HK1179963A - Amine salts of a crth2 antagonist - Google Patents

Abstract

Provided are amine salts of {4,6-bis(dimethylamino)-2-(4-(4- (trifluoromethyl)benzamido)benzyl)pyrimidin-5-yl} -acetic acid, processes for their preparation, pharmaceutical compositions containing them, and their use for treating, preventing, or ameliorating one or more symptoms of a CRTH2-mediated disorder or disease.

Description

Amine salts of a CRTH2 antagonist

The present application is a division of the invention application having application date of 2008/06/18, application number of 200880101962.9, entitled "an amine salt of a CRTH2 antagonist".

Technical Field

The present invention provides amine salts of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid, methods of preparation, and pharmaceutical compositions thereof. Also provided are methods of using the compounds for treating, preventing, or ameliorating one or more symptoms of a CRTH 2-mediated disorder or disease.

Background

CRTH2 is a G protein-coupled chemokine receptor expressed in Th2 cells, eosinophils, and basophils (Nagata et al, J.Immunol.1999,162, 1278-1286; Hirai et al, J.Exp.Med.,2001,193, 255-261). Prostaglandin D, the major inflammatory mediator produced by mast cells₂（PGD₂) Is a natural ligand of CRTH 2. Recently, it has been shown to pass PGD₂Activation of CRTH2 induces migration and activation of Th2 cells and eosinophils, suggesting that CRTH2 may play a pro-inflammatory role in allergic diseases (Hirai et al, j.exp.med.2001,193, 255-261; Gervais et al, j.allergy clin.immunol.2001,108, 982-988). It has also been shown that circulating T cells expressing CRTH2 are increased in atopic dermatitis patients, and this increase is correlated with the severity of the disease (Cosmi et al, Eur. J. Immunol.,2000,30, 2972-. PGD has been further shown in a mouse asthma model₂Role in the initiation and maintenance of allergic inflammation, the model being shown by PGD₂Synthetase produces excess PGD in vivo₂Airway inflammation can be exacerbated (Fujitani et al, J.Immunol.2002,168, 443-449). Thus, CRTH2 antagonists may be useful in the treatment of CRTH2 mediated disorders or diseases, such as allergic rhinitis, allergic asthma, bronchoconstriction, atopic dermatitis, or systemic inflammatory disorders.

Disclosure of Invention

The present invention provides amine salts of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid, said acid having the structural formula I:

or pharmaceutically acceptable hydrates and solvates thereof. The compound of structural formula I has been identified as a CRTH2 antagonist (WO 2004/0022218).

In one embodiment, the amine salt comprising an acid having structural formula I and a pharmaceutically acceptable amine is crystalline.

In another embodiment, the amine salt comprises an acid having structural formula I and a pharmaceutically acceptable amine, but excludes ethanolamine, triethylamine, and tris (hydroxymethyl) aminomethane.

In another embodiment, the amine salt of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid is a diamine salt comprising about 2 molar equivalents of an acid of formula I and about 1 molar equivalent of a diamine.

In another embodiment, the diamine salt of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid is crystalline.

In another embodiment, the amine salt of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid is a monoamine salt comprising about 1 molar equivalent of an acid of formula I and about 1 molar equivalent of a monoamine.

In another embodiment, the monoamine salt of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzoylamino) benzyl) pyrimidin-5-yl } acetic acid is crystalline.

Also provided are methods for the preparation of amine salts of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid or pharmaceutically acceptable hydrates or solvates thereof.

In one embodiment, the method comprises reacting {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid with an amine in a solvent at a first predetermined temperature.

In another embodiment, the method further comprises precipitating the amine salt at a second predetermined temperature.

In another embodiment, the method comprises the steps of: (a) reacting {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid with an amine in a solvent at a first predetermined temperature to produce the amine salt; and (b) precipitating the amine salt at a second predetermined temperature.

The invention further provides a pharmaceutical composition comprising an amine salt of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid, or a pharmaceutically acceptable hydrate or solvate thereof, and one or more pharmaceutically acceptable carriers or excipients.

In addition, the present invention provides a method of treating, preventing, or ameliorating one or more symptoms of a CRTH 2-mediated disorder or disease, the method comprising administering to a mammal a therapeutically effective amount of an amine salt of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid, or a pharmaceutically acceptable hydrate or solvate thereof.

Drawings

FIG. 1 shows proton nuclear magnetic resonance of crystalline ethylenediamine salt of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid of formula I ((III¹H NMR) spectrum.

Figure 2 shows an X-ray powder (XRP) diffraction pattern of a crystalline ethylenediamine salt of the acid of formula I.

FIG. 3 shows a Differential Scanning Calorimetry (DSC) thermogram of a crystalline ethylenediamine salt of an acid of formula I.

FIG. 4 shows the Thermogravimetric (TG) thermogram of crystalline ethylenediamine salt of acid of formula I.

Figure 5 shows a Scanning Electron Microscope (SEM) image of a crystalline ethylenediamine salt of the acid of formula I.

FIG. 6 shows crystallization of piperazine salt of acid of structural formula I¹H NMR spectrum.

Figure 7 shows an XRP diffraction pattern of a piperazine salt crystal of an acid of structural formula I.

Figure 8 shows a DSC thermogram of a piperazine salt crystal of an acid of structural formula I.

Fig. 9 shows TG thermogram of piperazine salt crystal of acid with structural formula I.

Fig. 10 depicts an SEM image of piperazine salt crystals of an acid having structural formula I.

FIG. 11 shows a benzathine salt crystal of an acid of formula I¹H NMR spectrum.

Figure 12 shows an XRP diffraction pattern of a benzathine salt crystal of an acid of structural formula I.

FIG. 13 shows a DSC thermogram of a benzathine salt crystal of an acid of structural formula I.

Figure 14 shows an SEM image of benzathine salt crystals of an acid of structural formula I.

FIG. 15 shows choline salt crystals of an acid of structural formula I¹H NMR spectrum.

Figure 16 shows an XRP diffraction pattern of a choline salt crystal of an acid of structural formula I.

Figure 17 shows a DSC thermogram of choline salt crystals of an acid of structural formula I.

Figure 18 shows TG thermogram of choline salt crystal of acid with structural formula I.

Figure 19 shows an SEM image of choline salt crystals of an acid of structural formula I.

FIG. 20 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 CRTH2 transfected cells.

FIG. 21 shows a dose response curve for {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid with structural formula I (a) compared to the DP1 selective antagonist BWA 868C (■) in a competitive radioligand binding assay using cells transfected with DP 1.

Detailed description of exemplary embodiments

To facilitate understanding of the disclosure of the present specification, certain terms are defined below.

As used herein, the singular forms "a", "an" and "the" may refer to a plurality of items unless specifically stated otherwise. In general, the nomenclature used herein and the organic chemical, pharmaceutical chemical, and pharmacological laboratory procedures 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. If a term in this document has multiple definitions, the definition in this section controls unless otherwise indicated.

The term "anti-solvent" refers to a liquid added to a solvent that reduces the solubility of a compound in the solvent, resulting in precipitation of the compound.

The term "subject" refers to an animal, including but not limited to a primate (e.g., human), a cow, sheep, goat, horse, dog, cat, rabbit, rat, or mouse. Generally, 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 reduce or eradicate the cause of the condition or disease itself.

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

The term "therapeutically effective amount" refers to an amount of a compound that, when administered, is sufficient to prevent the development of, or alleviate to some extent, one or more symptoms of, a disease, disorder, or condition being treated. The term "therapeutically effective amount" also means that amount of a compound that elicits the biological or medical response of a tissue, system, animal, or human that is being sought by a researcher, veterinarian, medical doctor, or outpatient physician.

The term "pharmaceutically acceptable carrier," "pharmaceutically acceptable excipient," "physiologically acceptable carrier," or "physiologically acceptable excipient" refers to a pharmaceutically acceptable material, composition, or carrier, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. Each ingredient must be pharmaceutically acceptable in the sense of being compatible with the other ingredients of the pharmaceutical formulation. Each component must also be suitable for use in contact with human or animal tissues or organs 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" when used with respect to biological materials, such as nucleic acid molecules, polypeptides, host cells, and the like, refers to materials that are found in nature and are not 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 altered or artificially synthesized.

The term "CRTH 2" refers to the CRTH2 receptor protein or a variant thereof, which mediates protection of PGD in vitro or in vivo₂The cellular response of (a). The CRTH2 variant includes a protein substantially homologous to native CRTH2, i.e., with the amino group of native CRTH2Proteins having one or more naturally or non-naturally occurring amino acid deletions, insertions, or substitutions (e.g., CRTH2 derivatives, homologs, and fragments) in comparison in sequence. The amino acid sequence of the CRTH2 variant is at least about 80% identical, at least about 90% identical, or at least about 95% identical to native CRTH 2.

The term "other PGD₂Receptor "refers to a prostaglandin receptor protein other than CRTH2 or a variant thereof that mediates binding to PGD in vivo or in vitro₂The cellular response of (a). "other PGD₂The receptor can be PGD₂Optionally, DP, for example, or one or more other prostaglandins. "other PGD₂Receptor "variants" include proteins substantially homologous to the corresponding native prostaglandin receptor except for 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 prostaglandin receptor except for CRTH 2). Natural "other PGD₂Receptor "variants" having amino acid sequences corresponding to the native "other PGDs₂The receptors are "at least about 80% identical, at least about 90% identical, or at least about 95% identical.

The term "CRTH 2 antagonist" refers to, for example, a compound that blocks, attenuates, prevents, or downregulates, partially or totally, 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, delays activation of, inactivates, or inactivates CRTH2 or one or more other PGD2 receptors. The mode of action of the CRTH2 antagonist may be interference with PGD₂With CRTH2 or one or more other PGDs₂Receptor interaction.

The terms "CRTH 2-mediated disorder or disease" and "CRTH 2-mediated abnormality, disorder or disease" refer to an abnormality, disorder or disease characterized by an abnormal, e.g., decreased or increased, CRTH2 activity compared to normal. Aberrant CRTH2 functional activity may result from CRTH2 expression, increased CRTH2 expression or increased degree of intracellular activation in cells that do not normally express CRTH2, resulting in, for example, inflammation or immune-related disorders or diseases(ii) a Or reduced expression of CRTH 2. The CRTH 2-mediated abnormality, disorder or disease may be mediated in whole or in part by aberrant CRTH2 activity. In particular, modulation of CRTH2 or one or more other PGDs in CRTH 2-mediated abnormalities, conditions or diseases₂Receptors may cause some effect on underlying abnormalities or conditions, e.g., CRTH2 antagonists or agonists, to achieve some improvement in at least some of the treated patients.

Amine salts

The present invention provides amine salts of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzoylamino) benzyl) pyrimidin-5-yl } acetic acid and solvates thereof; and a process for the preparation of said compounds. Also provided are pharmaceutical compositions of the amine salts or solvates thereof, and methods of using the compounds for treating, preventing, or ameliorating one or more symptoms of a CRTH 2-mediated disorder or disease.

According to one embodiment, the present invention provides a pharmaceutically acceptable amine salt of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid, the amine salt comprising an acid of formula I and a pharmaceutically acceptable amine. In another embodiment, the present invention provides a pharmaceutically acceptable solvate of a crystalline amine salt of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid, said solvate comprising an acid of formula I, a pharmaceutically acceptable amine, and a solvent.

In certain embodiments, the molar ratio of acid of formula I to amine in the amine salt or solvate thereof is from about 0.5 to about 10, from about 0.5 to about 5, from about 0.5 to about 3, from about 0.5 to about 2, or from about 0.8 to about 1.2, or about 1.

In certain embodiments, the present invention provides solvates of amine salts in which the molar ratio of the acid of formula I to the solvent is from about 0.1 to about 2, from about 0.2 to about 1, or from about 0.3 to about 0.5, or about 0.1, about 0.2, about 0.3, about 0.4, about 0.5, about 0.6, about 0.7, about 0.8, about 0.9, or about 1.

Amines suitable for use in the amine salts or solvates thereof provided herein include, but are not limited to, primary amines including methylamine, ethylamine, ethanolamine, tris (hydroxymethyl) aminomethane, and ethylenediamine; secondary amines including dimethylamine, diethylamine, diisopropylamine, dibutylamine, di-sec-butylamine, dicyclohexylamine, diethanolamine, meglumine, pyrrolidine, piperidine, piperazine, and benzathine; tertiary amines including trimethylamine, triethylamine, triethanolamine, and 1- (2-hydroxyethyl) pyrrolidine; quaternary amines including choline, tetramethylammonium, and tetraethylammonium. If other amines are to be understood, see "Handbook of Pharmaceutical Salts: Properties, Selection, and Use" by Stahl and Wermuth, Wiley-VCH, 2002.

In one embodiment, the pharmaceutically acceptable amine is a diamine. The pharmaceutically acceptable diamine has a first amino group and a second amino group, each independently being a primary, secondary, or tertiary amino group, or a quaternary ammonium group. Diamines suitable for use in the diamine salt include, but are not limited to, ethylenediamine, piperazine, and benzathine. The diamine salt of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid comprises about 1 to about 3, about 1.5 to about 2.5, about 1.75 to about 2.25, or about 2 molar equivalents of the acid of formula I relative to 1 molar equivalent of the diamine.

In one group of diamine salts of this embodiment, the first amino group of the diamine is a primary amino group, and the second amino group is independently a primary, secondary, or tertiary amino group, or a quaternary ammonium group. In another set of the diamine salts, the first amino group is independently a secondary amino group, and the second amino group is a primary, secondary, or tertiary amino group, or a quaternary ammonium group. In another group of the diamine salts, the first amino group is independently a tertiary amino group, and the second amino group is a primary, secondary, or tertiary amino group, or a quaternary ammonium group. In another group of the diamine salts, the first amino group is a quaternary ammonium group, and the second amino group is independently a primary, secondary, or tertiary amino group, or a quaternary ammonium group.

In another embodiment, the pharmaceutically acceptable amine is a diamine having 2 primary amino groups. The primary diamine salt of the acid of formula I comprises about 1 to about 3, about 1.5 to about 2.5, about 1.75 to about 2.25, or about 2 molar equivalents of the acid of formula I relative to 1 molar equivalent of the primary diamine.

In another embodiment, the primary diamine is ethylene diamine. Based on¹H NMR spectrum (fig. 1) estimated that the ethylenediamine salt of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid contained about 2 molar equivalents of the acid of formula I and about 1 molar equivalent of ethylenediamine.

In another embodiment, the ethylenediamine salt of the acid of formula I is crystalline. The XRP diffraction pattern of the crystalline ethylenediamine salt is substantially as shown in figure 2. In particular, the crystalline ethylenediamine salt has a characteristic XRP diffraction peak at about 5.9 degrees 2-theta. Further, the DSC thermogram of the crystalline ethylenediamine salt is substantially as shown in fig. 3. The crystalline ethylenediamine salt has an endothermic effect with a peak temperature of about 123 ℃ and an onset temperature of 114 ℃, or a peak temperature of about 216 ℃ and an onset temperature of about 212 ℃. Alternatively, the crystalline ethylenediamine salt has an endothermic effect with a peak temperature of about 123 ℃ and an onset temperature of 114 ℃, and a peak temperature of about 216 ℃ and an onset temperature of about 212 ℃.

In another embodiment, the present invention provides a crystalline ethylenediamine salt of an acid of formula I. The XRP diffraction pattern of the solvate is substantially as shown in figure 2. In particular, the solvate has a characteristic XRP diffraction peak at about 5.9 degrees 2-theta. Further, the DSC thermogram of the solvate is substantially as shown in figure 3. The solvate has an endothermic effect with a peak temperature of about 123 ℃ and an onset temperature of 114 ℃, or a peak temperature of about 216 ℃ and an onset temperature of about 212 ℃. Alternatively, the solvate has an endothermic effect with a peak temperature of about 123 ℃ and an onset temperature of 114 ℃, and a peak temperature of about 216 ℃ and an onset temperature of about 212 ℃.

In one embodiment, the solvate loses weight between about 1% and about 10% between 125 ℃ and 150 ℃, including, but not limited to, about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, and about 10%. In another embodiment, the solvate loses about 2%, about 3%, about 4%, or about 5% weight between 125 ℃ and 150 ℃.

In one embodiment, the solvate comprises from about 0.1 to about 2, from about 0.2 to about 1, or from about 0.3 to about 0.5; or about 0.1, about 0.2, about 0.3, about 0.4, about 0.5, about 0.6, about 0.7, about 0.8, about 0.9, or about 1 molar equivalent of solvent. In another embodiment, the solvate comprises from about 0.3 to about 0.5 molar equivalents of solvent per molar equivalent of amine salt. In another embodiment, the solvate comprises about 0.3 molar equivalents of solvent per molar equivalent of amine salt. In another embodiment, the solvate comprises about 0.4 molar equivalents of solvent per molar equivalent of amine salt. In another embodiment, the solvate comprises about 0.5 molar equivalents of solvent per molar equivalent of amine salt.

In one embodiment, the solvent in the solvate is an alcohol, including, but not limited to, 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. In another embodiment, the solvent is ethanol.

In another embodiment, the pharmaceutically acceptable amine is a diamine having 2 secondary amino groups. The secondary diamine salt of an acid of formula I comprises about 1 to about 3, about 1.5 to about 2.5, about 1.75 to about 2.25, or about 2 molar equivalents of an acid of formula I relative to 1 molar equivalent of a secondary diamine.

In another embodiment, the secondary diamine is piperazine. Based on¹H NMR Spectroscopy (FIG. 6) estimated that the piperazine salt of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid contained about 2 molar equivalents of the acid of formula I and about 1 molar equivalentPiperazine.

In another embodiment, the piperazine salt is crystalline. The crystalline piperazine salt has an XRP diffraction pattern substantially as shown in figure 7. In one embodiment, the piperazine salt has characteristic XRP diffraction peaks at about 10.7, 15.9, 22.3, and 24 degrees 2 Θ. In another embodiment, the piperazine salt has a characteristic XRP diffraction peak at about 10.7, 15.9, 22.3, or 24 degrees 2 Θ. In another embodiment, the crystalline piperazine salt has a DSC thermogram substantially as shown in figure 8. In another embodiment, the crystalline piperazine salt has an endothermic effect with a peak temperature of about 203 ℃ and an onset temperature of about 197 ℃, or with a peak temperature of about 223 ℃ and an onset temperature of about 219 ℃. In an alternative embodiment, the crystalline piperazine salt has an endothermic effect with a peak temperature of about 203 ℃ and an onset temperature of about 197 ℃, and a peak temperature of about 223 ℃ and an onset temperature of about 219 ℃.

In another embodiment, the secondary diamine is benzathine. Based on¹H NMR spectrum (fig. 11) estimated that the benzathine salt of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid contained about 2 molar equivalents of the acid of formula I and about 1 molar equivalent of benzathine.

In another embodiment, the benzathine salt is crystalline, which has an XRP diffraction pattern substantially as shown in figure 12. In one embodiment, the benzathine salt has characteristic XRP diffraction peaks at about 8.0, 11.5, 16.0, 17.5, and 23.4 degrees 2 Θ. In another embodiment, the benzathine salt has a characteristic XRP diffraction peak at about 8.0, 11.5, 16.0, 17.5, or 23.4 degrees 2 Θ. In another embodiment, the crystalline benzathine salt has a DSC thermogram substantially as shown in figure 13. In an alternative embodiment, the crystalline benzathine salt has an endothermic effect with a peak temperature of about 156 ℃ and an onset temperature of about 154 ℃.

In another embodiment, the amine is a monoamine. The monoamine salt of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid comprises about 0.5 to about 1.5, about 0.75 to about 1.25, or about 1 molar equivalent of the acid of formula I relative to 1 molar equivalent of the monoamine.

In one group of monoamine salts of this embodiment, the monoamine of said monoamine salt has one primary amino group. In another set of said monoamine salts, said monoamine has one secondary amino group. In another group of said monoamine salts, said monoamine has one tertiary amino group. In another group of said monoamine salts, said monoamine has a quaternary ammonium group.

In an alternative embodiment, the amine is choline. Based on¹H NMR spectrum (fig. 15) estimated that the choline salt of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid contained about 1 molar equivalent of the acid of formula I and about 1 molar equivalent of choline.

In another embodiment, the choline salt of an acid of structural formula I is crystalline, the crystalline choline salt having an XRP diffraction pattern substantially as shown in figure 16. In one embodiment, the choline salt has characteristic XRP diffraction peaks at about 6.5, 19.6, 20.0, 21.9, and 26.4 degrees 2 Θ. In another embodiment, the choline salt has a characteristic XRP diffraction peak at about 6.5, 19.6, 20.0, 21.9, or 26.4 degrees 2 Θ. In another embodiment, the crystalline choline salt has a DSC thermogram substantially as shown in figure 17. In an alternative embodiment, the crystalline choline salt has an endothermic effect with a peak temperature of about 195 ℃ and an onset temperature of about 193 ℃.

It will be appreciated that the peaks of the X-ray powder diffraction pattern may vary slightly from machine to machine or from sample to sample, and therefore the values quoted are not to be understood as absolute, but with appropriate variability, for example 0.1 degrees as recommended by the united states pharmacopoeia 2007,387-389.

Preparation method

The invention also provides a process for preparing amine salts of acids having structural formula I. In one embodiment, the method comprises reacting {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid with an amine in a solvent at a first predetermined temperature. In another embodiment, the method further comprises precipitating the amine salt at a second predetermined temperature.

In an alternative embodiment, the method comprises the steps of: (a) reacting {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid with an amine in a solvent at a first predetermined temperature to produce an amine salt; and (b) precipitating the amine salt at a second predetermined temperature.

Suitable solvents for preparing the amine salt of the acid of formula I include, but are not limited to, hydrocarbons including petroleum ether, pentane, hexane, heptane, octane, isooctane, cyclopentane, cyclohexane, methylcyclohexane, benzene, toluene, xylene, tetralin, and cumene; chlorinated hydrocarbons including Dichloromethane (DCM), 1, 2-dichloroethane, 1-dichloroethylene, 1, 2-dichloroethylene, chloroform, trichloroethane, trichloroethylene, carbon tetrachloride, chlorobenzene, and trifluorotoluene; 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-methoxy) ether, 1-dimethoxymethane, 2-dimethoxypropane, and anisole; ketones, including acetone, butanone, Methyl Ethyl Ketone (MEK), methyl isopropyl ketone, methyl butanone, and methyl isobutyl ketone (MD 3K); 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, trichloroacetic acid, and trifluoroacetic acid; phosphoramides, such as hexamethylphosphoramide; carbon disulfide; water; and mixtures of the foregoing.

In certain embodiments, the amine salt formation reaction (i.e., step a) is carried out at a temperature of from about-10 to about 150 ℃, from about 10 to about 110 ℃, or from about 20 to about 100 ℃. In one embodiment, the solvent is acetonitrile, acetone, methyl ethyl ketone, methyl isobutyl ketone, N-dimethylformamide, dimethyl sulfoxide, a lower alkanol (e.g., methanol, ethanol, N-propanol, isopropanol, sec-butanol, or 2-methoxyethanol), methyl acetate, ethyl formate, isopropyl acetate, isobutyl acetate, chloroform, dichloromethane, methyl tert-butyl ether, tetrahydrofuran, 1, 4-dioxane, petroleum ether, hexane, heptane, toluene, water, or a mixture thereof. In another embodiment, the solvent is a lower alkanol of 1 to 5 carbons, such as methanol, ethanol, propanol, isopropanol, sec-butanol, 2-methoxyethanol, or a mixture of the foregoing.

In certain embodiments, the amine salt formation reaction is carried out in the presence of excess amine to maximize the reaction yield. The molar ratio of the amino groups of the amine to the acid of formula I is not less than about 1.01, not less than about 1.05, not less than about 1.1, not less than about 1.2, about 1.05 to about 10, about 1.1 to 5, or about 1.2 to 2.5.

In certain embodiments, the amine salt formation reaction is carried out in solution, that is, both the acid of formula I and the amine are dissolved in the solvent. In certain embodiments, the amine salt formation reaction is carried out as a slurry mixture of an acid of formula I and an amine in the solvent. In this case, the acid of formula I is not completely dissolved, whereas the amine is completely dissolved.

In certain embodiments, the amine salt formed in the amine-forming reaction step may be precipitated from the reaction solution or slurry mixture using conventional methods including, but not limited to, cooling, quenching, solvent evaporation, addition of an anti-solvent, or reverse addition to an anti-solvent. The precipitation step may be carried out at a temperature of from about-50 to about 100 ℃, from about-30 to about 50 ℃, or from about-10 to about 30 ℃. To accelerate the precipitation (crystallization) step, the method may further comprise a step of adding a seed crystal to the reaction solution or mixture. The method further comprises a separation step in which the precipitate can be separated by conventional methods, such as filtration and centrifugation, and then washed with a solvent and dried.

In one embodiment, the amine salt is prepared by reacting (a) {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid with an amine in a low alkanol, such as ethanol, at elevated temperature to produce a clear reaction solution. When the amine is a diamine, the molar ratio of the diamine molecules to the acid of formula I in the reaction solution is not less than about 0.505, not less than about 0.525, not less than about 0.55, or not less than about 0.60; but not greater than about 10 to about 100. When the amine is a monoamine, the molar ratio of the monoamine molecule to the acid of formula I in the reaction solution is not less than about 1.01, not less than about 1.05, not less than about 1.1; but not greater than about 10 to about 100.

The amine salt may be precipitated by cooling the reaction solution to or below room temperature, or by solvent evaporation. The amine salt precipitate may also be formed by adding an anti-solvent to the reaction solution, or adding the reaction solution to an anti-solvent.

Suitable antisolvents include, but are not limited to, hydrocarbons including petroleum ether, pentane, hexane, heptane, octane, isooctane, cyclopentane, cyclohexane, methylcyclohexane, benzene, toluene, xylene, tetralin, and cumene; chlorinated hydrocarbons including dichloromethane, 1, 2-dichloroethane, 1-dichloroethylene, 1, 2-dichloroethylene, chloroform, trichloroethane, trichloroethylene, carbon tetrachloride, chlorobenzene, and trifluorotoluene; alcohols including isopropanol, 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, diphenyl ether, 1, 2-dimethoxyethane, bis (2-methoxy) ether, 1-dimethoxymethane, 2-dimethoxypropane, and anisole; ketones, including butanone, methyl isopropyl ketone, methyl butanone, and methyl isobutyl ketone; 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 such as nitromethane and nitrobenzene; heterocycles; carbon disulfide; water; and mixtures of the foregoing.

When two solvents are used as a solvent/anti-solvent pair, the amine salt of the acid of formula I is more soluble in the solvent than in the anti-solvent. Alternatively, the solvent and the anti-solvent of the solvent/anti-solvent pair are at least partially miscible.

In another embodiment, the amine salt is prepared by reacting a slurry of (a) {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid with an amine in a solvent such as ethanol at room temperature or elevated temperature. After the reaction, the amine salt may be recovered as a solid by cooling, solvent evaporation, or addition of an anti-solvent to the slurry reaction mixture.

To accelerate the precipitation (crystallization) step, the process may further comprise the step of adding seed crystals to the reaction solution or mixture, said step being performed before or during the initiation of the precipitation step. The amount of seed crystals added exceeds the saturation amount of the solvent used, and therefore undissolved seed crystals are present in the reaction solution.

Other salt generation methods may also be used in the present invention. For example, the amine salt of the acid of formula I may be prepared by converting a salt of the acid, such as a sodium or potassium salt, to an amine salt by cation exchange using a cation exchange column. The amine salt of the acid of formula I can also be prepared by physically milling solid {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid with an amine in the absence of a solvent.

In addition to precipitation and crystallization, the solid amine salts provided herein can be prepared using conventional methods well known to those skilled in the art, including spray drying, drum drying, lyophilization, and melt crystallization.

Pharmaceutical composition

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

The pharmaceutical compositions provided herein may be provided in unit dosage forms or multiple dosage forms. As used herein, unit dosage forms refer to physically discrete units suitable for administration to human and animal subjects and packaged individually as is well known in the art. Each unit dose contains a predetermined amount of the active ingredient sufficient to produce the desired therapeutic effect, in association with the necessary pharmaceutically acceptable carrier or excipient. Examples of unit dosage forms include ampoules, syringes, and individually packaged tablets and capsules. The unit dosage form may be administered in portions or in multiple portions. A multiple dosage form is a plurality of identical unit dosage forms packaged in a single container for administration as discrete unit dosage forms. Examples of multiple dosage forms include vials, tablet or capsule bottles, or pint or gallon bottles.

The amine salt of an 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 comprising the amine salts provided herein can be formulated in a variety of dosage forms for oral administration, parenteral administration, and topical administration. The pharmaceutical compositions may also be formulated as modified release dosage forms, such as delayed release dosage forms, extended release dosage forms, sustained release dosage forms, pulsatile release dosage forms, controlled release dosage forms, accelerated and rapid release dosage forms, targeted release dosage forms, programmed release dosage forms, and gastric retention dosage forms. Such dosage forms can be prepared according to conventional methods and techniques well known to those skilled in The art (see Remington: The Science and Practice of Pharmacy, Modified-Release Drug delivery Technology, Rathbone et al, eds., Drugs and The Pharmaceutical Science, Marcel Dekker, Inc.: New York, NY, 2002; Vol.126), supra).

The pharmaceutical compositions provided by the present invention may be administered in a single dose or in multiple doses separated by a time interval. It will be appreciated that the precise dosage and treatment time may vary with the age, weight, and health of the patient being treated, and may be determined empirically using known test records or by inference from in vivo or in vitro tests or diagnostic data. It will be further appreciated that for any particular individual, the specific dosage regimen will be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the formulation.

A. Is administered orally

The pharmaceutical compositions provided herein can be provided in solid, semi-solid, or liquid dosage forms for oral administration. When used in the present invention, oral administration also includes buccal, lingual, and sublingual administration. Suitable oral dosage forms include, but are not limited to, tablets, capsules, pills, lozenges, troches, cachets, pellets, medicated 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 contain one or more pharmaceutically acceptable carriers or excipients, including, but not limited to, binders, fillers, diluents, disintegrants, wetting agents, lubricants, glidants, colorants, dye migration inhibitors, sweeteners, and flavoring agents.

The binder or granule imparts cohesiveness to the tablet to ensure that the tablet remains intact after compression. Suitable binders or granules include, but are not limited to, starches, such as corn STARCH, potato STARCH, and pregelatinized STARCH (e.g., STARCH 1500); gelatin; sugars such as sucrose, glucose, dextrose, syrup, and lactose; natural and synthetic gums such as acacia, alginic acid, alginates, irish moss extract, panval gum, ghatti gum, psyllium mucilage, carboxymethyl cellulose, methyl cellulose, polyvinyl pyrrolidone (PVP), magnesium aluminum silicate, larch arabinogalactan, tragacanth gum powder, and guar gum; celluloses, such as ethyl cellulose, cellulose acetate, calcium carboxymethylcellulose, sodium carboxymethylcellulose, 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., Marcus Hook, Pa.); and mixtures of the foregoing. Suitable fillers include, but are not limited to, talc, calcium carbonate, microcrystalline cellulose, cellulose powder, dextrates, kaolin, mannitol, silicic acid, sorbitol, starch, pregelatinized starch, and mixtures of the foregoing. The binder or filler may be present in the pharmaceutical compositions provided herein in an amount of about 50 to about 99% by weight.

Suitable diluents include, but are not limited to, dicalcium 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 impart the property that certain compressed tablets can disintegrate in the mouth by chewing. Such compressed tablets may be used as chewable tablets.

Suitable disintegrants include, but are not limited to, agar; bentonite; 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 veegum 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 polacrilin; starches, such as corn starch, potato starch, tapioca starch, and pregelatinized starch; clay; align; and mixtures of the foregoing. The content of the disintegrant in the pharmaceutical composition provided by the present invention varies depending on the type of formulation, and is easily understood by those skilled in the art. The pharmaceutical compositions provided herein contain from about 0.5 to about 15% or from about 1 to about 5% by weight of disintegrant.

Suitable lubricants include, but are not limited to, calcium stearate; magnesium stearate; mineral oil; light mineral oil; glycerol; sorbitol; mannitol; glycols, such as glyceryl behenate and polyethylene glycol (PEG); stearic acid; sodium lauryl 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 (w.r.grace co., Baltimore, MD) and(Cabot co. of boston, MA); and mixtures of the foregoing. The pharmaceutical compositions provided herein contain about 0.1 to about 5% by weight of a lubricant.

Suitable glidants include colloidal silicon dioxide,(Cabot co. of Boston, MA), and asbestos-free talc. Colorants include any approved, certified, water-soluble pharmaceutical and cosmetic dyes, and suspensionsWater insoluble pharmaceutical and cosmetic dyes in hydrated alumina, and lakes, and mixtures of the foregoing. Lakes are water-soluble dyes that bind to the hydrated oxides of heavy metals by adsorption, producing an insoluble form of the dye. Fragrances include natural flavors extracted from plants, such as fruits, and synthetic mixtures of compounds that produce a pleasant taste sensation, such as peppermint and wintergreen oils. Sweetening agents 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 sorbitan monooleatePolyoxyethylene sorbitan monooleate 80And triethanolamine oleate. Suspending and dispersing agents include sodium carboxymethylcellulose, pectin, tragacanth, magnesium aluminum silicate, acacia, sodium carboxymethylcellulose, hydroxypropylmethylcellulose, 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 lauryl ether. Solvents include glycerol, sorbitol, 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 in the same formulation.

The pharmaceutical composition provided by the invention can be provided in the following dosage forms: compressed tablets, tablet triturates, chewable lozenges, fast-dissolving tablets, compressed tablets, or enteric-coated tablets, sugar-coated or film-coated tablets. Enteric coated tablets are compressed tablets coated with a substance that resists the action of gastric acid but dissolves or disintegrates in the intestine, thus preventing the acid environment of the stomach from destroying the active ingredient. 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 that may help to mask an objectionable taste or odor and protect the tablet from oxidation. Film coated tablets are compressed tablets coated with a thin layer or film of a water soluble material. Film coatings include, but are not limited to, hydroxyethyl cellulose, sodium carboxymethyl cellulose, polyethylene glycol 4000, and cellulose acetate phthalate. Film coatings provide the same general characteristics as sugar coatings. Compressed tablets are compressed tablets prepared by more than one compression cycle, including multilayer tablets, and compression-coated tablets or dry-coated tablets.

The tablet dosage form may be prepared from the active ingredient in powder, crystalline, or granular form, alone or in combination with one or more carriers or excipients as described herein, including binders, disintegrants, long acting polymers, lubricants, diluents, and/or colorants. Flavoring and sweetening agents are particularly useful in chewable tablet and lozenge formulations.

The pharmaceutical composition provided by the invention can be provided in a soft capsule or a hard capsule, and the capsule can be prepared from gelatin, methylcellulose, starch or calcium alginate. The hard gelatin capsules, also known as Dry Fill Capsules (DFC), consist of two parts, one overlapping the other, thereby completely packaging the active ingredient. The Soft Elastic Capsules (SEC) are soft spherical shells, such as gelatin shells, which can be plasticized by the addition of glycerol, sorbitol, or similar polyhydroxy compounds. The soft gelatin shell may contain a preservative to prevent microbial growth. Suitable preservatives are described herein, including methylparaben and propylparaben, and sorbic acid. The liquid, semi-solid, and solid dosage forms provided by the present invention can be encapsulated. Suitable liquid and semi-solid dosage forms include solutions and suspensions in propylene carbonate, vegetable oils, or triglycerides. Capsules containing such solutions can be prepared as described in U.S. patents 4,328,245, 4,409,239, and 4,410,545. The capsules may also be coated to alter or maintain the dissolution of the active ingredient, as is well known to those skilled in the art.

The pharmaceutical compositions provided herein can be provided in liquid and semi-solid dosage forms, including emulsions, solutions, suspensions, elixirs, and syrups. Emulsions are biphasic systems in which one liquid is dispersed in another in the form of globules, and the emulsion may be water-in-oil or oil-in-water. Emulsions may include pharmaceutically acceptable non-aqueous liquids or solvents, emulsifiers, and preservatives. Suspensions may include pharmaceutically acceptable suspending agents and preservatives. The aqueous alcoholic solution may comprise a pharmaceutically acceptable acetal, such as a di (lower alkyl) acetal or a lower alkyl aldehyde (the term "low" means an alkyl group having from 1 to 6 carbon atoms), such as acetaldehyde diethyl acetal; and water-soluble solvents having one or more hydroxyl groups, such as propylene glycol and ethanol. Elixirs are clear, sweetened, hydroalcoholic solutions. Syrups are concentrated aqueous solutions of sugars, such as sucrose, and may also contain preservatives. For liquid dosage forms, such as solutions in polyethylene glycol, the solution can be diluted with a sufficient amount of a pharmaceutically acceptable liquid carrier, such as water, to be weighed out at the time of administration.

Other useful liquid and semi-solid dosage forms include, but are not limited to, dosage forms comprising the active ingredients provided herein, and dialkyl mono-or polyethylene glycols including, 1, 2-dimethoxymethane, diglyme, triglyme, tetraglyme, polyethylene glycol-350-dimethyl ether, polyethylene glycol-550-dimethyl ether, polyethylene glycol-750-dimethyl ether, wherein 350, 550, 750 refer to the approximate average molecular weight of the polyethylene glycol. 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 composition for oral administration provided by the present invention 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 be provided as non-effervescent or effervescent granules or powders that can be reconstituted into liquid dosage forms. Pharmaceutically acceptable carriers and excipients for the non-foaming 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.

Colorants and fragrances may be used in all of the above dosage forms.

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

The pharmaceutical compositions provided by the present invention may be co-formulated with other active ingredients that do not impair the desired therapeutic effect, or substances that supplement the desired effect, including antacids, proton pump inhibiting factor, and H₂A receptor antagonist.

B. Parenteral administration

The pharmaceutical compositions provided by the present invention can be administered by injection, infusion, or implantation, for local or systemic administration, without passing through the alimentary canal. When used in the present invention, parenteral administration includes intravenous, intraarterial, intraperitoneal, intramembranous, intraventricular, intraurethral, intrasternal, intracranial, intramuscular, intrasynovial, and subcutaneous administration.

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

Pharmaceutical compositions intended for parenteral administration may comprise one or more pharmaceutically acceptable carriers and excipients, including, but not limited to, aqueous carriers, water-soluble carriers, anhydrous carriers, antimicrobial or preservative agents to inhibit 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 glucose injection, sterile water injection, dextrose and lactated ringer's injection. Anhydrous carriers include, but are not limited to, non-volatile 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 medium chain triglycerides of coconut oil, and palm seed oil. Water soluble carriers 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 agents or preservatives include, but are not limited to, phenol, cresol, mercurial, benzyl alcohol, chlorobutanol, methyl and propyl p-hydroxybenzenes, thimerosal, benzalkonium chloride, benzethonium chloride, methyl and propyl p-hydroxybenzoates, and sorbic acid. Suitable isotonic agents include, but are not limited to, sodium chloride, glycerol, and glucose. Suitable buffers include, but are not limited to, phosphate and citrate. Suitable antioxidants are described herein and include bisulfite and sodium metabisulfite. Suitable local anaesthetic packsIncluding, but not limited to procaine hydrochloride. Suitable suspending and dispersing agents are described herein and include sodium carboxymethylcellulose, hydroxypropylmethylcellulose, and polyvinylpyrrolidone. Suitable emulsifiers include those described herein, including polyoxyethylene sorbitan laurate, polyoxyethylene sorbitan monooleate 80, and triethanolamine oleate. Suitable masking 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 (c) ((r))CyDex,Lenexa,KS）。

The pharmaceutical compositions provided herein can be formulated for single or multiple dose administration. The single dose formulations are packaged in ampoules, vials, or syringes. The multi-dose parenteral formulation must contain a bacteriostatic or fungistatic concentration of an antimicrobial agent. As is well known and practiced in the art, all parenteral formulations must be sterile.

In one embodiment, the pharmaceutical composition may be provided as a ready-to-use sterile solution. In another embodiment, the pharmaceutical composition is provided as a sterile, dissolvable dry product that can be reconstituted with a carrier prior to use, including lyophilized powders and hypodermic tablets. 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, insoluble dry product that can be reconstituted with a carrier prior to use. In another embodiment, the pharmaceutical composition may be provided as a ready-to-use sterile emulsion.

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

The pharmaceutical composition may be formulated as a suspension, solid, semi-solid, or thixotropic liquid for administration as an implanted reservoir. In one embodiment, the pharmaceutical composition provided by the present invention is dispersed in a solid inner matrix surrounded by an outer polymeric membrane which is insoluble in body fluids but which allows the active ingredient in the pharmaceutical composition to diffuse through.

Suitable internal matrices include polymethylmethacrylate, polybutylmethacrylate, plasticized or unplasticized polyvinyl chloride, plasticized nylon, plasticized polyethylene terephthalate, natural rubber, polyisoprene, polyisobutylene, polybutadiene, polyethylene, ethylene vinyl acetate copolymers, silicone rubber, polydimethylsiloxane, silicone carbonate copolymers, hydrophilic polymers such as hydrogels of acrylates and methacrylates, collagen, crosslinked polyvinyl alcohol, and crosslinked partially 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, copolymers of vinyl chloride and vinyl acetate, vinylidene chloride, ethylene and propylene, polyethylene terephthalate ionomers, butyl rubber epichlorohydrin rubber, ethylene/vinyl alcohol copolymers, ethylene/vinyl acetate/vinyl alcohol terpolymers, and ethylene/ethyleneoxyethanol copolymers.

C. Surface application

The pharmaceutical compositions provided by the present invention can be topically applied to the skin, opening, or mucosa. When used in the present invention, topical administration includes dermal (intra), 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 for topical or systemic treatment including emulsions, solutions, suspensions, creams, gels, hydrogels, ointments, dusting powders, dressings, elixirs, lotions, suspensions, tinctures, pastes, foams, films, aerosols, rinses, sprays, suppositories, bandages, dermal ointments. The surface preparation of the pharmaceutical composition provided by the invention can also comprise liposome, micelle, microsphere, nano system and the mixture of the above.

Pharmaceutically acceptable carriers and excipients suitable for use in the surface formulations provided herein include, but are not limited to, aqueous carriers, water-soluble carriers, anhydrous carriers, antimicrobial agents or preservatives 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, permeation enhancers, cryoprotectants, lyoprotectants, thickening agents, and inert gases.

The pharmaceutical composition may also be topically administered by: electroporation, iontophoresis, sonophoresis, and micro-needle or needle-less injection, e.g. POWDERJECT^TM(Chiron Corp., Emeryville, Calif.), and BIOJECT^TM（Bioject Medical Technologies Inc.,Tualatin,OR）。

The pharmaceutical compositions provided by the present invention may be provided in the form of ointments, creams, and gels. Suitable ointment carriers include oleaginous or hydrocarbon bases including lard, benzoic lard, olive oil, cottonseed oil, white soft paraffin, and plastibase; emulsifiable or absorbent bases such as water-absorbent soft paraffin, sulfuric acid hydroxy glycerol tristearate, and anhydrous lanolin; water-removable bases, such as hydrophilic ointments; water-soluble ointment bases including polyethylene glycols of varying molecular weights; emulsion bases, oil-in-water (W/O) emulsions or water-in-oil (O/W) emulsions, including cetyl alcohol, glyceryl monostearate, lanolin, and stearic acid (see Remington: The Science and Practice of Pharmacy, supra). These carriers are emollients, but generally require the addition of antioxidants and preservatives.

Suitable cream bases may be water-in-oil or oil-in-water. The cream carrier may be water-washable and comprise an oil phase, an emulsifier, and an aqueous phase. The oil phase, also referred to as the "internal" phase, is typically composed of a soft paraffin and a fatty alcohol such as hexadecane or octadecanol. The aqueous phase is typically, although not necessarily, more voluminous than the oil phase and typically comprises a humectant. The emulsifier in the cream formulation may be a nonionic, anionic, cationic, or amphoteric surfactant.

The gel is a semi-solid suspension type system. Single phase gels comprise organic macromolecules distributed substantially uniformly in a liquid carrier. Suitable gelling agents include crosslinked acrylic acid polymers, such as carbomers, polycarboxylates, polyols,Hydrophilic polymers such as polyethylene oxide, polyoxyethylene-polyoxypropylene copolymer, and polyvinyl alcohol; cellulose polymers such as hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxypropyl methylcellulose phthalate, and methylcellulose; gums such as tragacanth and xanthan gum; sodium alginate; and gelatin. To prepare a homogeneous gel, a dispersing agent such as ethanol or glycerol 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 a suppository, pessary, stick, dressing, or paste, slurry, powder, dressing, cream, plaster, contraceptive, ointment, solution, emulsion, suspension, tampon, gel, foam, spray, or enema. These dosage forms can be prepared using conventional methods described in Remington, The Science and Practice of Pharmacy, supra.

Rectal, urethral, and vaginal suppositories are solid bodies that are inserted into the body openings, the suppositories being solid at ordinary temperatures but melting or softening at body temperature to release the active ingredient within the openings. Pharmaceutically acceptable carriers for rectal and vaginal use include carriers such as sclerosing agents that, when formulated in the pharmaceutical compositions provided herein, have a melting point near body temperature; also described herein are antioxidants including bisulfite and sodium metabisulfite. Suitable carriers include, but are not limited to, cocoa butter (theobroma oil), glycerol-gelatin, carbowax (polyoxyethylene glycol), spermaceti, paraffin, white and yellow waxes, and suitable mixtures of fatty acid mono-, di-and triglycerides, hydrogels, such as polyvinyl alcohol, hydroxyethyl methacrylate, polyacrylic acid; glycerol gelatin. Combinations of various carriers can be used. Rectal and vaginal suppositories may be prepared by compression methods or by molding. Rectal and vaginal suppositories typically weigh about 2 to 3 grams.

The pharmaceutical compositions provided by the present invention can be administered to the eye in the form of solutions, suspensions, ointments, emulsions, gel-forming solutions, powders for solutions, gels, ocular inserts, and implants.

The pharmaceutical compositions provided by the present invention may be administered intranasally or by inhalation to the respiratory tract. The pharmaceutical compositions may be provided in aerosol or solution form for administration using a pressurized container, pump, spray, nozzle, e.g., a nozzle that generates a fine aerosol using electrohydrodynamics, or a nebulizer, alone or in combination with a suitable propellant, e.g., 1,1,1, 2-tetrafluoroethane or 1,1,1,2,3,3, 3-heptafluoroethane. The pharmaceutical compositions may also be provided as a dry powder for insufflation, either alone or with an intercalating carrier such as lactose or a phospholipid; and provided as nasal drops. For intranasal use, the powder may contain a bioadhesive, including chitosan or cyclodextrin.

Solutions or suspensions for pressurized containers, pumps, sprays, nozzles, or sprayers can be formulated to contain the following ingredients: ethanol, aqueous ethanol, or suitable formulations for dispersing, dissolving, or prolonged release of the active ingredients provided herein, as a propellant for the solvent; and/or surfactants such as sorbitan trioleate, oleic acid, or oligolactic acid.

The pharmaceutical compositions provided herein can be micronized to a size suitable for inhalation delivery, for example 50 microns or less, or 10 microns or less. Particles of such size can be prepared using comminution methods well known to those skilled in the art, such as 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 or insufflator may be formulated as a powder mix comprising a pharmaceutical composition as provided by the present invention; suitable powder bases, such as lactose or starch; and a therapeutic effect modifier, such as l-leucine, mannitol, or magnesium stearate. The lactose may be in anhydrous or monohydrate form. Other suitable excipients include dextran, glucose, maltose, sorbitol, xylitol, fructose, sucrose, and trehalose. Pharmaceutical compositions provided by the present invention for inhalation/intranasal administration may further comprise suitable flavorants, such as menthol and levomenthol, or sweeteners, such as saccharin or saccharin sodium.

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

D. Modified release

The pharmaceutical compositions provided by the present invention can be formulated as modified release dosage forms. The term "modified release" as used herein refers to a dosage form in which the rate or site of release of the active ingredient is different from the immediate release dosage form when the route of administration is the same. Modified release dosage forms include delayed release, extended release, sustained release, pulsatile or pulsed release, controlled release, accelerated and rapid release, targeted release, programmed release, and gastric retention dosage forms. The pharmaceutical compositions of modified release dosage forms may be prepared using a variety of modified release devices and methods well known to those skilled in the art, including, but not limited to, matrix controlled release devices, osmotic controlled release 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 varied by varying the particle size and polymorphism of the active ingredient.

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

1. Matrix controlled release device

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

In one embodiment, the modified release dosage form pharmaceutical compositions provided herein are formulated using erodable matrix devices which are water-swellable, erodable, or soluble polymers, including synthetic polymers, and natural polymers and derivatives thereof, such as polysaccharides and proteins.

Materials that can be used to form the erodable matrix include, but are not limited to, chitin, chitosan, dextran, and pullulan; agar gum, gum arabic, karaya gum, locust bean 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; propylene glycol alginate; gelatin; collagen protein; and celluloses, e.g. Ethyl Cellulose (EC), methyl cellulose (MEC), carboxymethylCellulose Acetate (CA), Cellulose Propionate (CP), Cellulose Butyrate (CB), Cellulose Acetate Butyrate (CAB), CAP, CAT, Hydroxypropylmethylcellulose (HPMC), HPMCP, HPMCAS, hydroxypropylmethylcellulose acetate trimellitate (HPMCAT), and ethylhydroxyethylcellulose (EHEC); polyvinylpyrrolidone; polyvinyl alcohol; polyvinyl acetate; glycerin fatty acid ester; polyacrylamide; polyacrylic acid; copolymers of ethacrylic acid or methacrylic acid (EUDR)Rohm America, inc., Piscataway, NJ); poly (2-hydroxyethyl-methacrylate); a polylactide; copolymers of L-glutamic acid and ethyl-L-glutamate; degradable lactic acid-glycolic acid copolymers; poly-D- (-) -3-hydroxybutyric acid; and other acrylic acid derivatives such as butyl methacrylate, methyl methacrylate, ethyl acrylate, (2-dimethylaminoethyl) methacrylate, and chlorinated (trimethylaminoethyl) methacrylate.

In another embodiment, the pharmaceutical composition is formulated using a non-erodible matrix device. The active ingredient is dissolved or dispersed in an inert matrix and is released upon application primarily by diffusion within the inert matrix. Materials suitable for use as a non-erodible matrix device include, but are not limited to, insoluble plastics such as polyethylene, polypropylene, polyisoprene, polyisobutylene, polybutadiene, polymethylmethacrylate, polybutylmethacrylate, chlorinated polyethylene, polyvinyl chloride, methyl acrylate-methylmethacrylate copolymer, ethylene-vinyl acetate copolymer, ethylene/propylene copolymer, ethylene/ethyl acrylate copolymer, vinyl chloride-vinyl acetate copolymer, ethylene dicholoride, ethylene and propylene, polyethylene terephthalate ionomers, butyl rubber epichlorohydrin rubber, ethylene/vinyl alcohol copolymer, ethylene/vinyl acetate/vinyl alcohol terpolymer, and ethylene/ethyleneoxy alcohol copolymer, polyvinyl chloride, plasticized nylon, plasticized polyethylene terephthalate, natural rubber, silicone rubber, polydimethylsiloxane, silicone carbonate copolymers, and; hydrophilic polymers such as ethyl cellulose, cellulose acetate, povidone, and crosslinked partially hydrolyzed polyvinyl acetate; and also aliphatic compounds such as carnauba wax, microcrystalline wax, and triglycerides.

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

The modified release dosage form pharmaceutical compositions provided by the present invention may be prepared by methods well known to those skilled in the art, such as direct compression, dry or wet granulation followed by compression, and melt granulation followed by compression.

2. Osmotic controlled release device

The modified release dosage form pharmaceutical composition provided by the present invention can be prepared using an osmotic controlled release device comprising a one compartment system, a two compartment system, Asymmetric Membrane Technology (AMT), and Extruded Core System (ECS). Generally, such devices have at least two components: (a) a core comprising the active ingredient; and (b) a semi-permeable membrane having at least one delivery port encapsulating the core. The semipermeable membrane controls the flow of water from the aqueous environment to the core, thereby effecting drug release through the delivery port.

In addition to the active ingredient, the core of the osmotic engine optionally includes an osmotic agent that generates a driving force for the transfer of water from the environment to the core of the engine. One class of osmotic agents are water-swellable hydrophilic polymers, also referred to as "osmopolymers" and "hydrogels," including, but not limited to, hydrophilic ethylene and propylene polymers, polysaccharides such as calcium alginate, polyethylene oxide (PEO), polyethylene glycol (PEG), polypropylene glycol (PPG), poly (2-hydroxyethyl-methacrylate), polyacrylic acid, polymethacrylic acid, polyvinylpyrrolidone (PVP), crosslinked PVP, Polyethylene and Vinyl Acetate (PVA), PVA/PVP copolymers, copolymers of PVA/PVP with hydrophobic monomers such as methyl methacrylate and vinyl acetate, hydrophilic polyurethanes containing large PEO blocks, crosslinked sodium carboxymethylcellulose, carrageenan, hydroxyethyl cellulose (HEC), hydroxypropyl cellulose (HPC), hydroxypropyl methyl cellulose (HPMC), carboxymethyl cellulose (CMC), and carboxyethyl cellulose (CEC), Sodium alginate, polycarbophil, gelatin, xanthan gum, and sodium starch glycolate.

Another class of osmotic agents is osmogens, which absorb water to affect the osmotic pressure gradient across the barrier of the surrounding coating. Suitable osmogens 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 sulfite, 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, ethylenediaminetetraacetic acid, glutamic acid, p-toluenesulfonic acid, succinic acid, and tartaric acid; urea; and mixtures of the foregoing.

Different dissolution rates of the osmotic agent may be used to affect the rate at which the active ingredient is initially released from the dosage form. For example, amorphous sugars, such as those that can be delivered using Mannogeme EZ (SPI Pharma, Lewes, DE) provide rapid delivery over the first few hours to immediately produce the desired therapeutic effect and slowly and continuously release the remaining dose over an extended period of time to maintain the desired level of therapeutic or prophylactic effect. In this case, the rate of release of the active ingredient may replace the dosage of the active ingredient for metabolism and excretion.

The core may also include a variety of other excipients and carriers as described herein to enhance the performance of the dosage form or to enhance stability or advantageous processing.

Materials that may be used to form the semipermeable membrane include various grades of acrylic acid, ethylene, esters, polyamides, polyesters, and cellulose derivatives, which are permeable to water but not soluble in water at physiologically relevant pH values, or which may be rendered water insoluble by chemical changes such as cross-linking. Examples of suitable polymers that can be used to form the coating include plasticized, unplasticized, and reinforced Cellulose Acetate (CA), cellulose diacetate, cellulose triacetate, CA propionate, cellulose nitrate, Cellulose Acetate Butyrate (CAB), CA urethane, CAP, CA methyl carbamate, CA succinate, Cellulose Acetate Trimellitate (CAT), CA dimethylglycinate, CA ethyl carbonate, CA chloroacetate, CA ethyl oxalate, CA methyl sulfonate, CA butyl sulfonate, CA p-methylbenzenesulfonate, agar acetate, amylose triacetate, beta glucan acetate, beta glucan triacetate, dimethyl acetoacetate, locust bean triacetate, hydrated ethylene vinyl acetate, EC, PEG, PPG, PEG/PPG copolymer, PVP, HEC, HPC, CMC, CMEC, HPMC, HPMCP, HPMCAS, HPMCAT, polyacrylates and polymethacrylates and copolymers thereof, and mixtures thereof, Starches, dextrans, dextrins, chitosans, collagen, gelatin, polyalkenes, polyethers, polysulfones, polyethersulfones, polystyrenes, halogenated polyethylenes, polyvinyl esters and ethers, natural waxes, and synthetic waxes.

The semi-permeable membrane may also be a hydrophobic microporous membrane wherein the micropores are substantially gas filled and not wetted by an aqueous medium, 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, polytetrafluoroethylenes, polyacrylic acid derivatives, polyethers, polysulfones, polyethersulfones, polystyrenes, halogenated polyethylenes, polyvinylidene fluorides, polyvinylesters and ethers, natural waxes, and synthetic waxes.

The delivery port of the semi-permeable membrane may be formed by mechanical or laser drilling after coating. The delivery port may also be formed in situ by etching a plug of water soluble material or by rupturing a thinner portion of the film at the breach of the core. In addition, when the asymmetric membrane coating is of the type disclosed in U.S. Pat. Nos. 5,612,059 and 5,698,220, delivery openings may be formed during the coating process.

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

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

The osmotic controlled Release dosage forms can be prepared according to conventional methods and techniques well known to those skilled in The art (see Remington: The Science and Practice of Pharmacy; Santus and Baker, J.Controled Release 1995,55, 1-21; Verma et al., Drug Development and Industrial Pharmacy 2000,26, 695-.

In certain embodiments, the pharmaceutical compositions provided herein are formulated as AMT controlled release dosage forms comprising an asymmetrically permeable membrane coating a core comprising 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 forms may be prepared according to conventional methods and techniques well known to those skilled in the art, including direct compression, dry granulation, wet granulation, and dip coating.

In a certain embodiment, the pharmaceutical composition provided by the present invention is formulated as an ESC controlled release dosage form comprising a permeable membrane coating a core comprising the active ingredient, hydroxyethylcellulose, and other pharmaceutically acceptable excipients.

3. Multiparticulate controlled release device

The pharmaceutical composition of the modified release dosage form provided by the present invention may be prepared using a multiparticulate controlled release device comprising a plurality of microparticles, granules, or pellets having a diameter of about 10 μm to 3mm, about 50 μm to 2.5mm, or about 100 μm to 1 mm. Such multiparticulates can be prepared by methods well known to those skilled in the art, including wet and dry granulation, extrusion/spheronization, roller compaction, fusing, and spray coating of the core. See, e.g., Multiparticulate organic Drug Delivery, Marcel Dekker: 1994; and Pharmaceutical pelletisation Technology, Marcel Dekker, 1989.

Other excipients described herein may be mixed with the pharmaceutical composition to aid in processing and forming the multiparticulates. The resulting microparticles may themselves comprise the multiparticulate device or may be coated with a variety of 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 being treated, including liposomes, re-encapsulated red blood cells, and antibody-based delivery systems. Examples include, but are not limited to, U.S. patent 6,316,652; 6,274,552, respectively; 6,271,359; 6,253,872, respectively; 6,139,865, respectively; 6,131,570, respectively; 6,120,751, respectively; 6,071,495; 6,060,082, respectively; 6,048,736, respectively; 6,039,975, respectively; 6,004,534, respectively; 5,985,307, respectively; 5,972,366; 5,900,252, respectively; 5,840,674, respectively; 5,759,542, respectively; and 5,709,874.

Application method

In one embodiment, there is provided a method of treating, preventing, or ameliorating CRTH2 and/or one or more other PGDs₂A method of treating a disorder or disease associated with a receptor by administering to an individual having or suspected of having said disorder or disease a therapeutically effective amount of an amine salt of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid, or an amine salt thereofA pharmaceutically acceptable hydrate or solvate; or a pharmaceutical composition thereof.

In another embodiment, there is provided a method of treating, preventing, or ameliorating CRTH2 and/or one or more other PGDs₂A method of modulating one or more symptoms of a responsive disease or disorder by a receptor, the method comprising administering to an individual having or suspected of having the disease or disorder a therapeutically effective amount of one or more amine salts or compositions provided herein.

In another embodiment, there is provided a method of treating, preventing, or ameliorating CRTH2 and/or one or more other PGDs₂A method of treating one or more symptoms of a receptor-mediated disease or disorder, the method comprising administering to an individual having or suspected of having the abnormality or disease a therapeutically effective amount of one or more amine salts or compositions provided herein.

In another embodiment, there is provided a method of treating, preventing, or ameliorating one or more symptoms of an eosinophil-associated disease, comprising administering to an individual a therapeutically effective amount of an amine salt of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid, or a pharmaceutically acceptable hydrate or solvate thereof; or a pharmaceutical composition thereof.

In another embodiment, there is provided a method of treating, preventing, or ameliorating one or more symptoms of a basophil-related disease, the method comprising administering to an individual a therapeutically effective amount of an amine salt of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid, or a pharmaceutically acceptable hydrate or solvate thereof; or a pharmaceutical composition thereof.

In another embodiment, there is provided a method of treating, preventing, or ameliorating one or more symptoms of an inflammatory disease, the method comprising administering to an individual a therapeutically effective amount of an amine salt of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid, or a pharmaceutically acceptable hydrate or solvate thereof; or a pharmaceutical composition thereof.

Conditions and diseases that can be treated with one or more amine salts provided herein include, but are not limited to, (1) inflammatory or allergic diseases, including systemic allergic and hypersensitivity conditions, atopic dermatitis, nettle rash, drug allergies, insect sting allergies, food allergies (including celiac disease and the like), and mastocytosis; (2) inflammatory bowel diseases including Crohn's disease, ulcerative colitis, ileitis, and enteritis; (3) vasculitis, and behcet's syndrome; (4) psoriasis and inflammatory skin diseases including dermatitis, eczema, atopic dermatitis, allergic contact dermatitis, urticaria, viral skin lesions including lesions from human papilloma virus, HIV or RLV infections, bacteria, flugal, and other parasitic skin lesions, and cutaneous lupus erythematosus; (5) asthma and allergic diseases of the respiratory tract including allergic asthma, allergic rhinitis, otitis media, exercise-induced asthma, allergic conjunctivitis, hypersensitivity lung diseases, and chronic obstructive pulmonary disease; (6) autoimmune diseases, including arthritis (including rheumatoid and psoriatic), systemic lupus erythematosus, type I diabetes, myasthenia gravis, multiple sclerosis, hyperthyroidism, and glomerulonephritis; (7) transplant rejection (including allograft rejection and graft-versus-host disease), such as skin transplant rejection, solid organ transplant rejection, bone marrow transplant rejection; (8) generating heat; (9) cardiovascular disorders including acute heart failure, hypotension, hypertension, angina pectoris, myocardial infarction, cardiomyopathy, congestive heart failure, atherosclerosis, coronary heart disease, restenosis, and vascular stenosis; (10) cerebrovascular disorders including traumatic brain injury, stroke, ischemic reperfusion injury, and aneurysm; (11) breast cancer, skin cancer, prostate cancer, cervical cancer, uterine cancer, ovarian cancer, testicular cancer, bladder cancer, lung cancer, liver cancer, laryngeal cancer, oral cancer, colon cancer, and gastrointestinal cancer (e.g., esophageal cancer, gastric cancer, pancreatic cancer), brain cancer, thyroid cancer, leukemia, and lymphoma; (12) fibrosis, connective tissue disease, and sarcoidosis; (13) genital and reproductive abnormalities, including erectile dysfunction; (14) gastrointestinal disorders including gastritis, ulcers, nausea, pancreatitis, and vomiting; (15) neurological disorders including alzheimer's disease; (16) sleep disorders including insomnia, narcolepsy, sleep apnea syndrome, and Pickwick syndrome; (17) pain; (18) renal disorders; (19) eye disorders, including glaucoma; and (20) infectious diseases, including HIV.

In certain embodiments, the disease is selected from the group consisting of: asthma, allergic asthma, exercise-induced asthma, allergic rhinitis, perennial 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, allergic granulomatous vasculitis, 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 allergic granulomatous vasculitis or sinusitis.

Depending on the disease to be treated and the condition of the individual, the amine salts 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, intranasal, vaginal, rectal, sublingual, or topical (e.g., transdermal or topical) routes of administration, and can be formulated in suitable dosage units, either alone or in combination with pharmaceutically acceptable carriers, adjuvants, and vehicles appropriate for each route of administration. Also provided is the administration of the amine salts provided herein in a depot formulation, wherein the active ingredient is released over a predetermined period of time.

When treating, preventing, or ameliorating asthma, allergic rhinitis, eczema, psoriasis, atopic dermatitis, fever, sepsis, systemic lupus erythematosus, diabetes, rheumatoid arthritis, multiple sclerosis, atherosclerosis, transplant rejection, inflammatory bowel disease, cancer, or other conditions associated with CRTH2 and/or one or more other PGDs₂Where one or more symptoms of a subject-related disorder, condition, or disease are indicated, suitable dosage levels will generally be from about 0.001 to about 100mg per kilogram 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 dosage may be administered in single or multiple doses. Within this range the dose may be 0.005 to 0.05, 0.05 to 0.5, or 0.5 to 5.0, 1 to 15, 1 to 20, or 1 to 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 in the form of a tablet containing 1.0 to 1,000mg of the active ingredient, specifically 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 the active ingredient, the dosage being adjusted according to the symptoms of the patient being treated. The composition may be administered in a treatment regimen of 1 to 4 times per day, including 1,2,3, and 4 times per day.

It will be understood, however, that the specific dose level and frequency of dosage for any 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 disorder, and the recipient undergoing therapy.

Also provided is a method of modulating CRTH2 and/or one or more other PGDs₂A method of contacting a subject comprising contacting the subject with one or more amine salts or compositions provided herein. In one embodiment, the receptor is expressed by a cell.

The amine salts provided herein may also be used in combination or association with other agents useful in the treatment, prevention, or amelioration of one or more symptoms of diseases or disorders for which the amine salts 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 pathologies described above.

Such other formulations, or medicaments, may be administered simultaneously or sequentially with the amine salts provided herein by the usual routes and dosages. When the amine salts provided herein are used concurrently with one or more other drugs, pharmaceutical compositions containing such other drugs in addition to the amine salts provided herein may be used, but are not required. Accordingly, the pharmaceutical compositions provided herein include those that contain one or more other active ingredients or therapeutic agents in addition to the amine salts provided herein.

The weight ratio of the amine salt to the second active ingredient provided by the present invention may vary and is dependent upon the effective dosage of each ingredient. Generally, effective dosages of each ingredient are employed. Thus, for example, when the amine salt provided herein is combined with an NSAID, the weight ratio of amine salt provided herein to NSAID may range from about 1,000: 1 to about 1: 1,000, or about 200: 1 to about 1: 200. combinations of the amine salts provided by the present invention with other active ingredients are also generally within the above-described ranges, but in each case, an effective dose of each active ingredient should be used.

Examples

In the following examples, amine salts of acids of formula I were detected using nuclear magnetic resonance spectroscopy (NMR), X-ray powder diffraction (XRPD), Differential Scanning Calorimetry (DSC), Thermogravimetry (TGA), and Scanning Electron Microscopy (SEM).

Unless otherwise indicated, all¹H NMR spectra were recorded at 300MHz using Bruker Instruments NMR. Coupling constants (J) are given in Hertz (Hz) and peaks are listed relative to TMS (Δ 0.00 ppm).

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

Differential scanning calorimetry was performed using Mettler 850, TA 2920. The samples were analyzed in sealed aluminum pans, with empty aluminum pans as controls. A heating rate of 10 ℃/min was used in the temperature range of 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 in the temperature range of 20 ℃ to 600 ℃.

The average particle size of the solid amine salt was determined using a scanning electron microscope.

Example 1

Base selection of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetate

The free acid of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid was screened using 22 pharmaceutically acceptable bases and the results are summarized in table 1 together with its water solubility. Among the bases screened, ethylenediamine, piperazine, benzathine, and choline each produced crystalline solids suitable for the pharmaceutical formulations and uses described herein.

Example 2

Preparation of ethylenediamine salt of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid

A mixture of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid (25.2 mg, 0.05 mmol) and ethylenediamine (1.68 mg, 0.03 mmol) in ethanol (1.1 mL) was heated at reflux temperature to obtain a clear solution. The clear solution was cooled to room temperature and stirring was continued for 2 hours. The precipitate was collected by filtration, washed with 0.5mL ethanol/heptane (1: 1, v/v), and dried under vacuum at ambient temperature to give plate-like crystals (20.1 mg) (fig. 5).

Stoichiometric use of the ethylenediamine salt¹H NMR measurement (fig. 1). The ethylenediamine salt of the acid of formula I comprises about 2 molar equivalents of the acid and 1 molar equivalent of ethylenediamine. For 2 molar equivalents of ethanol, about 1 molar equivalent of ethanol is in the crystalline salt. The crystalline ethylenediamine salt contained about 4% ethanol. The ethylenediamine salt had an X-ray powder diffraction pattern as shown in figure 2, with a characteristic XRP diffraction peak at 5.9 degrees 2-theta. This confirmed that the ethylenediamine salt was a crystalline material.

The differential scanning calorimetry thermogram of the ethylenediamine salt is shown in figure 3. The ethylenediamine salt showed an endothermic effect with a peak temperature of 123.1 ℃ and an onset temperature of 114.1 ℃, and a peak temperature of 216.0 ℃ and an onset temperature of 212.1 ℃.

TABLE 1 alkali screening

The thermogravimetric analysis thermogram is shown in fig. 4. The ethylenediamine salt showed a slight weight loss at 75 ℃ and a weight loss of 3.951% from 125 ℃ to 150 ℃, which is comparable to¹H NMR determination (fig. 1) the crystalline ethylenediamine salt observed contained about 4% ethanol phase.

Other solvent systems for preparing crystalline ethylenediamine salts of the acids of formula I were also evaluated. The results obtained are summarized in table 2.

TABLE 2 solvent screening of ethylenediamine salts

Solvent system	Results
		Methyl Ethyl Ketone (MEK)	Soluble solid
Methanol	Soluble solid
		Ethanol	Crystalline solid
Acetone (II)	Insoluble solid
		Methyl isobutyl ketone (MIBK)	Soluble solid

Example 3

Preparation of piperazine salt of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid

A mixture of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid (25.0 mg, 0.05 mmol) and piperazine (2.38 mg, 0.03 mmol) in ethanol (1.1 mL) was heated at reflux temperature to obtain a clear solution. The clear solution was cooled to room temperature and stirring was continued for 2 hours. The precipitate was collected by filtration, washed with 0.5mL of tert-butyl methyl ether (TBME), and dried under vacuum at ambient temperature to give needle-shaped crystals (21.7 mg) (fig. 10).

Stoichiometric use of the piperazine salt¹H NMR (fig. 6). The piperazine salt of an acid of structural formula I comprises about 2 molar equivalents of the acid and 1 molar equivalent of piperazine.

The piperazine salt has an X-ray powder diffraction pattern as shown in fig. 7 with characteristic XRP diffraction peaks at 10.7, 15.9, 22.3, and 24.0 degrees 2-theta. This confirms that the piperazine salt is a crystalline material.

The DSC thermogram of the piperazine salt is shown in figure 8. The piperazine salt showed strong endotherm with a peak temperature of 223.2 ℃ and an onset temperature of 219.3 ℃, and weak endotherm with a peak temperature of 202.6 ℃ and an onset temperature of 198.4 ℃.

The thermogravimetric analysis thermogram of the piperazine salt is shown in fig. 9. The piperazine salt showed a slight weight loss at 75 ℃ and 3.951% weight loss at 125 ℃ to 150 ℃.

Other solvent systems for preparing crystalline piperazine salts of acids having structural formula I were also evaluated. The results obtained are summarized in table 3.

TABLE 3 solvent screening of piperazine salts

Solvent system	Results
		Ethanol	Crystalline solid
Acetone (II)	Insoluble in water
		THF	Crystalline solid
Ethanol/water	Crystalline solid
		Methyl Ethyl Ketone (MEK)	Insoluble in water
Isopropanol (IPA)	Insoluble in water
		Acetonitrile	Insoluble in water

Example 4

Preparation of benzathine salt of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid

A mixture of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid (250.0 mg, 0.5 mmol) and benzathine (134.4 mg, 0.56 mmol) in ethanol (3.0 mL) was heated at reflux temperature to obtain a clear solution. The clear solution was cooled to room temperature and stirring was continued for 2 hours. The precipitate was collected by filtration, washed with 2mL IPA/heptane (1: 1, v/v), and dried under vacuum at ambient temperature to give a crystalline solid in the form of a rod (256.8 mg) (FIG. 14).

The stoichiometric use of the benzathine salt¹H NMR (fig. 11). The benzathine salt of the acid of formula I comprises about 2 molar equivalents of the acid and 1 molar equivalent of benzathine.

The benzathine salt has an X-ray powder diffraction pattern as shown in fig. 12 with characteristic XRP diffraction peaks at 8.0, 11.5, 16.0, 17.5, and 23.4 degrees 2 Θ. This confirms that the benzathine salt is a crystalline material.

The DSC thermogram of the benzathine salt is shown in figure 13. The piperazine salt showed a strong endothermic effect with a peak temperature of 155.8 ℃ and an onset temperature of 154.2 ℃.

Example 5

Preparation of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzoylamino) benzyl) pyrimidin-5-ylCholine salt of acetic acid

A mixture of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid (249.5 mg, 0.5 mmol) and choline (69.4 mg, 0.58 mmol) in IPA (3.5 mL) was heated at reflux temperature to obtain a clear solution. The clear solution was cooled to room temperature and stirring was continued for 2 hours. The precipitate was collected by filtration, washed with 2mL IPA/heptane (1: 1, v/v), and dried under vacuum at ambient temperature to yield a rod-like and rhombohedral crystalline solid (232 mg) (FIG. 19).

Stoichiometric use of the choline salt¹H NMR (fig. 15). The choline salt of an acid of structural formula I comprises about 1 molar equivalent of the acid and 1 molar equivalent of choline.

The choline salt has an X-ray powder diffraction pattern as shown in figure 16 with characteristic XRP diffraction peaks at 6.5, 19.6, 20.0, 21.9, and 26.4 degrees 2 Θ angles. This confirms the choline salt as a crystalline material.

The DSC thermogram of the choline salt is shown in figure 17. The choline salt showed a strong endothermic effect with a peak temperature of 194.8 ℃ and an onset temperature of 192.6 ℃.

The thermogravimetric analysis thermogram of the choline salt is shown in fig. 18. The choline salt showed almost no weight loss up to 200 ℃.

Other solvent systems for preparing crystalline choline salts of acids having structural formula I were also evaluated. The results obtained are summarized in Table 4.

TABLE 4 solvent screening of choline salts

Solvent system	Results
		IPA/water	Soluble solid
IPA	Crystalline solid
		Ethanol	Soluble solid
Ethanol/water	Soluble solid

Example 6

Competitive radioligand binding assay

Competitive radioligand binding assays using stably transfected cell lines expressing CRTH2 or DPI evaluated 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 invention.

Prostaglandin D₂And 13, 14-dihydro-15-one PGD₂Obtained from Cayman Chemical (Ann Arbor, MI). Radiolabelled PGD with a specific activity of 160Ci/mmol₂（5，6，8，9，12，14，15-³H (N)) from PerkinElmer (Boston, MA). Cell culture media RPMI 1640, HEPES buffer, Phosphate Buffered Saline (PBS), L-glutamine, and penicillin-streptomycin solutions were obtained from Mediatech Inc. Calf Serum (FCS) was obtained from Cambrex (Walkersville, MD). Puromycin was obtained from Invitrogen (San Diego, Calif.). Polyethyleneimine (PEI) obtained fromAcros Organics (Morris Plains, NJ). Purified monoclonal rat anti-human CRTH2 antibody (BM 16), rat IgG2a, κ, (clone R35-95), and goat anti-rat Ig-FITC were obtained from Becton Dickinson Biosciences (San Diego, CA). BSA (fraction V) and sodium azide were obtained from Sigma Chemical Company (St. Louis).

The CRTH2 and DPI stable cell lines were prepared according to the methods 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 (25 mM), FCS (10%), L-glutamine (2 mM), penicillin (10 IU/mL), streptomycin (100. mu.g/mL), and puromycin (1. mu.g/mL).

The surface expression of CRTH2 on transfected cell lines was monitored periodically during culture and prior to each competitive radioligand binding assay. The CRTH2 expressing cells and untransfected cells were incubated with purified anti-CRTH 2 antibody (clone BM 16) or isotype control antibody (rat IgG2a, κ) on ice for 15 minutes. The cells were washed in 2mL FACS buffer (PBS containing 1% BSA (fraction V) and 0.1% sodium azide). The cells were then incubated with FITC-conjugated anti-rat antibody (goat anti-rat Ig-FITC) on ice. After washing with cold FACS buffer, the cells were analyzed using a FACScan II analyzer and CellQuest software, both from Becton Dickinson (Mountain View, CA).

Also by direct determination of the response PGD₂The functionality of the receptor transfectants was analyzed by calcium mobilization (Sugimoto et al, J.Pharm.Exp.Therap.2003,305, 347-352; Sugimoto et al, Eur.J.Pharmacol.2005,524, 30-37).

Radioligand binding assays were performed according to the method of Sugimoto et al (J.Pharm.Exp.Therap.2003,305, 347-352; Eur.J.Pharmacol.2005,524, 30-37). The test compounds were dissolved in DMSO at a concentration of 100mM as stock solutions. Initial serial dilutions of 10 μ M were prepared in binding buffer prior to analysis. Transfected cells were at 4X 10 at room temperature⁶Concentrate in/mLThe suspension was resuspended in binding buffer (50 mM Tris-HCl, 40mM MgCl)₂And 0.1% bovine serum albumin, pH 7.4). Transfected cells were seeded on U-bottom polypropylene 96-well plates (Fisher) by adding 50. mu.L of the cell suspension followed by 10. mu.L³H-PGD₂10 μ L of the test compound or control, 30 μ L of the binding buffer, to give a final volume of 100 μ L. In each hole³H-PGD₂Was 1 nM. After incubation for 1 hour with gentle shaking at room temperature, the cell suspension was transferred to a filter plate (Millipore, MA) pre-wetted with 0.5% PEI buffer. The cell pellet was washed 3 times with the binding buffer and the filter discs were air dried. Add fluorescer (50. mu.L; Microscint) to each well^TMRadioactivity was counted using a TopCount (Packard Bioscience, Meriden, CT) 20, Perkin Elmer, Boston, Mass.). Using Prizm^TMData analysis was performed by a graphical program (GraphPad Software inc., San Diego, CA). As shown in fig. 20 and 21, {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid of structural formula I and its pharmaceutically acceptable salts have high affinity and binding selectivity for CRTH 2.

The examples set forth above are provided herein to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the embodiments, and are not intended to limit the scope of the disclosure. Modifications of the above-described methods for implementing the disclosure that are obvious to those skilled in the art are intended to be included within the scope of the appended claims. All publications, patents, and patent applications cited in this specification are herein incorporated by reference as if each such publication, patent, or patent application were specifically and individually indicated to be incorporated by reference.

Claims (22)

1. A crystalline benzathine salt of {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid of formula I:

wherein the crystalline salt has an X-ray powder diffraction pattern substantially as shown in figure 12.

2. A pharmaceutical composition comprising the salt of claim 1 and a pharmaceutically acceptable excipient.

3. The pharmaceutical composition of claim 2, wherein the composition is formulated for oral, intranasal, bronchial, or topical administration.

4. The pharmaceutical composition of claim 2, wherein the composition is formulated as a single dosage form.

5. Use of a salt of claim 1 in the manufacture of a medicament for treating, preventing, or ameliorating one or more symptoms of a CRTH 2-mediated disease.

6. Use of a salt according to claim 1 in the manufacture of a medicament for treating, preventing, or ameliorating one or more symptoms of an eosinophil-related disease.

7. Use of a salt of claim 1 in the manufacture of a medicament for treating, preventing, or ameliorating one or more symptoms of a basophil-related disease.

8. Use of a salt of claim 1 in the manufacture of a medicament for treating, preventing, or ameliorating one or more symptoms of an inflammatory disease.

9. The use of claim 5, wherein the disease is selected from: asthma, allergic rhinitis, atopic dermatitis, contact hypersensitivity, contact dermatitis, 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, allergic granulomatous vasculitis, sinusitis, basophilic leukemia, chronic urticaria, basophilic leukocytosis, eczema, chronic obstructive pulmonary disease, and arthritis.

10. The use according to claim 5, wherein the disease is asthma, allergic rhinitis, atopic dermatitis, chronic obstructive pulmonary disease, or allergic conjunctivitis.

11. The use according to claim 5, wherein the disease is allergic granulomatous vasculitis or sinusitis.

12. The use as claimed in claim 5, wherein the disease is allergic asthma or exercise-induced asthma.

13. The use as claimed in claim 5, wherein the disease is persistent allergic rhinitis or seasonal allergic rhinitis.

14. The use as claimed in claim 5, wherein the disease is rheumatoid arthritis, psoriatic arthritis or osteoarthritis.

15. A process for preparing the salt of claim 1, comprising the steps of:

(a) reacting {4, 6-bis (dimethylamino) -2- (4- (4- (trifluoromethyl) benzamido) benzyl) pyrimidin-5-yl } acetic acid with the amine in a solvent at a first predetermined temperature to form the amine salt; and

(b) precipitating the amine salt at a second predetermined temperature.

16. The method of claim 15, wherein the first temperature is from-10 to 150 ℃.

17. The method of claim 15, wherein the first temperature is 20 to 100 ℃.

18. The method of claim 15, wherein the second temperature is from-50 to 50 ℃.

19. The method of claim 15, wherein the second temperature is-23 to 35 ℃.

20. The method of claim 15, wherein the solvent is selected from the group consisting of: methanol, ethanol, isopropanol, propanol, tetrahydrofuran, water, or a mixture of the above solvents.

21. The method of claim 15, wherein an anti-solvent is added in the precipitation step to effect precipitation.

22. The method of claim 21, wherein the anti-solvent is selected from the group consisting of: water, alkanes, ethers, aromatic hydrocarbons, or mixtures of the above solvents.