HK1168354A - Crystalline forms of 3-[5-(2-fhjorophenyl)-[1,2,4]oxadiazol-3-yl]-benzoic acid - Google Patents

Abstract

The present invention relates to crystalline forms of 3-[5-(2-fluorophenyl)-[1,2,4]oxadiazol-3-yl]-benzoic acid, pharmaceutical compositions and dosage forms comprising the crystalline forms, methods of making the crystalline forms and methods for their use for the treatment, prevention or management of diseases ameliorated by modulation of premature translation termination or nonsense-mediated mRNA decay.

Description

Crystalline forms of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid

The application is a division of the invention application with the application date of 24.09.2007 and the application number of 200780043582.X and the name of 'the crystal form of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazole-3-yl ] -benzoic acid'.

This application claims priority from U.S. provisional application No. 60/847,326 filed on 25.9.2006, which is hereby incorporated by reference in its entirety.

1. Field of the invention

The present invention relates to crystalline forms of the compound 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid, pharmaceutical compositions and dosage forms comprising the crystalline forms, methods of preparing the crystalline forms, and methods of using the same for the treatment, prevention or control of diseases ameliorated by the modulation of premature translation termination or nonsense-mediated mRNA degradation.

2. Background of the invention

As described in U.S. patent No. 6,992,096B2, granted on 31/2006 (which is incorporated herein by reference in its entirety), 1,2, 4-oxadiazole compounds are useful in the treatment, prevention, or control of diseases ameliorated by the modulation of premature translation termination or nonsense-mediated mRNA degradation. One such compound is 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid.

It is known in the pharmaceutical art that solid forms of a compound, such as salts, crystalline forms (e.g., polymorphs), affect, for example, the solubility, stability, flowability, fractionability, and compressibility of the compound, as well as the safety and efficacy of pharmaceutical products based on the compound (see, e.g., Knapman, k. modern Drug discovery, 2000: 53). The potential impact of solid forms in a single drug product on the safety and efficacy of the corresponding drug product is so important that the U.S. food and drug administration requires the identification and control of the solid form, e.g., crystalline form, of each compound used in each drug product sold in the U.S. Thus, the novel crystalline forms of 1,2, 4-oxadiazole benzoic acid may advance the development of formulations for the treatment, prevention or control of diseases ameliorated by modulation of premature translation termination or nonsense-mediated mRNA degradation. The present invention provides such novel crystalline forms, for example, a crystalline form of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid.

Citation of any reference in the second section of this application is not to be construed as an admission that such reference is prior art to the present application.

3. Summary of the invention

The present invention provides a novel crystalline form of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid, which compound has the following chemical structure (I):

in particular, crystalline forms of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid are useful in the treatment, prevention or control of diseases ameliorated by the modulation of premature translation termination or nonsense-mediated mRNA degradation as described in U.S. patent No. 6,992,096B2, granted on month 1 and 31 of 2006, which is hereby incorporated by reference in its entirety. In addition, the present invention provides substantially pure crystalline forms of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid, i.e., having a purity of greater than about 90%.

Certain embodiments of the present invention provide pharmaceutical dosage forms and compositions comprising the crystalline form of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid and a pharmaceutically acceptable diluent, excipient or carrier. The invention further provides methods of using them to treat, prevent or control diseases ameliorated by modulation of premature translation termination or nonsense-mediated mRNA degradation. In certain embodiments, the present invention provides methods of making, isolating and/or characterizing the crystalline forms of the present invention. The crystalline forms of the invention are useful as active pharmaceutical ingredients for the preparation of formulations for animals or humans. Thus, the present invention encompasses the use of these crystalline forms as the final pharmaceutical product. The crystalline forms and final pharmaceutical products of the invention are useful, for example, in the treatment, prevention or management of the diseases described herein.

4. Detailed description of the invention

4.1 brief description of the drawings

Figure 1 provides an X-ray powder diffraction (XRPD) pattern of a sample comprising form a of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid.

Figure 2 provides Differential Scanning Calorimetry (DSC) and thermogravimetric analysis (TGA) temperature profiles of a sample comprising crystalline form a of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid.

Figure 3 provides dynamic gas phase adsorption (DVS) isotherms for samples comprising form a of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid.

FIG. 4 provides a peptide composition comprising 3- [5- (2-fluorophenyl) - [1,2,4]]Oxadiazol-3-yl]-solid state of a sample of benzoic acid form a¹³C NMR spectrum.

Figure 5 provides an XRPD pattern of a sample comprising form B of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid.

Figure 6 provides DSC and TGA temperature profiles for a sample comprising form B of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid.

Figure 7 provides DVS isotherms of a sample comprising form B of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid.

Figure 8 provides an overlay of experimental XRPD patterns showing a set of characteristic peaks for form a (top) relative to several samples containing form B of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid (second from top to bottom), illustrating peak migration in certain samples of form B.

Fig. 9 provides a crystal packing diagram of form a of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid, viewed down the b-axis of the crystal diagram, showing the outline of the unit cell.

Figure 10 provides an XRPD pattern of form a of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid that mimics a single crystal X-ray diffraction crystal structure obtained from a representative form a single crystal.

FIG. 11 provides an ORTEP plot of single crystal XRD crystal structure asymmetric units of form A of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid. The atoms are represented as thermal ellipsoids with 50% probability anisotropy.

4.2 terminology

Crystalline forms equivalent to those described below and claimed herein may exhibit similar, but non-identical analytical properties within reasonable margins of error, depending on test conditions, purity, equipment and other common variables known to those skilled in the art or reported in the literature. The term "crystalline" and related terms used herein, when used to describe a substance, ingredient or product, means that the substance, ingredient or product is substantially crystalline, as determined by X-ray diffraction, microscopy, polarization microscopy, or other known analytical methods known to those skilled in the art. See, e.g., Remington's Pharmaceutical Sciences, 18 th edition, Mack Publishing, Easton PA, 173 (1990); the United States Pharmacopeia, 23 rd edition, 1843-1844 (1995).

Thus, it will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the scope and spirit of the invention. Other embodiments of the invention will be apparent to those skilled in the art from consideration of the specification and practice of the invention disclosed herein. It is intended that the specification and examples be considered as exemplary only, with no limitation on the scope.

The crystalline forms of the present invention can be characterized using single crystal data, powder X-ray diffraction (PXRD), Differential Scanning Calorimetry (DSC), and thermogravimetric analysis (TGA). It is to be understood that the numerical values set forth and claimed herein are approximations. Variations in the values may be due to equipment calibration, equipment errors, material purity, crystal size, and sample size, among other factors. Moreover, variations are possible while still achieving the same results. For example, X-ray diffraction values are typically accurate to within 0.2 degrees, while intensities (including relative intensities) in an X-ray diffraction pattern may fluctuate depending on the measurement conditions employed. Similarly, DSC results are typically accurate to within about 2 ℃. Accordingly, it is to be understood that the crystalline forms of the present invention are not limited to crystalline forms providing the exact same profiles (i.e., one or more of PXRD, DSC, and TGA) as those depicted in the figures of the present disclosure. Any crystalline form having a characteristic pattern substantially the same as those described in the figures falls within the scope of the present invention. The ability to determine substantially identical feature maps is within the ability of one of ordinary skill in the art.

The embodiments provided by the present invention can be more fully understood by reference to the following detailed description and illustrative examples, which are intended to illustrate non-limiting embodiments.

U.S. patent No. 6,992,096B2, granted on 31.2006, and U.S. patent application No. 11/899,813, filed 9.2007, both of which are incorporated herein by reference in their entirety, describe methods for the preparation of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid.

4.33- [5- (2-fluorophenyl) - [1,2,4]]Oxadiazol-3-yl]-benzoic acid in crystal form A

In one embodiment, the present invention provides form a crystalline form of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid. In certain embodiments, form a may be obtained by crystallization from different solvents including, but not limited to: methanol, t-butanol (t-BuOH), 1-butanol (1-BuOH), acetonitrile, Isopropanol (IPA), isopropyl ether, dimethylformamide, heptane, isopropyl acetate (IPOAc), toluene and/or water. Figure 1 provides a representative XRPD pattern of form a of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid. In certain embodiments, form a of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid has an XRPD pattern substantially similar to the pattern presented in figure 1.

Representative thermal properties of form a of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid are shown in figure 2. The representative DSC temperature profile presented in fig. 2 shows an endothermic event with a peak temperature of about 244 ℃. The representative TGA temperature profile, also presented in figure 2, exhibits a mass loss of less than about 1% of the total mass of the sample when heated from about 33 ℃ to about 205 ℃. These thermal data indicate that form a of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid contains substantially no water or solvent in its crystal lattice. In certain embodiments, form a exhibits TGA weight loss phenomena beginning at about 212 ℃, which corresponds to sublimation prior to melting.

Single crystal X-ray diffraction (XRD) crystal structure obtained from 3- [5- (2-fluorophenyl) - [1,2,4]]Oxadiazol-3-yl]-a representative single crystal of form a of benzoic acid. Using XRD data collected at about 150K, the following unit cell parameters were obtained:α＝90°；β＝92.9938(15)°；γ＝90°； figure 9 provides a crystallographic stacking diagram of a single crystal XRD structure from form a, viewed down the crystallographic b axis. A simulated XRPD pattern for Cu radiation was generated using Powdercell 2.3(Powdercell for Windows Version 2.3 Kraus, W.; Nolze, G.Federal Institute for Materials Research and Testing, Berlin Germany, EU, 1999) and the atomic coordinates, space groups and cell parameters from the single crystal data. FIG. 10 provides 3- [5- (2-fluorophenyl) - [1,2,4]]Oxadiazol-3-yl]-a simulated XRPD pattern of form a of benzoic acid.

In certain embodiments, crystalline form a of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid is characterized by its physical stability when subjected to certain processing conditions. In certain embodiments, form a is physically stable when stored for 6 days under one or more of the following Relative Humidity (RH) conditions: 40 ℃ at 53% RH; 75% RH at 40 ℃; 60 ℃ 50% RH; and 60 ℃ at 79% RH. In other embodiments, form a is physically stable when milled at ambient and sub-ambient temperatures. In other embodiments, form a is physically stable when slurried under one or more of the following conditions: 4 days in 1-butanol at room temperature; at 50 ℃ in chloroform for 2 days; and 50 ℃ in dichloromethane for 2 days.

The hygroscopicity of the a crystalline form of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid was evaluated. Dynamic gas phase sorption (DVS) analysis of moisture uptake and moisture release as a function of Relative Humidity (RH) was obtained by cycling between 5% and 95% RH. As indicated by the representative form a DVS isotherm in fig. 3, the maximum uptake was about 0.06% of the total mass of the sample. Thus, in certain embodiments, form a is non-hygroscopic.

FIG. 4 provides 3- [5- (2-fluorophenyl) - [1,2,4]]Oxadiazol-3-yl]Representative of-benzoic acids¹³C solid state NMR spectrum. In certain embodiments, 3- [5- (2-fluorophenyl) - [1,2,4] when the external control glycine is 176.5ppm]Oxadiazol-3-yl]-the A form of benzoic acid passing through one or more of the following approximate sites¹³C CP/MAS solid-state NMR signals: 172.6, 167.0, 131.3, 128.4 and 117.1 ppm.

In certain embodiments, form a of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid exhibits properties desirable for processing and/or manufacture of a pharmaceutical product containing 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid. For example, in certain embodiments, form a of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid has a relatively high melting point, a property important for processing and manufacturing, and the like. Furthermore, in certain embodiments, form a of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid is found to be substantially non-hygroscopic. A non-hygroscopic solid form is desirable for a variety of reasons, including, for example, for processing and storage considerations. Furthermore, in certain embodiments, form a of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid is found to be physically and chemically stable after undergoing micronization, a method of reducing particle size. Physical stability is an important property of pharmaceutical materials during manufacture, processing and storage.

4.43- [5- (2-fluorophenyl) - [1,2,4]]Oxadiazol-3-yl]-benzoic acid in form B

In one embodiment, the present invention provides crystalline form B of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid. In certain embodiments, form B may be obtained by crystallization from various solvents including, but not limited to, Tetrahydrofuran (THF), hexane, Isopropanol (IPA), ethyl acetate (EtOAc), acetic acid, 1-butyl acetate, acetone, dimethyl ether, diethyl ether, dioxane, water, methyl isobutyl ketone (MIBK), Methyl Ethyl Ketone (MEK), nitromethane, and/or water.

In certain embodiments of the present invention, form B of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid has an amount of solvent in the crystal lattice that depends on one or more conditions, such as, but not limited to, crystallization, handling, processing, formulation, manufacturing or storage conditions. In certain embodiments of the invention, form B has a solvent in the crystal lattice. In certain embodiments, form B is substantially free of solvent in the crystal lattice. In certain embodiments, the maximum solvent bound molar equivalent per mole of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid in the sample form B is less than 6, less than 5, less than 4, less than 3, less than 2, less than 1.5, less than 1, less than 0.75, less than 0.5, or less than 0.25 molar equivalents. Without intending to be limited by theory, it is believed that the variable nature in the solvent content of form B arises from the presence of lattice channels that can accommodate different types and/or amounts of solvents and, depending on the particular conditions, allow for the addition and/or removal of solvents. In certain embodiments, the structure of form B represents the basis of a family of polymorphic forms. In certain embodiments, form B is a desolvated solvent crystalline form.

Figure 5 provides a representative XRPD pattern of form B of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid. In certain embodiments, form a of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid is characterized by XRPD peaks located at one or more of the following positions: about 6.4, about 8.0, about 14.1, about 15.9, about 17.2, and about 20.1 degrees 2 θ. It will be appreciated by those skilled in the art that when solvent and/or water is added to or removed from the crystal lattice, the crystal lattice will expand or contract slightly, resulting in a slight shift in the XRPD peak position. In certain embodiments of the present invention, there is provided crystalline form B of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid, the crystalline form being characterized by an XRPD pattern substantially similar to the pattern presented in figure 5. In certain embodiments, form B exhibits an XRPD pattern substantially similar to the pattern presented in fig. 5, but exhibits a slight shift in peak position due to the presence or absence of a particular solvent or water in the crystal lattice. Certain representative XRPD patterns of form B of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid (second from top to bottom) are compared with form a (top) in figure 8. In certain embodiments, form B has an XRPD pattern substantially similar to one or more of the XRPD patterns presented in figure 8.

The thermal properties of a sample of form B of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid crystallized from a 2.5: 1 THF: hexane mixture are shown in FIG. 6. The TGA temperature profile of this crystalline form B sample in figure 6 shows two mass loss phenomena: once at 5% of the total mass of the sample when heated from about 25 c to about 165 c and a second at the beginning at about 220 c. Hot stage microscopy showed that the first mass loss phenomenon resulted from loss of solvent and/or water from the crystal lattice, while the second mass loss phenomenon resulted from sublimation of the form B. XRPD analysis of the obtained sublimate indicated that form a of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid was formed. The DSC temperature profile of the form B sample in figure 6 shows a sharp endotherm with a peak temperature at about 243 ℃, which corresponds to the melting of the sublimate of form a. The DSC of this form B sample also exhibits at least one additional endotherm below a temperature of about 220 ℃. These thermal data indicate that a sample of form B of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid contains water and/or solvent in the crystal lattice. Due to the variable water and/or solvent content in certain samples of form B of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid, in certain embodiments of the present invention, the thermal properties of form B will exhibit certain variations. For example, in particular embodiments of the present invention, a sample of form B that is substantially free of water and solvent does not exhibit substantial TGA mass loss or DSC thermal behavior below about 220 ℃. Since form B sublimes and crystallizes into form a, the endothermic heat of fusion in fig. 6 occurs after the sample is transformed into form a.

In one embodiment of the invention, the TGA and¹analysis by H NMR A sample of crystalline form B crystallized from IPA had 3- [5- (2-fluorophenyl) - [1,2,4] per mole]Oxadiazol-3-yl]Benzoic acid about 0.1 molar equivalents of IPA and about 1 molar equivalent of water. In a particular embodiment of the invention, each molar equivalent of 3- [5- (2-fluorophenyl) - [1,2,4]]Oxadiazol-3-yl]Samples of form B with benzoic acid having about 1 molar equivalent of water are called monohydrate. In another embodiment of the invention, a sample of form B that has been treated with vacuum drying at 105 ℃ for 10 minutes exhibits a total weight loss of 2% of the sample mass when subsequently analyzed by TGA at about 25 to about 185 ℃. In certain embodiments, depending on the amount and/or nature of solvent and/or water in the crystal lattice (e.g., mass loss upon heating or drying), the form B will exhibit variations based on the total amount or type of solvent and/or water in the crystal lattice. In certain embodiments, regardless of the amount and/or type of solvent and/or water in the crystal lattice, the XRPD pattern of form B will exhibit the peak characteristics of form B as described above, but with a slight shift in the peaks due to differences in the amount and/or type of solvent and/or water in the crystal lattice of form B. Representative XRPD patterns exhibiting peak shifts in certain form B samples overlap in fig. 8 (second from top to bottom)Individually).

In certain embodiments of the invention, a conversion of form B to form a is observed after milling at ambient or sub-ambient temperatures. In other embodiments of the invention, form B is physically stable after 6 days of storage under one of the following Relative Humidity (RH) conditions: 40 ℃ at 53% RH; 75% RH at 40 ℃; and 60 ℃ 50% RH. In other embodiments of the invention, form B is substantially non-hygroscopic as shown by the representative form B DVS isotherm plot in figure 7. In other embodiments of the invention, form B exhibits a partial conversion to form a after 6 days of storage at 60 ℃, 79% RH. In other embodiments of the invention, form B is physically stable when compressed only, and when compressed in the presence of a 1: 1 mixture of t-butanol and water. In other embodiments of the invention, form B is physically stable when slurried in a 1: 1 mixture of THF and heptane at room temperature for 1 day. In other embodiments, the conversion of form B to form a is observed after slurrying form B in methyl isobutyl ketone or in a 1: 1 mixture of dioxane and water.

4.5 methods of use

The present invention provides methods of treating, preventing and managing a disease or disorder ameliorated by inhibiting premature translational termination and/or nonsense-mediated mRNA degradation in a patient, comprising administering to a patient in need thereof an effective amount of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid in solid form.

In one embodiment, the invention provides a method of treating, preventing or managing any disease associated with a gene exhibiting premature translation termination and/or nonsense-mediated mRNA degradation. In one embodiment, the disease is caused in part by a lack of gene expression due to a premature stop codon. Specific examples of genes that can exhibit premature translation termination and/or nonsense-mediated mRNA degradation and diseases associated with premature translation termination and/or nonsense-mediated mRNA degradation are described in U.S. patent application publication No. 2005-0233327, entitled: methods For Identifying Small Molecules, which is disclosed in the third document data transfer Termination And non sensitive mRNA Decay, which is incorporated herein by reference in its entirety.

Diseases or disorders associated with or ameliorated by inhibiting premature translation termination and/or nonsense-mediated mRNA degradation include, but are not limited to: genetic diseases, cancer, autoimmune diseases, hematologic diseases, collagen diseases, diabetes, neurodegenerative diseases, proliferative diseases, cardiovascular diseases, pulmonary diseases, inflammation or central nervous system diseases.

Specific genetic diseases within the scope of the methods of the invention include, but are not limited to, multiple endocrine tumors (types 1,2 and 3), amyloidosis, mucopolysaccharidosis (types I and III), congenital adrenal insufficiency, colonic adenocarcinoma, Von Hippel Landau disease, Menkes syndrome, hemophilia A, hemophilia B, collagen VII, Alagille syndrome, Townes-Brocks syndrome, rhabdoid tumor, epidermoid bullous disease, Heller's syndrome, Coffin-Lowry syndrome, aniridia, fibular muscle atrophy, myotubular myopathy, X-linked myomyopathy, X-linked chondrodysplasia, X-linked agammaglobulinemia, polycystic kidney disease, spinal muscular atrophy, familial colonic adenocarcinoma, pyruvate dehydrogenase deficiency, phenylketonuria, neurofibroma type 1, neurofibroma type 2, Alzheimer's disease, Crohn's disease, Tay Sachs disease, Rett syndrome, Hermansky-Pudlak syndrome, ectodermal dysplasia/dermfragile syndrome, Leri-Weill chondrogenesis disorder, rickets, hypophosphatemia, adrenoleukodystrophy, atrophy of convolution, atherosclerosis, sensorineural deafness, dystonia, Dent's disease, acute intermittent porphyria, Cowden disease, Herlitz epidermolysis bullosa, Wilson's disease, Treacher-Collins syndrome, pyruvate kinase deficiency, giant disease, dwarfism, hypothyroidism, hyperthyroidism, aging, obesity, parkinson's disease, Niemann Pick's disease type C, cystic fibrosis, muscular dystrophy, heart disease, kidney stones, ataxia telangiectasia, familial hypercholesterolemia, retinitis pigmentosa, lysosomal storage diseases, tuberous sclerosis, Duchenne muscular dystrophy, and Marfan syndrome.

In another embodiment, the genetic disease is an autoimmune disease. In a preferred embodiment, the autoimmune disease is rheumatoid arthritis or graft versus host disease.

In another embodiment, the genetic disease is a hematological disease. In specific embodiments, the hematologic disease is hemophilia a, Von Willebrand disease (type 3), ataxia telangiectasia, b-thalassemia, or kidney stones.

In another embodiment, the genetic disease is a collagen disease. In a specific embodiment, the collagen disease is osteogenesis imperfecta or liver cirrhosis.

In another embodiment, the genetic disease is diabetes.

In another embodiment, the genetic disease is inflammation. In a particular embodiment, the inflammation is arthritis.

In another embodiment, the genetic disease is a central nervous system disease. In one embodiment, the central nervous system disorder is a neurodegenerative disorder. In specific embodiments, the central nervous system disease is multiple sclerosis, muscular dystrophy, Duchenne muscular dystrophy, alzheimer's disease, tayssachs disease, advanced infant neuronal cerotic lipofuscinosis (LINCL), or parkinson's disease.

In another embodiment, the genetic disease is cancer. In particular embodiments, the cancer is a cancer of the head and neck, eye, skin, mouth, throat, esophagus, chest, bone, lung, colon, sigmoid, rectum, stomach, prostate, breast, ovary, kidney, liver, pancreas, brain, intestine, heart, or adrenal gland. Cancer may be primary or metastatic. Cancer includes solid tumors, hematologic cancers, and other neoplasias.

In another specific embodiment, the cancer is associated with a tumor suppressor gene (see, e.g., Garrinis et al, 2002, Hum Gen 111: 115-. Such tumor suppressor genes include, but are not limited to, APC, ATM, BRAC1, BRAC2, MSH1, pTEN, Rb, CDKN2, NF1, NF2, WT1, and p 53.

In a particularly preferred embodiment, the tumor suppressor gene is the p53 gene. Nonsense mutations have been identified in the p53 gene and are involved in cancer. Several nonsense mutations in the p53 gene have been identified (see, e.g., Masuda et al, 2000, Tokai J Exp Clin Med.25 (2): 69-77; Oh et al, 2000, Mol Cells 10 (3): 275-80; Li et al, 2000, Lab invest.80 (4): 493-9; Yang et al, 1999, Zhonghua Zhong Liu Zhi 21 (2): 114-8; Finkelstein et al, 1998, Mol Diagn.3 (1): 37-41; Kajiyamaera et al, 1998, Dis Esophagus.11 (4): 279-83; Kawamura et al, 1999, Leuk Res.23 (2): 115-26; Raddige et al, 1998, Hum Pathol.29 (11): 1310-6; intuyer et al, 1998, 76, Leu J.1997; Leu J.14, 76, J.14; Rockwell et al; Rockwell J.14; 76, J.14, 76; Rockwell et al, 1996, Cancer Lett.100 (1-2): 107-13; rall et al, 1996, Pancreas.12 (l): 10-7; fukutomi et al, 1995, Nippon Rinsho.53 (11): 2764-8; frebourg et al, 1995, Am J Hum Genet.56 (3): 608-15; dove et al, 1995, Cancer Surv.25: 335-55; adamson et al, 1995, Br J Haematol.89 (l): 61-6; grayson et al, 1994, Am J Pediatr Hematol Oncol.16 (4): 341-7; lepellety et al, 1994, Leukemia.8 (8): 1342-9; mclntyre et al, 1994, J Clin Oncol.12 (5): 925-30; horio et al, 1994, oncogene.9 (4): 1231-5; nakamura et al, 1992, Jpn J Cancer Res.83 (12): 1293-8; davidoff et al, 1992, oncogene.7 (l): 127-33; and Ishioka et al, 1991, Biochem Biophys Res Commun.177 (3): 901-6; the disclosure of which is hereby incorporated by reference in its entirety).

In other embodiments, diseases treated, prevented or controlled by administering an effective amount of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid in solid form to a patient in need thereof include, but are not limited to, solid tumors, sarcomas, malignancies, fibrosarcomas, myxosarcomas, liposarcomas, chondrosarcomas, osteogenic sarcomas, chordomas, angiosarcomas, endotheliosarcomas, lymphangiosarcomas, lymphangioendotheliosarcomas, synoviomas, mesotheliomas, ewing's tumors, leiomyosarcomas, rhabdomyosarcomas, colon cancers, pancreatic cancers, breast cancers, ovarian cancers, prostate cancers, squamous cell carcinomas, basal cell carcinomas, adenocarcinomas, sweat gland cancers, sebaceous gland cancers, papillary carcinomas, papillary adenocarcinomas, cystadenocarcinoma, medullary cancer, bronchial cancer, renal cell carcinoma, and combinations thereof, Hepatoma, cholangiocarcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilms' tumor, cervical carcinoma, testicular tumor, lung cancer, small-cell lung cancer, bladder cancer, epithelial cancer, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, cutaneous multiple hemorrhagic sarcoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, melanoma, neuroblastoma, retinoblastoma, blood-borne tumor, acute lymphoblastic leukemia, acute lymphoblastic B-cell leukemia, acute lymphoblastic T-cell leukemia, acute myeloblastic leukemia, acute promyelocytic leukemia, acute myelomonocytic leukemia, acute erythroleukemia, acute megakaryoblastic leukemia, acute myelomonocytic leukemia, acute nonlymphocytic leukemia, acute myelocytic leukemia, acute nonlymphocytic leukemia, small-cell lung cancer, bladder cancer, epithelial cancer, glioma, astrocytoma, Acute undifferentiated leukemia, chronic myelogenous leukemia, chronic lymphocytic leukemia, hairy cell leukemia, or multiple myeloma. See, for example, Harrison's Principles of Internal Medicine, ed by Eugene Braunwald et al, 491-.

4.6 pharmaceutical compositions

Pharmaceutical compositions and single unit dosage forms comprising a compound of the invention, or a pharmaceutically acceptable polymorph, prodrug, salt, solvate, hydrate, or clathrate thereof, are also encompassed by the invention. The individual dosage forms of the present invention may be adapted for oral, mucosal (including sublingual, buccal, rectal, nasal, or vaginal), parenteral (including subcutaneous, intramuscular, bolus injection, intraarterial, or intravenous), transdermal, or topical administration.

The single unit dosage forms of the invention are suitable for oral, mucosal (e.g., nasal, sublingual, vaginal, buccal, or rectal), parenteral (e.g., subcutaneous, intravenous, bolus injection, intramuscular, or intraarterial), or transdermal administration to a patient.

The composition, shape and type of dosage forms of the present invention will generally vary depending on their use. All manner in which the particular dosage forms encompassed by the present invention will differ from one another will be apparent to those skilled in the art. See, for example, Remington's Pharmaceutical Sciences, 18 th edition, Mack Publishing, Easton PA (1995).

Typical pharmaceutical compositions and dosage forms contain one or more carriers, excipients or diluents. Suitable excipients are well known to those skilled in the art of pharmacy, and the present invention provides non-limiting examples of suitable excipients. Whether a particular excipient is suitable for incorporation into a pharmaceutical composition or dosage form depends on a variety of factors well known in the art, including, but not limited to, the method by which the dosage form is administered to a patient. For example, oral dosage forms such as tablets may contain excipients not suitable for use in parenteral dosage forms. The suitability of a particular excipient may also depend on the particular active ingredient in the dosage form.

5. Examples of the embodiments

5.13- [5- (2-fluorophenyl) - [1,2,4]]Oxadiazol-3-yl]Synthesis of benzoic acid in solid form

The 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid product obtained from the synthesis described above may be crystallized or recrystallized by various methods to yield the solid forms of the present invention. Some non-limiting examples are provided below.

5.1.1 Synthesis of form A

5.1.1.1 Slow Evaporation

The 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid product obtained as described in the present invention is crystallized as form a by slow evaporation from the following solvents: acetonitrile, t-butanol, isopropanol, and isopropyl ether. A solution of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid is prepared in the indicated solvent and sonicated between aliquots to aid dissolution. Once complete dissolution of the mixture was achieved, the solution was filtered through a 0.2 μm filter, as judged by visual inspection. The filtered solution was placed in a vial covered with aluminum foil with small holes and evaporated at 60 ℃ (50 ℃ in the case of t-butanol). The solid formed was isolated and identified as form a by XRPD.

5.1.1.2 rapid evaporation

The 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid product obtained as described in the present invention is crystallized as form a by means of flash evaporation from the following solvents or solvent systems: 1-butanol, bismethoxy ether, tert-butanol, a mixture of dimethylformamide and water, isopropyl ether, and a mixture of tert-butanol and water (3: 2 ratio), 1 molar equivalent of methanol and 1 molar equivalent of sodium chloride. Solutions were prepared in the indicated solvent or solvent system and sonicated between aliquots to aid dissolution. Once complete dissolution of the mixture was achieved, the solution was filtered through a 0.2 μm filter, as judged by visual inspection. The filtered solution was placed in an open vial and evaporated at a temperature of 60 ℃ (50 ℃ in the case of t-butanol and isopropyl ether; 81 ℃ in the case of t-butanol/water/methanol/NaCl system). The solid formed was isolated and identified as form a by XRPD.

5.1.1.3 slurry conversion

Form B of the free acid of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid obtained as described in the present invention is converted to form a by slurrying in a 1: 1 dioxane: water solvent system. The slurry is prepared by adding sufficient form B solid to a given solvent such that an excess of solid occurs. The mixture was then placed in a closed vial and stirred at a temperature of 60 ℃. After 2 days, the solid was isolated by vacuum filtration and identified as form a by XRPD, with a small amount of form B.

5.1.1.4 sublimation and heating

The crystalline form B of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid obtained as described in the present invention is converted to crystalline form a by sublimation and heating. In one experiment, form B was sublimed at 160-. In another trial, form B melted at 255 ℃ and was then placed directly into liquid nitrogen to produce a crystalline material, which was identified as form a by XRPD. In another trial, form B melted at 255 ℃ and then cooled slowly to yield a crystalline material, which was identified as form a by XRPD.

5.1.2 Synthesis of Crystal form B

5.1.2.1 slow evaporation

The 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid product obtained as described in the present invention is crystallized in form B by slow evaporation from the following solvents: acetone, dimethyl ether, and methyl ethyl ketone. A solution of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid is prepared in a given solvent and sonicated between aliquots to aid dissolution. Once complete dissolution of the mixture was achieved, the solution was filtered through a 0.2 μm filter, as judged by visual inspection. The filtered solution was placed in a vial covered with aluminum foil with small holes and evaporated at a temperature of 50 ℃ (60 ℃ in the case of t-butanol).

In one embodiment, 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid is dissolved in bismethoxy ether. The solution was placed in a clean vial. The vial was filtered through a 0.2 μm filter covered with aluminum foil perforated with small holes and the solvent was evaporated. The solid formed was isolated and identified as form B by XRPD. XRPD analysis is presented in table 8 (P.O.).

5.1.2.2 fast Evaporation

The 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid product obtained as described in the present invention is crystallized as form B by a process of flash evaporation from the following solvents or solvent systems: acetone, acetic acid, 1-butyl acetate, dimethyl ether, THF and diethyl ether, dioxane, methyl ethyl ketone, nitromethane, methyl isobutyl ketone, THF: hexane (2.5: 1), and dioxane: water (3: 2). Solutions were prepared in a given solvent or solvent system and sonicated between aliquots to aid dissolution. Once complete dissolution of the mixture was achieved, the solution was filtered through a 0.2 μm filter, as judged by visual inspection. The filtered solution was placed in an open vial and evaporated at elevated temperature. The solid formed was isolated and identified as form B by XRPD.

5.1.2.3 slurry conversion

The crystalline form a of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid obtained as described in the present invention is converted into crystalline form B by a process of slurrying in the following solvents: acetic acid, 1-butyl acetate, and nitromethane. In one embodiment, 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid is slurried in 1-butyl acetate (13mL) on a rotary shaker at room temperature for 3 days. After 3 days, the solvent was removed with a pipette, dried and identified as form B by XRPD (table 5).

5.1.2.4 rotary oscillator conversion

Form a of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid obtained as described herein is converted to form B by heating in 1-propanol (10mL) for 1 day at 60 ℃ on a rotary shaker. The resulting solution was filtered through a 0.2 μm nylon filter into a clean vial. After 1 day, the solvent was decanted and the sample dried under nitrogen. XRPD analysis of form B is shown in table 4.

5.1.2.5 other embodiments

3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid (20mg, form B) was slurried at ambient temperature in a mixture of tetrahydrofuran/heptane 1/1 (2mL) for 1 day. After 1 day, the slurry was inoculated with form a (10mg) and form B (9mg) and reslurried for one day, after which additional form a (30mg) was added. After slurrying the samples for a total of 7 days, additional form a (30mg) was added and the temperature was increased to 50 ℃. The solids were collected after one day of slurrying at 50 ℃. The solid formed was isolated and identified as form B by XRPD. XRPD analysis is shown in table 6.

3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid (unmeasured amount; form B) is resistant to a relative humidity of 75% at 40 ℃ for 6 days. The solid formed was isolated and identified as form B by XRPD. XRPD analysis is shown in table 7.

5.2 analytical methods

The following method of solid state analysis provides an example of how to characterize the solid forms of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid of the present invention. The solid state property data according to the present invention was obtained using a specific method as described below.

5.2.1X-ray powder diffraction method (XRPD)

Some XRPD analysis was performed using a Shimadzu XRD-6000X-ray powder diffractometer using Cu ka radiation. The instrument is fitted with a long, precision focus X-ray tube. The voltage and amperage of the tube were set to 40kV and 40mA, respectively. The divergence and scattering slits are set at 1 deg. and the receiving slits are set at 0.15 mm. The diffracted radiation was detected by a NaI scintillation detector. Continuous scans from 2.5 to 40 ° 2 θ using 3 °/minute (0.4 seconds/0.02 ° step) of θ -2 θ were used. The silicon standards were analyzed to check the calibration of the instrument. Data were collected and analyzed using XRD-6100/7000 v.5.0. The sample is prepared for analysis by placing the sample in a sample holder.

Some XRPD analysis was performed using an Inel XRG-3000 diffractometer equipped with a CPS (bend placement sensitive) detector with a 2 θ range of 120 °. Real-time data was collected using Cu-ka radiation at 0.03 ° 2 θ resolution. The voltage and amperage of the tube were set to 40kV and 30mA, respectively. The monochromator slit was set to be 5mm high and 160 μm wide (5mm by 160 μm). The spectra are shown from 2.5-40 ° 2 θ. Samples were prepared for analysis using an aluminum sample holder inserted into silicon or by loading it into a thin-walled glass capillary. Each capillary is fixed on a goniometer head, which is motorized to allow the capillary to rotate during data acquisition. The sample was analyzed for 300 seconds. The instrument calibration was performed using a silicon control standard.

Certain XRPD patterns were collected using a Bruker D-8 Discover diffractometer and Bruker's General Area diffraction detection system (GADDS, v.4.1.20). Using a precision-focusing tube (40kV, 40mA),the mirror and 0.5mm dual-aperture collimator produced an incident beam of Cu ka radiation. A sample of the sample is loaded into the capillary and secured to the transfer stage. The region of interest is positioned to intersect the incident beam in transmission geometry using a camera and a laser. The incident beam is scanned to optimize the directional statistics. Beam-termination is used to minimize air scattering from low-angle incident beams. Diffraction patterns were collected using a Hi-Star area detector positioned 15cm from the sample and treated with GADDS. The intensities in the GADDS image of the diffractogram were integrated with a 0.04 ° 2 θ step. The integrated plot shows the diffraction intensity as a function of 2 θ. Prior to this analysis, the silicon standard was analyzed to verify the peak position of Si 111.

Some XRPD files generated from Inel XRPD instruments were converted to shimadzu. Raw file was processed through Shimadzu XRD-6000 software version 2.6 to automatically find peak locations. "Peak position" refers to the maximum intensity of the peak intensity profile. The parameters used in the peak selection are shown in the lower half of the data in each parameter set. The following processing was used with Shimadzu XRD-6000 "Basic Process" version 2.6 algorithm:

smoothing all maps.

Background was subtracted to determine the net relative intensity of the peaks.

From 50% intensity for all spectra by Cu K α 1Resulting peak minus from Cu Ka 2The peak of the wavelength.

5.2.2 Differential Scanning Calorimetry (DSC)

Differential Scanning Calorimetry (DSC) was performed using a differential scanning calorimeter 2920 from TA Instruments. The samples were placed in an aluminum DSC pan and the weight was accurately recorded. The tray is covered with a lid and then crimped. The sample cell was equilibrated at 25 ℃ and heated at a rate of 10 ℃/min under a nitrogen purge until the final temperature was 350 ℃. Indium metal was used as a calibration standard. The reported temperature is at the transition maximum.

5.2.3 thermogravimetric analysis (TGA)

Thermogravimetric (TG) analysis was performed using a 2950 thermogravimetric analyzer from TA Instruments. Each sample was placed in an aluminum sample pan and inserted into a TG furnace. The furnace (first equilibrated at 35 ℃ followed by) was heated at a rate of 10 ℃/min under nitrogen until the final temperature was 350 ℃. Nickel and Alumel^TMIs used byAs a calibration standard.

5.2.4 dynamic vapor sorption/Desorption (DVS)

Moisture sorption/desorption data was collected on a VTI SGA-100 vapor sorption analyzer. Adsorption and desorption data were collected at 10% Relative Humidity (RH) intervals in the range of 5% to 95% RH under a nitrogen purge. The samples were not dried prior to analysis. The equilibrium criterion used for the analysis was less than 0.0100% weight change in 5 minutes, and if this weight criterion was not met, the maximum equilibration time was 3 hours. No data correction was made for the initial moisture content of the sample. NaCl and PVP were used as calibration standards.

5.2.5 Karl method (Karl Fischer, KF)

Coulomb-Karsc (KF) analysis for water determination was performed using a Mettler Toledo DL39 Karsc meter. Approximately 21mg of the sample was placed in a KF titration vessel containing Hydranal-Coulomat AD and mixed for 42-50 seconds to ensure dissolution. The sample is then titrated through a generator electrode that, by electrochemical oxidation: 2I- ═ I₂+2e iodine is produced. Three replicates were performed to ensure reproducibility.

5.2.6 Hot stage microscopy

Hot stage microscopy was performed using a Linkam FTIR 600 hot stage with a TMS93 controller mounted on a Leica DM LP microscope equipped with a Spot light color Camera for image acquisition. Unless indicated, images were acquired using the 4.5.9 version Spot Advanced software created on 9.6.2005. The camera is white balanced before use. The samples were observed and obtained using a 20 x 0.40 n.a. long working distance objective with orthogonal polarization and a primary red compensator. The sample was placed on a cover glass. Another coverslip was then placed over the sample. Each sample was visually observed as the stage was heated. The hot stage was calibrated using USP melting point standards.

5.2.7 solid state cross-polarized magic angle spins ¹³ C nuclear magnetic resonance spectroscopy ( ¹³ C CP/MAS ssNMR)

Samples for solid-state nuclear magnetic resonance spectroscopy were prepared by loading the samples into a 4mm pensil-type zirconia rotor. Scans were acquired at room temperature with a relaxation delay of 120.000s, a pulse width of 2.2 μ s (90.0 degrees), an acquisition time of 0.030s, and a spectral width of 44994.4Hz (447.520 ppm). A total of 100 scans were acquired. Use of¹³C as an observation nucleus and¹h as a decoupling nucleus achieved cross polarization with a contact time of 10.0 ms. A magic angle spin rate of 12000Hz was used. The spectral outer control glycine is at 176.5 ppm.

Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are encompassed by the following claims. All publications, patents and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated by reference.

5.2.8 Single Crystal X-ray diffraction

Sample preparation

Crystals for structural determination of form a were prepared by sublimating form a. The crystals were removed from the condenser after heating the sample at 155-206 ℃ for approximately 90 minutes. (Table 3 experiment)

Data acquisition

C having a size of about 0.44X 0.13X 0.03mm₁₅H₉FN₂O₃The colorless needles were fixed in random directions on the glass fiber. By Mo Ka radiationPreliminary examination and data acquisition was performed on a Nonius KappaCCD diffractometer. Refinement was performed on LINUX PC using SHELX97 (Sheldrick, G.M. SHELX97, A Program for Crystal Structure reference, University of Gottingen, Germany, 1997).

The cell constants and orientation matrix for data acquisition were obtained by least squares refinement using set angles in the range of 13862 reflections 2 ° < θ < 24 °. The mosaic (mosaicity) after refinement from DENZO/SCALEPACK (Otwinowski, Z.; Minor, W.methods enzymol.1997, 276, 307) was 0.33 deg., which indicates good crystal quality. The space groups are determined by the program XPREP (Bruker, SHELXTL 6.12. XPREP in edition, Bruker AXS Inc., Madison, Wis., USE, 2002). The system exists according to the following conditions: h0l h + l 2 n; and determining the space group as P2 according to the following least square refinement method₁/n(#14)。

Data were collected to a maximum 2 θ value of 2469 ° at a temperature of 150 ± 1K.

Data subtraction

The map (frame) was integrated using DENZO-SMN (Otwinowski, Z.; Minor, W.methods enzymol.1997, 276, 307). A total of 13862 reflections were acquired, 3201 of which were unique. Data were Lorentz and polarization corrected. The linear absorption coefficient of the radiation of Mo Ka is 0.110mm^-1. Empirical absorption correction using SCALEPACK (Otwinowski, z.; Minor, w. methods enzymol.1997, 276, 307) was applied. The transmission coefficient ranged from 0.951 to 0.997. Secondary extinction correction was applied (Shell drick, G.M. SHELX97, A Program for Crystal Structure reference, University of Gottingen, Germany, 1997). The final coefficient refined in the least squares method was 0.0046 (absolute units). The intensities of the equivalent reflections are averaged. The agreement factor for the averaging was 10.1% based on intensity.

Structure analysis and refinement

The structure was resolved by direct assay using SIR2004 (Burla, m.c., et al, j.appl.cryst.2005, 38, 381). The remaining atoms are located in the subsequent differential fourier synthesis method. Refinements include hydrogen atoms, but the hydrogen atoms are constrained to be on the atoms to which they are attached. The structure is refined in a full matrix least squares method by minimizing the following function:

∑w(|F_o|²-|F_c|²)²。

the weight w is defined as 1/[ sigma ]²(F₀ ²)+(0.0975P)²+(0.0000P)]Wherein P ═ F₀ ²+2F_c ²)/3。

Scattering factors were taken from The International Table of Crystallography (International Tables for Crystallography, Vol.C., Kluwer Academic Publishers: Dordrecht, The Netherlands, 1992, Tables 4.2.6.8 and 6.1.1.4). Of the 3201 reflections used for refinement, only F₀ ²＞2σ(F₀ ²) Is used to calculate R. A total of 2010 reflections were used for the calculation. The final round of refinement included 382 variable parameters and converged with unweighted and weighted conformity factors (maximum parameter offset < 0.01 times the estimated standard deviation):

R＝∑|F_o-F_c|/∑F_o＝0.062

the standard deviation of the observations per weight was 1.01. The highest peak in the final differential Fourier method hasOf (c) is measured. The lowest negative peak hasOf (c) is measured.

Calculated X-ray powder diffraction (XRPD) pattern

Calculated XRPD patterns of Cu radiation were generated using PowderCell 2.3(PowderCell for Windows Version 2.3 Kraus, w.; Nolze, g. federal Institute for Materials Research and Testing, Berlin Germany, EU, 1999) and atomic coordinates, space groups and cell parameters from single crystal data.

ORTEP and stacking maps

An ORTEP map was made using ORTEP III (Johnson, C.K. ORTEPIII, Report ORNL-6895, Oak Ridge National Laboratory, TN, U.S. A.1996, and OPTEP-3for Windows V1.05, Farrugia, L.J., J.Appl.Cryst.1997, 30, 565). Atoms are represented by thermal ellipsoids with 50% probability anisotropy. Stack maps were prepared using CAMERON (Watkin, D.J., et al, CAMERON, Chemical Crystallography Laboratory, University of Oxford, Oxford, 1996) modeling.

Results and discussion

The monoclinic cell parameters and calculated volume for form a are:α＝90.00°，β＝92.9938(15)°，γ＝90.00°， molecular weight is 284.25g/mol^-1And Z ═ 8 (where Z is the number of drug molecules per asymmetric unit), resulting in a calculated density of the crystal structure (d)_Computing，g cm^-3) Is 1.517g cm^-3. Space group is determined as P2₁And/n (#14), which is an achiral space group. A summary of the crystal data and crystallization data acquisition parameters is provided below:

1/[s²(F₀ ²)+(0.0975P)²+0.0000P]wherein P ═ F₀ ²+2F_c ²)/3

The quality of the structure obtained was excellent to moderate, as indicated by an R-value of 0.062 (6.2%). R-values, typically in the range of 0.02 to 0.06, are recognized as the most reliably determined structures. Although the quality of the crystal structure is slightly outside the acceptable range for the most reliably defined structure, the quality of this data is sufficient to ensure that the positioning of the atomic sites in the molecular structure is correct.

The ORTEP map of form a is shown in figure 11. The asymmetric unit shown in the figure comprises a dimer of two molecules arranged to form a possible hydrogen bond between adjacent carboxyl groups. Since acidic protons are not located in the fourier map, the molecule is assumed to be neutral. Fig. 9 shows the stacking pattern of form a, viewed down the axis of crystal b.

The simulated XRPD pattern for form a shown in fig. 10 was generated from single crystal data and was in good agreement with the experimental XRPD pattern for form a (see, e.g., fig. 1). The intensity difference may result from a preferred orientation. Preferred orientation is the tendency of crystals (usually plate-like or needle-like crystals) to align with a degree of order. Preferred orientations may affect peak intensities in the XRPD pattern, but not peak positions. The slight shift in peak position may be caused by the fact that experimental powder spectra were acquired at room temperature, while single crystal data were acquired at 150K. Low temperatures are used for single crystal analysis to improve the quality of the structure.

Table 1 shows the fractional atomic coordinates of the asymmetric units of form a.

Table 1: position parameter of crystal form A and estimation standard deviation thereof

U_eq＝(1/3)∑_i∑_jU_ija_ia_ja_i.a_j

Hydrogen atoms are included in the calculation of the structural factors without being refined

Table 2: peak position of form a in calculated XRPD pattern obtained from single crystal data

Position (° 2 θ)a	d-spacing	I/Ioc
			4.74	18.63	3.24
4.99	17.69	20.99
			6.44	13.72	4.46
7.30	12.10	6.46
			10.15	8.70	32.47
10.51	8.41	1.90
			11.27	7.85	6.14
11.59	7.63	13.97
			12.90	6.86	15.05
14.25	6.21	100.00
			14.50	6.10	8.25
14.64	6.05	75.70
			15.17	5.84	65.12
15.69	5.64	47.56
			16.31	5.43	8.61
16.37	5.41	8.11
			16.74	5.29	14.82
18.44	4.81	2.04
			18.78	4.72	3.13
19.04	4.66	4.05
			19.07	4.65	3.81
19.40	4.57	2.85
			20.03	4.43	11.28
20.06	4.42	5.41
			20.30	4.37	1.92
20.39	4.35	10.87
			21.11	4.20	21.30
21.20	4.19	7.07
			22.03	4.03	4.07
22.64	3.92	4.72
			23.16	3.84	4.71
23.86	3.73	2.64

[0152]

23.95	3.71	9.76
			24.21	3.67	12.14
24.27	3.67	32.98
			24.61	3.61	61.89
24.84	3.58	3.05
			24.86	3.58	8.00
24.94	3.57	7.15
			25.00	3.56	2.17
25.02	3.56	2.09
			25.13	3.54	10.36
25.61	3.48	1.67
			25.79	3.45	3.04
25.87	3.44	25.14
			26.02	3.42	15.19
26.20	3.40	3.41
			26.48	3.36	10.64
26.87	3.31	3.11
			26.87	3.32	5.65
27.08	3.29	5.60
			27.10	3.29	33.71
27.16	3.28	93.68
			27.26	3.27	82.52
27.45	3.25	4.42
			27.92	3.19	5.61
28.05	3.18	3.96
			28.20	3.16	59.41
28.28	3.15	3.04
			28.53	3.13	6.29
28.83	3.09	13.36
			28.93	3.08	15.74
28.96	3.08	6.42
			29.05	3.07	3.93
29.18	3.06	2.42
			29.24	3.05	2.10
29.42	3.03	2.64
			29.52	3.02	2.19
29.57	3.02	15.65
			29.94	2.98	2.66
30.00	2.98	4.98
			30.43	2.94	1.68
30.58	2.92	1.21
			30.79	2.90	1.79
30.93	2.89	1.07
			31.07	2.88	3.23
31.18	2.87	7.65
			31.42	2.84	2.68
31.97	2.80	2.16
			32.46	2.76	1.99
32.65	2.74	1.23

[0153]

32.88	2.72	1.02
			33.13	2.70	2.89
33.17	2.70	4.30
			33.40	2.68	2.97
33.64	2.66	2.39
			33.90	2.64	1.46
34.25	2.62	2.54
			34.74	2.58	1.40
35.18	2.55	1.60
			35.59	2.52	1.21
35.96	2.50	1.50
			36.64	2.45	7.44

a.I/I₀Relative strength

b. Having I/I₀Peaks with relative intensity less than 1 and peak position greater than 36.6 ° 2 θ do not show

Table 3: peak position of crystal form A experimental XRPD pattern

a.I/I₀Relative strength

b. The characteristic peak groups are shown in bold (no peaks within 0.2 ° 2 θ relative to PTC 124B crystal form files 169490, 172972, 172173, 170901, 169284, and 168717).

TABLE 4 peak position of XRPD pattern of form B (document 169490)

a.I/I₀Relative intensity.

b. Bold represents the set of characteristic peaks compared to form a.

TABLE 5 Peak position of form B (offset 1) XRPD pattern (File 168717)

a.I/I₀Relative intensity.

b. Bold represents the set of characteristic peaks compared to form a.

TABLE 6 Peak position of form B (offset 2) XRPD pattern (File 172972)

a.I/I₀Relative intensity.

b. Bold represents the set of characteristic peaks compared to form a.

TABLE 7 Peak position of type B (offset 3) XRPD pattern (File 172173)

a.I/I₀Relative intensity.

b. Bold represents the set of characteristic peaks compared to form a.

TABLE 8 Peak position of form B (PO) XRPD pattern (document 170901)

a.I/I₀Relative intensity.

b. Bold represents the set of characteristic peaks compared to form a.

TABLE 9 Peak position of XRPD pattern of form B shift (document 169284)

a.I/I₀Relative intensity.

b. Bold represents the set of characteristic peaks compared to form a.

Claims (19)

1. A crystalline form of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid having at least one of the following characteristics:

(a) an X-ray powder diffraction pattern comprising at least three peak positions (° 2 θ ± 0.2) when measured using Cu ka radiation, the peak positions selected from: 6.14, 6.39, 6.96, 7.92, 10.78, 12.44, 12.61, 12.88, 13.52, 13.78, 13.97, 14.30, 15.46, 15.68, 15.89, 16.33, 16.76, 17.03, 20.10, 21.03, 23.34, 23.86, 24.18, 24.42, 24.64, 26.62, 26.96, 27.29, 27.64, 27.96, 28.81, 31.05, 32.38, 32.58, 36.23, 37.81, 38.28, 38.44 and 39.16;

(b) a thermogravimetric analysis temperature profile with less than 5% mass loss of the total mass of the sample when heated from 25 ℃ to 165 ℃; and

(c) a differential scanning calorimetry temperature profile having an endotherm with a peak temperature at 243 ℃.

2. The crystalline form of claim 1 having a solvent in the crystal lattice, wherein the solvent is 1-propanol.

A crystalline form of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid characterized by an X-ray powder diffraction pattern including at least three peak positions (° 2 Θ ± 0.2) when measured using Cu ka radiation, the peak positions being selected from the group consisting of: 6.42, 7.00, 7.89, 10.85, 12.61, 12.92, 13.47, 13.97, 15.81, 16.45, 17.12, 20.05, 21.05, 23.92, 24.28, 27.00, 27.39, 27.84, 28.04, 28.94, 31.10, 32.58, 36.11, 37.71, 38.15 and 38.61.

4. The crystalline form of claim 3 having a solvent in the crystal lattice, wherein the solvent is 1-butyl acetate, acetic acid, or nitromethane.

A crystalline form of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid characterized by an X-ray powder diffraction pattern including at least three peak positions (° 2 θ ± 0.2) when measured using Cu Ka radiation, the peak positions being selected from the group consisting of: 6.10, 6.38, 6.54, 7.10, 8.02, 10.91, 12.71, 13.50, 13.62, 13.86, 14.10, 15.56, 15.70, 15.91, 16.55, 16.96, 17.22, 17.50, 19.82, 20.08, 20.34, 21.15, 23.78, 23.93, 24.38, 24.56, 26.88, 27.16, 27.48, 27.88, 28.04, 28.78, 29.02, 32.71, 36.01, 38.10, 38.56 and 39.38.

6. The crystalline form of claim 5 having a solvent in the crystal lattice, wherein the solvent is an 1/1 mixture of tetrahydrofuran and heptane.

A crystalline form of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid characterized by an X-ray powder diffraction pattern including at least three peak positions (° 2 θ ± 0.2) when measured using Cu Ka radiation, the peak positions being selected from: 1.79, 2.30, 2.57, 2.78, 3.29, 3.59, 3.89, 4.07, 4.34, 4.49, 4.76, 5.06, 6.47, 6.91, 7.96, 10.89, 12.87, 13.58, 13.99, 15.97, 16.48, 17.10, 20.00, 20.36, 21.04, 23.40, 24.29, 24.89, 26.87, 27.49, 27.80, 28.07, 29.08 and 38.61.

A crystalline form of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid characterized by an X-ray powder diffraction pattern including at least three peak positions (° 2 θ ± 0.2) when measured using Cu Ka radiation, the peak positions being selected from the group consisting of: 6.22, 6.51, 7.13, 8.17, 10.91, 12.87, 13.80, 14.12, 14.28, 15.78, 16.23, 16.54, 17.15, 20.33, 21.22, 21.36, 23.94, 24.30, 27.30, 27.58, 28.00, 28.74, 28.96, 32.70, 36.74, 38.18, 38.38, 38.52 and 39.31.

9. The crystalline form of claim 8 having a solvent in the crystal lattice, wherein the solvent is dimethoxy ether.

A crystalline form of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid characterized by an X-ray powder diffraction pattern including at least three peak positions (° 2 θ ± 0.2) when measured using Cu Ka radiation, the peak positions being selected from: 6.04, 6.49, 7.91, 10.92, 12.61, 12.92, 13.10, 13.42, 13.82, 13.99, 15.40, 15.76, 16.51, 17.15, 19.92, 20.04, 21.01, 23.92, 24.28, 24.48, 26.77, 27.14, 27.40, 27.74, 28.09, 28.82, 28.99, 31.03, 32.58, 35.64, 35.85, 37.48, 37.66 and 38.62.

11. The crystalline form of any one of claims 2,4, 6, or 9 having less than about 6 molar equivalents of solvent per mole of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid in the crystal lattice.

12. The crystalline form of any one of claims 1, 3, 5, 7, 8, or 10, wherein the compound is partially converted to a non-hygroscopic crystalline form.

13. The crystalline form of any one of claims 1, 3, 5, 7, 8, or 10, wherein the compound is converted to a non-hygroscopic crystalline form.

14. A pharmaceutical composition comprising the crystalline form of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid of any one of claims 1 to 13 together with one or more carriers, excipients or diluents for use as a single unit dosage form.

15. Use of a crystalline form of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid as defined in any one of claims 1 to 13 to modulate a premature stop codon in a cell by contacting the cell with the crystalline form.

16. Use of a crystalline form of 3- [5- (2-fluorophenyl) - [1,2,4] oxadiazol-3-yl ] -benzoic acid according to any one of claims 1 to 13 in the manufacture of a medicament for the treatment or prevention of a disease or condition associated with a premature stop codon in a patient in need thereof.

17. The use of claim 16, wherein the disease or disorder associated with premature stop codons is multiple endocrine tumors (types 1,2, and 3), amyloidosis, mucopolysaccharidosis (types I and III), congenital adrenal insufficiency, colonic adenomatous polyposis, Von Hippel Landau disease, Menkes syndrome, hemophilia a, hemophilia b, collagen VII, Alagille syndrome, Townes-Brocks syndrome, rhabdoid tumor, epidermoid bullous disease, heller's syndrome, Coffin-Lowry syndrome, aniridia, peroneal muscular atrophy, myotubular myopathy, X-linked chondroplasia, X-linked gammoproteinemia, polycystic kidney disease, spinal muscular atrophy, familial colonic adenocarcinoma, pyruvate dehydrogenase deficiency, ketonuria, type 1 neurofibroma, type 1 neurofibromatosis, and/or multiple sclerosis, Neurofibroma type 2, Alzheimer's disease, Tay Sachs disease, Rett syndrome, Hermansky-Pudlak syndrome, ectodermal dysplasia/cutaneous fragility syndrome, Leri-Weill chondrogenesis disorder, rickets, hypophosphatemia, adrenoleukodystrophy, cyclotron atrophy, atherosclerosis, sensorineural deafness, dystonia, Dent disease, acute intermittent porphyria, Cowden disease, Herlitz epidermolysis bullosa, Wilson disease, Treacher-Collins syndrome, pyruvate kinase deficiency, megaly disease, dwarfism, hypothyroidism, hyperthyroidism, obesity, Parkinson's disease, Niemann Pick's disease type C, cystic fibrosis, muscular dystrophy, heart disease, kidney stones, ataxia telangiectasia, familial hypercholesterolemia, retinitis pigmentosa, lysosomes, diseases, Tuberous sclerosis, Duchenne muscular dystrophy, diabetes, cancer, rheumatoid arthritis, graft versus host disease, Von Willebrand disease, b-thalassemia or kidney stones, osteogenesis imperfecta, cirrhosis, multiple sclerosis, advanced stage infant neuronal cerotic lipofuscin, and Marfan syndrome.

18. The use of claim 17, wherein the cancer is a cancer of the head and neck, eye, skin, mouth, throat, esophagus, chest, bone, lung, colon, sigmoid colon, rectum, stomach, prostate, breast, ovary, kidney, liver, pancreas, brain, intestine, heart or adrenal gland, wherein the cancer is a solid tumor selected from the group consisting of sarcoma, malignancy, fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, synovioma, mesothelioma, ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary adenocarcinoma, cystadenocarcinoma, medullary carcinoma, bronchial carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, bladder carcinoma, colon carcinoma, seminoma, embryonic carcinoma, Wilms 'tumor, cervical cancer, testicular tumor, lung cancer, small cell lung cancer, bladder cancer, epithelial cancer, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, Kaposi's sarcoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, melanoma, neuroblastoma, retinoblastoma, blood-borne tumor, acute lymphoblastic leukemia, acute lymphoblastic B-cell leukemia, acute lymphoblastic T-cell leukemia, acute myeloblastic leukemia, acute promyelocytic leukemia, acute monocytic leukemia, acute erythroleukemia, acute megakaryoblastic leukemia, acute myelomonocytic leukemia, acute nonlymphocytic leukemia, acute undifferentiated leukemia, chronic myelogenous leukemia, human leukemia, Chronic lymphocytic leukemia, hairy cell leukemia, or multiple myeloma.

19. The use of claim 17, wherein the cancer is associated with a premature stop codon in a tumor suppressor gene, wherein the tumor suppressor gene is APC, ATM, BRAC1, BRAC2, MSH1, pTEN, Rb, or p 53.