

In certain embodiments, the particulates of the acid of formula I in form II have a residual organic solvent content of no greater than about 5%, no greater than about 2%, no greater than about 1%, no greater than about 0.9%, no greater than about 0.8%, no greater than about 0.7%, no greater than about 0.6%, no greater than about 0.5%, no greater than about 0.4%, no greater than about 0.3%, no greater than about 0.2%, no greater than about 0.1%, no greater than about 0.05%, or no greater than about 0.01% by weight.

In another embodiment, the microparticles of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid are amorphous. The amorphous particles have an X-ray powder diffraction pattern substantially as shown in figure 9, which is free of characteristic XRP diffraction peaks of form I and/or form II particles. In one embodiment, the amorphous microparticles may comprise not less than about 95%, not less than about 97%, not less than about 98%, not less than about 99%, or not less than about 99.5% by weight of said acid of formula I. In another embodiment, the microparticles may further comprise not less than about 90%, not less than about 95%, not less than about 98%, not less than about 99%, or not less than about 99.5% by weight of the amorphous microparticles.

In certain embodiments, the amorphous particulates of the acid of formula I have a residual organic solvent content of no greater than about 5%, no greater than about 2%, no greater than about 1%, no greater than about 0.9%, no greater than 0.8%, no greater than about 0.7%, no greater than about 0.6%, no greater than about 0.5%, no greater than about 0.4%, no greater than about 0.3%, no greater than about 0.2%, no greater than about 0.1%, no greater than about 0.05%, or no greater than about 0.01% by weight.

It should be understood that the values of the peaks of the X-ray powder diffraction pattern may differ slightly from machine to machine or from sample to sample, and therefore the cited values should not be interpreted as absolute, but with the permissible variability, for example 0.1 °, recommended by the united states pharmacopeia (2007 edition, page 387) -389).

Preparation method

The invention also provides a process for the preparation of microparticles of said acid of formula I in amorphous form, in crystalline form I or II. The method comprises the step of contacting the acid of formula I with a solvent, wherein particles of the acid of formula I in amorphous form or in crystalline form I or II can be formed from a solution or can be converted from one solid form to another solid form. The method may further comprise a separation step wherein the microparticles are separated by conventional means, such as filtration and centrifugation followed by washing with a solvent followed by drying (e.g., vacuum oven drying, air drying or desiccant drying).

Suitable solvents for preparing microparticles in amorphous form or in crystalline form I or II include, but are not limited to: hydrocarbons including petroleum ether, pentane, hexane, heptane, octane, isooctane, cyclopentane, cyclohexane, methylcyclohexane, benzene, toluene, xylene, tetrahydronaphthalene, and cumene; chlorinated hydrocarbons including Dichloromethane (DCM), 1, 2-dichloroethane, 1-dichloroethylene, 1, 2-dichloroethylene, chloroform, trichloroethane, trichloroethylene, carbon tetrachloride, chlorobenzene, and trifluoromethylbenzene; alcohols including methanol, ethanol, Isopropanol (IPA), 1-propanol, 1-butanol, 2-butanol, t-butanol, 3-methyl-1-butanol, 1-pentanol, 2-methoxyethanol, 2-ethoxyethanol, and ethylene glycol; ethers including diethyl ether, diisopropyl ether, methyl tert-butyl ether (MTBE), diphenyl ether, 1, 2-dimethoxyethane, bis (2-methoxyethyl) ether, 1-dimethoxymethane, 2-dimethoxypropane, and anisole; ketones, including acetone, butanone, Methyl Ethyl Ketone (MEK), methyl isopropyl ketone, methyl butyl ketone, and methyl isobutyl ketone (MIBK); esters including methyl acetate, ethyl formate, ethyl acetate, propyl acetate, isopropyl acetate, isobutyl acetate and butyl acetate; carbonates, including ethylene carbonate and propylene carbonate; amides, including formamide, N-Dimethylformamide (DMF), and N, N-dimethylacetamide; nitriles, including Acetonitrile (ACN); sulfoxides, such as dimethyl sulfoxide (DMSO); sulfones, such as sulfolane; nitro compounds such as nitromethane and nitrobenzene; heterocycles, such as N-methylpyrrolidone, 2-methyltetrahydrofuran, Tetrahydrofuran (THF), dioxane and pyridine; carboxylic acids such as acetic acid, tetrachloroacetic acid and trifluoroacetic acid; phosphoramides, such as hexamethylphosphoramide; carbon disulfide; water; and mixtures thereof.

The microparticles of the acid of formula I in crystalline form I can be prepared from a solution or slurry of the acid of formula I in a solvent by conventional methods including, but not limited to: cooling, freezing, solvent evaporation or addition of an anti-solvent.

In one embodiment, the method of making the microparticles of form I comprises the steps of: (a) preparing a solution of the acid of formula I in a solvent at a first temperature; and (b) forming the microparticles of form I at a second temperature. To accelerate the formation of microparticles of form I, the method may further comprise a seeding step prior to step (b) of seeding said solution with said form I crystals. The method further comprises the separation step according to the invention.

The solution may be prepared from any form of the acid of formula I, including but not limited to an oil, a semi-solid, a solid (e.g., amorphous form or form I or II), or a mixture thereof. The solution of step (a) may be prepared as a saturated or near saturated solution at a first temperature. The saturated or near saturated solution may be prepared by dissolving a sufficient amount of the acid of formula I in the solvent at a temperature above the first temperature such that, when the solution is cooled to the first temperature, a saturated or near saturated solution is obtained. The sufficient amount of the acid of formula I can be estimated based on the solubility of the microparticles of form I in the solvent at the first temperature, which can be determined by methods known to those skilled in the art.

The first temperature may range from room temperature to the boiling point of the solvent, for example, from about 20 to about 200 ℃, from about 20 to about 150 ℃, or from about 20 to about 100 ℃. The second temperature may range from about-100 to about 100 ℃, about-50 to about 50 ℃, about-10 to about 30 ℃, about 20 to about 200 ℃, about 20 to about 150 ℃, or about 20 to about 100 ℃. The first temperature may be higher or lower than or equal to the second temperature. The second temperature is typically set lower than the first temperature in order to maximize the throughput and efficiency of the process.

In one embodiment, the microparticles of form I can be formed by evaporating the solvent from the solution at a second temperature. Evaporation of the solvent may be facilitated by heating the solution and/or subjecting the solution to a vacuum. In one embodiment, the solvent is acetonitrile, dichloromethane, DMF, 1, 4-dioxane, methanol, 2-methoxyethanol, MIBK, acetone, 1-butanol, MTBE, DMSO, ethanol, ethyl acetate, isobutyl acetate, isopropyl acetate, 1-propanol, IPA, MEK, THF, or a mixture thereof.

In another embodiment, the particulates of form I can be formed by cooling the solution to the second temperature. In this case, the second temperature is set to be lower than the first temperature. In one embodiment, the solvent is DMF, 1, 4-dioxane, methanol, 2-methoxyethanol, 1-butanol, 1-propanol, IPA, MEK, THF, or a mixture thereof.

In another embodiment, the microparticles of form I can be formed by adding an anti-solvent to the solution at the second temperature.

Suitable anti-solvents include, but are not limited to: hydrocarbons including petroleum ether, pentane, hexane, heptane, octane, isooctane, cyclopentane, cyclohexane, methylcyclohexane, benzene, toluene, xylene, tetrahydronaphthalene, and cumene; chlorinated hydrocarbons including Dichloromethane (DCM), 1, 2-dichloroethane, 1-dichloroethylene, 1, 2-dichloroethylene, chloroform, trichloroethane, trichloroethylene, carbon tetrachloride, chlorobenzene and trifluoromethylbenzene; alcohols including isopropanol (EPA), 1-propanol, 1-butanol, 2-butanol, tert-butanol, 3-methyl-1-butanol, 1-pentanol, 2-ethoxyethanol, and ethylene glycol; ethers including diethyl ether, diisopropyl ether, methyl tert-butyl ether (MTBE), diphenyl ether, 1, 2-dimethoxyethane, bis (2-methoxyethyl) ether, 1-dimethoxymethane, 2-dimethoxypropane and anisole; ketones, including butanone, methyl isopropyl ketone, methyl butyl ketone, and methyl isobutyl ketone (MIBK); esters, including methyl acetate, ethyl formate, ethyl acetate, propyl acetate, isopropyl acetate, isobutyl acetate, and butyl acetate; carbonates, including ethylene carbonate and propylene carbonate; nitro compounds including nitromethane and nitrobenzene; heterocycles; carbon disulfide; water; and mixtures thereof.

When two solvents are used as a solvent/anti-solvent pair, the acid of formula I has a higher solubility in the solvent than in the anti-solvent. Optionally, the solvent and the anti-solvent in the solvent/anti-solvent pair are at least partially miscible. In one embodiment, the solvent is THF, methoxyethanol, DMSO, DMF, or a mixture thereof; and the anti-solvent is hexane, MTBE, toluene water or a mixture thereof. In another embodiment, the solvent/anti-solvent pair is THF/hexane, THF/water, DME/MTBE, or DMF/water.

In another embodiment, the microparticles of form I can be formed by adding the solution to an anti-solvent at a second temperature. In one embodiment, the solvent is THF, methoxyethanol, DMSO, DMF, or a mixture thereof; and the anti-solvent is hexane, MTBE, toluene, isopropyl acetate, water, or a mixture thereof. In another embodiment, the solvent/anti-solvent pair is THF/hexane, THF/water, 2-methoxyethanol/hexane, DMSO/water, or DMF/water.

In another embodiment, the method of making the microparticles of form I comprises the steps of: (a) preparing a slurry of an acid of formula I in a solvent at a first temperature; and (b) forming the particulates of form I by exposing the slurry to a second temperature. The slurry may be prepared from any form of the acid of formula I, including, but not limited to, an oil, a semi-solid, a solid (e.g., amorphous form or form I or II), or a mixture thereof. The method further comprises the inoculation step and/or the separation step described herein.

The first and second temperatures and the solvent are as defined herein. In one embodiment, the solvent is acetonitrile, chloroform, dichloromethane, DMF, 1, 4-dioxane, methanol, 2-methoxyethanol, MIBK, toluene, hexane, acetone, 1-butanol, MTBE, DMSO, ethanol, ethyl acetate, ethyl formate, heptane, isobutyl acetate, isopropyl acetate, 1-propanol, IPA, MEK, THF, water, or a mixture thereof.

In another embodiment, the method of making the microparticles of form I comprises the steps of: (a) preparing a solution of the acid of formula I in a solvent at a first temperature; (b) forming a slurry by cooling the solution to a second temperature; and (c) generating the particulates of form I by exposing the slurry to one or more heating and cooling cycles. The method further comprises the inoculation step and/or the separation step described herein.

The first and second temperatures and the solvent are as defined herein. In one embodiment, the solvent is acetonitrile, 1, 4-dioxane, 2-methoxyethanol, MIBK, acetone, 1-butanol, ethanol, 1-propanol, IPA, MEK, THF, water/THF (3: 7, v/v) or a mixture thereof. The heating and cooling cycle is conducted at a temperature in the range of from about-50 to about 120 deg.C, from about-50 to about 100 deg.C, from about-20 to about 80 deg.C, from about 0 to about 80 deg.C, from about 10 to about 80 deg.C, from 20 to about 80 deg.C, from about 20 to about 60 deg.C, or from about 20 to about 50 deg.C.

The microparticles of the acid of formula I in crystalline form II can be prepared from a solution or slurry of the acid of formula I in a solvent by conventional methods including, but not limited to: cooling, freezing, solvent evaporation or addition of an anti-solvent.

In one embodiment, the method of making the microparticles of form II comprises the steps of: (a) preparing a solution of the acid of formula I in a solvent at a first temperature; and (b) forming the microparticles of form II at a second temperature. To accelerate the formation of microparticles of form II, the method may further comprise a seeding step of seeding the solution with the form II crystals prior to step (b). The method further comprises the separation step according to the invention.

The solution may be prepared from any form of the acid of formula I, including but not limited to an oil, a semi-solid, a solid (e.g., amorphous form or form I or II), or a mixture thereof. The solution of step (a) may be prepared as a saturated or near saturated solution at a first temperature. The saturated or near saturated solution may be prepared by dissolving a sufficient amount of said acid of formula I in said solvent at a temperature above a first temperature, such that, upon cooling of the solution to the first temperature, the saturated or near saturated solution is obtained. The sufficient amount of the acid of formula I can be estimated based on the solubility of the type II particles in the solvent at the first temperature, which can be determined by methods known to those skilled in the art.

In one embodiment, the microparticles of form II can be formed by evaporating the solvent from the solution at the second temperature. Solvent evaporation may be facilitated by heating the solution and/or subjecting the solution to a vacuum. In one embodiment, the solvent is DMF, 1, 4-dioxane, methanol, 2-methoxyethanol, 1-butanol, 1-propanol, IPA, MEK, THF or mixtures thereof.

In another embodiment, the microparticles of form II can be formed by cooling the solution to a second temperature.

In another embodiment, the microparticles of form II can be formed by adding an anti-solvent to the solution at a second temperature. In one embodiment, the solvent is THF, methoxyethanol, DMSO, DMF, or a mixture thereof; and the anti-solvent is hexane, MTBE, toluene water or a mixture thereof.

In another embodiment, the microparticles of form II can be formed by adding the solution to an anti-solvent at a second temperature. In one embodiment, the solvent is THF, methoxyethanol, DMSO, DMF, and mixtures thereof; and the anti-solvent is hexane, MTBE, toluene water and mixtures thereof. In another embodiment, the solvent/anti-solvent pair is DMF/MTBE.

In another embodiment, the method of making the microparticles of form II comprises the steps of: (a) preparing a slurry of the acid of formula I in a solvent at a first temperature; and (b) forming the particulates of form II by exposing the slurry to a second temperature. The slurry may be prepared from any form of the acid of formula I including, but not limited to, an oil, a semi-solid, a solid (e.g., amorphous form or form I or II), or a mixture thereof. The method further comprises the inoculation step and/or the separation step described herein.

The first and second temperatures and the solvent are as defined herein. In one embodiment, the solvent is acetonitrile, chloroform, dichloromethane, DMF, 1, 4-dioxane, methanol, 2-methoxyethanol, MIBK, toluene, hexane, acetone, 1-butanol, MTBE, DMSO, ethanol, ethyl acetate, ethyl formate, heptane, isobutyl acetate, isopropyl acetate, 1-propanol, IPA, MEK, THF, water, or a mixture thereof.

The microparticles of the amorphous acid of formula I can be prepared from a solution or slurry of the acid of formula I in a solvent by conventional methods including, but not limited to: cooling, freezing, solvent evaporation or addition of an anti-solvent.

In one embodiment, the method of making the amorphous microparticles of the acid of formula I comprises the steps of: (a) preparing a solution of the acid of formula I in a solvent at a first temperature; and (b) forming the amorphous microparticles at a second temperature. The method may further comprise a separation step as described herein.

The solution may be prepared from any form of the acid of formula I including, but not limited to, an oil, a semi-solid, a solid (e.g., amorphous form or form I or II), or a mixture thereof. The solution of step (a) may be prepared as a saturated or near saturated solution at a first temperature. The saturated or near saturated solution may be prepared by dissolving a sufficient amount of said acid of formula I in said solvent at a temperature above a first temperature such that, when the solution is cooled to the first temperature, the saturated or near saturated solution is obtained. The sufficient amount of the acid of formula I can be estimated based on the solubility of the amorphous microparticles in the solvent at the first temperature, which can be determined by methods known to those skilled in the art.

In one embodiment, the amorphous microparticles may be formed by evaporating a solvent from the solution at a second temperature. Solvent evaporation may be facilitated by heating the solution and/or subjecting the solution to a vacuum. In one embodiment, the solvent is ethyl formate, isobutyl acetate, MTBE or a mixture thereof.

In another embodiment, the amorphous particulates may be formed by cooling the solution to a second temperature. In one embodiment, the solvent is MEK.

In another embodiment, the amorphous microparticles may be formed by adding an anti-solvent to the solution at a second temperature. In one embodiment, the solvent is THF, methoxyethanol, DMSO, DMF, or a mixture thereof; and the anti-solvent is hexane, MTBE, toluene, water or a mixture thereof. In another embodiment, the solvent/anti-solvent pair is THF/hexane or DMSO/water.

In another embodiment, the amorphous microparticles may be formed by adding the solution to an anti-solvent at a second temperature. In one embodiment, the solvent is THF, methoxyethanol, DMSO, DMF, or a mixture thereof; and the anti-solvent is hexane, MTBE, toluene water or a mixture thereof. In another embodiment, the solvent/anti-solvent pair is THF/hexane or DMSO/water.

In another embodiment, the method of making the microparticles of the amorphous acid of formula I comprises the steps of: (a) preparing a slurry of the acid of formula I in a solvent at a first temperature; and (b) forming the amorphous microparticles by phase inversion at a second temperature. The slurry may be prepared from any form of the acid of formula I, including but not limited to an oil, a semi-solid, a solid (e.g., amorphous form or form I or II), or a mixture thereof. The first and second temperatures and the solvent are as defined herein. In one embodiment, the solvent is chloroform, hexane, MTBE, ethyl formate, heptane, IPA, water or mixtures thereof.

Other particle forming methods may also be used to prepare the particles of the acid of formula I in amorphous form or crystalline form I or II, including spray drying, drum drying, freeze drying and melt crystallization.

Pharmaceutical composition

The invention also provides a pharmaceutical composition comprising as an active pharmaceutical ingredient microparticles of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid, or a pharmaceutically acceptable hydrate or solvate thereof, in amorphous form or in crystalline form I or II, together with one or more pharmaceutically acceptable carriers or excipients. The choice of excipients depends to a large extent on factors such as the particular mode of administration, the influence of the excipients on the solubility and stability of the active ingredient, and the nature of the dosage form.

The pharmaceutical compositions provided herein can be provided as a unit dosage form or a multiple dosage form. Unit dosage forms, as used herein, refer to physically discrete units known in the art that are suitable for administration to individual human and animal subjects and are packaged separately. Each unit dosage form contains a predetermined amount of the active ingredient sufficient to produce the desired therapeutic effect, in association with the required pharmaceutical carrier or excipient. Examples of unit dosage forms include ampoules and syringes and individually packaged tablets and capsules. The unit dosage form may be administered in portions or in multiple portions. Multiple dosage forms are a plurality of identical unit dosage forms packaged in a single container for administration as separate unit dosage forms. Exemplary multiple dosage forms include vials, bottles of tablets or capsules, or pints or gallon bottles.

The microparticles of said acid of formula I provided herein can be administered alone or in combination with one or more other compounds, one or more other active ingredients provided herein. The pharmaceutical compositions provided by the present invention can be formulated in a variety of dosage forms for oral, parenteral, and topical administration. The pharmaceutical compositions may also be formulated as modified release dosage forms, including delayed, extended, prolonged, sustained, pulsatile, controlled, accelerated and rapid, targeted, programmed release, and gastric retention. Such dosage forms can be prepared by conventional methods and techniques known to those skilled in the art (see Remington: the Science and Practice of Pharmacy, supra; Modified-Release Drug delivery technology, Cathbone et al, Drugs and the Pharmaceutical Science, Marcel Dekker, Inc.: New York, NY, 2002; Vol. 126).

The pharmaceutical compositions provided herein may be administered once or multiple times at intervals. It will be understood that the exact dosage and treatment time may vary with the age, weight and condition of the patient to be treated, and may be determined empirically using known test protocols or may be inferred from in vivo or in vitro test or diagnostic data. It will be further understood that for a particular individual, the particular dosage regimen will be adjusted over time according to the individual requirements and the professional judgment of the person administering or instructing the administration of the dosage form.

A. Oral administration

The pharmaceutical compositions provided herein can be provided as solid, semi-solid, or liquid dosage forms for oral administration. Oral administration as used herein also includes buccal, lingual and sublingual administration. Suitable oral dosage forms include, but are not limited to: tablets, capsules, pills, lozenges, troches, pastilles, cachets, bolus, medicated chewing gums, granules, bulk powders, effervescent or non-effervescent powders or granules, solutions, emulsions, suspensions, solutions, wafers, sprays, elixirs, and syrups. In addition to the active ingredient, the pharmaceutical composition may also include one or more pharmaceutically acceptable carriers or excipients, including, but not limited to, binding agents, fillers, diluents, disintegrants, wetting agents, lubricants, glidants, coloring agents, dye migration inhibitors, sweetening agents, and flavoring agents.

The binding agent and granulating agent bind the tablets together to ensure that the tablets remain intact after compression. Suitable binders or granulating agents include, but are not limited to: starches, such as corn STARCH, potato STARCH, and pre-milled STARCH (e.g., STARCH 1500); gelatin; sugars such as sucrose, glucose, dextran, molasses, and lactose; natural and synthetic gums such as acacia, alginic acid, alginates, Irish moss extract, Panwar gum, ghatti gum, elsinan shell gum, carboxymethylcellulose, methylcellulose, polyvinylpyrrolidone (PVP), magnesium aluminum silicate, larch arabinogalactans, powdered tragacanth, and guar gum; celluloses such as ethyl cellulose, cellulose acetate, calcium carboxymethyl cellulose, sodium carboxymethyl cellulose, methyl cellulose, hydroxyethyl cellulose (HEC), hydroxypropyl cellulose (HPC), hydroxypropyl methyl cellulose (HPMC); microcrystalline celluloses, such as AVICEL-PH-101, AVICEL-PH-103, AVICELRC-581, AVICEL-PH-105(FMC Corp., Marcushook, Pa.); and mixtures thereof. Suitable fillers include, but are not limited to, talc, calcium carbonate, microcrystalline cellulose, powdered cellulose, dextran, kaolin, mannitol, silicic acid, sorbose, starch, pregelatinized starch, and mixtures thereof. The binding agent and filler may be present in an amount of about 50 to about 99% by weight of the pharmaceutical composition provided herein.

Suitable diluents include, but are not limited to, calcium phosphate, calcium sulfate, lactose, sorbitol, sucrose, inositol, cellulose, kaolin, mannitol, sodium chloride, dry starch, and powdered sugar. Certain diluents, such as mannitol, lactose, sorbitol, sucrose and inositol, when present in sufficient amounts, can enable the partially compressed tablet to disintegrate in the mouth by chewing. The compressed tablet can be used as a chewable tablet.

Suitable disintegrants include, but are not limited to: agar; soil removal; cellulose such as methyl cellulose and carboxymethyl cellulose; wood products; a natural sponge; a cation exchange resin; alginic acid; gums such as guar gum and magnesium aluminum silicate HV; citrus pulp; crosslinked cellulose, such as crosslinked carboxy methyl cellulose; crosslinked polymers, such as crospovidone; cross-linked starch; calcium carbonate; microcrystalline cellulose, such as sodium starch glycolate; potassium polycrystallin; starches, such as corn starch, potato starch, tapioca starch, and pregelatinized starch; clay; alginic acid (aligns); and mixtures thereof. The amount of disintegrant in the pharmaceutical compositions provided herein may vary with the type of formulation and can be readily identified by one of ordinary skill in the art. The pharmaceutical compositions provided herein may comprise from about 0.5 to about 15% or from about 1 to about 5% by weight of a disintegrant.

Suitable lubricants include, but are not limited to: calcium stearate; magnesium stearate; mineral oil; light mineral oil; glycerol; sorbitol; glycols, such as glyceryl behenate and polyethylene glycol (PEG); stearic acid; sodium laurate sulfate; talc; hydrogenated vegetable oils including peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, and soybean oil; zinc stearate; ethyl oleate; ethyl laurate; agar; starch; stone pine nuts; silica or silica gels, e.g.200(WG. Grace Co., Baltimore, MD) and CAB-O-(Cabot co., Boston, MA); and mixtures thereof. The pharmaceutical compositions provided herein may comprise from about 0.1 to about 5% by weight of a lubricant.

Suitable glidants include colloidal silicon dioxide, CAB-O-(Cabot co., Boston, MA) and asbestos-free talc. Colorants include any approved, certified, water-soluble FD&C dye and water insoluble FD suspended on hydrated alumina&C dyes and lakes and mixtures thereof. Lakes adsorb a water-soluble dye onto an aqueous heavy metal oxide, which forms an insoluble form of the dye. Flavoring agents include natural flavors extracted from plants such as fruits, and mixtures of synthetic compounds that produce a pleasant taste sensation, such as peppermint and methyl salicylate. Sweeteners include sucrose, lactose, mannitol, syrups, glycerol and artificial sweeteners, such as saccharin and aspartame. Suitable emulsifying agents include gelatin, acacia, tragacanth, bentonite and surfactants, for example polyoxyethylene sorbitol monooleate (I: (I) (II))20) Polyoxyethylene sorbitol monooleate (A), (B), (C)80) And triethanolamine oleate. Suspending and dispersing agents include sodium carboxymethylcellulose, pectin, tragacanth, magnesium aluminum silicate, acacia, sodium carboxymethylcellulose, hydroxypropyl methylcellulose, and polyvinylpyrrolidone. Preservatives include glycerol, methyl and propyl parabens, benzoic acid, sodium benzoate and ethanol. Wetting agents include propylene glycol monostearate, sorbitan monooleate, diethylene glycol monolaurate and polyoxyethylene lauric ether. The solvent comprises glycerol and fructus Pyri CalleryanaeAlcohol, ethanol and syrup. Examples of non-aqueous liquids for emulsions include mineral oil and cottonseed oil. Organic acids include citric acid and tartaric acid. Sources of carbon dioxide include sodium bicarbonate and sodium carbonate.

It will be appreciated that many carriers and excipients may serve multiple functions, even within the same formulation.

The pharmaceutical compositions provided by the present invention may be provided as compressed tablets, triturated tablets, chewable lozenges, fast dissolving tablets, double compressed tablets or enteric coated tablets, sugar coated or film coated tablets. Enteric-coated tablets are compressed tablets coated with a substance that is resistant to the action of gastric acid but dissolves or disintegrates in the intestine, thereby protecting the active ingredient from the acidic environment of the stomach. Enteric coatings include, but are not limited to, fatty acids, fats, phenyl salicylate, waxes, shellac, ammoniated shellac, and cellulose acetate phthalate. Sugar-coated tablets are compressed tablets coated with a sugar coating which helps to mask off-tastes or odors and prevent oxidation of the tablet. Film-coated tablets are compressed tablets coated with a thin layer or film of water-soluble substance. Film coatings include, but are not limited to, hydroxyethyl cellulose, sodium carboxymethyl cellulose, polyethylene glycol 4000, and cellulose acetate phthalate. Film coatings can bring about the same general characteristics as sugar coatings. A tabletted tablet is a compressed tablet prepared by more than one compression cycle, including layered tablets and compression-or dry-coated tablets.

The tablet dosage form may be prepared with the active ingredient in powdered, crystalline or granular form alone or in combination with one or more of the carriers or excipients described herein, including binders, disintegrants, controlled release polymers, lubricants, diluents and/or colorants. Flavoring and sweetening agents are particularly useful for forming chewable tablets and lozenges.

The pharmaceutical composition provided by the invention can be provided as a soft or hard capsule, and the soft or hard capsule can be prepared from gelatin, methylcellulose, starch or calcium alginate. The hard gelatin capsule, also known as a Dry Fill Capsule (DFC), consists of two parts, one sliding over the other, thereby completely encapsulating the active ingredient. Soft Elastic Capsules (SEC) are soft, spherical shells, such as gelatin shells, which are plasticized by the addition of glycerol, sorbitol or similar polyols. The soft gelatin shell may contain a preservative to prevent microbial growth. Suitable preservatives have been described herein and include methyl and propyl parabens, and sorbic acid. The liquid, semi-solid and solid dosage forms provided by the present invention can be encapsulated in a capsule. Suitable liquid and semi-solid dosage forms include solutions and suspensions in propylene carbonate, vegetable oils or triglycerides. Capsules containing such solutions were prepared as described in us patent 4,328,245, 4,409,239 and 4,410,545. The capsules may also be coated by means known to those skilled in the art to limit or maintain the dissolution of the active ingredient.

The pharmaceutical compositions provided herein can be provided in liquid and semi-solid dosage forms, including emulsions, solutions, suspensions, elixirs and syrups. An emulsion is a two-phase system in which one liquid is dispersed throughout another liquid in the form of fine particles, which may be oil-in-water or water-in-oil. The emulsion may comprise a pharmaceutically acceptable non-aqueous liquid or solvent, an emulsifier and a preservative. Suspensions may contain pharmaceutically acceptable suspending agents and preservatives. The aqueous alcoholic solution may comprise pharmaceutically acceptable acetals, for example di (lower alkyl) acetals of lower alkyl aldehydes (the term "lower" refers to alkyl groups having 1 to 6 carbon atoms), for example acetaldehyde diethyl acetal; and water-miscible solvents having one or more hydroxyl groups, such as propylene glycol and ethanol. Elixirs are clear, sweetened hydroalcoholic solutions. A syrup is a concentrated aqueous solution of a sugar (e.g., sucrose), and may also contain preservatives. For liquid dosage forms, for example, solutions in polyethylene glycol, can be diluted with a sufficient amount of a pharmaceutically acceptable liquid carrier (e.g., water) to facilitate dosing.

Other useful liquid and semi-solid dosage forms include, but are not limited to, dosage forms comprising the active ingredients provided herein, as well as dialkylated mono-or polyalkylene glycols, including 1, 2-dimethoxymethane, diglyme, triglyme, tetraglyme, polyethylene glycol-350-dimethyl ether, polyethylene glycol-550-dimethyl ether, polyethylene glycol-750-dimethyl ether (where 350, 550, and 750 refer to approximate average molecular weights of polyethylene glycols). These formulations may further comprise one or more antioxidants, such as Butylated Hydroxytoluene (BHT), Butylated Hydroxyanisole (BHA), propyl gallate, vitamin E, hydroquinone, hydroxycoumarins, ethanolamine, lecithin, cephalin, ascorbic acid, malic acid, sorbitol, phosphoric acid, bisulfite, sodium metabisulfite, thiodipropionic acid and its esters, and dithiocarbamates.

The pharmaceutical compositions provided by the present invention for oral administration may also be provided in the form of liposomes, micelles, microspheres or nanosystems. The micellar dosage form can be prepared as described in U.S. Pat. No. 6,350,458.

The pharmaceutical compositions provided herein may also be provided as non-effervescent or effervescent granules and powders to be reconstituted into liquid dosage forms. Pharmaceutically acceptable carriers and excipients for the non-effervescent granules or powders may include diluents, sweeteners and wetting agents. Pharmaceutically acceptable carriers and excipients for the effervescent granules or powders may include organic acids and a source of carbon dioxide.

Coloring and flavoring agents may be used in all of the above dosage forms.

The pharmaceutical compositions provided herein may be formulated as direct or modified release dosage forms, including delayed, sustained, pulsed, controlled, targeted, and programmed release forms.

The pharmaceutical compositions provided by the present invention may be formulated in combination with other active ingredients which do not impair the desired therapeutic effect or with substances which supplement the desired effect, such as antacids, proton pump inhibitors and H2-receptor antagonists.

B. Parenteral administration

The pharmaceutical compositions provided herein may also be administered by parenteral injection, infusion or infusion, for local or systemic administration. Parenteral administration as used herein includes intravenous, intraarterial, intraperitoneal, intravesicular, intraventricular, intraurethral, intrasternal, intracranial, intramuscular, intrasynovial and subcutaneous administration.

The pharmaceutical compositions provided herein may be formulated in any dosage form suitable for parenteral administration, including solutions, suspensions, emulsions, micelles, liposomes, microspheres, nanosystems and solid forms suitable for forming solutions or suspensions in liquids prior to injection. Such dosage forms can be prepared by conventional methods known to those skilled in The art of pharmaceutical Science (see Remington: The Science and practice of Pharmacy, supra).

Pharmaceutical compositions for parenteral administration may include one or more pharmaceutically acceptable carriers and excipients, including but not limited to aqueous vehicles, water-miscible vehicles, non-aqueous vehicles, antibacterial or preservative agents against microbial growth, stabilizers, co-solvents, isotonic agents, buffers, antioxidants, local anesthetics, suspending and dispersing agents, wetting or emulsifying agents, complexing agents, sequestering or chelating agents, cryoprotectants, lyoprotectants, thickening agents, pH adjusting agents, and inert gases.

Suitable aqueous carriers include, but are not limited to, water, saline, physiological saline or Phosphate Buffered Saline (PBS), sodium chloride injection, ringer's injection, isotonic dextran injection, sterile water injection, dextran, and lactated ringer's injection. Non-aqueous vehicles include, but are not limited to, fixed oils of vegetable origin, castor oil, corn oil, cottonseed oil, olive oil, peanut oil, peppermint oil, safflower oil, sesame oil, soybean oil, hydrogenated vegetable oils, hydrogenated soybean oil, and the medium chain triglycerides of coconut oil and palm seed oil. Water-miscible vehicles include, but are not limited to, ethanol, 1, 3-butanediol, liquid polyethylene glycols (e.g., polyethylene glycol 300 and polyethylene glycol 400), propylene glycol, glycerol, N-methyl-2-pyrrolidone, dimethylacetamide, and dimethylsulfoxide.

Suitable antimicrobial or preservative agents include, but are not limited to, phenol, cresol, mercuryFormulations, benzyl alcohol, chlorobutanol, methyl and propyl parabens, thimerosal, benzalkonium chloride, benzethonium chloride, methyl and propyl parabens, and sorbic acid. Suitable isotonic agents include, but are not limited to, sodium chloride, glycerol, and dextran. Suitable buffers include, but are not limited to, phosphate and citrate. Suitable antioxidants are described herein and include bisulfite and sodium metabisulfite. Suitable local anesthetics include, but are not limited to, procaine hydrochloride. Suitable suspending and dispersing agents, as described herein, include sodium carboxymethylcellulose, hydroxypropyl methylcellulose and polyvinylpyrrolidone. Suitable emulsifiers include those described herein, including polyoxyethylene sorbitol monolaurate, polyoxyethylene sorbitol monolaurate 80, and triethanolamine oleate. Suitable sequestering or chelating agents include, but are not limited to, EDTA. Suitable pH adjusters include, but are not limited to, sodium hydroxide, hydrochloric acid, citric acid, and lactic acid. Suitable complexing agents include, but are not limited to, cyclodextrins, including alpha-cyclodextrin, beta-cyclodextrin, hydroxypropyl-beta-cyclodextrin, sulfobutylether-beta-cyclodextrin, and sulfobutylether-7-beta-cyclodextrin(s) ((S))CyDex，Lenexa，KS)。

The pharmaceutical compositions provided herein can be formulated for single or multiple dose administration. Single dose formulations may be packaged in ampoules, vials or syringes. Multiple dose parenteral formulations must contain bacteriostatic or fungistatic concentrations of the antibacterial agent. As is known and practiced in the art, all parenteral formulations must be sterile.

In one embodiment, the pharmaceutical composition is provided as a ready-to-use sterile solution. In another embodiment, the pharmaceutical composition is provided as a sterile dry soluble product, including lyophilized powder and subcutaneous tablets, which can be reconstituted with a carrier prior to use. In another embodiment, the pharmaceutical composition is provided as a ready-to-use sterile suspension. In another embodiment, the pharmaceutical composition is provided as a sterile dry insoluble product that can be reconstituted with a carrier prior to use. In another embodiment, the pharmaceutical composition is provided as a ready-to-use sterile emulsion.

The pharmaceutical compositions provided herein may be formulated as direct or modified release dosage forms, including delayed, sustained, pulsed, controlled, targeted, and programmed release forms.

The pharmaceutical composition may be formulated as a suspension, solid, semi-solid, or thixotropic liquid for administration as an implant depot. In one embodiment, the pharmaceutical composition provided herein is dispersed in a solid phase internal matrix surrounded by an outer polymeric membrane that is insoluble in body fluids, but allows diffusion of the active ingredient in the pharmaceutical composition therethrough.

Suitable internal matrices include polymethylmethacrylate, polybutylacrylate, plasticized or unplasticized polyvinyl chloride, plasticized nylon, plasticized polyethoxylated terephthalate, natural rubber, polyisoprene, polyisobutylene, polybutadiene, polyethylene, ethylene vinyl acetate copolymers, silicone rubber, polydimethylsiloxane, carbonate rock silicone copolymers, hydrogels of hydrophilic polymers such as acrylates and methacrylates, collagen, crosslinked polyvinyl alcohol, and partially crosslinked hydrolyzed polyvinyl acetate.

Suitable outer polymeric films include polyethylene, polypropylene, ethylene/propylene copolymers, ethylene/ethyl acrylate copolymers, ethylene/vinyl acetate copolymers, silicone rubber, polydimethylsiloxane, neoprene, chlorinated polyethylene, polyvinyl chloride, vinyl chloride and vinyl acetate copolymers, vinylidene chloride, ethylene and propylene, ionic polyethylene terephthalate, butyl rubber, epichlorohydrin rubber, ethylene/vinyl alcohol copolymers, ethylene/vinyl acetate/vinyl alcohol terpolymers, and ethylene/ethyleneoxy-alcohol copolymers.

C. Topical application

The pharmaceutical compositions provided herein can be topically applied to the skin, the mouth, or the mucosa. Topical administration as used herein includes cutaneous (intra-cutaneous), conjunctival, intracorneal, intraocular, ocular, otic, transdermal, nasal, vaginal, urethral, respiratory and rectal administration.

The pharmaceutical compositions provided herein can be formulated in any dosage form suitable for topical application to produce a topical or systemic effect, including emulsions, solutions, suspensions, creams, gels, hydrogels, ointments, powders, dressings, elixirs, lotions, suspensions, tinctures, pastes, foams, films, aerosols, lavages, sprays, suppositories, bandages, skin patches. Topical formulations of the pharmaceutical compositions provided herein can also include liposomes, micelles, microspheres, nanosystems, and mixtures thereof.

Pharmaceutically acceptable carriers and excipients for use in the topical formulations provided herein include, but are not limited to, aqueous vehicles, water-miscible vehicles, non-aqueous vehicles, antimicrobial or preservative agents to combat microbial growth, stabilizers, co-solvents, isotonic agents, buffers, antioxidants, local anesthetics, suspending and dispersing agents, wetting or emulsifying agents, complexing agents, sequestering or chelating agents, permeation enhancers, cryoprotectants, lyoprotectants, thickening agents, and inert gases.

The pharmaceutical composition can also be administered by electroporation, iontophoresis, sonophoresis, and microneedle or needle-free injection such as POWDERJECT^TM(Chiron Corp., Emeryville, Calif.) and BIOJECT^TM(Bioject Medical Technologies Inc., Tualatin, OR) topical administration.

The pharmaceutical compositions provided by the present invention may also be provided in the form of ointments, creams and gels. Suitable ointment carriers include oily or hydrocarbon bases including, for example, lard benzoate, olive oil, cottonseed oil and other oils, white petrolatum; emulsifiable or absorbable bases such as hydrophilic petrolatum, hydroxystearic acid sulfate and anhydrous lanolin; water-removable bases, such as hydrophilic ointments; water-soluble ointment bases including polyethylene glycols of various molecular weights; an emulsifying base which is a water-in-oil (W/O) or oil-in-water (O/W) emulsion comprising cetyl alcohol, glyceryl monostearate, lanolin and stearic acid (see Remington: The Science and Practice of Pharmacy). These vehicles are emollients, but generally require the addition of antioxidants and preservatives.

Suitable creams may be oil-in-water or water-in-oil. Cream carriers may be water-washable, and may include an oil phase, an emulsifier, and an aqueous phase. The oil phase, also referred to as the "internal" phase, is typically composed of petrolatum and a fatty alcohol such as cetyl or stearyl alcohol. The aqueous phase typically (although not necessarily) exceeds the oil phase by volume and typically includes a humectant. The emulsifier in the cream formulation may be a nonionic, anionic, cationic or zwitterionic surfactant.

Gels are semi-solid suspension type systems. Single phase gels contain organic macromolecules distributed substantially uniformly in a liquid carrier. Suitable gelling agents include cross-linked acrylic acid polymers, such as carbomers, carboxypolyalkylene, polyethylene glycol,hydrophilic polymers such as polyethylene oxide, polyoxyethylene-polyoxypropylene copolymer, and polyvinyl alcohol; cellulosic polymers such as hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxypropyl methyl cellulose, phthalic acid and methyl cellulose; gums, such as tragacanth and xanthan gum; sodium alginate; and gelatin. To prepare a homogeneous gel, a dispersing agent such as an alcohol or glycerin may be added, or the gelling agent may be dispersed by grinding, mechanical mixing, and/or stirring.

The pharmaceutical compositions provided herein can be administered rectally, urethrally, vaginally, or perivaginally in the form of suppositories, pessaries, rods, poultices, or poultices, pastes, powders, dressings, creams, plasters, contraceptives, ointments, solutions, emulsions, suspensions, tampons, gels, foams, sprays, or enemas. These dosage forms may employ Remington: the conventional method described in The Science and practice of Pharmacy.

Rectal, urethral and vaginal suppositories are solid bodies inserted into body orifices, which are solid at ordinary temperatures but dissolve or soften at body temperature to release the active ingredient within the orifice. Pharmaceutically acceptable carriers that may be used in rectal and vaginal suppositories include carriers such as sclerosing agents which, when formulated with the pharmaceutical compositions provided herein, form a melting point around body temperature; and the antioxidant of the invention comprises bisulfite and sodium metabisulfite. Suitable carriers include, but are not limited to, cocoa butter (cocoa butter), glycerol-gelatin, carbowax (polyethylene glycol), sperm oil, paraffin, white and yellow waxes and suitable mixtures of mono-, di-and triglycerides of fatty acids, hydrogels, such as polyvinyl alcohol, hydroxyethyl methacrylate, polyacrylic acid; glycerol gelatin. Combinations of various carriers may be used. Rectal and vaginal suppositories may be prepared by compression or by molding. The typical weight of rectal and vaginal suppositories is 2-3 g.

The pharmaceutical compositions provided herein can be administered ocularly in the form of solutions, suspensions, ointments, creams, gel-forming solutions, powders for solutions, gels, ocular inserts and implants.

The pharmaceutical compositions provided herein can be administered to the respiratory tract by intranasal or inhalation. The pharmaceutical compositions may be provided in the form of an aerosol or solution for delivery using a pressurized container, pump, spray, nebulizer (e.g., one that utilizes electrohydrodynamic generation of a fine mist) or nebulizer, alone or in combination with a suitable propellant (e.g., 1, 1, 1, 2-tetrafluoroethane or 1, 1, 1, 2,3, 3, 3-heptafluoropropane). The pharmaceutical compositions may also be provided as dry powders for inhalation, and nasal drops, either alone or in combination with inert carriers (lactose or phospholipids). For intranasal use, the powder may comprise a bioadhesive agent, including chitosan or cyclodextrin.

The solutions or suspensions that can be used in the pressurized container, pump, sprayer, atomizer or nebulizer can be formulated to contain ethanol, aqueous ethanol or suitable alternative agents for dispersing, dissolving or prolonged release of the active ingredients provided herein, a propellant as a solvent; and/or surfactants such as sorbitan trioleate, oleic acid or oligomeric lactic acid.

The pharmaceutical compositions provided herein can be micronized to a suitable size for delivery by inhalation, for example 50 microns or less, or 10 microns or less. Particles of this size can be prepared by grinding methods known to those skilled in the art, including spiral jet milling, fluidized bed jet milling, supercritical fluid processing to form nanoparticles, high pressure homogenization, or spray drying.

Capsules, blisters and cartridges for use in an inhaler, insufflator may be formulated to contain a powder mix of a pharmaceutical composition provided by the present invention; suitable powder bases, such as lactose or starch; and performance modifiers, such as 1-leucine, mannitol, or magnesium stearate. The lactose may be anhydrous lactose or lactose monohydrate. Other suitable excipients include dextran, glucose, maltose, sorbitol, xylitol, fructose, sucrose and trehalose. The pharmaceutical compositions provided by the present invention for inhalation/intranasal administration may further comprise suitable flavouring agents such as menthol and levomenthol, or sweetening agents such as saccharin or saccharin sodium.

The pharmaceutical compositions provided herein for topical administration may be formulated as direct or modified release dosage forms, including delayed, sustained, pulsed, controlled, targeted, and programmed release forms.

D. Improved release

The pharmaceutical compositions provided herein can be formulated as modified release dosage forms. The term "modified release" as used herein refers to a dosage form in which the rate and location of release of the active ingredient is different from the immediate dosage form administered by the same route. Modified release dosage forms include delayed, extended, sustained, pulsatile or pulsed, controlled, accelerated or rapid, targeted, programmed release, and gastric retention dosage forms. The pharmaceutical compositions in modified release dosage forms may be prepared by a variety of modified release devices (modified release devices) and methods known to those skilled in the art, including but not limited to: matrix controlled release devices, osmotic controlled devices, multiparticulate controlled release devices, ion exchange resins, enteric coatings, multilayer coatings, microspheres, liposomes, and combinations thereof. The release rate of the active ingredient can also be modified by varying the particle size and polymorph of the active ingredient.

Examples of modified release include, but are not limited to, those described in U.S. Pat. nos. 3,845,770, 3,916,899, 3,536,809, 3,598,123, 4,008,719, 5,674,533, 5,059,595, 5,591,767, 5,120,548, 5,073,543, 5,639,476, 5,354,556, 5,639,480, 5,733,566, 5,739,108, 5,891,474, 5,922,356, 5,972,891, 5,980,945, 5,993,855, 6,045,830, 6,087,324, 6,113,943, 6,197,350, 6,248,363, 6,264,970, 6,267,981, 6,376,461, 6,419,961, 6,589,548, 6,613,358, and 6,699,500.

1. Matrix controlled release device

The pharmaceutical compositions provided herein in modified release dosage forms can be prepared using matrix controlled release devices known to those skilled in the art (see Takada et al in "Encyclopedia of controlled Drug Delivery," Vol.2, Mathiowitz, Inc., Wiley, 1999).

In one embodiment, the pharmaceutical compositions provided herein in a modified release dosage form may be formulated with an erodible matrix device that is a water-swellable, erodible or soluble polymer, including synthetic polymers and naturally occurring polymers and derivatives, such as polysaccharides and proteins.

Materials that can be used to form the erodible matrix include, but are not limited to: chitin, chitosan, dextran, and pullulan; agar gum, gum arabic, karaya gum, tragacanth gum, carrageenan, ghatti gum, guar gum, xanthan gum and scleroglucan; starches, such as dextrin and maltodextrin; hydrocolloids, such as pectin; phospholipids, such as lecithin; an alginate; algae (Saccharum sinensis Roxb.)Propylene glycol acid ester; gelatin; collagen; and cellulosic materials such as cellulose acetate (EC), methyl vinyl cellulose (MEC), carboxymethyl cellulose (CMC), CMEC, hydroxyethyl cellulose (HEC), hydroxypropyl cellulose (HPC), Cellulose Acetate (CA), Cellulose Propionate (CP), Cellulose Butyrate (CB), Cellulose Acetate Butyrate (CAB), CAP, CAT, hydroxypropyl methyl cellulose (HPMC), HPMCP, HPMCAS, hydroxypropyl methyl cellulose acetate trimellitate (HPMCAT), and ethyl hydroxyethyl cellulose (EHEC); polyvinylpyrrolidone; polyvinyl alcohol; polyvinyl acetate; glycerin fatty acid ester; polyacrylamide; polyacrylic acid; ethyl acrylic acid or methacrylic acid copolymer (Rohm America, inc., Piscataway, NJ); poly (2-hydroxyethyl-methacrylate); polylactic acid; copolymers of L-glutamic acid and ethyl-L-glutamine; degradable lactic acid-glycolic acid copolymers; poly-D- (-) -3-hydroxybutyric acid; and other acrylic acid derivatives such as butyl methacrylate, methyl methacrylate, ethyl acrylate, homopolymers and copolymers of (2-dimethylaminoethyl) methacrylate and (trimethylaminoethyl) chlorinated methacrylate.

In another embodiment, the pharmaceutical composition is formulated with a non-erodible matrix device. The active ingredient is dissolved or dispersed in an inert matrix and is released upon application primarily by diffusion through the inert matrix. Materials suitable for use as the non-erodable matrix means include, but are not limited to: insoluble plastics such as polyethylene, polypropylene, polyisoprene, polyisobutylene, polybutadiene, polymethyl methacrylate, polybutyl methacrylate, chlorinated polyethylene, polyvinyl chloride, methacrylate-methyl methacrylate copolymer, ethylene-vinyl acetate copolymer, ethylene/propylene copolymer, ethylene/ethyl acrylate copolymer, vinyl chloride-vinyl acetate copolymer, vinylidene chloride, ethylene and propylene, ionic polyethylene terephthalate, butyl rubber, epichlorohydrin rubber, ethylene/vinyl alcohol copolymer, ethylene/vinyl acetate/vinyl alcohol terpolymer, and ethylene/ethyleneoxy alcohol copolymer, polyvinyl chloride, plasticized nylon, plasticized polyvinyl alcohol terephthalate, natural rubber, silicone rubber, polydimethylsiloxane, polyisoprene, polybutadiene, styrene-acrylonitrile copolymer, Carbonate rock silicone copolymers, as well as hydrophilic polymers such as vinyl cellulose, cellulose acetate, crospovidone, and partially crosslinked hydrolyzed polyvinyl acetate; and fatty compounds such as mace wax, microcrystalline wax, and triglycerides.

In matrix controlled release systems, the desired release kinetics can be controlled, for example, by the type of polymer employed, the viscosity of the polymer, the particle size of the polymer and/or active ingredient, the ratio of active ingredient to polymer, and other excipients in the composition.

The pharmaceutical compositions provided herein in modified release dosage forms may be prepared by methods known to those skilled in the art including direct compression, compression after dry or wet granulation, compression after melt granulation.

2. Osmotic controlled release device

The pharmaceutical compositions provided by the present invention in controlled release dosage forms can be prepared using osmotic controlled release devices including single compartment systems, dual compartment systems, Asymmetric Membrane Technology (AMT) and Extruded Core Systems (ECS). Generally, such devices have at least two components: (a) a core comprising an active ingredient; and (b) a semipermeable membrane having at least one delivery end encapsulating the core. The semipermeable membrane controls the flow of water from the aqueous environment of use into the core, thereby releasing the drug by extrusion through the delivery tip.

In addition to the active ingredient, the core of the osmotic engine optionally contains an osmotic agent that provides a driving force for the transport of water from the environment of use to the core of the engine. One class of osmotic agents are water swellable hydrophilic polymers, also known as "osmopolymers" and "hydrogels," including but not limited to: hydrophilic vinyl and acrylic polymers, polysaccharides such as calcium alginate, polyethylene oxide (PEO), polyethylene glycol (PEG), polypropylene glycol (PPG), poly (2-hydroxyethyl methacrylate), poly (acrylic) acid, poly (methacrylic) acid, polyvinylpyrrolidone (PVP), cross-linked PVP, polyvinyl alcohol (PVA), PVA/PVP copolymers, copolymers of PVA/PVP with hydrophobic monomers such as methyl methacrylate and vinyl acetate, hydrophilic polyurethanes containing large PEO blocks, cross-linked sodium carboxymethylcellulose, carrageenan, hydroxyethyl cellulose (HEC), hydroxypropyl cellulose (HPC), hydroxypropyl methylcellulose (HPMC), carboxymethyl cellulose (CMC), and carboxyethyl cellulose (CEC), sodium alginate, polycarbophil, gelatin, xanthan gum, and sodium starch glycolate.

Other classes of osmotic agents are zymogens (osmogens) that are capable of imbibing water to form an osmotic pressure gradient across the peripheral coating barrier. Suitable zymogens include, but are not limited to: inorganic salts such as magnesium sulfate, magnesium chloride, calcium chloride, sodium chloride, lithium chloride, potassium sulfate, potassium phosphate, sodium carbonate, sodium sulfate, lithium sulfate, potassium chloride, and sodium sulfate; sugars such as dextran, fructose, glucose, inositol, lactose, maltose, mannitol, raffinose, sorbitol, sucrose, trehalose, and xylitol; organic acids such as ascorbic acid, benzoic acid, fumaric acid, citric acid, maleic acid, sebacic acid, sorbic acid, adipic acid, edetic acid, glutamic acid, p-toluenesulfonic acid, succinic acid and tartaric acid; urea; and mixtures thereof.

Different dissolution rates of the osmotic agent may be used to affect the initial rate of delivery of the active ingredient from the dosage form. For example, amorphous sugars, such as Mannogeme EZ (SPI Pharma, Lewes, DE), can be used to provide a more rapid delivery during the first hours to immediately produce the desired therapeutic effect and to gradually and continuously release the remaining amount over a longer period of time to maintain the desired level of therapeutic or prophylactic effect. In this case, the active ingredient is released at a rate that replaces the amount of active ingredient that is metabolized or excreted.

The core may also contain various other excipients and carriers as described herein to enhance the performance or to promote processing stability of the dosage form.

Materials that can be used to form the semipermeable membrane include various grades of acrylic, vinyl, ether, polyamide, polyester, and cellulosic derivatives that are water permeable and water insoluble at physiologically relevant phs or that are susceptible to being altered to water insoluble by chemical changes (e.g., crosslinking). Examples of suitable polymers that may be used to form the coating include plasticized, non-plasticized, and reinforced Cellulose Acetate (CA), cellulose diacetate, cellulose triacetate, CA propionate, cellulose nitrate, Cellulose Acetate Butyrate (CAB), CA ethylcarbamate, CAP, CA methylcarbamate, CA succinate, Cellulose Acetate Trimellitate (CAT), CA dimethylaminoacetate, CA ethylcarbonate, CA chloroacetate, CA oxalate, CA methyl sulfonate, CA butyl sulfonate, CA p-toluenesulfonate, agar acetate, amylose triacetate, beta glucan acetate, beta glucan triacetate, acetaldehyde methyl diacetate, triacetate of Carobu gum, hydroxylated ethylene vinyl acetate, EC, PEG, PPG, PEG/PPG copolymer, PVP, HEC, HPC, CMC, CMEC, HPMC, HPMCP, MCAS, HPMCAT, poly (propylene) acids and esters, and poly (methacrylic) acids and esters and copolymers thereof, Starches, dextrans, dextrins, chitosans, collagen, gelatin, polyolefins, polyethers, polysulfones, polyethersulfones, polystyrenes, polyvinyl halides, polyvinyl esters and ethers, natural and artificial waxes.

The semi-permeable membrane may also be a hydrophobic microporous membrane in which the pores are substantially gas filled and not wetted by aqueous media, but permeable to water vapor, as described in U.S. patent 5,798,119. Such hydrophobic but water vapor permeable membranes are typically composed of hydrophobic polymers such as polyolefins, polyethylene, polypropylene, polytetrafluoroethylene, polyacrylic derivatives, polyethers, polysulfones, polyethersulfones, polystyrenes, polyhaloethylenes, polyvinylidene fluorides, polyvinylesters and ethers, natural and artificial waxes.

The delivery port on the semi-permeable membrane may be formed by mechanical or laser drilling after coating. The delivery tip may also be formed in situ by etching a plug of water soluble material or by disrupting a thinner portion of the membrane over the breach in the core. Further, for asymmetric membrane coating types disclosed in U.S. Pat. Nos. 5,612,059 and 5,698,220, the transfer tips may be formed during the coating process.

The total amount of active ingredient released and the rate of release can be adjusted substantially by the thickness and porosity of the semi-permeable membrane, the composition of the core and the number, size and location of the delivery ends.

The pharmaceutical composition in an osmotic controlled release dosage form may further comprise additional conventional excipients as described herein to facilitate the performance or processing of the formulation.

The osmotic controlled Release dosage forms can be prepared by conventional methods and techniques known to those skilled in The art (see Remington: The Science and Practice of Pharmacy; Santus and Baker, J.Controled Release 1995, 35, 1-21; Verma et al, Drug Development and industry Pharmacy 2000, 26, 695-; Verma et al, J.Controled Release2002, 79, 7-27).

In certain embodiments, the pharmaceutical compositions provided herein are formulated as AMT controlled release dosage forms comprising an asymmetrically permeable membrane covering a core containing the active ingredient and other pharmaceutically acceptable excipients. See U.S. Pat. No. 5,612,059 and WO 2002/17918. The AMT controlled release dosage form may be prepared according to conventional methods and techniques known to those skilled in the art, including direct compression, dry granulation, wet granulation, and dip coating methods.

In certain embodiments, the pharmaceutical compositions provided herein are formulated as an ESC controlled release dosage form comprising a permeable membrane covering a core containing the active ingredient, hydroxyethylcellulose and other pharmaceutically acceptable excipients.

3. Multi-particle controlled release device

The pharmaceutical compositions provided by the present invention in modified release dosage forms can be prepared by a multiparticulate controlled release device comprising a plurality of particles, granules or pellets having a diameter ranging from about 10 μm to about 3mm, from about 50 μm to about 2.5mm or from about 100 μm to about 1 mm. Such a plurality of particles may be prepared by methods known to those skilled in the art, including by wet and dry granulation, extrusion/spheronization, roller compaction, melt-freezing and spray seed nucleation. See, for example, multiparticulate organic Drug Delivery; marcel Dekker: 1994 and Pharmaceutical pelletification technology, Marcel Dekker: 1989.

other excipients described herein may be mixed with the pharmaceutical composition to aid in the processing and formation of the multiparticulates. The resulting particles may themselves constitute the multi-particle device or may be coated with various film-forming materials, such as enteric polymers, water swellable and water soluble polymers. The multiparticulates can be further processed into capsules or tablets

4. Targeted delivery

The pharmaceutical compositions provided herein can also be formulated to target specific tissues, receptors or other areas of the body of the individual to be treated, including liposomes, re-encapsulated red blood cells and antibody-based delivery systems. Examples include, but are not limited to, U.S. patents 6,316,652, 6,274,552, 6,271,359, 6,253,872, 6,139,865, 6,131,570, 6,120,751, 6,071,495, 6,060,082, 6,048,736, 6,039,975, 6,004,534, 5,985,307, 5,972,366, 5,900,252, 5,840,674, 5,759,542, and 5,709,874.

Application method

In one embodiment, the invention provides methods for treating or preventing diseases caused by or susceptible to CRTH2 and/or one or more other PGDs₂A method of treating, preventing or ameliorating one or more symptoms of a receptor-associated disorder or disease in a subject by administering a therapeutically effective amount of microparticles of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid in amorphous form or in crystalline form I or II.

In another embodiment, the invention provides a method of treating, preventing or ameliorating CRTH2 and/or one or more other PGDs₂A method of modulating one or more symptoms of a disease or disorder responsive to a receptor, comprising administering to a subject suffering from or susceptible to such a disorderIndividuals suffering from this condition or disease are administered a therapeutically effective amount of microparticles of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl) acetic acid in amorphous form or in crystalline form I or II.

In another embodiment, the invention provides a method of treating, preventing or ameliorating CRTH2 and/or one or more other PGDs₂A method of one or more symptoms of a receptor-mediated disease or disorder comprising administering to an individual having or susceptible to the disorder or disease a therapeutically effective amount of microparticles of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl) acetic acid in amorphous form or in crystalline form I or II.

In another embodiment, the invention provides a method of treating, preventing or ameliorating one or more symptoms of a disease associated with eosinophils, comprising administering to an individual having or susceptible to the condition or disease a therapeutically effective amount of microparticles of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl) acetic acid in amorphous form or in crystalline form I or II.

In another embodiment, the invention provides a method of treating, preventing or ameliorating one or more symptoms of a disease associated with basophils, comprising administering to an individual having or susceptible to the disorder or disease a therapeutically effective amount of microparticles of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl) acetic acid in amorphous form or in crystalline form I or form II.

In another embodiment, the invention provides a method of treating, preventing or ameliorating one or more symptoms of an inflammatory disease, comprising administering to an individual having or susceptible to the disorder or disease a therapeutically effective amount of microparticles of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) -benzoylamino) benzyl) pyrimidin-5-yl } acetic acid in amorphous form or in crystalline form I or form II.

Conditions and diseases that can be treated by the microparticles provided herein include, but are not limited to: (1) inflammatory or allergic diseases including systemic anaphylaxis and hypersensitivity diseases, atopic dermatitis, urticaria, drug allergy, insect bite allergy, food allergy (including celiac disease, etc.), and mast cell disease; (2) inflammatory bowel diseases including Crohn's disease, ulcerative colitis, ileitis, and enteritis; (3) vasculitis and Behcet syndrome (Behcet syndrome); (4) psoriasis and inflammatory dermatoses including dermatitis, eczema, atopic dermatitis, allergic contact dermatitis, urticaria; including viral skin lesions from human papillomavirus, HIV or RLV infection; bacterial, fungal and other parasitic skin lesions and cutaneous lupus erythematosus; (5) asthma and allergic diseases of the respiratory tract including allergic asthma, exercise-induced asthma, allergic rhinitis, otitis media, allergic conjunctivitis, hypersensitivity lung disease and chronic obstructive pulmonary disease; (6) autoimmune diseases, including arthritis (including rheumatic and psoriasis), systemic lupus erythematosus, type I diabetes, myasthenia gravis, multiple sclerosis, Graves' disease, and glomerulonephritis; (7) transplant rejection (including allograft rejection and graft-versus-host disease) such as graft rejection, parenchymal organ transplant rejection, bone marrow transplant rejection; (8) fever is caused; (9) cardiovascular diseases including acute heart failure, hypotension, hypertension, angina pectoris, myocardial infarction, cardiomyopathy, congestive heart failure, arteriosclerosis, coronary artery disease, restenosis, and vascular stenosis; (10) cerebrovascular disorders including traumatic brain injury, stroke, ischemic reperfusion injury, and aneurysm; (11) cancers of the breast, skin, prostate, cervix, uterus, ovary, testis, bladder, lung, liver, larynx, oral cavity, colon, and gastrointestinal tract (e.g., esophagus, stomach, pancreas), brain, thyroid, blood, and lymphatic system; (12) fibrosis, connective tissue disease and sarcoidosis, (13) genital and reproductive disorders, including erectile dysfunction; (14) gastrointestinal disorders including gastritis, ulcers, nausea, pancreatitis and vomiting; (15) neurological diseases including alzheimer's disease; (16) sleep disorders including insomnia, narcolepsy, sleep apnea syndrome, and Pickwick syndrome; (17) pain; (18) renal disease; (19) ocular diseases including glaucoma; and (20) infectious diseases, including HIV.

In certain embodiments, the disease is selected from asthma, allergic asthma, exercise-induced asthma, allergic rhinitis, chronic allergic rhinitis, seasonal allergic rhinitis, atopic dermatitis, contact hypersensitivity, contact dermatitis, conjunctivitis, allergic conjunctivitis, eosinophilic bronchitis, food allergy, eosinophilic gastroenteritis, inflammatory bowel disease, ulcerative colitis, Crohn's disease, mastocytosis, hyper IgE syndrome, systemic lupus erythematosus, psoriasis, acne, multiple sclerosis, allograft rejection, reperfusion injury, chronic obstructive pulmonary disease, Churg-Strauss syndrome, sinusitis, basophilic leukemia, chronic urticaria, basophilic leukocytosis, psoriasis, eczema, COPD (chronic obstructive pulmonary disease), arthritis, rheumatoid arthritis, psoriatic arthritis and osteoarthritis.

In certain embodiments, the disease is asthma, exercise-induced asthma, allergic rhinitis, atopic dermatitis, chronic obstructive pulmonary disease, or allergic conjunctivitis.

In certain embodiments, the disease is Churg-Strauss syndrome or sinusitis.

The microparticles or compositions provided herein can be administered by oral, parenteral (e.g., intramuscular, intraperitoneal, intravenous, ICV, intracisternal injection or infusion, subcutaneous injection, or implant), inhalation, nasal, vaginal, rectal, sublingual, or topical (e.g., transdermal or topical) routes of administration, or formulated as suitable dosage units, either alone or in combination with pharmaceutically acceptable carriers, adjuvants, and vehicles appropriate for each route of administration, depending on the disease to be treated and the condition of the individual. Also provided are microparticles provided herein in the form of a depot, wherein the active ingredient is released over a predetermined period of time.

In asthma, allergic rhinitis, eczema, psoriasis, atopic dermatitis, fever, septicemia, systemic lupus erythematosusSores, diabetes, rheumatoid arthritis, multiple sclerosis, arteriosclerosis, transplant rejection, inflammatory bowel disease, cancer or other conditions associated with CRTH2 and/or one or more other PGDs₂In the treatment, prevention, or amelioration of a receptor-associated disorder, condition, or disease, suitable dosage levels will generally be from about 0.001 to about 100mg/kg of patient body weight per day (mg/kg per day), from about 0.01 to about 75mg/kg per day, from about 0.1 to about 50mg/kg per day, from about 0.5 to about 25mg/kg per day, or from about 1 to about 20mg/kg per day, which may be administered in single or multiple doses. Within this range, the dose may be 0.005-0.05, 0.05-0.5, or 0.5-5.0, 1-15, 1-20, or 1-50mg/kg per day. In certain embodiments, the dosage level is from about 0.001 to about 100mg/kg per day. In certain embodiments, the dosage level is from about 0.01 to about 75mg/kg per day. In certain embodiments, the dosage level is from about 0.1 to about 50mg/kg per day. In certain embodiments, the dosage level is from about 0.5 to about 25mg/kg per day. In certain embodiments, the dosage level is from about 1 to about 20mg/kg per day.

For oral administration, the pharmaceutical composition may be provided as a tablet containing 1.0-1,000mg of active ingredient, particularly about 1, about 5, about 10, about 15, about 20, about 25, about 50, about 75, about 100, about 150, about 200, about 250, about 300, about 400, about 500, about 600, about 750, about 800, about 900 and about 1,000mg of active ingredient to obtain a dose that modulates the symptoms of the patient to be treated. The composition may be administered on a regimen of 1-4 times per day, including once, twice, three times and four times per day.

It will be understood, however, that the specific dose level and frequency of dosage for a particular patient may be varied and will depend upon a variety of factors including the activity of the specific compound employed, the metabolic stability and length of action of that compound, the age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, drug combination, the severity of the particular condition, and the host undergoing therapy.

Also provided are methods of modulating CRTH2 and/or one or more other PGDs₂The method of receptor comprises the step of combining the receptor with the compound provided by the inventionOne or more solid forms of the particles are contacted. In one embodiment, the receptor is expressed by a cell.

The microparticles provided herein may also be combined or used in combination with other agents useful for treating, preventing or ameliorating one or more symptoms of a disease or condition for which the microparticles provided herein are useful (including asthma, allergic rhinitis, eczema, psoriasis, atopic dermatitis, fever, sepsis, systemic lupus erythematosus, diabetes, rheumatoid arthritis, multiple sclerosis, atherosclerosis, transplant rejection, inflammatory bowel disease, cancer, and the like).

Such other agents or drugs may be administered by their usual routes and amounts, either simultaneously or sequentially with the microparticles provided by the present invention. When the microparticles provided by the present invention are used simultaneously with one or more other drugs, a pharmaceutical composition containing such other drugs in addition to the microparticles provided by the present invention may be used, but this is not essential. Accordingly, the pharmaceutical compositions provided herein include pharmaceutical compositions that include one or more additional active ingredients or therapeutic agents in addition to the microparticles provided herein.

The weight ratio of the microparticles provided by the present invention to the second active ingredient can vary and depends on the effective dose of each ingredient. Generally, an effective dose of each ingredient is used. Thus, for example, when the microparticles provided herein are combined with an NSAID, the weight ratio of the microparticles to the NSAID can range from about 1000: 1 to about 1: 1000 or from about 200: 1 to about 1: 200. Combinations of microparticles with other active ingredients provided by the present invention are also generally within the aforementioned ranges, but in each case an effective dose of each active ingredient should be used.

Examples

The particles of the acid of formula I are characterized in the following examples by X-ray powder diffraction (XRPD), Differential Scanning Calorimetry (DSC), thermogravimetric analysis (TGA) and Scanning Electron Microscopy (SEM).

X-ray powder diffraction data were recorded using a Rigaku MiniFlex X-ray powder diffractometer (Rigaku America, The Woodlands, TX). The radiation was CuKa (40kV, 40 mA). Data were collected at room temperature over a 2 theta angle range of 3-45 degrees at 0.02 degrees per step and 0.6 seconds per step. The samples were prepared as thin layers of solvent-free powdered material on glass sample holders.

Differential scanning calorimetry was performed using Mettler 850, TA 2920. The samples were placed in sealed aluminum pans and analyzed with an empty aluminum pan as a reference. A heating rate of 10 ℃/min was used, with a temperature range from 30 ℃ to 280 ℃.

Thermogravimetric analysis was also performed on Mettler 850, TA 2920. The samples were placed in ceramic or aluminum sample trays. A heating rate of 20 ℃/min was used, with a temperature range from 20 ℃ to 600 ℃.

Example 1

{4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzoylamino) benzyl) pyrimidin-5-yl } acetic acid Solubility determination

A saturated solution of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) -benzoylamino) benzyl) pyrimidin-5-yl } acetic acid of formula I in the solvent listed in Table 1 can be prepared with this acid predominantly in form I at 25 and 50 deg.C simultaneously. The concentration of each saturated solution was determined by HPLC and the results are summarized in table 1.

Example 2

{ 4.6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzoylamino) benzyl) pyrimidin-5-yl } of formula I Production of acetic acid

A slurry of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid in methanol was stirred at room temperature overnight. The microparticles were then filtered, washed with hexane, and dried.

The X-ray powder diffraction pattern of the microparticles is shown in FIG. 1. The particles exhibited characteristic XRP diffraction peaks at about 9.8, 13.1, 22.0 and 26.4 ° 2 θ, indicating that the particles were crystalline. As shown in fig. 4, the particles are long needle crystals with a size of about 1 to about 5 μm.

The differential scanning calorimetry thermogram of the microparticles is shown in figure 2. The particles showed an endotherm with a peak temperature of about 223.8 ℃ and an onset temperature of about 220.2 ℃.

The thermogravimetric thermogram of the microparticle of formula I is shown in figure 3. The microparticles are very stable and show no significant weight loss before decomposition at about 200 ℃.

The average particle size and particle size distribution of the microparticles of formula I were determined using Microtrac S3000(Microtrac inc., Montgomeryville, PA). The results are shown in fig. 5 and summarized in table 2. The average particle size (MV) of the microparticles of formula I was 6.495 μm, with a uniformity coefficient of 3.24.

TABLE 1 solubility

TABLE 2 particle size distribution

Percentile	Particle size (. mu.m)
		10	1.807
20	2.393
		30	3.004
40	3.713
		50	4.627
60	5.852
		70	7.498
80	9.825
		90	13.70
95	17.57

Example 3

{4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzoylamino) benzyl) pyrimidine-5- Preparation of a phenyl acetic acid

A solution of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzoylamino) benzyl) pyrimidin-5-yl } acetic acid (50mg) in DMF (0.5mL) was prepared with the acid predominantly form I. The solution was added to 8ml of mtbe at room temperature. After stirring for a further 15 minutes at room temperature, the mixture became a slurry. The microparticles were then filtered, washed with hexane, and dried.

The X-ray powder diffraction pattern of the microparticles is shown in fig. 6. The particles showed a characteristic XRP diffraction peak at about 31.7 ° 2 θ, indicating that the particles are in a crystalline form other than said form I (form II). The crystal morphology of the form II fine particles is shown in fig. 7.

Example 4

Phase equilibrium

Based on the solubility of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid in different solvents as determined in example 1, microparticles of predominantly the acid of form I were exposed to the solvents listed in table 3 at 25 or 50 ℃ for a sufficient period of time. The microparticles were filtered and dried, then analyzed by XRPD. The results are summarized in table 3.

Example 5

Phase equilibrium between form I and form II

The form II particles were prepared as a slurry as described in example 2. Briefly, a solution of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzoylamino) benzyl) -pyrimidin-5-yl } acetic acid (50mg) in DMF (0.5mL) was added to 8mL MTBE to generate a slurry of microparticles of form II. Microparticles of form I (20mg) were then added to the slurry, and the resulting slurry was stirred at room temperature overnight. The microparticles were then filtered, washed with hexane, and dried. The microparticles were analyzed using XRPD. The XRP diffractogram is shown in figure 8, showing that the particles after phase equilibrium are form I.

Example 6

Formation of microparticles by solvent evaporation

Based on the solubility of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid in different solvents as determined in example 1, the acid, predominantly form I, was dissolved in the solvent system listed in table 4. The solvent is evaporated from the solution at 25 or 50 ℃. The microparticles were collected by filtration and analyzed using XRPD. The results are summarized in table 4.

TABLE 3 crystalline forms after phase equilibrium

a, NA: is not obtained. The sample produces any or a sufficient amount of solid.

Example 7

Forming microparticles by adding anti-solvent

Saturated solutions of the acid of formula I in the solvents listed in table 5 were prepared with the acid predominantly in form I. Then an anti-solvent is added to the saturated solution to induce precipitation. THF, 2-methoxyethanol, DMSO and DMF were used as solvents for dissolving the type I microparticles, while hexane, MTBE, toluene and water were used as anti-solvents. The microparticles were collected by filtration and analyzed using XRPD. The results are summarized in table 5.

Example 8

Forming microparticles by adding reverse antisolvent

Saturated or nearly saturated solutions of the acid of formula I in the solvents listed in table 6 were prepared with the acid predominantly in form I. The solutions are then each added to a larger volume of miscible anti-solvent. THF, 2-methoxyethanol, DMSO and DMF were used as solvents for dissolving the microparticles, while hexane, MTBE, toluene and water were used as anti-solvents. The microparticles were collected by filtration and analyzed using XRPD. The results are summarized in table 6.

Example 9

Formation of microparticles by Quench Cooling (Quench Cool)

A saturated solution of the acid of formula I in the solvent listed in table 7 was prepared with the acid predominantly in form I at about 30-50 ℃. The solution was quenched and cooled in an acetone/ice bath. The microparticles were collected by filtration and analyzed using XRPD. The results are summarized in table 7.

TABLE 4 crystalline forms formed by solvent evaporation

a, NA: is not obtained. The sample produces any or a sufficient amount of solid.

TABLE 5 crystalline forms formed by addition of antisolvent

Solvent(s)	Anti-solvent	Polymorphic substance
			THF	Hexane (C)	Type I

THF	MTBE	NAa
			THF	Toluene	NA
THF	Water (W)	Type I
			2-methoxyethanol	Hexane (C)	NA
2-methoxyethanol	MTBE	NA
			2-methoxyethanol	Toluene	NA
2-methoxyethanol	Water (W)	NA
			DMSO	Hexane (C)	NA
DMSO	MTBE	NA
			DMSO	Toluene	NA
DMSO	Water (W)	Mostly amorphous
			DMF	Hexane (C)	NA
DMF	MTBE	A small number (
			DMF	Toluene	NA
DMF	Water (W)	Type I

a, NA: is not obtained. The sample did not produce any or sufficient amount of solids.

Example 10

Formation of particulate crystals by heating and cooling cycles

A saturated solution of the acid of formula I in the solvent listed in table 8 was prepared with the acid predominantly in form I at about 30-50 ℃. The solution was slowly cooled using a programmed circulation bath to form a slurry. The resulting slurry was then heated to 50 ℃ over 2 hours and then cooled to 25 ℃ over 2 hours. The heating and cooling process was repeated overnight. The microparticles were then collected by filtration and analyzed using XRPD. The results are summarized in table 8.

TABLE 6 crystalline forms formed by addition of reverse antisolvent

Solvent(s)	Anti-solvent	Polymorphic substance
			THF(1mL)	Hexane (4mL)	Type I
THF(0.5mL)	MTBE(8mL)	NAa
			THF(0.5mL)	Toluene (8mL)	NA
THF(0.5mL)	Water (4mL)	Type I
			2-methoxyethanol (0.5mL)	Hexane (8mL)	Type I

2-methoxyethanol (0.5mL)	MTBE(8mL)	NA
			2-methoxyethanol (0.5mL)	Toluene (8mL)	NA
2-methoxyethanol (0.5mL)	Water (8mL)	NA
			DMSO(0.5mL)	Hexane (8mL)	NA
DMSO(0.5mL)	MTBE(8mL)	A small number (
			DMSO(0.5mL)	Toluene (8mL)	NA
DMSO(1mL)	Water (4mL)	Type I
			DMF(0.5mL)	Hexane (8mL)	NA
DMF(0.5mL)	MTBE(8mL)	Type II
			DMF(0.5mL)	Toluene (8mL)	NA
DMF(1mL)	Water (4mL)	Type I

a, NA: is not obtained. The sample did not produce any or sufficient amount of solids.

TABLE 7 Rapid Cooling crystalline forms

Solvent(s)	Polymorphic substance
		DMF	NAa
1, 4-dioxane	NA
		2-methoxyethanol	NA
Acetonitrile	Type I
		Methanol	Type I
N-butanol	NA
		1-propanol	Type I
Isopropanol (I-propanol)	NA
		MEK	Type I
THF	NA

a, NA: is not obtained. The sample produces any or a sufficient amount of solid.

TABLE 8 evaporative crystalline forms

Solvent(s)	Polymorphic substance
		Acetonitrile	Type I
1, 4-dioxane	Type I
		2-methoxyethanol	Type I
MIBK	Type I
		Acetone (II)	Type I
N-butanol	Type I
		Ethanol	Type I
1-propanol	Type I
		Isopropanol (I-propanol)	Type I
Water/THF (3: 7, v/v)	Type I

MEK	Type I
		THF	Type I

Example 11

Competitive radioligand binding assay

The binding affinity and selectivity of the amine salts of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid provided by the present invention were evaluated by a competitive radioligand binding assay using stably transfected cell lines expressing CRTH2 or DPI.

Prostaglandin D₂And 13, 14-dihydro-15-one PGD₂Obtained from Cayman Chemical (annorbor, MI). Radiolabeled PGD having a specific Activity of 160 Ci/mmol₂(5，6，8，9，12，14，15-³H (N)) from PerkinElmer (Boston, MA). Cell culture medium RPMI1640. HEPES buffer, Phosphate Buffered Saline (PBS), L-glutamine, and the avidin-streptomycin solution were obtained from Mediatech Inc (Herndon, VA). Fetal Calf Serum (FCS) was obtained from Cambrex (walker, MD). Puromycin was obtained from Invitrogen (San Diego, Calif.). Poly (ethylenimine) (PEI) was obtained from Acros Organics (Morris Plains, NJ). Purified monoclonal rat anti-human CRTH2 antibody (BM 16), rat IgG2 α, κ (clone R35-95) and goat anti-rat Ig-FITC were obtained from becton dickinson Biosciences (San Diego, CA). BSA (V moiety) and sodium azide were obtained from sigma chemical Company (st.

CRTH2 and DPI stable cell lines were generated as described by Sugimoto et al (J.Pharm.Exp.Therap.2003, 305, 347-352). These cell lines were maintained in RPMI 1640 medium containing HEPES buffer (25mM), FCS (10%), L-glutamine (2mM), penicillin (10IU/mL), streptomycin (100. mu.g/mL) and puromycin (1. mu.g/mL).

Surface expression of CRTH2 on transfected cell lines was monitored periodically during culture and prior to each competitive radioligand binding assay. CRTH2 expressing cells and untransfected cells were incubated on ice for 15 min with purified anti-CRTH 2 antibody (clone BM16) or isotype control antibody (rat IgG2 α, κ). Cells were then washed with 2mL FACS buffer (PBS with 1% BSA (V fraction) and 0.1% sodium azide). Cells were then incubated on ice with FITC-conjugated anti-rat antibody (goat anti-rat Ig-FITC). After washing with cold FACS buffer, cells were analyzed using a FACScan II analyzer and CellQuest software (both from Becton Dickinson (Mountain View, CA)).

Receptor transfectants may also function by, for example, Sugimoto et al, J.Pharm.Exp.Therap.2003, 305, 347-352; direct measurement of response to PGD as described by Sugimoto et al, Eur.J.Pharmacol.2005, 524, 30-37₂Calcium metabolism of (a).

Radioligand binding assays were performed as described by Sugimoto et al (J.Pharm.Exp.Therap.2003, 505, 347-352; Eur.J.Pharmacol.2005, 524, 30-37). Test compounds were dissolved in DMSO as a reservoir at a concentration of 100mMA solution is prepared. Serial dilutions were made in binding buffer at 10 μ M prior to assay. Transfected cells were plated at 4X 10 at room temperature⁶Resuspended in binding buffer (50mM Tris-HCl, 40mM MgCl) at a concentration of/mL₂And 0.1% bovine serum albumin, pH 7.4). Transfected cells were then seeded on U-bottom polypropylene 96-well plates (Fisher) by adding 50. mu.L of cell suspension, followed by 10. mu.L³H-PGD₂10 μ L of test compound solution or control, 30 μ L of binding buffer, to give a final volume of 100 μ L.³H-PGD₂The final concentration in each well was 1 nM. After incubation at room temperature for 1 hour with gentle shaking, the cell suspension was transferred to a filter plate (Millipore, MA) pre-wetted with 0.5% PEI buffer. The cell pellet was washed three times with binding buffer and the plate was allowed to air dry. Add scintillator (50. mu.L; Microscint) to each microwell^TM20, Perkin Elmer, Boston, MA) and radioactivity was measured on TopCount (PackardBioscience, Meriden, CT). Using Prizm^TMThe data was analyzed by a graphical program (graphpad software inc., San Diego, CA). As shown in fig. 10 and 11, form I {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid has high affinity and binding selectivity for CRTH 2.

The above examples are provided to fully disclose and describe to those of ordinary skill in the art how to make and use these embodiments, and are not intended to limit the scope of the present disclosure. Modifications of the above-described modes for carrying out the disclosure that are obvious to those skilled in the art are intended to be within the scope of the following claims. All publications, patents, and patent applications cited in this specification are herein incorporated by reference as if each individual publication, patent, or patent application were specifically and individually indicated to be incorporated by reference.

Claims (37)

1. Microparticles of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid of formula I in form I crystal form:

   wherein the microparticles have an average particle size of 0.1 to 150 μm; and is

   Wherein the form I crystalline form has an X-ray powder diffraction pattern substantially as shown in figure 1.
2. Microparticles of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid of formula I in form I crystal form:

   wherein the microparticles have a uniformity coefficient of 1 to 20; and is

   Wherein the form I crystalline form has an X-ray powder diffraction pattern substantially as shown in figure 1.
3. Microparticles of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid of formula I in form I crystal form:

   wherein the microparticles have a tapping bulk density of 0.1 to 1.0 g/mL; and is

   Wherein the form I crystalline form has an X-ray powder diffraction pattern substantially as shown in figure 1.
4. Microparticles of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid of formula I in form I crystal form:

   wherein the microparticles comprise not less than 95% by weight of an acid of formula I; and is

   Wherein the form I crystalline form has an X-ray powder diffraction pattern substantially as shown in figure 1.
5. Microparticles of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid of formula I in form I crystal form:

   wherein the microparticles have a residual organic solvent content of no greater than 5% by weight; and is

   Wherein the form I crystalline form has an X-ray powder diffraction pattern substantially as shown in figure 1.
6. Microparticles of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid of formula I in form I crystal form I:

   wherein the particles have a residual methanol content of no greater than 100,000 ppm; and is

   Wherein the form I crystalline form has an X-ray powder diffraction pattern substantially as shown in figure 1.
7. Microparticles of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid of formula I in form I crystal form I:

   wherein the microparticles comprise not less than 90% by weight of form I crystal form; and is

   Wherein the form I crystalline form has an X-ray powder diffraction pattern substantially as shown in figure 1.
8. 8. The microparticle of claim 2, wherein the microparticle has an average particle size of 0.1 to 150 μ ι η.
9. 9. The microparticle of claim 1, wherein the microparticle has a tapped bulk density in the range of 0.1 to 1.0 g/mL.
10. 10. The microparticle of claim 1, wherein the microparticle comprises not less than 95% by weight of the acid of formula I.
11. 11. The microparticle of claim 1, wherein the microparticle has a residual organic solvent content of no greater than 5% by weight.
12. 12. The microparticles of claim 1, wherein the microparticle has a residual methanol content of no greater than 100,000 ppm.
13. 13. The microparticle of claim 1, wherein the microparticle comprises not less than 90% by weight of the form I crystalline form.
14. 14. The microparticles of claim 1, wherein the microparticle has a differential scanning calorimetry thermogram as shown in figure 2.
15. 15. The microparticles of claim 1, wherein the microparticle has a differential scanning calorimetry thermogram with an endotherm having a peak temperature of 224 ℃ and an onset temperature of 220 ℃.
16. 16. The microparticles of claim 1, wherein the crystalline morphology of the form I crystal form is needle-like.
17. 17. The microparticle of claim 1, having a water solubility of 3 μ g/mL at room temperature.
18. 18. A pharmaceutical composition comprising the microparticle of claim 1 and a pharmaceutically acceptable carrier.
19. 19. The pharmaceutical composition of claim 18, wherein the composition is formulated for oral or topical administration.
20. 20. The pharmaceutical composition of claim 18, wherein the composition is formulated as a single dosage form.
21. 21. Use of the microparticles of claim 1 in the manufacture of a medicament for treating, preventing, or ameliorating one or more symptoms of a CRTH 2-mediated disease.
22. 22. Use of microparticles according to claim 1 in the preparation of a medicament for the treatment, prevention or amelioration of one or more symptoms of an eosinophil-related disease.
23. 23. Use of the microparticles of claim 1 in the manufacture of a medicament for treating, preventing, or ameliorating one or more symptoms of a basophil-related disease.
24. 24. Use of the microparticles of claim 1 in the manufacture of a medicament for treating, preventing, or ameliorating one or more symptoms of an inflammatory disease.
25. 25. The use according to claim 21, wherein the disease is selected from asthma, allergic rhinitis, atopic dermatitis, contact hypersensitivity, contact dermatitis, conjunctivitis, eosinophilic bronchitis, food allergy, inflammatory bowel disease, crohn's disease, mastocytosis, hyper IgE syndrome, systemic lupus erythematosus, psoriasis, acne, multiple sclerosis, allograft rejection, reperfusion injury, Churg-Strauss syndrome, sinusitis, basophilic leukemia, chronic urticaria, basophilic leukocytosis, eczema, COPD (chronic obstructive pulmonary disease) and arthritis.
26. 26. The use according to claim 21, wherein the disease is allergic asthma, exercise-induced asthma, allergic rhinitis, atopic dermatitis, chronic obstructive pulmonary disease or allergic conjunctivitis.
27. 27. The use according to claim 21, wherein the disease is Churg-Strauss syndrome or sinusitis.
28. 28. A method of making the microparticle of claim 1, comprising the steps of: (a) preparing a solution of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid in a solvent at a first temperature; and (b) evaporating said solvent from said solution at a second temperature to produce the microparticles; wherein the solvent is selected from acetonitrile, DMF, 1, 4-dioxane, methanol, 2-methoxyethanol, MIBK, acetone, DMSO, ethanol, ethyl acetate, isobutyl acetate, isopropyl acetate, 1-propanol, IPA, MEK and THF.
29. 29. The method of claim 28, wherein the second temperature is 25 or 50 ℃.
30. 30. A method of making the microparticle of claim 1, comprising the steps of: (a) preparing a solution of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid in a solvent at a first temperature; and (b) evaporating said solvent from said solution at a second temperature to produce the microparticles; wherein the solvent is selected from acetonitrile, DMF, 1, 4-dioxane, methanol, 2-methoxyethanol, MIBK, acetone, 1-butanol, MTBE, DMSO, ethanol, ethyl acetate, isobutyl acetate, isopropyl acetate, 1-propanol, IPA, MEK and THF; wherein the second temperature is 50 ℃.
31. 31. A method of making the microparticle of claim 1, comprising the steps of: (a) preparing a solution of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid in a solvent at a first temperature; and (b) generating the microparticles by cooling the solution to a second temperature; wherein the solvent is selected from acetonitrile, methanol, 1-propanol, and MEK.
32. 32. A method of making the microparticle of claim 1, comprising the steps of: (a) preparing a solution of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid in a solvent at a first temperature; and (b) generating said microparticles by adding an anti-solvent to said solution; wherein (i) the solvent is THF and the anti-solvent is hexane or water; or (ii) the solvent is DMF and the anti-solvent is water.
33. 33. A method of making the microparticle of claim 1, comprising the steps of: (a) preparing a solution of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid in a solvent at a first temperature; and (b) generating the microparticles by adding the solution to an anti-solvent; wherein (i) the solvent is THF or 2-methoxyethanol, and the anti-solvent is hexane; or (ii) the solvent is THF, DMSO or DMF and the anti-solvent is water.
34. 34. A method of making the microparticle of claim 1, comprising the steps of: (a) preparing a slurry of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid in a solvent at a first temperature; and (b) forming the microparticles by exposing the slurry to a second temperature, wherein the solvent is (i) methanol or (ii) a mixture of DMF and MTBE.
35. 35. A method of making the microparticle of claim 1, comprising the steps of: (a) preparing a solution of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid in a solvent at a first temperature; (b) forming a slurry by cooling the solution to a second temperature; and (c) producing a slurry by treating the slurry with one or more heating and cooling cyclesForming the microparticles, wherein the solvent is acetonitrile, 1, 4-bisAlkyl, 2-methoxyethanol, MIBK, acetone, 1-butanol, ethanol, 1-propanol, IPA, MEK, THF or a mixture of water and THF.
36. 36. The method of claim 35, wherein the heating and cooling cycle is performed at a temperature in the range of 20 to 50 ℃.
37. 37. The method of any one of claims 28-36, further comprising the step of isolating the microparticles.