A solid form of n-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide

TECHNICAL FIELD OF THE INVENTION

0001The present invention relates to crystalline forms of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide as defined in the claims, pharmaceutical compositions thereof, and the forms and compositions for use in methods therewith.

BACKGROUND OF THE INVENTION

0002CFTR is a cAMP/ATP-mediated anion channel that is expressed in a variety of cells types, including absorptive and secretory epithelia cells, where it regulates anion flux across the membrane, as well as the activity of other ion channels and proteins. In epithelia cells, normal functioning of CFTR is critical for the maintenance of electrolyte transport throughout the body, including respiratory and digestive tissue. CFTR is composed of approximately 1480 amino acids that encode a protein made up of a tandem repeat of transmembrane domains, each containing six transmembrane helices and a nucleotide binding domain. The two transmembrane domains are linked by a large, polar, regulatory (R)-domain with multiple phosphorylation sites that regulate channel activity and cellular trafficking.

0003The gene encoding CFTR has been identified and sequenced (See <nplcit id="ncit0001" npl-type="s"><text>Gregory, R. J. et al. (1990) Nature 347:382-386</text></nplcit>; <nplcit id="ncit0002" npl-type="s"><text>Rich, D. P. et al. (1990) Nature 347:358-362</text></nplcit>), (<nplcit id="ncit0003" npl-type="s"><text>Riordan, J. R. et al. (1989) Science 245: 1066-1073</text></nplcit>). A defect in this gene causes mutations in CFTR resulting in cystic fibrosis ("CF"), the most common fatal genetic disease in humans. Cystic fibrosis affects approximately one in every 2,500 infants in the United States. Within the general United States population, up to 10 million people carry a single copy of the defective gene without apparent ill effects. In contrast, individuals with two copies of the CF associated gene suffer from the debilitating and fatal effects of CF, including chronic lung disease.

0004In patients with cystic fibrosis, mutations in CFTR endogenously expressed in respiratory epithelia leads to reduced apical anion secretion causing an imbalance in ion and fluid transport. The resulting decrease in anion transport contributes to enhanced mucus accumulation in the lung and the accompanying microbial infections that ultimately cause death in CF patients. In addition to respiratory disease, CF patients typically suffer from gastrointestinal problems and pancreatic insufficiency that, if left untreated, results in death. In addition, the majority of males with cystic fibrosis are infertile and fertility is decreased among females with cystic fibrosis. In contrast to the severe effects of two copies of the CF associated gene, individuals with a single copy of the CF associated gene exhibit increased resistance to cholera and to dehydration resulting from diarrhea - perhaps explaining the relatively high frequency of the CF gene within the population.

0005Sequence analysis of the CFTR gene of CF chromosomes has revealed a variety of disease causing mutations (<nplcit id="ncit0004" npl-type="s"><text>Cutting, G. R. et al. (1990) Nature 346:366-369</text></nplcit>; <nplcit id="ncit0005" npl-type="s"><text>Dean, M. et al. (1990) Cell 61:863:870</text></nplcit>; and <nplcit id="ncit0006" npl-type="s"><text>Kerem, B-S. et al. (1989) Science 245:1073-1080</text></nplcit>; <nplcit id="ncit0007" npl-type="s"><text>Kerem, B-S et al. (1990) Proc. Natl. Acad. Sci. USA 87:8447-8451</text></nplcit>). To date, > 1000 disease causing mutations in the CF gene have been identified (<u>http://www.genet.sickkids.on.ca/cftr/</u>). The most prevalent mutation is a deletion of phenylalanine at position 508 of the CFTR amino acid sequence, and is commonly referred to as ΔF508-CFTR. This mutation occurs in approximately 70% of the cases of cystic fibrosis and is associated with a severe disease.

0006The deletion of residue 508 in ΔF508-CFTR prevents the nascent protein from folding correctly. This results in the inability of the mutant protein to exit the ER, and traffic to the plasma membrane. As a result, the number of channels present in the membrane is far less than observed in cells expressing wild-type CFTR. In addition to impaired trafficking, the mutation results in defective channel gating. Together, the reduced number of channels in the membrane and the defective gating lead to reduced anion transport across epithelia leading to defective ion and fluid transport. (<nplcit id="ncit0008" npl-type="s"><text>Quinton, P. M. (1990), FASEB J. 4: 2709-2727</text></nplcit>). Studies have shown, however, that the reduced numbers of ΔF508-CFTR in the membrane are functional, albeit less than wild-type CFTR. (<nplcit id="ncit0009" npl-type="s"><text>Dalemans et al. (1991), Nature Lond. 354: 526-528</text></nplcit>; Denning et al., supra; <nplcit id="ncit0010" npl-type="s"><text>Pasyk and Foskett (1995), J. Cell. Biochem. 270: 12347-50</text></nplcit>). In addition to ΔF508-CFTR, other disease causing mutations in CFTR that result in defective trafficking, synthesis, and/or channel gating could be up- or down-regulated to alter anion secretion and modify disease progression and/or severity.

0007Although CFTR transports a variety of molecules in addition to anions, it is clear that this role (the transport of anions) represents one element in an important mechanism of transporting ions and water across the epithelium. The other elements include the epithelial Na⁺ channel, ENaC, Na⁺/2Cl^-/K⁺ co-transporter, Na⁺-K⁺-ATPase pump and the basolateral membrane K⁺ channels, that are responsible for the uptake of chloride into the cell.

0008These elements work together to achieve directional transport across the epithelium via their selective expression and localization within the cell. Chloride absorption takes place by the coordinated activity of ENaC and CFTR present on the apical membrane and the Na⁺-K⁺-ATPase pump and Cl- channels expressed on the basolateral surface of the cell. Secondary active transport of chloride from the luminal side leads to the accumulation of intracellular chloride, which can then passively leave the cell via Cl^- channels, resulting in a vectorial transport. Arrangement of Na⁺/2Cl^-/K⁺ co-transporter, Na⁺-K⁺-ATPase pump and the basolateral membrane K⁺ channels on the basolateral surface and CFTR on the luminal side coordinate the secretion of chloride via CFTR on the luminal side. Because water is probably never actively transported itself, its flow across epithelia depends on tiny transepithelial osmotic gradients generated by the bulk flow of sodium and chloride.

0009In addition to cystic fibrosis, modulation of CFTR activity may be beneficial for other diseases not directly caused by mutations in CFTR, such as secretory diseases and other protein folding diseases mediated by CFTR. These include, but are not limited to, chronic obstructive pulmonary disease (COPD), dry eye disease, and Sjögren's Syndrome. COPD is characterized by airflow limitation that is progressive and not fully reversible. The airflow limitation is due to mucus hypersecretion, emphysema, and bronchiolitis. Activators of mutant or wild-type CFTR offer a potential treatment of mucus hypersecretion and impaired mucociliary clearance that is common in COPD. Specifically, increasing anion secretion across CFTR may facilitate fluid transport into the airway surface liquid to hydrate the mucus and optimized periciliary fluid viscosity. This would lead to enhanced mucociliary clearance and a reduction in the symptoms associated with COPD. Dry eye disease is characterized by a decrease in tear aqueous production and abnormal tear film lipid, protein and mucin profiles. There are many causes of dry eye, some of which include age, Lasik eye surgery, arthritis, medications, chemical/thermal burns, allergies, and diseases, such as cystic fibrosis and Sjögrens's syndrome. Increasing anion secretion via CFTR would enhance fluid transport from the corneal endothelial cells and secretory glands surrounding the eye to increase corneal hydration. This would help to alleviate the symptoms associated with dry eye disease. Sjögrens's syndrome is an autoimmune disease in which the immune system attacks moisture-producing glands throughout the body, including the eye, mouth, skin, respiratory tissue, liver, vagina, and gut. Symptoms, include, dry eye, mouth, and vagina, as well as lung disease. The disease is also associated with rheumatoid arthritis, systemic lupus, systemic sclerosis, and polymypositis/dermatomyositis. Defective protein trafficking is believed to cause the disease, for which treatment options are limited. Modulators of CFTR activity may hydrate the various organs afflicted by the disease and help to elevate the associated symptoms.

0010As discussed above, it is believed that the deletion of residue 508 in ΔF508-CFTR prevents the nascent protein from folding correctly, resulting in the inability of this mutant protein to exit the ER, and traffic to the plasma membrane. As a result, insufficient amounts of the mature protein are present at the plasma membrane and chloride transport within epithelial tissues is significantly reduced. Infact, this cellular phenomenon of defective ER processing of ABC transporters by the ER machinery, has been shown to be the underlying basis not only for CF disease, but for a wide range of other isolated and inherited diseases. The two ways that the ER machinery can malfunction is either by loss of coupling to ER export of the proteins leading to degradation, or by the ER accumulation of these defective/misfolded proteins [<nplcit id="ncit0011" npl-type="s"><text>Aridor M, et al., Nature Med., 5(7), pp 745- 751 (1999</text></nplcit>);<nplcit id="ncit0012" npl-type="s"><text> Shastry, B.S., et al., Neurochem. International, 43, pp 1-7 (2003</text></nplcit>); <nplcit id="ncit0013" npl-type="s"><text>Rutishauser, J., et al., Swiss Med Wkly, 132, pp 211-222 (2002</text></nplcit>);<nplcit id="ncit0014" npl-type="s"><text> Morello, JP et al., TIPS, 21, pp. 466- 469 (2000</text></nplcit>); <nplcit id="ncit0015" npl-type="s"><text>Bross P., et al., Human Mut., 14, pp. 186-198 (1999</text></nplcit>)]. The diseases associated with the first class of ER malfunction are cystic fibrosis (due to misfolded ΔF508-CFTR as discussed above), hereditary emphysema (due to a1-antitrypsin; non Piz variants), hereditary hemochromatosis, hoagulation-fibrinolysis deficiencies, such as protein C deficiency, Type 1 hereditary angioedema, lipid processing deficiencies, such as familial hypercholesterolemia, Type 1 chylomicronemia, abetalipoproteinemia, lysosomal storage diseases, such as I-cell disease/pseudo-Hurler, Mucopolysaccharidoses (due to lysosomal processing enzymes), Sandhof/Tay-Sachs (due to β-hexosaminidase), Crigler-Najjar type II (due to UDP-glucuronyl-sialyc-transferase), polyendocrinopathy/hyperinsulemia, Diabetes mellitus (due to insulin receptor), Laron dwarfism (due to growth hormone receptor), myleoperoxidase deficiency, primary hypoparathyroidism (due to preproparathyroid hormone), melanoma (due to tyrosinase). The diseases associated with the latter class of ER malfunction are Glycanosis CDG type 1, hereditary emphysema (due to α1-Antitrypsin (PiZ variant), congenital hyperthyroidism, osteogenesis imperfecta (due to Type I, II, IV procollagen), hereditary hypofibrinogenemia (due to fibrinogen), ACT deficiency (due to α1-antichymotrypsin), Diabetes insipidus (DI), neurophyseal DI (due to vasopvessin hormone/V2-receptor), neprogenic DI (due to aquaporin II), Charcot-Marie Tooth syndrome (due to peripheral myelin protein 22), Perlizaeus-Merzbacher disease, neurodegenerative diseases such as Alzheimer's disease ( due to βAPP and presenilins), Parkinson's disease, amyotrophic lateral sclerosis, progressive supranuclear plasy, Pick's disease, several polyglutamine neurological disorders asuch as Huntington, spinocerebullar ataxia type I, spinal and bulbar muscular atrophy, dentatorubal pallidoluysian, and myotonic dystrophy, as well as spongiform encephalopathies, such as hereditary Creutzfeldt-Jakob disease (due to prion protein processing defect), Fabry disease (due to lysosomal α-galactosidase A) and Straussler-Scheinker syndrome (due to Prp processing defect).

0011In addition to up-regulation of CFTR activity, reducing anion secretion by CFTR modulators may be beneficial for the treatment of secretory diarrheas, in which epithelial water transport is dramatically increased as a result of secretagogue activated chloride transport. The mechanism involves elevation of cAMP and stimulation of CFTR.

0012Although there are numerous causes of diarrhea, the major consequences of diarrheal diseases, resulting from excessive chloride transport are common to all, and include dehydration, acidosis, impaired growth and death.

0013Acute and chronic diarrheas represent a major medical problem in many areas of the world. Diarrhea is both a significant factor in malnutrition and the leading cause of death (5,000,000 deaths/year) in children less than five years old.

0014Secretory diarrheas are also a dangerous condition in patients of acquired immunodeficiency syndrome (AIDS) and chronic inflammatory bowel disease (IBD). 16 million travelers to developing countries from industrialized nations every year develop diarrhea, with the severity and number of cases of diarrhea varying depending on the country and area of travel.

0015Diarrhea in barn animals and pets such as cows, pigs and horses, sheep, goats, cats and dogs, also known as scours, is a major cause of death in these animals. Diarrhea can result from any major transition, such as weaning or physical movement, as well as in response to a variety of bacterial or viral infections and generally occurs within the first few hours of the animal's life.

0016The most common diarrheal causing bacteria is enterotoxogenic E.coli (ETEC) having the K99 pilus antigen. Common viral causes of diarrhea include rotavirus and coronavirus. Other infectious agents include cryptosporidium, giardia lamblia, and salmonella, among others.

0017Symptoms of rotaviral infection include excretion of watery feces, dehydration and weakness. Coronavirus causes a more severe illness in the newborn animals, and has a higher mortality rate than rotaviral infection. Often, however, a young animal may be infected with more than one virus or with a combination of viral and bacterial microorganisms at one time. This dramatically increases the severity of the disease.

0018Accordingly, there is a need for stable polymorphic forms of modulators of CFTR activity, such as Compound 1, that can be used to modulate the activity of CFTR in the cell membrane of a mammal.

0019There is a need for methods of treating CFTR-mediated diseases using such modulators of CFTR activity.

SUMMARY OF THE INVENTION

0020The present invention relates to a crystal form of of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide, characterized by: an X-ray powder diffraction pattern having one or more peaks (2θ) at 6.2 ± 0.2, 7.6 ± 0.2, 12.3 ± 0.2, and 18.0 ± 0.2 degrees obtained using Cu K alpha radiation; and/or a monoclinic crystal system, a P2₁ space grouping, and the following unit cell dimensions: <tables id="tabl0001" num="0001"><table frame="none"><tgroup cols="2" colsep="0" rowsep="0"><colspec colnum="1" colname="col1" colwidth="28mm" /><colspec colnum="2" colname="col2" colwidth="26mm" /><tbody><row><entry>a = 11.8011(7) Å</entry><entry>α= 90°</entry></row><row><entry>b = 5.9819(3) Å</entry><entry>β= 105.110(4)°</entry></row><row><entry>c = 14.7974(8) Å</entry><entry>γ = 90°.</entry></row></tbody></tgroup></table></tables> N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide (hereinafter "Compound 1") has the structure below: <chemistry id="chem0001" num="0001"><img file="EP3708564B1_D0001.tif" /></chemistry>

0021The crystal forms of Compound <b>1</b> and pharmaceutically acceptable compositions thereof are useful for treating or lessening the severity of a variety of CFTR mediated diseases. Compound <b>1</b> is known as both N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide and N-(5-hydroxy-2,4-di-<i>tert-</i>butyl-phenyl)-4-oxo-1H-quinoline-3-carboxamide.

0022Disclosed herein is solid amorphous N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide. In some embodiments, the solid amorphous N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide comprises less than about 15% crystalline N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide.

0023Disclosed herein is a preparation of amorphous N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide substantially free of crystalline N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide.

0024In some embodiments, the preparation further comprises a surfactant, polymer, or inert pharmaceutically acceptable substance.

0025In some embodiments, the preparation comprises a solid dispersion, a mixture or a liquid dispersion.

0026In some embodiments, the preparation comprises solid particles.

0027In some embodiments, the preparation comprises less than about 15% of crystalline N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide.

0028In some embodiments, the amorphous N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide has a particle size distribution of D10, less than 5 µm. In some embodiments, the amporphous N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide have a particle size distribution of D50, less than 17 µm. In some embodiments, the amporphous N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide have a particle size distribution of D90, less than 100 µm.

0029Disclosed herein is a solid dispersion comprising amorphous N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide.

0030In some embodiments, the solid dispersion comprises less than about 40% of crystalline N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide. In some embodiments, the solid dispersion is substantially free of crystalline N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide.

0031In some embodiments, the solid dispersion further comprises a surfactant, polymer, or inert pharmaceutically acceptable substance. For example, the solid dispersion comprises a polymer, and the polymer is one or more than one water-soluble polymer or partially water-soluble polymer.

0032In some embodiments, the N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide has improved physical or chemical stability relative to amorphous N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide without being in the presence of polymer.

0033In some embodiments,the solid dispersion has a higher glass transition temperature than the glass transition temperature of neat amorphous N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide.

0034In some embodiments, the polymer is hydroxypropylmethylcellulose (HPMC). In some embodiments, the polymer is hydroxypropylmethylcellulose acetate succinate (HPMCAS). In some embodiments, the polymer is vinylpyrrolidone/vinyl acetate copolymer (PVP/VA). In some embodiments, the polymer is present in an amount of from about 10% by weight to about 80% by weight, for example, the polymer is present in an amount of less than about 70% by weight, the polymer is present in an amount of about 50% by weight, or the polymer is present in an amount of about 49.5% by weight.

0035In some embodiments, the N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is present in an amount of from about 10% by weight to about 80% by weight, for example, the N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is present in an amount of less than about 70% by weight or the N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is present in an amount of about 50% by weight.

0036In some embodiments, the solid dispersion comprises a surfactant, for example, sodium lauryl sulfate. In some embodiments, the surfactant is present in an amount from about 0.1 to about 5%, for example, the surfactant is present in 0.5%.

0037In some embodiments, at least about 80% by weight of the N N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is in an amorphous form. In some embodiments, substantially all the N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is in an amorphous form.

0038In some embodiments, the solid dispersion is obtained by spray drying.

0039Disclosed herein is a pharmaceutical composition comprising amorphous N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide. In some embodiments, the amorphous N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is substantially free of crystalline N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide.

0040Disclosed herein is a pharmaceutical composition comprising an amorphous N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide as a solid dispersion and one or more of a surfactant, polymer, inert pharmaceutically acceptable substance, or pharmaceutically acceptable carrier.

0041In some embodiments, the solid dispersion comprises a polymer and wherein the polymer is one or more than one water-soluble polymer or partially water-soluble polymer.

0042In some embodiments, the solid dispersion has a higher glass transition temperature than the glass transition temperature of neat amorphous N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide.

0043In some embodiments, the polymer is HPMC. In some embodiments, the polymer is HPMCAS. In some embodiments, the polymer is PVP/VA.

0044Disclosed herein is a pharmaceutical composition comprising: an amorphous solid dispersion of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide wherein said N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide comprises about 30-75% wt/wt of the pharmaceutical composition, one or more polymer selected from the group of HPMC and HPMCAS, wherein said polymer is comprises about 30-75% wt/wt of the pharmaceutical composition, and a surfactant, wherein said surfactant comprises about 0.25-1% wt/wt of the pharmaceutical composition.

0045In some embodiments, the polymer is HPMCAS. In some embodiments, the polymer is HPMC.

0046In some embodiments, the surfactant is sodium laurel sulfate.

0047In some embodiments, said N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide comprises about 50% wt/wt of the pharmaceutical composition, said polymer is HPMCAS and comprises about 49.5% wt/wt of the pharmaceutical composition, and a said surfactant is sodium laurel sulfate and comprises about 0.5 % wt/wt of the pharmaceutical composition.

0048Disclosed herein is a pharmaceutical composition comprising;

0049an aqueous suspension comprising amorphous N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide particles and a pharmaceutically acceptable carrier.

0050In some embodiments, the pharmaceutically acceptable carrier is a polymer in solution selected from the group of HPMC and HPMCAS. In some embodiments, the pharmaceutically acceptable carrier is a polymer in solution is PVP/VA.

0051In some embodiments, the amorphous compound is in the form of a solid dispersion.

0052In some embodiments, the pharmaceutical composition further comprises a surfactant, either in the solution or as a component of the solid dispersion, for example, SLS. In some embodiments, the polymer is either in the solution or as a component of the solid dispersion particles or both. In some embodiments, the aqueous suspension comprises from about 0.1% to about 20% by weight of the surfactant. In some embodiments, the aqueous suspension comprises from about 0.1% to about 2.0% by weight of polymer, for example, about 1% by weight of polymer.

0053Disclosed herein is a process for preparing an amorphous form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide comprising spray-drying N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide to provide an amorphous form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide.

0054In some embodiments, the method comprises combining N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide and a suitable solvent to form a mixture and then spray-drying the mixture to obtain the amorphous form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide.

0055In some embodiments, the mixture is a solution N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide and the suitable solvent. In some embodiments, the suitable solvent comprises acetone or MEK. In some embodiments, the suitable solvent comprises a mixture of solvents, for example, a mixture of acetone and water or a mixture of MEK and water. In some embodiments, the water in the solvent mixture is present at about 10% wt.

0056In some embodiments, the method comprises a) forming a mixture comprising N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide, a polymer, and a solvent; and b) spray-drying the mixture to form a solid dispersion comprising N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide.

0057In some embodiments, the mixture comprises a solution of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide, the polymer, and the solvent. In some embodiments, the polymer is selected from HPMC and HPMCAS. In some embodiments, the polymer is PVP/VA. In some embodiments, the polymer is present in an amount of from about 30% to about 70% by weight in the solid dispersion. In some embodiments, the mixture further comprises a surfactant, for example, SLS.

0058In some embodiments, the solvent comprises acetone, for example, a mixture of acetone and water. In some embodiments, the solvent comprises from about 0% to about 20% water and from about 70% to about 100% acetone.

0059Disclosed herein is a solid dispersion prepared according a process described herein.

0060Disclosed herein is a method for treating a CFTR-mediated disease in a mammal comprising administering amorphous N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide. In some embodiments, the method comprises administering an amorphous solid dispersion of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide. In some embodiments, the method comprises administering an additional therapeutic agent.

0061Disclosed herein is a pharmaceutical pack or kit comprising amorphous N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide and a pharmaceutically acceptable carrier.

0062Disclosed herein is a crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide, characterized by one or more peaks at from about 4.8 to about 5.2 degrees, for example, about 5.0 degrees, and from about 15.4 to about 15.8 degrees, for example, about 15.6 degrees in an X-ray powder diffraction pattern obtained using Cu K alpha radiation. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak at from about 7.6 to about 8.0, e.g., 7.8. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak at from about 8.3 to about 8.7, for example, about 8.5. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak at from about 9.0 to about 9.4, for example, about 9.2. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak at from about 9.7 to about 10.1, for example about 9.9. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak at from about 11.7 to about 12.1, for example, about 11.9. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak at from about 12.4 to about 12.8, for example, about 12.6. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak at from about 13.7 to about 14.1, for example about 13.9. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak at from about 14.7 to about 15.1, for example, about 14.9. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak from about 16.3 to about 16.7, for example about 16.5. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak from about 17.9 to about 18.3, for example, about 18.1. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak from about 18.3 to about 18.7, for example, about 18.5. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak from about 20.5 to about 20.9, for example, about 20.7. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak from about 21.8 to about 22.2, for example, about 22.0. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak from about 23.1 to about 23.7, for example, about 23.5. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak from about 25.1 to about 25.5, for example, about 25.3. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak from about 27.8 to about 28.2, for example, about 28.0. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak from about 29.2 to about 29.6, for example, about 29.4. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak from about 30.7 to about 31.1, for example, about 30.9. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is characterized by an X-ray powder diffraction pattern obtained using Cu K alpha radiation as shown in <figref idref="f0003"><b>FIGURE 4</b></figref><b>.</b>

0063Disclosed herein is a pharmaceutical composition comprising the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide having the characteristics of Form A, for example as described above, and a pharmaceutically acceptable adjuvant or carrier.

0064Disclosed herein is a process for preparing a crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide of Form A, for example as as characterized above, wherein said process comprises the step of heating N N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide to about 250 °C and cooling to room temperature.

0065Disclosed herein is a method for treating a CFTR mediated disease in a mammal comprising administering N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide of Form A, for example as as characterized above. In some embodiments, the N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is a component of a pharmaceutical composition. In some embodiments, the method comprises administering an additional therapeutic agent.

0066Disclosed herein is a pharmaceutical pack or kit comprising crystalline N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide of Form A, for example as as characterized above and a pharmaceutically acceptable carrier.

0067In one aspect, the invention features a crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide, characterized by one or more peaks at from about 6.2 to about 6.6, for example, about 6.4, from about 7.5 to about 7.9, for example, about 7.7, from about 12.5 to about 12.9, for example, about 12.7, and from about 17.9 to about 18.3, for example, about 18.1 degrees in an X-ray powder diffraction pattern obtained using Cu K alpha radiation.

0068In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak from about 8.2 to about 8.6, for example, about 8.4. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak from about 10.8 to about 11.2, for example, about 11.0. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak from about 14.6 to about 15.0, for example, about 14.8. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak from about 15.9 to about 16.3, for example, about 16.1. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak from about 16.9 to about 17.3, for example, about 17.1. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak from about 18.4 to about 18.8, for example, about 18.6. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak from about 19.2 to about 19.6, for example, about 19.4. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak from about 20.9 to about 21.3, for example, about 21.1. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak from about 22.4 to about 22.8, for example, about 22.6. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak from about 23.2 to about 23.6, for example, about 23.4. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak from about 23.7 to about 24.1, for example, about 23.9. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak from about 24.7 to about 25.1, for example, about 24.9. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak from about 25.3 to about 25.7, for example, about 25.5. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak from about 26.5 to about 26.9, for example, about 26.7. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak from about 27.3 to about 27.7, for example, about 27.5. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is further characterized by the following peak from about 29.4 to about 29.8, for example, about 29.6. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro 4-oxoquinoline-3-carboxamide is further characterized by the following peak from about 33.3 to about 33.7, for example, about 33.5. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro 4-oxoquinoline-3-carboxamide is further characterized by the following peak from about 36.6 to about 37.0, for example, about 36.8. In some embodiments, the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide is characterized by an X-ray powder diffraction pattern obtained using Cu K alpha radiation as shown in <figref idref="f0006"><b>FIGURE 7</b></figref><b>.</b>

0069In one aspect, the invention features a crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide, having a monoclinic crystal system, a P21 space grouping, and the following unit cell dimensions: <tables id="tabl0002" num="0002"><table frame="none"><tgroup cols="2" colsep="0" rowsep="0"><colspec colnum="1" colname="col1" colwidth="28mm" /><colspec colnum="2" colname="col2" colwidth="26mm" /><tbody><row><entry>a = 11.8011(7) Å</entry><entry>α= 90°</entry></row><row><entry>b = 5.9819(3) Å</entry><entry>β= 105.110(4)°</entry></row><row><entry>c = 14.7974(8) Å</entry><entry>γ = 90°.</entry></row></tbody></tgroup></table></tables>

0070In one aspect, the invention features a pharmaceutical composition comprising the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide according to Form B, for example, as characterized above, and a pharmaceutically acceptable adjuvant or carrier.

0071In one aspect, the invention features a process for preparing the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide according to Form B, for example, as characterized above, wherein said process comprises the steps of alternatively heating and cooling a slurry of Compound <b>1</b> and acetonitrile. In some embodiments, the process comprises heating said slurry at about 50 C for about 12 hours. In some embodiments, said cooling step comprises placing said slurry at room temperature for about 12 hours, followed by cooling at about 0 °C overnight.

0072The crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide of the invention, or a pharmaceutical composition comprising the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide according to Form B, for example, as characterized above can be used in a method of treating a CFTR mediated disease in a patient, wherein the method comprises the step of administering to said patient a crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide of the invention or a pharmaceutical composition comprising a crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide according to Form B, for example, as characterized above.

0073The CFTR mediated disease is selected from cystic fibrosis, hereditary emphysema, hereditary hemochromatosis, coagulation-fibrinolysis deficiencies, such as protein C deficiency, Type 1 hereditary angioedema, lipid processing deficiencies, such as familial hypercholesterolemia, Type 1 chylomicronemia, abetalipoproteinemia, lysosomal storage diseases, such as I-cell disease/pseudo-Hurler, mucopolysaccharidoses, Sandhof/Tay-Sachs, Crigler-Najjar type II, polyendocrinopathy/hyperinsulemia, Diabetes mellitus, Laron dwarfism, myleoperoxidase deficiency, primary hypoparathyroidism, melanoma, glycanosis CDG type 1, hereditary emphysema, congenital hyperthyroidism, osteogenesis imperfecta, hereditary hypofibrinogenemia, ACT deficiency, Diabetes insipidus (DI), neurophyseal DI, neprogenic DI, Charcot-Marie Tooth syndrome, Perlizaeus-Merzbacher disease, neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, progressive supranuclear plasy, Pick's disease, several polyglutamine neurological disorders asuch as Huntington, spinocerebullar ataxia type I, spinal and bulbar muscular atrophy, dentatorubal pallidoluysian, and myotonic dystrophy, as well as spongiform encephalopathies, such as hereditary Creutzfeldt-Jakob disease, Fabry disease, Straussler-Scheinker syndrome, COPD, dry-eye disease, and Sjogren's disease. In some embodiments, the disease is cystic fibrosis.

0074Processes described herein can be used to prepare the compositions of this invention. The amounts and the features of the components used in the processes would be as described herein.

0075As used herein, the term "amorphous" refers to a solid material having no long range order in the position of its molecules Amorphous solids are generally supercooled liquids in which the molecules are arranged in a random manner so that there is no well-defined arrangement, e.g., molecular packing, and no long range order. Amorphous solids are generally isotropic, i.e. exhibit similar properties in all directions and do not have definite melting points. For example, an amorphous material is a solid material having no sharp characteristic crystalline peak(s) in its X-ray power diffraction (XRPD) pattern (i.e., is not crystalline as determined by XRPD). Instead, one or several broad peaks (e.g., halos) appear in its XRPD pattern. Broad peaks are characteristic of an amorphous solid. See, <patcit id="pcit0001" dnum="US20040006237A"><text>US 2004/0006237</text></patcit> for a comparison of XRPDs of an amorphous material and crystalline material.

0076As used herein, the phrase "substantially amorphous Compound <b>1</b>" is used interchangeably with the phrase "amorphous Compound <b>1</b> substantially free of crystalline Compound <b>1</b>." In some embodiments, substantially amorphous Compound <b>1</b> has less than about 30% crystalline Compound <b>1,</b> for example, less than about 30% of crystalline Compound <b>1,</b> e.g., less than about 25% crystalline Compound <b>1,</b> less than about 20% crystalline Compound <b>1,</b> less than about 15% crystalline Compound <b>1,</b> less than about 10% crystalline Compound <b>1,</b> less than about 5% crystalline Compound <b>1,</b> less than about 2% crystalline Compound <b>1.</b> In some preferred embodiments, Compound <b>1</b> has less than about 15% crystalline compound <b>1.</b> Some embodiments include a preparation of substantially amorphous Compound <b>1,</b> for example having the degree of crystalline Compound <b>1</b> as described above.

0077As used herein "crystalline solids" refers to compounds or compositions where the structural units are arranged in fixed geometric patterns or lattices, so that crystalline solids have rigid long range order. The structural units that constitute the crystal structure can be atoms, molecules, or ions. Crystalline solids show definite melting points.

0078As used herein, a "dispersion" refers to a disperse system in which one substance, the dispersed phase, is distributed, in discrete units, throughout a second substance (the continuous phase or vehicle). The size of the dispersed phase can vary considerably (e.g. colloidal particles of nanometer dimension, to multiple microns in size). In general, the dispersed phases can be solids, liquids, or gases. In the case of a solid dispersion, the dispersed and continuous phases are both solids. In pharmaceutical applications, a solid dispersion can include a crystalline drug (dispersed phase) in an amorphous polymer (continuous phase), or alternatively, an amorphous drug (dispersed phase) in an amorphous polymer (continuous phase). In some embodiments an amorphous solid dispersion includes the polymer constituting the dispersed phase, and the drug constitute the continuous phase. In some embodiments, the dispersion includes amorphous Compound <b>1</b> or substantially amorphous Compound <b>1.</b>

0079The term "solid amorphous dispersion" generally refers to a solid dispersion of two or more components, usually a drug and polymer, but possibly containing other components such as surfactants or other pharmaceutical excipients, where Compound <b>1</b> is amorphous or substantially amorphous (e.g., substantially free of crystalline Compound <b>1</b>), and the physical stability and/or dissolution and/or solubility of the amorphous drug is enhanced by the other components.

0080Solid dispersions typically include a compound dispersed in an appropriate carrier medium, such as a solid state carrier. In one embodiment, a carrier disclosed herein comprises a polymer, preferably, a water-soluble polymer or a partially water-soluble polymer. It would be understood that one or more than one water-soluble polymer could be used in a solid dispersion disclosed herein.

0081An exemplary solid dispersion is a co-precipitate or a co-melt of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide with at least one polymer. A "Co-precipitate" is a product after dissolving a drug and a polymer in a solvent or solvent mixture followed by the removal of the solvent or solvent mixture. Sometimes the polymer can be suspended in the solvent or solvent mixture. The solvent or solvent mixture includes organic solvents and supercritical fluids. A "co-melt" is a product after heating a drug and a polymer to melt, optionally in the presence of a solvent or solvent mixture, followed by mixing, removal of at least a portion of the solvent if applicable, and cooling to room temperature at a selected rate. In some cases, the solid dispersions are prepared by adding a solution of a drug and a solid polymer followed by mixing and removal of the solvent. To remove the solvent, vacuum drying, spray drying, tray drying, lyophilization, and other drying procedures may be applied. Applying any of these methods using appropriate processing parameters, according to this invention, would provide N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide in an amorphous state in the final solid dispersion product.

BRIEF DESCRIPTION OF THE DRAWINGS

0082<ul id="ul0001" list-style="none" compact="compact"><li><figref idref="f0001"><b>FIGURE 1</b></figref> is an X-Ray powder diffraction pattern of Compound <b>1.</b></li><li><figref idref="f0001"><b>FIGURE 2</b></figref> is the ¹H NMR spectrum of Compound <b>1.</b></li><li><figref idref="f0002"><b>FIGURE 3</b></figref> is the DSC trace of Compound <b>1.</b></li><li><figref idref="f0003"><b>FIGURE 4</b></figref> is the X-Ray powder diffraction pattern of Form A.</li><li><figref idref="f0004"><b>FIGURE 5</b></figref> is the DSC trace of Form A.</li><li><figref idref="f0005"><b>FIGURE 6</b></figref> is the TGA trace of Form A.</li><li><figref idref="f0006"><b>FIGURE 7</b></figref> is the X-Ray powder diffraction pattern of Form B.</li><li><figref idref="f0007"><b>FIGURE 8</b></figref> is the DSC trace of Form B.</li><li><figref idref="f0008"><b>FIGURE 9</b></figref> is the TGA trace of Form B.</li><li><figref idref="f0009"><b>FIGURE 10</b></figref> is a conformational picture of Form B, based on single crystal X-Ray analysis.</li><li><figref idref="f0010"><b>FIGURE 11</b></figref> is the X-Ray powder diffraction pattern of the Amorphous Form.</li><li><figref idref="f0011"><b>FIGURE 12</b></figref> is the TGA trace of the Amorphous Form.</li><li><figref idref="f0012"><b>FIGURE 13</b></figref> is the DSC trace of the Amorphous Form.</li></ul>

DETAILED DESCRIPTION OF THE INVENTION

<u>Solid forms of Compound <b>1</b></u>

Form A (not according to the invention)

0083Form A of Compound <b>1</b> is characterized by one or more peaks at from about 4.8 to about 5.2, for example, about 5.0, e.g., 4.99, and from about 15.4 to about 15.8, for example, about 15.6 e.g., 15.58 degrees in an X-ray powder diffraction pattern obtained using Cu K alpha radiation (2Θ). Other peaks (2Θ), which can be characteristic of Form A, include the following: from about 7.6 to about 8.0, for example, about 7.8, e.g., 7.75; from about 8.3 to about 8.7, for example, about 8.5, e.g., 8.46; from about 9.0 to about 9.4, for example, about 9.2, e.g., 9.21; from about 9.7 to about 10.1, for example, about 9.9, e.g., 9.92; from about 11.7 to about 12.1, for example, about 11.9, e.g., 11.93; from about 12.4 to about 12.8, for example, about 12.6, e.g., 12.64; from about 13.7 to about 14.1, for example, about 13.9, e.g., 13.88; from about 14.7 to about 15.1, for example, about 14.9, e.g., 14.91; from about 16.3 to about 16.7, for example, about 16.5, e.g., 16.46; from about 17.9 to about 18.3, for example, about 18.1, e.g., 18.09; from about 18.3 to about 18.7, for example, about 18.5, e.g., 18.52; from about 21.5 to about 21.9, for example, about 21.7, e.g., 20.65; from about 21.8 to about 22.2, for example, about 22.0, e.g., 21.95; from about 23.1 to about 23.7, for example, about 23.5, e.g., 23.49; from about 25.1 to about 25.5, for example, about 25.3, e.g., 25.26; from about 27.8 to about 28.2 for example, about 28.0, e.g., 28.02; from about 29.2 to about 29.6, for example, about 29.4, e.g., 29.35; and about from about 30.7 to about 31.1, for example, 30.9, e.g., 30.85. For example, Form A can be characterized by an X-ray powder diffraction pattern obtained using Cu K alpha radiation substantially similar to <figref idref="f0003"><b>FIGURE 4</b></figref><b>.</b>

0084Pharmaceutical compositions including Form A and a pharmaceutically acceptable adjuvant or carrier, such as a polymer or surfactant are also described. Form A can be formulated in a pharmaceutical composition, in some instances, with another therapeutic agent, for example another therapeutic agent for treating cystic fibrosis or a symptom thereof.

0085Processes for preparing Form A are exemplified herein.

0086Form A or a pharmaceutical composition comprising Form A can be used in methods of treating a CFTR mediated disease, such as cystic fibrosis, in a patient, wherein the methods include administering to said patient Form A or a pharmaceutical composition comprising Form A.

Form B

0087The solid state crystal Form B of Compound <b>1</b> is characterized by one or more peaks at from about 6.0 to about 6.4 for example, about 6.2, e.g., 6.17, from about 7.4 to about 7.8 for example, about 7.6, e.g., 7.61, from about 12.1 to about 12.5 for example, about 12.3, e.g., 12.33, and from about 17.8 to about 18.2 for example, about 18.0, e.g., 17.96 degrees in an X-ray powder diffraction pattern obtained using Cu K alpha radiation (2Θ). Other peaks (2Θ), which can be characteristic of Form B, include the following: from about 8.2 to about 8.6 for example, about 8.4, e.g., 8.40; from about 10.8 to about 11.2 for example, about 11.0, e.g., 11.02; from about 14.6 to about 15.0 for example, about 14.8, e.g., 14.83; from about 15.9 to about 16.3 for example, about 16.1, e.g., 16.14; from about 16.9 to about 17.3 for example, about 17.1, e.g., 17.11; from about 18.4 to about 18.8 for example, about 18.6, e.g., 18.55; from about 19.2 to about 19.6 for example, about 19.4, e.g., 19.43; from about 20.9 to about 21.3 for example, about 21.1, e.g., 21.05; from about 22.4 to about 22.8 for example, about 22.6, e.g., 22.56; from about 23.2 to about 23.6 for example, about 23.4, e.g., 23.37; from about 23.7 to about 24.1 for example, about 23.9, e.g., 23.94; from about 24.7 to about 25.1 for example, about 24.9, e.g., 24.86; from about 25.3 to about 25.7 for example, about 25.5, e.g., 25.50; from about 26.5 to about 26.9 for example, about 26.7, e.g., 26.72; from about 27.3 to about 27.7 for example, about 27.5, e.g., 27.51; from about 29.4 to about 29.8 for example, about 29.6, e.g., 29.60; from about 33.3 to about 33.7 for example, about 33.5, e.g., 33.48; and from about 36.6 to about 37.0 for example, about 36.8, e.g., 36.78. Form B can be further characterized, for example, by an X-ray powder diffraction pattern obtained using Cu K alpha radiation substantially similar to <figref idref="f0006"><b>FIGURE 7</b></figref><b>.</b>

0088Applicants have determined crystal structure dimensions of Form B by analysis of single crystal data. Form B is a monoclinic crystal system having a P2₁ space grouping, and the following unit cell dimentions: a = 11.8011(7) Å, α= 90°; b = 5.9819(3) Å, β= 105.110(4)°l; c = 14.7974(8) Å, γ = 90°. Additional details about the structure and packing of Form B are provided in the Examples.

0089Pharmaceutical compositions including Form B and a pharmaceutically acceptable adjuvant or carrier, such as a polymer or surfactant are also described. Form B can be formulated in a pharmaceutical composition, in some instances, with another therapeutic agent, for example another therapeutic agent for treating cystic fibrosis or a symptom thereof.

Processes for preparing Form B are exemplified herein.

0090Methods of treating a CFTR mediated disease, such as cystic fibrosis, in a patient include administering to said patient Form B or a pharmaceutical composition comprising Form B.

Amorphous Compound <b>1</b> (not according to the invention)

0091Compound <b>1</b> can be present as an amorpous solid, for example amorphous Compound <b>1</b> as a substantially neat preparation, or amorphous compound <b>1</b> as a component as a dispersion such as a solid amorphous dispersion.

0092An amorphous form of Compound <b>1</b> is substantially free of crystalline Compound <b>1</b> (e.g., Form A, Form B or any crystalline form of Compound <b>1</b>), for example Compound 1 has less than about 30% of crystalline Compound <b>1,</b> e.g., less than about 25% crystalline Compound <b>1,</b> less than about 20% crystalline Compound <b>1,</b> less than about 15% crystalline Compound <b>1,</b> less than about 10% crystalline Compound <b>1,</b> less than about 5% crystalline Compound <b>1,</b> less than about 2% crystalline. Compound <b>1,</b> preferably less than about 15% crystalline compound <b>1.</b> Compound <b>1</b> can be characterized by an X-ray powder diffraction pattern obtained using Cu K alpha radiation as shown in <figref idref="f0010"><b>FIGURE 11</b></figref><b>.</b> For example, the substantially amorphous form of Compound <b>1</b> can be characterized as having an XRPD having no sharp characteristic crystalline peak(s) in its X-ray power diffraction (XRPD) pattern (i.e., is not crystalline as determined by XRPD). Instead, one or several broad peaks (e.g., halos) appear in its XRPD pattern.

Polymers

0093Solid dispersions including amorphous Compound <b>1</b> and a polymer (or solid state carrier) also are included herein. For example, Compound <b>1</b> is present as an amorphous compound as a component of a solid amorphous dispersion. The solid amorphous dispersion, generally includes Compound <b>1</b> and a polymer. Exemplary polymers include cellulosic polymers such as HPMC or HPMCAS and pyrrolidone containing polymers such as PVP/VA. In some embodiments, the solid amporphous dispersion includes one or more additional exipients, such as a surfactant.

0094In one embodiment, a polymer is able to dissolve in aqueous media. The solubility of the polymers may be pH-independent or pH-dependent. The latter include one or more enteric polymers. The term "enteric polymer" refers to a polymer that is preferentially soluble in the less acidic environment of the intestine relative to the more acid environment of the stomach, for example, a polymer that is insoluble in acidic aqueous media but soluble when the pH is above 5-6. An appropriate polymer should be chemically and biologically inert. In order to improve the physical stability of the solid dispersions, the glass transition temperature (T_g) of the polymer should be as high as possible. For example, preferred polymers have a glass transition temperature at least equal to or greater than the glass transition temperature of the drug (e.g., Compound <b>1</b>). Other preferred polymers have a glass transition temperature that is within about 10 to about 15 °C of the drug (e.g., Compound <b>1</b>). Examples of suitable glass transition temperatures of the polymers include at least about 90 °C, at least about 95 °C, at least about 100 °C, at least about 105 °C, at least about 110 °C, at least about 115 °C, at least about 120 °C, at least about 125 °C, at least about 130 °C, at least about 135 °C, at least about 140 °C, at least about 145 °C, at least about 150 °C, at least about 155 °C, at least about 160 °C, at least about 165 °C, at least about 170 °C, or at least about 175 °C (as measured under dry conditions). Without wishing to be bound by theory, it is believed that the underlying mechanism is that a polymer with a higher T_g generally has lower molecular mobility at room temperature, which can be a crucial factor in stabilizing the physical stability of the amorphous solid dispersion.

0095Additionally, the hygroscopicity of the polymers should be as low, e.g., less than about 10%. For the purpose of comparison in this application, the hygroscopicity of a polymer or composition is characterized at about 60% relative humidity. The polymer may have less than about 10% water absorption, for example less than about 9%, less than about 8%, less than about 7%, less than about 6%, less than about 5%, less than about 4%, less than about 3%, or less than about 2% water absorption. The hygroscopicity can also affect the physical stability of the solid dispersions. Generally, moisture adsorbed in the polymers can greatly reduce the T_g of the polymers as well as the resulting solid dispersions, which will further reduce the physical stability of the solid dispersions as described above.

0096In one embodiment, the polymer is one or more water-soluble polymer(s) or partially water-soluble polymer(s). Water-soluble or partially water-soluble polymers include but are not limited to, cellulose derivatives (e.g., hydroxypropylmethylcellulose (HPMC), hydroxypropylcellulose (HPC)) or ethylcellulose; polyvinylpyrrolidones (PVP); polyethylene glycols (PEG); polyvinyl alcohols (PVA); acrylates, such as polymethacrylate (e.g., Eudragit^® E); cyclodextrins (e.g., β-cyclodextin) and copolymers and derivatives thereof, including for example PVP-VA (polyvinylpyrollidone-vinyl acetate).

0097The polymer may be hydroxypropylmethylcellulose (HPMC), such as HPMC E50, HPMCE15, or HPMC60SH50).

0098As discussed herein, the polymer can be a pH-dependent enteric polymer. Such pH-dependent enteric polymers include, but are not limited to, cellulose derivatives (e.g., cellulose acetate phthalate (CAP)), hydroxypropyl methyl cellulose phthalates (HPMCP), hydroxypropyl methyl cellulose acetate succinate (HPMCAS), carboxymethylcellulose (CMC) or a salt thereof (e.g., a sodium salt such as (CMC-Na)); cellulose acetate trimellitate (CAT), hydroxypropylcellulose acetate phthalate (HPCAP), hydroxypropylmethyl-cellulose acetate phthalate (HPMCAP), and methylcellulose acetate phthalate (MCAP), or polymethacrylates (e.g., Eudragit^® S). The polymer may be hydroxypropyl methyl cellulose acetate succinate (HPMCAS).

0099In yet another embodiment, the polymer is a polyvinylpyrrolidone co-polymer, for example, avinylpyrrolidone/vinyl acetate co-polymer (PVP/VA).

0100In embodiments where Compound <b>1</b> forms a solid dispersion with a polymer, for example with an HPMC, HPMCAS or PVP/VA polymer, the amount of polymer relative to the total weight of the solid dispersion ranges from about 0.1% to 99% by weight. Unless otherwise specified, percentages of drug, polymer and other excitpients as described within a dispersion are given in weight percentages. The amount of polymer is typically at least about 20%, and preferably at least about 30%, for example, at least about 35%, at least about 40%, at least about 45%, or about 50% (e.g., 49.5%). The amount is typically about 99% or less, and preferably about 80% or less, for example about 75% or less, about 70% or less, about 65% or less, about 60% or less, or about 55% or less. In one embodiment, the polymer is in an amount of up to about 50% of the total weight of the dispersion (and even more specifically, between about 40% and 50%, such as about 49%, about 49.5%, or about 50%). HPMC and HPMCAS are available in a variety of grades from ShinEtsu, for example, HPMCAS is available in a number of varieties, including AS-LF, AS-MF, AS-HF, AS-LG, AS-MG, AS-HG. Each of these grades vary with the percent substitution of acetate and succinate.

0101In some preferred embodiments, Compound <b>1</b> and polymer are present in roughly equal amounts, for example each of the polymer and the drug make up about half of the percentage weight of the dispersion. For example, the polymer is present in about 49.5% and the drug is present in about 50%.

0102The dispersion may further include other minor ingredients, such as a surfactant (e.g., SLS). The surfactant may be present in less than about 10% of the dispersion, for example less than about 9%, less than about 8%, less than about 7%, less than about 6%, less than about 5%, less than about 4%, less than about 3%, less than about 2%, about 1%, or about 0.5%.

0103In embodiments including a polymer, the polymer should be present in an amount effective for stabilizing the solid dispersion. Stabilizing includes inhibiting or preventing, the crystallization of Compound <b>1.</b> Such stabilizing would inhibit the conversion Compound <b>1</b> from amorphous to crystalline form. For example, the polymer would prevent at least a portion (e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, or greater) of Compound <b>1</b> from converting from an amorphous to a crystalline form. Stabilization can be measured, for example, by measuring the glass transition temperature of the solid dispersion, measuring the rate of relaxation of the amorphous material, or by measuring the solubility or bioavailability of Compound <b>1.</b>

0104Suitable polymers for use in combination with Compound <b>1,</b> for example to form a solid dispersion such as an amorphous solid dispersion, should have one or more of the following properties:

0105The glass transition temperature of the polymer should have a temperature of no less than about 10-15 °C lower than the glass transition temperature of Compound <b>1.</b> Preferably, the glass transition temperature of the polymer is greater than the glass transition temperature of Compound <b>1,</b> and in general at least 50°C higher than the desired storage temperature of the drug product. For example, at least about 100 °C, at least about 105 °C, at least about 105 °C, at least about 110 °C, at least about 120 °C, at least about 130 °C, at least about 140 °C, at least about 150 °C, at least about 160 °C, at least about 160 °C, or greater.

0106The polymer should be relatively non-hygroscopic. For example, the polymer should, when stored under standard conditions, absorb less than about 10% water, for example, less than about 9%, less than about 8%, less than about 7%, less than about 6%, or less than about 5%, less than about 4%, or less than about 3% water. Preferably the polymer will, when stored under standard conditions, be substantially free of absorbed water.

0107The polymer should have similar or better solubility in solvents suitable for spray drying processes relative to that of Compound <b>1.</b> The polymer may dissolve in one or more of the same solvents or solvent systems as Compound <b>1.</b> It is preferred that the polymer is soluble in at least one non-hydroxy containing solvent such as methylene chloride, acetone, or a combination thereof.

0108The polymer, when combined with Compound <b>1,</b> for example in a solid dispersion or in a liquid suspension, should increase the solubility of Compound <b>1</b> in aqueous and physiologically relative media either relative to the solubility of Compound <b>1</b> in the absence of polymer or relative to the solubility of Compound <b>1</b> when combined with a reference polymer. For example, the polymer could increase the solubility of amorphous Compound <b>1</b> by reducing the amount of amorphous Compound <b>1</b> that converts to crystalline Compound <b>1,</b> either from a solid amorphous dispersion or from a liquid suspension.

0109The polymer should decrease the relaxation rate of the amorphous substance.

0110The polymer should increase the physical and/or chemical stability of Compound <b>1.</b>

0111The polymer should improve the manufacturability of Compound <b>1.</b>

0112The polymer should improve one or more of the handling, administration or storage properties of Compound <b>1.</b>

0113The polymer should not interact unfavorably with other pharmaceutical components, for example excipients.

0114The suitability of a candidate polymer (or other component) can be tested using the spray drying methods (or other methods) described herein to form an amorphous composition. The candidate composition can be compared in terms of stability, resistance to the formation of crystals, or other properties, and compared to a reference preparation, e.g., a preparation of neat amorphous Compound <b>1</b> or crystalline Compound <b>1.</b> E.g., a candidate composition could be tested to determine whether it inhibits the time to onset of solvent mediated crystallization, or the percent conversion at a given time under controlled conditions, by at least 50 %, 75 %, 100%, or 110% as well as the reference preparation, or a candidate composition could be tested to determine if it has improved bioavailability or solubility relative to crystalline Compound <b>1.</b>

Surfactants

0115A solid dispersion or other composition may include a surfactant. A surfactant or surfactant mixture would generally decrease the interfacial tension between the solid dispersion and an aqueous medium. An appropriate surfactant or surfactant mixture may also enhance aqueous solubility and bioavailability of Compound <b>1</b> from a solid dispersion. The surfactants for use in connection with the present invention include, but are not limited to, sorbitan fatty acid esters (e.g., Spans^®), polyoxyethylene sorbitan fatty acid esters (e.g., Tweens^®), sodium lauryl sulfate (SLS), sodium dodecylbenzene sulfonate (SDBS) dioctyl sodium sulfosuccinate (Docusate), dioxycholic acid sodium salt (DOSS), Sorbitan Monostearate, Sorbitan Tristearate, hexadecyltrimethyl ammonium bromide (HTAB), Sodium N-lauroylsarcosine, Sodium Oleate, Sodium Myristate, Sodium Stearate, Sodium Palmitate, Gelucire 44/14, ethylenediamine tetraacetic acid (EDTA), Vitamin E d-alpha tocopheryl polyethylene glycol 1000 succinate (TPGS), Lecithin, MW 677-692, Glutanic acid monosodium monohydrate, Labrasol, PEG 8 caprylic/capric glycerides, Transcutol, diethylene glycol monoethyl ether, Solutol HS-15, polyethylene glycol/hydroxystearate, Taurocholic Acid, Pluronic F68, Pluronic F108, and Pluronic F127 (or any other polyoxyethylene-polyoxypropylene co-polymers (Pluronics^®) or saturated polyglycolized glycerides (Gelucirs^®)). Specific example of such surfactants that may be used in connection with this invention include, but are not limited to, Span 65, Span 25, Tween 20, Capryol 90, Pluronic F108, sodium lauryl sulfate (SLS), Vitamin E TPGS, pluronics and copolymers. SLS is generally preferred.

0116The amount of the surfactant (e.g., SLS) relative to the total weight of the solid dispersion may be between 0.1-15%. Preferably, it is from about 0.5% to about 10%, more preferably from about 05 to about 5%, e.g., about 1%, about 2%, about 3%, about 4%, or about 5%.

0117In certain embodiments, the amount of the surfactant relative to the total weight of the solid dispersion is at least about 0.1, preferably about 0.5%. In these embodiments, the surfactant would be present in an amount of no more than about 15%, and preferably no more than about 12%, about 11%, about 10%, about 9%, about 8%, about 7%, about 6%, about 5%, about 4%, about 3%, about 2% or about 1%. The surfactant may be in an amount of about 0.5% by weight.

0118Candidate surfactants (or other components) can be tested for suitability for use in the invention in a manner similar to that described for testing polymers.

<u>Methods of making solid forms of Compound <b>1</b></u>

0119The solid form of Compound <b>1</b> can vary depending on the method used to prepare Compound <b>1.</b> For example, Compound <b>1</b> can be prepared using a method to provide crystalline Compound <b>1,</b> such as Form A or Form B, or Compound <b>1</b> can be prepared using a method to provide amorphous Compound <b>1,</b> for example as a neat preparation or where Compound <b>1</b> is a component in a dispersion such as a solid amorphous dispersion (e.g., a dispersion of Compound <b>1</b> and a polymer such as a cellulosic polymer e.g., HPMC or HPMCAS or a pyrrolidone polymer such as PVP/VA).

Form A (not according to the invention)

0120Form A of Compound <b>1</b> can be prepared, for example, by heating Compound <b>1</b> to at or above its melting point, for example to about 250 °C and then cooling the compound, thereby providing Compound <b>1</b> having a solid state of Form A. Form A can be characterized by one or more characteristic peaks as determined using XRPD. For example, Compound <b>1</b> as Form A can be identified by the presence of one or peaks at 2θ, including one or more of the following peaks at or about: about 5.0 e.g. 4.99; about 7.8, e.g., 7.75; about 8.5, e.g., 8.46; about 9.2, e.g., 9.21; about 9.9, e.g., 9.92; about 11.9, e.g., 11.93; about 12.6, e.g., 12.64; about 13.9, e.g., 13.88; about 14.9, e.g., 14.91; about 15.6, e.g., 15.58; about 16.5, e.g., 16.46; about 18.1, e.g., 18.09; about 18.5, e.g., 18.52; about 21.7, e.g., 20.65; about 22.0, e.g., 21.95; about 23.5, e.g., 23.49; about 25.3, e.g., 25.26; about 28.0, e.g., 28.02; about 29.4, e.g., 29.35; and about 30.9, e.g., 30.85.

Form B

0121Form B of Compound <b>1</b> can be prepared, for example, by subjecting a slurry of compound 1 in a solvent to heating and cooling cycles.

0122In some preferred embodiments, the solvent is a solvent where Compound <b>1</b> has limited solubility at room temperature, for example, acetone.

0123The slurry is subject to a plurality of heat/cool cycles, where the slurry is generally warmed to a temperature above room temperature but below the boiling point of the solvent, for example about 40 °C to about 60 °C , e.g., about 50 °C. The slurry is generally subjected to at least 2 heat/cool cycles, for example, 2, 3, 4, 5, or 6, preferably 5 cycles. Each cycle was timed to last at least about 8 hours (e.g., 4 hours of heating followed by 4 hours at room temperature, 6 hours of heating followed by 6 hours at room temperature, 8 hours of heating followed by 8 hours at room temperature, preferably 12 hours of heating followed by 12 hours at room temperature).

0124In an alternative embodiment crude Compound <b>1</b> can be refluxed in as a slurry in acetonitrile (e.g., 27 volumes of acetonitrile) for 24 hours. The mixture is then cooled, e.g., to about room temperature, e.g., about 20 °C. Form B is then isolated, for example, by filtration as a white to off-white. The resulting wet cake is rinsed with acetonitrile (e.g., 5 volumes) and dried under vacuum at 50 °C until a constant weight is attained.

0125Form B can be characterized by one or more characteristic peaks as determined using XRPD. For example, Compound <b>1</b> as Form B can be identified by the presence of one or peaks at 2θ, including one or more of the following peaks at or about: about 6.2, e.g., 6.17; about 7.6, e.g., 7.61; about 8.4, e.g., 8.40; about 11.0, e.g., 11.02; about 12.3, e.g., 12.33; about 14.8, e.g., 14.83; about 16.1, e.g., 16.14; about 17.1, e.g., 17.11; about 18.0, e.g., 17.96; about 18.6, e.g., 18.55; about 19.4, e.g., 19.43; about 21.1, e.g., 21.05; about 22.6, e.g., 22.56; about 23.4, e.g., 23.37; about 23.9, e.g., 23.94; about 24.9, e.g., 24.86; about 25.5, e.g., 25.50; about 26.7, e.g., 26.72; about 27.5, e.g., 27.51; about 29.6, e.g., 29.60; about 33.5, e.g., 33.48; and about 36.8, e.g., 36.78.

Amorphous Compound <b>1</b> (not according to the invention)

0126Amorphous compound <b>1</b> can be made using a variety of techniques, including, for example spray drying a solution of Compound <b>1</b> to provide amorphous Compound <b>1,</b> e.g., as a neat solid or as a component of a solid dispersion, said method utilizing spray-drying means to effect said conversion. For example, Amorphous Compound <b>1</b> can be made by converting a form of Compound <b>1,</b> e.g., a crystalline form of Compound 1, such as Form A or Form B, into a substantially amorphous form of Compound <b>1</b> by dissolving Compound into a solution and spray drying the solution of Compound <b>1,</b> thereby converting a form of Compound 1, such as crystalline Compound <b>1,</b> into amorphous Compound <b>1.</b> An exemplary process for making amorphous Compound <b>1</b> by converting Form B into a substantially amorphous form of Compound <b>1</b> is recited in the examples.

0127Any method for obtaining amorphous forms of Compound <b>1,</b> including neat amorphous Compound <b>1</b> and solid amorphous dispersions of Compound <b>1,</b> can be used including, for example, those described in <patcit id="pcit0002" dnum="US20030186952A"><text>US 2003/0186952</text></patcit> (see the documents cited therein at paragraph 1092) and <patcit id="pcit0003" dnum="US20030185891A"><text>US 2003/0185891</text></patcit>). In general, methods that could be used include those that involve rapid removal of solvent from a mixture or cooling a molten sample. Such methods include, but are not limited to, rotational evaporation, freeze-drying (i.e., lyophilization), vacuum drying, melt congealing, and melt extrusion. An embodiment includes amorphous Compound <b>1,</b> such as a neat preparation or a solid dispersion obtained by spray-drying. Accordingly, in some embodiments, the amorphous product obtained by spray-drying is further dried, for example, to remove residual solvent.

0128Preparations disclosed herein, e.g., a pharmaceutical composition, can be obtained by spray-drying a mixture comprising Compound <b>1,</b> a suitable polymer, and an appropriate solvent. Spray drying is a method that involves atomization of a liquid mixture containing, e.g., a solid and a solvent, and removal of the solvent. Atomization can be done, for example, through a nozzle or on a rotating disk.

0129Spray drying is a process that converts a liquid feed to a dried particulate form. Optionally, a secondary drying process such as fluidized bed drying or vacuum drying, may be used to reduce residual solvents to pharmaceutically acceptable levels. Typically, spray-drying involves contacting a highly dispersed liquid suspension or solution, and a sufficient volume of hot air to produce evaporation and drying of the liquid droplets. The preparation to be spray dried can be any solution, coarse suspension, slurry, colloidal dispersion, or paste that may be atomized using the selected spray-drying apparatus. In a standard procedure, the preparation is sprayed into a current of warm filtered air that evaporates the solvent and conveys the dried product to a collector (e.g., a cyclone). The spent air is then exhausted with the solvent, or alternatively the spent air is sent to a condenser to capture and potentially recycle the solvent. Commercially available types of apparatus may be used to conduct the spray-drying. For example, commercial spray dryers are manufactured by Buchi Ltd. and Niro (e.g., the PSD line of spray driers manufactured by Niro) (see, <patcit id="pcit0004" dnum="US20040105820A"><text>US 2004/0105820</text></patcit>; <patcit id="pcit0005" dnum="US20030144257A"><text>US 2003/0144257</text></patcit>).

0130Spray-drying typically employs solids loads of material from about 3% to about 30% by weight, (i.e., drug plus and excipients), for example about 4% to about 20% by weight, preferably at least about 10%. In general, the upper limit of solids loads is governed by the viscosity of (e.g., the ability to pump) the resulting solution and the solubility of the components in the solution. Generally, the viscosity of the solution can determine the size of the particle in the resulting powder product.

0131Techniques and methods for spray-drying may be found in <nplcit id="ncit0016" npl-type="b"><text>Perry's Chemical Engineering Handbook, 6th Ed., R.H. Perry, D.W. Green &amp; J.O. Maloney, eds.), McGraw-Hill book co. (1984</text></nplcit>); and <nplcit id="ncit0017" npl-type="s"><text>Marshall "Atomization and Spray-Drying" 50, Chem. Eng. Prog. Monogr. Series 2 (1954</text></nplcit>). In general, the spray-drying is conducted with an inlet temperature of from about 60 °C to about 200 °C, for example, from about 95 °C to about 185 °C, from about 110 °C to about 182 °C, from about 96 °C to about 108 °C, e.g., about 175 °C. The spray-drying is generally conducted with an outlet temperature of from about 30 °C to about 80 °C, for example from about 31 °C to about 72 °C, about 37 °C to about 41 °C e.g., about 60 °C. The atomization flow rate is generally from about 4 kg/h to about 12 kg/h, for example, from about 4.3 kg/h to about 10.5 kg/h, e.g., about 6 kg/h or about 10.5 kg/h. The feed flow rate is generally from about 3 kg/h to about 10 kg/h, for example, from about 3.5 kg/h to about 9.0 kg/h, e.g., about 8 kg/h or about 7.1 kg/h. The atomization ratio is generally from about 0.3 to 1.7, e.g., from about 0.5 to 1.5, e.g., about 0.8 or about 1.5.

0132Removal of the solvent may require a subsequent drying step, such as tray drying, fluid bed drying (e.g., from about room temperature to about 100 °C), vacuum drying, microwave drying, rotary drum drying or biconical vacuum drying (e.g., from about room temperature to about 200 °C).

0133In one embodiment, the solid dispersion is fluid-bed dried.

0134In preferred processes, the solvent includes a volatile solvent, for example a solvent having a boiling point of less than about 100 °C. In some embodiments, the solvent includes a mixture of solvents, for example a mixture of volatile solvents or a mixture of volatile and non-volatile solvents. Where mixtures of solvents are used, the mixture can include one or more non-volatile solvents, for example, where the non-volatile solvent is present in the mixture at less than about 15%, e.g., less than about 12%, less than about 10%, less than about 8%, less than about 5%, less than about 3%, or less than about 2%.

0135Preferred solvents are those solvents where Compound <b>1</b> has a solubility of at least about 10 mg/ml (e.g., at least about 15 mg/ml, 20 mg/ml, 25 mg/ml, 30 mg/ml, 35 mg/ml, 40 mg/ml, 45 mg/ml, 50 mg/ml, or greater). More preferred solvents include those where Compound 1 has a solubility of at least about 50 mg/ml.

0136Exemplary solvents that could be tested include acetone, cyclohexane, dichloromethane, N,N-Dimethylacetamide (DMA), N,N-Dimethylformamide (DMF), 1,3 Dimethyl-2-imidazolidinone (DMI), dimethyl sulfoxide (DMSO), dioxane, ethyl acetate, ethyl ether, glacial acetic acid (HAc), methyl ethyl ketone (MEK), N-methyl-2-pyrrolidinone (NMP), methyl tert-butyl ether, tetrahydrofuran (THF) and pentane. Exemplary co-solvents include acetone/DMSO, acetone/DMF, acetone/water, MEK/water, THF/water, dioxane/water. In a two solvent system, the solvents can be present in of from about 0.1% to about 99.9%. In some preferred embodiments, water is a co-solvent with acetone where water is present from about 0.1% to about 15%, for example about 9% to about 11%, e.g., about 10%. In some preferred embodiments, water is a co-solvent with MEK where water is present from about 0.1% to about 15%, for example about 9% to about 11%, e.g., about 10%. In some embodiments the solvent solution include three solvents. For example, acetone and water can be mixed with a third solvent such as DMA, DMF, DMI, DMSO, or HAc. In instances where amorphous Compound <b>1</b> is a component of a solid amorphous dispersion, preferred solvents dissolve both Compound <b>1</b> and the polymer. Suitable solvents include those described above, for example, MEK, acetone, water, and mixtures thereof.

0137The particle size and the temperature drying range may be modified to prepare an optimal solid dispersion. As would be appreciated by skilled practitioners, a small particle size would lead to improved solvent removal. Applicants have found however, that smaller particles can lead to fluffy particles that, under some circumstances do not provide optimal solid dispersions for downstream processing such as tabletting. At higher temperatures, crystallization or chemical degradation of Compound <b>1</b> may occur. At lower temperatures, a sufficient amount of the solvent may not be removed. The methods herein provide an optimal particle size and an optimal drying temperature.

0138In general, particle size is such that D10 (µm) is less than about 5, e.g., less than about 4.5, less than about 4.0, or less than about 3.5, D50 (µm) is generally less than about 17, e.g., less than about 16, less than about 15, less than about 14, less than about 13, and D90 (µm) is generally less than about 175, e.g., less than about 170, less than about 170, less than about 150, less than about 125, less than about 100, less than about 90, less than about 80, less than about 70, less than about 60, or less than about less than about 50. In general bulk density of the spray dried particles is from about 0.08 g/cc to about 0.20 g/cc, e.g., from about 0.10 to about 0.15 g/cc, e.g., about 0.11 g/cc or about 0.14 g/cc. Tap density of the spray dried particles generally ranges from about 0.08 g/cc to about 0.20 g/cc, e.g., from about 0.10 to about 0.15 g/cc, e.g., about 0.11 g/cc or about 0.14 g/cc, for 10 taps; 0.10 g/cc to about 0.25 g/cc, e.g., from about 0.11 to about 0.21 g/cc, e.g., about 0.15 g/cc, about 0.19 g/cc, or about 0.21 g/cc for 500 taps; 0.15 g/cc to about 0.27 g/cc, e.g., from about 0.18 to about 0.24 g/cc, e.g., about 0.18 g/cc, about 0.19 g/cc, about 0.20 g/cc, or about 0.24 g/cc for 1250 taps; and 0.15 g/cc to about 0.27 g/cc, e.g., from about 0.18 to about 0.24 g/cc, e.g., about 0.18 g/cc, about 0.21 g/cc, about 0.23 g/cc, or about 0.24 g/cc for 2500 taps.

0139Unless otherwise stated, structures depicted herein are also meant to include all isomeric (e.g., enantiomeric, diastereomeric, and geometric (or conformational)) forms of the structure; for example, the R and S configurations for each asymmetric center, (Z) and (E) double bond isomers, and (Z) and (E) conformational isomers. Therefore, single stereochemical isomers as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of the present compounds are within the scope of the invention. All tautomeric forms of the Compound <b>1</b> are included herein. E.g., Compound <b>1</b> may exist as tautomers, both of which are included herein: <chemistry id="chem0002" num="0002"><img file="EP3708564B1_D0002.tif" /></chemistry>

0140Additionally, unless otherwise stated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds of formula (I), wherein one or more hydrogen atoms are replaced deuterium or tritium, or one or more carbon atoms are replaced by a 13C- or 14C-enriched carbon are within the scope of this invention. Such compounds are useful, for example, as analytical tools, probes in biological assays, or compounds with improved therapeutic profile.

Uses, Formulation and Administration

Pharmaceutically acceptable compositions

0141In another aspect of the present invention, pharmaceutically acceptable compositions comprising the crystal form of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide according to Form B, for example, as characterized above, are provided, wherein these compositions optionally further comprise one or more additional therapeutic agents.

0142As used herein, the term "pharmaceutically acceptable salt" refers to those salts which are, within the scope of sound medical judgement, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit/risk ratio. A "pharmaceutically acceptable salt" means any non-toxic salt or salt of an ester of a compound of this invention that, upon administration to a recipient, is capable of providing, either directly or indirectly, a compound of this invention or an inhibitorily active metabolite or residue thereof.

0143Pharmaceutically acceptable salts are well known in the art. For example, S. M. <nplcit id="ncit0018" npl-type="s"><text>Berge, et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19</text></nplcit>. Pharmaceutically acceptable salts of the compounds of this invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable, nontoxic acid addition salts are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and the like. Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N⁺(C_1-4alkyl)₄ salts. This invention also envisions the quaternization of any basic nitrogen-containing groups of the compounds disclosed herein. Water or oil-soluble or dispersable products may be obtained by such quaternization. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, loweralkyl sulfonate and aryl sulfonate.

0144As described above, the pharmaceutically acceptable compositions of the present invention additionally comprise a pharmaceutically acceptable carrier, adjuvant, or vehicle, which, as used herein, includes any and all solvents, diluents, or other liquid vehicle, dispersion or suspension aids, surface active agents, isotonic agents, thickening or emulsifying agents, preservatives, solid binders, lubricants and the like, as suited to the particular dosage form desired.<nplcit id="ncit0019" npl-type="b"><text> Remington's Pharmaceutical Sciences, Sixteenth Edition, E. W. Martin (Mack Publishing Co., Easton, Pa., 1980</text></nplcit>) discloses various carriers used in formulating pharmaceutically acceptable compositions and known techniques for the preparation thereof. Except insofar as any conventional carrier medium is incompatible with the compounds of the invention, such as by producing any undesirable biological effect or otherwise interacting in a deleterious manner with any other component(s) of the pharmaceutically acceptable composition, its use is contemplated to be within the scope of this invention. Some examples of materials which can serve as pharmaceutically acceptable carriers include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, or potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, wool fat, sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil; safflower oil; sesame oil; olive oil; corn oil and soybean oil; glycols; such a propylene glycol or polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol, and phosphate buffer solutions, as well as other non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, releasing agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the composition, according to the judgment of the formulator.

Uses of Compounds and Pharmaceutically Acceptable Compositions

0145A pharmaceutical composition of the invention, or a crystal form of the invention (Form B) can be used in a method of treating a condition, disease, or disorder implicated by CFTR. In certain embodiments, the method of treating a condition, disease, or disorder implicated by a deficiency of CFTR activity comprises administering a composition comprising a pharmaceutical composition of the invention or a crystal form of the invention (Form B) to a subject, preferably a mammal, in need thereof.

0146A "CFTR-mediated disease" as used herein is a disease selected from cystic fibrosis, Hereditary emphysema, Hereditary hemochromatosis, Coagulation-Fibrinolysis deficiencies, such as Protein C deficiency, Type 1 hereditary angioedema, Lipid processing deficiencies, such as Familial hypercholesterolemia, Type 1 chylomicronemia, Abetalipoproteinemia, Lysosomal storage diseases, such as I-cell disease/Pseudo-Hurler, Mucopolysaccharidoses, Sandhof/Tay-Sachs, Crigler-Najjar type II, Polyendocrinopathy/Hyperinsulemia, Diabetes mellitus, Laron dwarfism, Myleoperoxidase deficiency, Primary hypoparathyroidism, Melanoma, Glycanosis CDG type 1, Hereditary emphysema, Congenital hyperthyroidism, Osteogenesis imperfecta, Hereditary hypofibrinogenemia, ACT deficiency, Diabetes insipidus (DI), Neurophyseal DI, Neprogenic DI, Charcot-Marie Tooth syndrome, Perlizaeus-Merzbacher disease, neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, Amyotrophic lateral sclerosis, Progressive supranuclear plasy, Pick's disease, several polyglutamine neurological disorders asuch as Huntington, Spinocerebullar ataxia type I, Spinal and bulbar muscular atrophy, Dentatorubal pallidoluysian, and Myotonic dystrophy, as well as Spongiform encephalopathies, such as Hereditary Creutzfeldt-Jakob disease, Fabry disease, Straussler-Scheinker syndrome, COPD, dry-eye disease, and Sjogren's disease.

0147A pharmaceutical composition of the invention, or a crystal form of the invention (Form B) can be used in a method of treating cystic fibrosis, hereditary emphysema, hereditary hemochromatosis, coagulation-fibrinolysis deficiencies, such as protein C deficiency, Type 1 hereditary angioedema, lipid processing deficiencies, such as familial hypercholesterolemia, Type 1 chylomicronemia, abetalipoproteinemia, lysosomal storage diseases, such as I-cell disease/pseudo-Hurler, mucopolysaccharidoses, Sandhof/Tay-Sachs, Crigler-Najjar type II, polyendocrinopathy/hyperinsulemia, Diabetes mellitus, Laron dwarfism, myleoperoxidase deficiency, primary hypoparathyroidism, melanoma, glycanosis CDGtype 1, congenital hyperthyroidism, osteogenesis imperfecta, hereditary hypofibrinogenemia, ACT deficiency, Diabetes insipidus (DI), neurophyseal DI, neprogenic DI, Charcot-Marie Tooth syndrome, Perlizaeus-Merzbacher disease, neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, progressive supranuclear plasy, Pick's disease, several polyglutamine neurological disorders asuch as Huntington, spinocerebullar ataxia type I, spinal and bulbar muscular atrophy, dentatorubal pallidoluysian, and myotonic dystrophy, as well as spongiform encephalopathies, such as hereditary Creutzfeldt-Jakob disease (due to prion protein processing defect), Fabry disease, Straussler-Scheinker syndrome, COPD, dry-eye disease, or Sjogren's disease in a mammal, wherein the method comprises the step of administering to said mammal an effective amount of a composition comprising a pharmaceutical composition of the invention, or a crystal form of the invention (Form B).

0148A pharmaceutical composition of the invention, or a crystal form of the invention (Form B) can be used in a method of treating cystic fibrosis in a mammal comprising the step of administering to said mammal a composition comprising a pharmaceutical composition of the invention, or a crystal form of the invention (Form B).

0149According to the invention an "effective amount" of a crystal form of Compound <b>1</b> (Form B) or a pharmaceutically acceptable composition thereof is that amount effective for treating or lessening the severity of any of the diseases recited above.

0150A crystal form of Compound <b>1</b> (Form B) or a pharmaceutically acceptable composition thereof may be administered using any amount and any route of administration effective for treating or lessening the severity of one or more of the diseases recited above.

0151In certain embodiments, a crystal form of Compound <b>1</b> described herein (Form B) or a pharmaceutically acceptable composition thereof is useful for treating or lessening the severity of cystic fibrosis in patients who exhibit residual CFTR activity in the apical membrane of respiratory and non-respiratory epithelia. The presence of residual CFTR activity at the epithelial surface can be readily detected using methods known in the art, e.g., standard electrophysiological, biochemical, or histochemical techniques. Such methods identify CFTR activity using <i>in vivo</i> or <i>ex vivo</i> electrophysiological techniques, measurement of sweat or salivary Cl^- concentrations, or <i>ex vivo</i> biochemical or histochemical techniques to monitor cell surface density. Using such methods, residual CFTR activity can be readily detected in patients heterozygous or homozygous for a variety of different mutations, including patients homozygous or heterozygous for the most common mutation, ΔF508.

0152In one embodiment, a crystal form of Compound <b>1</b> described herein (Form B) or a pharmaceutically acceptable composition thereof is useful for treating or lessening the severity of cystic fibrosis in patients within certain genotypes exhibiting residual CFTR activity, e.g., class III mutations (impaired regulation or gating), class IV mutations (altered conductance), or class V mutations (reduced synthesis) (<nplcit id="ncit0020" npl-type="s"><text>Lee R. Choo-Kang, Pamela L., Zeitlin, Type I, II, III, IV, and V cystic fibrosis Tansmembrane Conductance Regulator Defects and Opportunities of Therapy; Current Opinion in Pulmonary Medicine 6:521 - 529, 2000</text></nplcit>). Other patient genotypes that exhibit residual CFTR activity include patients homozygous for one of these classes or heterozygous with any other class of mutations, including class I mutations, class II mutations, or a mutation that lacks classification.

0153In one embodiment, a crystal form of Compound <b>1</b> described herein (Form B) or a pharmaceutically acceptable composition thereof is useful for treating or lessening the severity of cystic fibrosis in patients within certain clinical phenotypes, e.g., a moderate to mild clinical phenotype that typically correlates with the amount of residual CFTR activity in the apical membrane of epithelia. Such phenotypes include patients exhibiting pancreatic insufficiency or patients diagnosed with idiopathic pancreatitis and congenital bilateral absence of the vas deferens, or mild lung disease.

0154The exact amount required will vary from subject to subject, depending on the species, age, and general condition of the subject, the severity of the infection, the particular agent, its mode of administration, and the like. The compounds of the invention are preferably formulated in dosage unit form for ease of administration and uniformity of dosage. The expression "dosage unit form" as used herein refers to a physically discrete unit of agent appropriate for the patient to be treated. It will be understood, however, that the total daily usage of the compounds and compositions of the present invention will be decided by the attending physician within the scope of sound medical judgment. The specific effective dose level for any particular patient or organism will depend upon a variety of factors including the disorder being treated and the severity of the disorder; the activity of the specific compound employed; the specific composition employed; the age, body weight, general health, sex and diet of the patient; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidental with the specific compound employed, and like factors well known in the medical arts. The term "patient", as used herein, means an animal, preferably a mammal, and most preferably a human.

0155The pharmaceutically acceptable compositions of this invention can be administered to humans and other animals orally, rectally, parenterally, intracisternally, intravaginally, intraperitoneally, topically (as by powders, ointments, or drops), bucally, as an oral or nasal spray, or the like, depending on the severity of the infection being treated. In certain embodiments, the compounds of the invention may be administered orally or parenterally at dosage levels of about 0.01 mg/kg to about 50 mg/kg and preferably from about 1 mg/kg to about 25 mg/kg, of subject body weight per day, one or more times a day, to obtain the desired therapeutic effect.

0156Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof. Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

0157Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution, suspension or emulsion in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, U.S.P. and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil can be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectables.

0158The injectable formulations can be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.

0159In order to prolong the effect of a compound of the present invention, it is often desirable to slow the absorption of the compound from subcutaneous or intramuscular injection. This may be accomplished by the use of a liquid suspension of crystalline or amorphous material with poor water solubility. The rate of absorption of the compound then depends upon its rate of dissolution that, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered compound form is accomplished by dissolving or suspending the compound in an oil vehicle. Injectable depot forms are made by forming microencapsule matrices of the compound in biodegradable polymers such as polylactide-polyglycolide. Depending upon the ratio of compound to polymer and the nature of the particular polymer employed, the rate of compound release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the compound in liposomes or microemulsions that are compatible with body tissues.

0160Compositions for rectal or vaginal administration are preferably suppositories which can be prepared by mixing the compounds of this invention with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound.

0161Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and/or a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, c) humectants such as glycerol, d) disintegrating agents such as agar--agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) solution retarding agents such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as, for example, cetyl alcohol and glycerol monostearate, h) absorbents such as kaolin and bentonite clay, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also comprise buffering agents.

0162Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulating art. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polethylene glycols and the like.

0163The active compounds can also be in microencapsulated form with one or more excipients as noted above. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings, release controlling coatings and other coatings well known in the pharmaceutical formulating art. In such solid dosage forms the active compound may be admixed with at least one inert diluent such as sucrose, lactose or starch. Such dosage forms may also comprise, as is normal practice, additional substances other than inert diluents, e.g., tableting lubricants and other tableting aids such a magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes.

0164Dosage forms for topical or transdermal administration of a compound of this invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches. The active component is admixed under sterile conditions with a pharmaceutically acceptable carrier and any needed preservatives or buffers as may be required. Ophthalmic formulation, eardrops, and eye drops are also contemplated as being within the scope of this invention. Additionally, the present invention contemplates the use of transdermal patches, which have the added advantage of providing controlled delivery of a compound to the body. Such dosage forms are prepared by dissolving or dispensing the compound in the proper medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate can be controlled by either providing a rate controlling membrane or by dispersing the compound in a polymer matrix or gel.

0165It will also be appreciated that the crystal form of Compound 1 described herein (Form B) or a pharmaceutically acceptable composition thereof can be employed in combination therapies, that is, Form B or a pharmaceutically acceptable composition thereof can be administered concurrently with, prior to, or subsequent to, one or more other desired therapeutics or medical procedures. The particular combination of therapies (therapeutics or procedures) to employ in a combination regimen will take into account compatibility of the desired therapeutics and/or procedures and the desired therapeutic effect to be achieved. It will also be appreciated that the therapies employed may achieve a desired effect for the same disorder (for example, an inventive compound may be administered concurrently with another agent used to treat the same disorder), or they may achieve different effects (e.g., control of any adverse effects). As used herein, additional therapeutic agents that are normally administered to treat or prevent a particular disease, or condition, are known as "appropriate for the disease, or condition, being treated".

0166In one embodiment, the additional agent is selected from a mucolytic agent, bronchodialator, an anti-biotic, an anti-infective agent, an anti-inflammatory agent, a CFTR modulator other than a compound of the present invention, or a nutritional agent.

0167The amount of additional therapeutic agent present in the compositions of this invention will be no more than the amount that would normally be administered in a composition comprising that therapeutic agent as the only active agent. Preferably the amount of additional therapeutic agent in the presently disclosed compositions will range from about 50% to 100% of the amount normally present in a composition comprising that agent as the only therapeutically active agent.

0168A crystal form of Compound <b>1</b> described herein (Form B) or a pharmaceutically acceptable composition thereof may also be incorporated into compositions for coating an implantable medical device, such as prostheses, artificial valves, vascular grafts, stents and catheters. Accordingly, the present invention, in another aspect, includes a composition for coating an implantable device comprising a crystal form of Compound <b>1</b> described herein (Form B) or a pharmaceutically acceptable composition thereof, and in classes and subclasses herein, and a carrier suitable for coating said implantable device. In still another aspect, the present invention includes an implantable device coated with a composition comprising a crystal form of Compound <b>1</b> described herein (Form B) or a pharmaceutically acceptable composition thereof, and a carrier suitable for coating said implantable device. Suitable coatings and the general preparation of coated implantable devices are described in <patcit id="pcit0006" dnum="US6099562A"><text>US Patents 6,099,562</text></patcit>; <patcit id="pcit0007" dnum="US5886026A"><text>5,886,026</text></patcit>; and <patcit id="pcit0008" dnum="US5304121A"><text>5,304,121</text></patcit>. The coatings are typically biocompatible polymeric materials such as a hydrogel polymer, polymethyldisiloxane, polycaprolactone, polyethylene glycol, polylactic acid, ethylene vinyl acetate, and mixtures thereof. The coatings may optionally be further covered by a suitable topcoat of fluorosilicone, polysaccarides, polyethylene glycol, phospholipids or combinations thereof to impart controlled release characteristics in the composition.

0169In order that the invention described herein may be more fully understood, the following examples are set forth. It should be understood that these examples are for illustrative purposes only and are not to be construed as limiting this invention (which is defined in the appended claims) in any manner.

EXAMPLES

<u>Methods &amp; Materials</u>

Differential Scanning Calorimetry (DSC)

0170The Differential scanning calorimetry (DSC) data of Form A, Form B, and amorphous Compound <b>1</b> were collected using a DSC Q100 V9.6 Build 290 (TA Instruments, New Castle, DE). Temperature was calibrated with indium and heat capacity was calibrated with sapphire. Samples of 3-6 mg were weighed into aluminum pans that were crimped using lids with 1 pin hole. The samples were scanned from 25°C to 350°C at a heating rate of 10°C/min and with a nitrogen gas purge of 50 ml/min. Data were collected by Thermal Advantage Q SeriesTM version 2.2.0.248 software and analyzed by Universal Analysis software version 4.1D (TA Instruments, New Castle, DE). The reported numbers represent single analyses.

Thermogravimetric analysis (TGA)

0171Thermal gravimetric analysis (TGA) was performed with a TGA Q500 V6.3 Build 189 (TA Instruments, New Castle, DE) was used for TGA measurement. Temperature was equilibrated by Curie point with nickel. Samples of 10-20 mg were scanned from 25°C to 350°C at a heating rate of 10°C/min. A nitrogen gas balance purge of 10 ml/min and a sample purge of 90 ml/min were used. Data were collected by Thermal Advantage Q SeriesTM software version 2.2.0.248 and analyzed by Universal Analysis software version 4.1D (TA Instruments, New Castle, DE). The reported numbers represent single analyses.

XRPD (X-ray Powder Diffraction)

0172The X-Ray diffraction (XRD) data of Form A, Form B, and amorphous Compound 1 were collected on a Bruker D8 DISCOVER with GADDS powder diffractometer with HI-STAR 2-dimensional detector and a flat graphite monochromator. Cu sealed tube with K□ radiation was used at 40 kV, 35mA. The samples were placed on zero-background silicon wafers at 25°C. For each sample, two data frames were collected at 120 seconds each at 2 different 2□ angles: 8° and 26°. The frames data were integrated with GADDS software and merged with DIFFRACT^plusEVA software.

Synthesis of N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide (Compound 1):

0173<chemistry id="chem0003" num="0003"><img file="EP3708564B1_D0003.tif" /></chemistry>

2-Phenylaminomethylene-malonic acid diethyl ester

0174A mixture of aniline (25.6 g, 0.275 mol) and diethyl 2-(ethoxymethylene)malonate (62.4 g, 0.288 mol) was heated at 140-150 °C for 2 h. The mixture was cooled to room temperature and dried under reduced pressure to afford 2-phenylaminomethylene-malonic acid diethyl ester as a solid, which was used in the next step without further purification. ¹H NMR (DMSO-<i>d₆</i>) δ 11.00 (d, 1H), 8.54 (d, <i>J</i> = 13.6 Hz, 1H), 7.36-7.39 (m, 2H), 7.13-7.17 (m, 3H), 4.17-4.33 (m, 4H), 1.18-1.40 (m, 6H).

4-Hydroxyquinoline-3-carboxylic acid ethyl ester

0175A 1 L three-necked flask fitted with a mechanical stirrer was charged with 2-phenylaminomethylene-malonic acid diethyl ester (26.3 g, 0.100 mol), polyphosphoric acid (270 g) and phosphoryl chloride (750 g). The mixture was heated to 70 °C and stirred for 4 h. The mixture was cooled to room temperature and filtered. The residue was treated with aqueous Na₂CO₃ solution, filtered, washed with water and dried. 4-Hydroxyquinoline-3-carboxylic acid ethyl ester was obtained as a pale brown solid (15.2 g, 70%). The crude product was used in next step without further purification.

4-Oxo-1,4-dihydroquinoline-3-carboxylic acid

01764-Hydroxyquinoline-3-carboxylic acid ethyl ester (15 g, 69 mmol) was suspended in sodium hydroxide solution (2N, 150 mL) and stirred for 2 h at reflux. After cooling, the mixture was filtered, and the filtrate was acidified to pH 4 with 2N HCl. The resulting precipitate was collected via filtration, washed with water and dried under vacuum to give 4-oxo-1,4-dihydroquinoline-3-carboxylic acid as a pale white solid (10.5 g, 92 %). ¹H NMR (DMSO-<i>d₆</i>) δ 15.34 (s, 1 H), 13.42 (s, 1 H), 8.89 (s, 1H), 8.28 (d, <i>J</i> = 8.0 Hz, 1H), 7.88 (m, 1H), 7.81 (d, J = 8.4 Hz, 1H), 7.60 (m, 1H). <chemistry id="chem0004" num="0004"><img file="EP3708564B1_D0004.tif" /></chemistry>

Carbonic acid 2,4-di-<i>tert</i>-butyl-phenyl ester methyl ester

0177Methyl chloroformate (58 mL, 750 mmol) was added dropwise to a solution of 2,4-di-tert-butyl-phenol (103.2 g, 500 mmol), Et₃N (139 mL, 1000 mmol) and DMAP (3.05 g, 25 mmol) in dichloromethane (400 mL) cooled in an ice-water bath to 0 °C. The mixture was allowed to warm to room temperature while stirring overnight, then filtered through silica gel (approx. 1L) using 10% ethyl acetate - hexanes (~ 4 L) as the eluent. The combined filtrates were concentrated to yield carbonic acid 2,4-di-<i>tert</i>-butyl-phenyl ester methyl ester as a yellow oil (132 g, quant.). ¹H NMR (400 MHz, DMSO-<i>d</i>₆) δ 7.35 (d, J = 2.4 Hz, 1H), 7.29 (dd, J = 8.5, 2.4 Hz, 1H), 7.06 (d, J = 8.4 Hz, 1H), 3.85 (s, 3H), 1.30 (s, 9H), 1.29 (s, 9H).

Carbonic acid 2,4-di-<i>tert</i>-butyl-5-nitro-phenyl ester methyl ester and Carbonic acid 2,4-di-<i>tert</i>-butyl-6-nitro-phenyl ester methyl ester

0178To a stirring mixture of carbonic acid 2,4-di-<i>tert</i>-butyl-phenyl ester methyl ester (4.76 g, 180 mmol) in conc. sulfuric acid (2 mL), cooled in an ice-water bath, was added a cooled mixture of sulfuric acid (2 mL) and nitric acid (2 mL). The addition was done slowly so that the reaction temperature did not exceed 50 °C. The reaction was allowed to stir for 2 h while warming to room temperature. The reaction mixture was then added to ice-water and extracted into diethyl ether. The ether layer was dried (MgSO₄), concentrated and purified by column chromatography (0 - 10% ethyl acetate - hexanes) to yield a mixture of carbonic acid 2,4-di-<i>tert</i>-butyl-5-nitro-phenyl ester methyl ester and carbonic acid 2,4-di-<i>tert</i>-butyl-6-nitro-phenyl ester methyl ester as a pale yellow solid (4.28 g), which was used directly in the next step.

2,4-Di-<i>tert</i>-butyl-5-nitro-phenol and 2,4-Di-<i>tert</i>-butyl-6-nitro-phenol

0179The mixture of carbonic acid 2,4-di-<i>tert</i>-butyl-5-nitro-phenyl ester methyl ester and carbonic acid 2,4-di-<i>tert</i>-butyl-6-nitro-phenyl ester methyl ester (4.2 g, 14.0 mmol) was dissolved in MeOH (65 mL) before KOH (2.0 g, 36 mmol) was added. The mixture was stirred at room temperature for 2 h. The reaction mixture was then made acidic (pH 2-3) by adding conc. HCl and partitioned between water and diethyl ether. The ether layer was dried (MgSO₄), concentrated and purified by column chromatography (0 - 5 % ethyl acetate - hexanes) to provide 2,4-di-<i>tert</i>-butyl-5-nitro-phenol (1.31 g, 29% over 2 steps) and 2,4-di-<i>tert</i>-butyl-6-nitrophenol. 2,4-Di-<i>tert</i>-butyl-5-nitro-phenol: ¹H NMR (400 MHz, DMSO-<i>d</i>₆) δ 10.14 (s, 1H, OH), 7.34 (s, 1H), 6.83 (s, 1H), 1.36 (s, 9H), 1.30 (s, 9H). 2,4-Di-<i>tert</i>-butyl-6-nitro-phenol: ¹H NMR (400 MHz, CDCl₃) δ 11.48 (s, 1H), 7.98 (d, J = 2.5 Hz, 1H), 7.66 (d, J = 2.4 Hz, 1H), 1.47 (s, 9H), 1.34 (s, 9H).

5-Amino-2,4-di-<i>tert</i>-butyl-phenol

0180To a reluxing solution of 2,4-di-<i>tert</i>-butyl-5-nitro-phenol (1.86 g, 7.40 mmol) and ammonium formate (1.86 g) in ethanol (75 mL) was added Pd-5% wt. on activated carbon (900 mg). The reaction mixture was stirred at reflux for 2 h, cooled to room temperature and filtered through Celite. The Celite was washed with methanol and the combined filtrates were concentrated to yield 5-amino-2,4-di-<i>tert</i>-butyl-phenol as a grey solid (1.66 g, quant.). ¹H NMR (400 MHz, DMSO-<i>d</i>₆) δ 8.64 (s, 1H, OH), 6.84 (s, 1H), 6.08 (s, 1H), 4.39 (s, 2H, NH₂), 1.27 (m, 18H); HPLC ret. time 2.72 min, 10-99 % CH₃CN, 5 min run; ESI-MS 222.4 m/z [M+H]⁺.

N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide

0181<chemistry id="chem0005" num="0005"><img file="EP3708564B1_D0005.tif" /></chemistry>

0182To a suspension of 4-oxo-1,4-dihydroquinolin-3-carboxylic acid (35.5 g, 188 mmol) and HBTU (85.7 g, 226 mmol) in DMF (280 mL) was added Et₃N (63.0 mL, 451 mmol) at ambient temperature. The mixture became homogeneous and was allowed to stir for 10 min before 5-amino-2,4-di-<i>tert</i>-butyl-phenol (50.0 g, 226 mmol) was added in small portions. The mixture was allowed to stir overnight at ambient temperature. The mixture became heterogeneous over the course of the reaction. After all of the acid was consumed (LC-MS analysis, MH+ 190, 1.71 min), the solvent was removed <i>in vacuo.</i> EtOH was added to the orange solid material to produce a slurry. The mixture was stirred on a rotovap (bath temperature 65 °C) for 15 min without placing the system under vacuum. The mixture was filtered and the captured solid was washed with hexanes to provide a white solid that was the EtOH crystalate. Et₂O was added to the material obtained above until a slurry was formed. The mixture was stirred on a rotovapor (bath temperature 25 °C) for 15 min without placing the system under vacuum. The mixture was filtered and the solid captured. This procedure was performed a total of five times. The solid obtained after the fifth precipitation was placed under vacuum overnight to provide 8 N-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide as a white powdery solid (38 g, 52%).

0183HPLC ret. time 3.45 min, 10-99% CH₃CN, 5 min run; ¹H NMR (400 MHz, DMSO-<i>d</i>₆) δ 12.88 (s, 1H), 11.83 (s, 1H), 9.20 (s, 1H), 8.87 (s, 1H), 8.33 (dd, J = 8.2, 1.0 Hz, 1H), 7.83-7.79 (m, 1H), 7.76 (d, J = 7.7 Hz, 1H), 7.54-7.50 (m, 1H), 7.17 (s, 1H), 7.10 (s, 1H), 1.38 (s, 9H), 1.37 (s, 9H); ESI-MS 393.3 m/z [M+H]⁺.

0184Set forth below is the characterizing data for Compound 1: <tables id="tabl0003" num="0003"><table frame="all"><title><b>Table 2</b></title><tgroup cols="3"><colspec colnum="1" colname="col1" colwidth="19mm" align="center" /><colspec colnum="2" colname="col2" colwidth="30mm" align="center" /><colspec colnum="3" colname="col3" colwidth="21mm" align="center" /><thead valign="middle"><row><entry><b>Cmd No.</b></entry><entry><b>LC-MS 0236] M+1</b></entry><entry><b>LC-RT min</b></entry></row></thead><tbody valign="middle"><row><entry><b>1</b></entry><entry>393.2</entry><entry>3.71</entry></row></tbody></tgroup></table></tables>

0185The XRPD spectrum of Compound <b>1</b> is shown in <figref idref="f0001"><b>FIG. 1</b></figref><b>.</b>

0186¹H NMR data for Compound <b>1</b> in shown in <figref idref="f0001"><b>FIG. 2</b></figref><b>.</b>

0187The DSC trace of Compound <b>1</b> is shown in <figref idref="f0002"><b>FIG. 3</b></figref><b>.</b>

<u>Preparation of Form A</u>

0188Form A was obtained by heating Compound <b>1</b> as a solid to 250°C and cooling to room temperature. The DSC thermogram on Compound <b>1</b> (See <figref idref="f0005"><b>FIG. 6</b></figref>) shows that the compound undergoes a melt with an onset temperature of 195°C, followed by a recrystallisation with onset at 220°C.

0189The XRPD spectrum of Form A is shown in <figref idref="f0003"><b>FIG. 4</b></figref><b>.</b>

0190The DSC data for Form A is shown in <figref idref="f0004"><b>FIG. 5</b></figref><b>.</b>

0191The TGA trace for Form A is shown in <figref idref="f0005"><b>FIG. 6</b></figref><b>.</b>

<u>Preparation of Form B</u>

0192Crude Compound <b>1</b> was a slurry in refluxing acetonitrile (27 volumes) for 24 hours. After 24 hours, the mixture was allowed to cool to 20 °C. Form B was isolated by filtration as a white to off-white. The wet cake was rinsed with acetonitrile (5 volumes) and dried under vacuum at 50°C until a constant weight is attained, thereby providing Form B.

0193The XRPD spectrum of Form B is shown in <figref idref="f0006"><b>FIG. 7</b></figref><b>.</b>

0194The DSC trace of Form B is shown in <figref idref="f0007"><b>FIG. 8</b></figref><b>.</b>

0195The TGA trace for Form B is shown in <figref idref="f0008"><b>FIG. 9</b></figref><b>.</b>

0196Single crystal data was obtained for Form B, providing detail additional detail about the crystal structure, including lattice size and packing.

Crystal preparation:

01971 g of Compound <b>1</b> was added to 10 ml of isopropanol acetate. The suspension was heated and remained at 60°C for 3 hours. The suspension was cooled to room temperature and remained stirred overnight. The suspended solid was filtered and washed with isopropanol acetate. The collected solid was dried under vacuum at room temperature. 300 mg of the dried solid was dissolved in 5 ml of 10% aqueous ethyl acetate solution. The solution was heated to 70°C for 10 minutes before it was cooled to room temperature. Over time, crystals grew in the vial.

Experimental:

0198A single crystal of Form B was mounted on a MicroMount loop and centered on a Bruker Apex II diffractometer that was equipped with a sealed copper X-ray tube and Apex II CCD detector. Initially, 3 sets of 40 frames were collected to determine a preliminary unit cell. Subsequently a full data set consisting of 15 scans and 6084 frames was acquired. Data collection was performed at 100 K. Data were integrated and scaled using Apex II software from Bruker AXS. Integration and scaling resulted in 7528 reflections, 3071 of which were unique [R(int)) = 0.0466]. Structure was solved by direct methods in space group P2iusing SHELXTL software. Refinement was performed with full-matrix least-square method on F² using SHELXTL software as well. Altogether 375 parameters were used in refinement resulting in reflection to parameter ratio of 8.19. The final refinement afforded a chiral structure with a Flack parameter of 0.0(3). The final refinement index was wR2 = 0.1242 and R1 = 0.0663 (wR2 = 0.1137 and R1 = 0.0482 for reflections with I > 2 sigma (I)).

0199A conformational picture of Form B is provided in <figref idref="f0009"><b>FIGURE 10</b></figref><b>,</b> which is in color. <tables id="tabl0004" num="0004"><table frame="none"><title>Table 1. Crystal data and structure refinement for Compound <b>1.</b></title><tgroup cols="3" colsep="0" rowsep="0"><colspec colnum="1" colname="col1" colwidth="48mm" /><colspec colnum="2" colname="col2" colwidth="41mm" /><colspec colnum="3" colname="col3" colwidth="27mm" /><tbody><row><entry>Identification code</entry><entry>Compound <b>1</b></entry><entry /></row><row><entry>Empirical formula</entry><entry>C24 H28 N2 O3</entry><entry /></row><row><entry>Formula weight</entry><entry>392.48</entry><entry /></row><row><entry>Temperature</entry><entry>100(2) K</entry><entry /></row><row><entry>Wavelength</entry><entry>1.54178 Å</entry><entry /></row><row><entry>Crystal system</entry><entry>monoclinic</entry><entry /></row><row><entry>Space group</entry><entry>P2₁</entry><entry /></row><row><entry>Unit cell dimensions</entry><entry>a = 11.8011(7) Å</entry><entry>α= 90°.</entry></row><row><entry /><entry>b = 5.9819(3) Å</entry><entry>β= 105.110(4)°</entry></row><row><entry /><entry>c = 14.7974(8) Å</entry><entry>γ = 90°.</entry></row><row><entry>Volume</entry><entry>1008.48(10) ų</entry><entry /></row><row><entry>Z</entry><entry>2</entry><entry /></row><row><entry>Density (calculated)</entry><entry>1.293 Mg/m³</entry><entry /></row><row><entry>Absorption coefficient</entry><entry>0.681 mm^-1</entry><entry /></row><row><entry>F(000)</entry><entry>420</entry><entry /></row><row><entry>Crystal size</entry><entry>0.20 x 0.08 x 0.08 mm³</entry><entry /></row><row><entry>Theta range for data collection</entry><entry>3.09 to 68.67°.</entry><entry /></row><row><entry>Index ranges</entry><entry namest="col2" nameend="col3" align="left">-14<=h<=14, -7<=k<=7, -14<=1<=17</entry></row><row><entry>Reflections collected</entry><entry>7528</entry><entry /></row><row><entry>Independent reflections</entry><entry>3071 [R(int) = 0.0466]</entry><entry /></row><row><entry>Completeness to theta = 68.67°</entry><entry>94.6 %</entry><entry /></row><row><entry>Max. and min. transmission</entry><entry>0.9475 and 0.8758</entry><entry /></row><row><entry>Refinement method</entry><entry namest="col2" nameend="col3" align="left">Full-matrix least-squares on F²</entry></row><row><entry>Data / restraints / parameters</entry><entry>3071 / 1 / 375</entry><entry /></row><row><entry>Goodness-of-fit on F²</entry><entry>1.001</entry><entry /></row><row><entry>Final R indices [I>2sigma(I)]</entry><entry>R1 = 0.0482, wR2 = 0.1137</entry><entry /></row><row><entry>R indices (all data)</entry><entry>R1 = 0.0663, wR2 = 0.1242</entry><entry /></row><row><entry>Absolute structure parameter</entry><entry>0.0(3)</entry><entry /></row><row><entry>Extinction coefficient</entry><entry>0.0008(6)</entry><entry /></row><row><entry>Largest diff. peak and hole</entry><entry>0.200 and -0.218 e.Å^-3</entry><entry /></row></tbody></tgroup></table></tables><tables id="tabl0005" num="0005"><table frame="topbot"><title>Table 2. Atomic coordinates ( x 10⁴) and equivalent isotropic displacement parameters (Å²× 10³) for Compound <b>1</b>. U(eq) is defined as one third of the trace of the orthogonalized U^ij tensor.</title><tgroup cols="5" colsep="0"><colspec colnum="1" colname="col1" colwidth="32mm" /><colspec colnum="2" colname="col2" colwidth="35mm" align="center" /><colspec colnum="3" colname="col3" colwidth="35mm" align="center" /><colspec colnum="4" colname="col4" colwidth="33mm" align="center" /><colspec colnum="5" colname="col5" colwidth="32mm" align="center" /><thead valign="top"><row><entry /><entry><b>x</b></entry><entry><b>y</b></entry><entry><b>z</b></entry><entry><b>U(eq)</b></entry></row></thead><tbody><row rowsep="0"><entry>N(1)</entry><entry>11624(2)</entry><entry>-530(5)</entry><entry>8667(2)</entry><entry>33(1)</entry></row><row rowsep="0"><entry>C(3)</entry><entry>10407(3)</entry><entry>2650(6)</entry><entry>8133(2)</entry><entry>31(1)</entry></row><row rowsep="0"><entry>C(4)</entry><entry>10335(3)</entry><entry>2115(6)</entry><entry>7158(2)</entry><entry>30(1)</entry></row><row rowsep="0"><entry>C(5)</entry><entry>11006(3)</entry><entry>150(6)</entry><entry>7013(2)</entry><entry>31(1)</entry></row><row rowsep="0"><entry>C(2)</entry><entry>11034(3)</entry><entry>1269(6)</entry><entry>8830(2)</entry><entry>32(1)</entry></row><row rowsep="0"><entry>C(8)</entry><entry>12233(3)</entry><entry>-3643(7)</entry><entry>6701(3)</entry><entry>38(1)</entry></row><row rowsep="0"><entry>C(7)</entry><entry>11610(3)</entry><entry>-2371(7)</entry><entry>5947(3)</entry><entry>37(1)</entry></row><row rowsep="0"><entry>C(6)</entry><entry>11015(3)</entry><entry>-530(6)</entry><entry>6093(2)</entry><entry>35(1)</entry></row><row rowsep="0"><entry>C(9)</entry><entry>12255(3)</entry><entry>-3039(7)</entry><entry>7618(2)</entry><entry>36(1)</entry></row><row rowsep="0"><entry>C(11)</entry><entry>9786(3)</entry><entry>4549(6)</entry><entry>8431(2)</entry><entry>32(1)</entry></row><row rowsep="0"><entry>C(15)</entry><entry>6468(3)</entry><entry>9414(6)</entry><entry>7354(2)</entry><entry>30(1)</entry></row><row rowsep="0"><entry>C(19)</entry><entry>7049(3)</entry><entry>7025(6)</entry><entry>6127(2)</entry><entry>31(1)</entry></row><row rowsep="0"><entry>C(18)</entry><entry>8461(3)</entry><entry>8580(6)</entry><entry>8706(2)</entry><entry>32(1)</entry></row><row rowsep="0"><entry>C(13)</entry><entry>8275(3)</entry><entry>7448(6)</entry><entry>7859(2)</entry><entry>29(1)</entry></row><row rowsep="0"><entry>C(16)</entry><entry>6578(3)</entry><entry>10442(6)</entry><entry>8223(2)</entry><entry>31(1)</entry></row><row rowsep="0"><entry>C(17)</entry><entry>7639(3)</entry><entry>10044(6)</entry><entry>8889(2)</entry><entry>30(1)</entry></row><row rowsep="0"><entry>C(14)</entry><entry>7271(3)</entry><entry>7965(6)</entry><entry>7130(2)</entry><entry>30(1)</entry></row><row rowsep="0"><entry>C(23)</entry><entry>5586(3)</entry><entry>11841(6)</entry><entry>8438(2)</entry><entry>34(1)</entry></row><row rowsep="0"><entry>C(21)</entry><entry>8075(3)</entry><entry>7722(7)</entry><entry>5705(2)</entry><entry>35(1)</entry></row><row rowsep="0"><entry>C(22)</entry><entry>5943(3)</entry><entry>8034(7)</entry><entry>5474(2)</entry><entry>35(1)</entry></row><row rowsep="0"><entry>C(20)</entry><entry>6879(3)</entry><entry>4481(7)</entry><entry>6096(3)</entry><entry>37(1)</entry></row><row rowsep="0"><entry>C(24)</entry><entry>4478(3)</entry><entry>11888(7)</entry><entry>7605(3)</entry><entry>39(1)</entry></row><row rowsep="0"><entry>C(25)</entry><entry>5981(3)</entry><entry>14254(7)</entry><entry>8672(3)</entry><entry>39(1)</entry></row><row rowsep="0"><entry>C(26)</entry><entry>5207(3)</entry><entry>10760(7)</entry><entry>9264(3)</entry><entry>37(1)</entry></row><row rowsep="0"><entry>N(12)</entry><entry>9082(2)</entry><entry>5775(5)</entry><entry>7752(2)</entry><entry>31(1)</entry></row><row rowsep="0"><entry>O(11)</entry><entry>9923(2)</entry><entry>4910(4)</entry><entry>9289(2)</entry><entry>37(1)</entry></row><row rowsep="0"><entry>O(4)</entry><entry>9748(2)</entry><entry>3206(4)</entry><entry>6485(2)</entry><entry>35(1)</entry></row><row rowsep="0"><entry>O(17)</entry><entry>7888(2)</entry><entry>11078(5)</entry><entry>9758(2)</entry><entry>37(1)</entry></row><row><entry>C(10)</entry><entry>11644(3)</entry><entry>-1165(6)</entry><entry>7761(2)</entry><entry>32(1)</entry></row></tbody></tgroup></table></tables><tables id="tabl0006" num="0006"><table frame="all"><title>Table 3. Bond lengths [Å] and angles [°] for Compound <b>1</b>.</title><tgroup cols="8"><colspec colnum="1" colname="col1" colwidth="22mm" /><colspec colnum="2" colname="col2" colwidth="22mm" /><colspec colnum="3" colname="col3" colwidth="8mm" /><colspec colnum="4" colname="col4" colwidth="32mm" /><colspec colnum="5" colname="col5" colwidth="23mm" /><colspec colnum="6" colname="col6" colwidth="8mm" /><colspec colnum="7" colname="col7" colwidth="31mm" /><colspec colnum="8" colname="col8" colwidth="23mm" /><thead valign="middle"><row><entry align="center"><b>Bond</b></entry><entry align="center"><b>Length (°A)</b></entry><entry rowsep="0" align="center" /><entry align="center"><b>Bond</b></entry><entry align="center"><b>Angle (deg)</b></entry><entry rowsep="0" align="center" /><entry align="center"><b>Bond</b></entry><entry align="center"><b>Angle (deg)</b></entry></row></thead><tbody valign="middle"><row><entry align="center">N(1)-C(2)</entry><entry align="center">1.337(5)</entry><entry morerows="30" align="center" /><entry align="center">C(2)-N(1)-C(10)</entry><entry align="center">122.0(3)</entry><entry morerows="30" align="center" /><entry align="center">C(15)-C(16)-C(23)</entry><entry align="center">122.4(3)</entry></row><row><entry align="center">N(1)-C(10)</entry><entry align="center">1.400(5)</entry><entry align="center">C(2)-C(3)-C(4)</entry><entry align="center">119.4(3)</entry><entry align="center">C(18)-C(17)-O(17)</entry><entry align="center">118.2(3)</entry></row><row><entry align="center">C(3)-C(2)</entry><entry align="center">1.377(5)</entry><entry align="center">C(2)-C(3)-C(11)</entry><entry align="center">116.6(3)</entry><entry align="center">C(18)-C(17)-C(16)</entry><entry align="center">120.9(3)</entry></row><row><entry align="center">C(3)-C(4)</entry><entry align="center">1.458(4)</entry><entry align="center">C(4)-C(3)-C(11)</entry><entry align="center">123.9(3)</entry><entry align="center">O(17)-C(17)-C(16)</entry><entry align="center">120.8(3)</entry></row><row><entry align="center">C(3)-C(11)</entry><entry align="center">1.481(5)</entry><entry align="center">O(4)-C(4)-C(3)</entry><entry align="center">123.8(3)</entry><entry align="center">C(15)-C(14)-C(13)</entry><entry align="center">116.3(3)</entry></row><row><entry align="center">C(4)-O(4)</entry><entry align="center">1.240(4)</entry><entry align="center">O(4)-C(4)-C(5)</entry><entry align="center">120.9(3)</entry><entry align="center">C(15)-C(14)-C(19)</entry><entry align="center">120.1(3)</entry></row><row><entry align="center">C(4)-C(5)</entry><entry align="center">1.465(5)</entry><entry align="center">C(3)-C(4)-C(5)</entry><entry align="center">115.3(3)</entry><entry align="center">C(13)-C(14)-C(19)</entry><entry align="center">123.5(3)</entry></row><row><entry align="center">C(5)-C(10)</entry><entry align="center">1.406(5)</entry><entry align="center">C(10)-C(5)-C(6)</entry><entry align="center">117.2(3)</entry><entry align="center">C(25)-C(23)-C(16)</entry><entry align="center">110.9(3)</entry></row><row><entry align="center">C(5)-C(6)</entry><entry align="center">1.423(5)</entry><entry align="center">C(10)-C(5)-C(4)</entry><entry align="center">122.2(3)</entry><entry align="center">C(25)-C(23)-C(24)</entry><entry align="center">107.9(3)</entry></row><row><entry align="center">C(8)-C(7)</entry><entry align="center">1.391(5)</entry><entry align="center">C(6)-C(5)-C(4)</entry><entry align="center">120.6(3)</entry><entry align="center">C(16)-C(23)-C(24)</entry><entry align="center">112.1(3)</entry></row><row><entry align="center">C(8)-C(9)</entry><entry align="center">1.398(5)</entry><entry align="center">N(1)-C(2)-C(3)</entry><entry align="center">123.4(3)</entry><entry align="center">C(25)-C(23)-C(26)</entry><entry align="center">110.5(3)</entry></row><row><entry align="center">C(7)-C(6)</entry><entry align="center">1.353(5)</entry><entry align="center">C(7)-C(8)-C(9)</entry><entry align="center">120.6(4)</entry><entry align="center">C(16)-C(23)-C(26)</entry><entry align="center">109.2(3)</entry></row><row><entry align="center">C(9)-C(10)</entry><entry align="center">1.379(5)</entry><entry align="center">C(6)-C(7)-C(8)</entry><entry align="center">120.3(4)</entry><entry align="center">C(24)-C(23)-C(26)</entry><entry align="center">106.1(3)</entry></row><row><entry align="center">C(11)-O(11)</entry><entry align="center">1.255(4)</entry><entry align="center">C(7)-C(6)-C(5)</entry><entry align="center">121.2(3)</entry><entry align="center">C(11)-N(12)-C(13)</entry><entry align="center">127.2(3)</entry></row><row><entry align="center">C(11)-N(12)</entry><entry align="center">1.343(4)</entry><entry align="center">C(10)-C(9)-C(8)</entry><entry align="center">118.7(3)</entry><entry align="center">C(9)-C(10)-N(1)</entry><entry align="center">120.5(3)</entry></row><row><entry align="center">C(15)-C(14)</entry><entry align="center">1.387(5)</entry><entry align="center">O(11)-C(11)-N(12)</entry><entry align="center">123.7(3)</entry><entry align="center">C(9)-C(10)-C(5)</entry><entry align="center">121.9(3)</entry></row><row><entry align="center">C(15)-C(16)</entry><entry align="center">1.399(5)</entry><entry align="center">O(11)-C(11)-C(3)</entry><entry align="center">119.4(3)</entry><entry align="center">N(1)-C(10)-C(5)</entry><entry align="center">117.6(3)</entry></row><row><entry align="center">C(19)-C(22)</entry><entry align="center">1.531(4)</entry><entry align="center">N(12)-C(11)-C(3)</entry><entry align="center">117.0(3)</entry><entry namest="col7" nameend="col8" morerows="13" align="center" /></row><row><entry align="center">C(19)-C(20)</entry><entry align="center">1.534(5)</entry><entry align="center">C(14)-C(15)-C(16)</entry><entry align="center">126.1(3)</entry></row><row><entry align="center">C(19)-C(14)</entry><entry align="center">1.544(4)</entry><entry align="center">C(22)-C(19)-C(20)</entry><entry align="center">106.8(3)</entry></row><row><entry align="center">C(19)-C(21)</entry><entry align="center">1.558(5)</entry><entry align="center">C(22)-C(19)-C(14)</entry><entry align="center">111.5(3)</entry></row><row><entry align="center">C(18)-C(17)</entry><entry align="center">1.385(5)</entry><entry align="center">C(20)-C(19)-C(14)</entry><entry align="center">112.2(3)</entry></row><row><entry align="center">C(18)-C(13)</entry><entry align="center">1.390(5)</entry><entry align="center">C(22)-C(19)-C(21)</entry><entry align="center">105.4(3)</entry></row><row><entry align="center">C(13)-C(14)</entry><entry align="center">1.413(4)</entry><entry align="center">C(20)-C(19)-C(21)</entry><entry align="center">111.3(3)</entry></row><row><entry align="center">C(13)-N(12)</entry><entry align="center">1.418(4)</entry><entry align="center">C(14)-C(19)-C(21)</entry><entry align="center">109.4(3)</entry></row><row><entry align="center">C(16)-C(17)</entry><entry align="center">1.397(4)</entry><entry align="center">C(17)-C(18)-C(13)</entry><entry align="center">122.0(3)</entry></row><row><entry align="center">C(16)-C(23)</entry><entry align="center">1.539(5)</entry><entry align="center">C(18)-C(13)-C(14)</entry><entry align="center">119.0(3)</entry></row><row><entry align="center">C(17)-O(17)</entry><entry align="center">1.387(4)</entry><entry align="center">C(18)-C(13)-N(12)</entry><entry align="center">119.4(3)</entry></row><row><entry align="center">C(23)-C(25)</entry><entry align="center">1.528(5)</entry><entry align="center">C(14)-C(13)-N(12)</entry><entry align="center">121.5(3)</entry></row><row><entry align="center">C(23)-C(24)</entry><entry align="center">1.546(5)</entry><entry align="center">C(17)-C(16)-C(15)</entry><entry align="center">115.1(3)</entry></row><row><entry align="center">C(23)-C(26)</entry><entry align="center">1.548(5)</entry><entry align="center">C(17)-C(16)-C(23)</entry><entry align="center">122.4(3)</entry></row></tbody></tgroup></table></tables>

0200Symmetry transformations used to generate equivalent atoms: <tables id="tabl0007" num="0007"><table frame="topbot"><title>Table 4. Anisotropic displacement parameters (Å²× 10³)for Compound <b>1</b>. The anisotropic displacement factor exponent takes the form: -2π²[ h²a*²U¹¹ + ... + 2 h k a* b* U¹² ]</title><tgroup cols="7" colsep="0"><colspec colnum="1" colname="col1" colwidth="25mm" /><colspec colnum="2" colname="col2" colwidth="24mm" align="center" /><colspec colnum="3" colname="col3" colwidth="24mm" align="center" /><colspec colnum="4" colname="col4" colwidth="24mm" align="center" /><colspec colnum="5" colname="col5" colwidth="24mm" align="center" /><colspec colnum="6" colname="col6" colwidth="24mm" align="center" /><colspec colnum="7" colname="col7" colwidth="24mm" align="center" /><thead valign="top"><row><entry /><entry><b>U¹¹</b></entry><entry><b>U²²</b></entry><entry><b>U³³</b></entry><entry><b>U²³</b></entry><entry><b>U¹³</b></entry><entry><b>U¹²</b></entry></row></thead><tbody><row rowsep="0"><entry>N(1)</entry><entry>42(1)</entry><entry>41(2)</entry><entry>14(2)</entry><entry>5(1)</entry><entry>4(1)</entry><entry>3(1)</entry></row><row rowsep="0"><entry>C(3)</entry><entry>34(2)</entry><entry>40(2)</entry><entry>16(2)</entry><entry>-1(1)</entry><entry>4(1)</entry><entry>-4(1)</entry></row><row rowsep="0"><entry>C(4)</entry><entry>34(2)</entry><entry>38(2)</entry><entry>17(2)</entry><entry>0(1)</entry><entry>4(1)</entry><entry>-1(1)</entry></row><row rowsep="0"><entry>C(5)</entry><entry>34(2)</entry><entry>42(2)</entry><entry>17(2)</entry><entry>-2(1)</entry><entry>6(1)</entry><entry>-6(1)</entry></row><row rowsep="0"><entry>C(2)</entry><entry>37(2)</entry><entry>42(2)</entry><entry>16(2)</entry><entry>1(1)</entry><entry>5(1)</entry><entry>1(2)</entry></row><row rowsep="0"><entry>C(8)</entry><entry>44(2)</entry><entry>41(2)</entry><entry>30(2)</entry><entry>-4(2)</entry><entry>10(1)</entry><entry>5(2)</entry></row><row rowsep="0"><entry>C(7)</entry><entry>46(2)</entry><entry>44(2)</entry><entry>22(2)</entry><entry>-4(1)</entry><entry>9(1)</entry><entry>-5(2)</entry></row><row rowsep="0"><entry>C(6)</entry><entry>41(2)</entry><entry>40(2)</entry><entry>23(2)</entry><entry>1(2)</entry><entry>9(1)</entry><entry>-1(2)</entry></row><row rowsep="0"><entry>C(9)</entry><entry>41(2)</entry><entry>40(2)</entry><entry>24(2)</entry><entry>5(1)</entry><entry>4(1)</entry><entry>3(2)</entry></row><row rowsep="0"><entry>C(11)</entry><entry>35(2)</entry><entry>41(2)</entry><entry>18(2)</entry><entry>1(1)</entry><entry>4(1)</entry><entry>-4(2)</entry></row><row rowsep="0"><entry>C(15)</entry><entry>37(2)</entry><entry>37(2)</entry><entry>15(2)</entry><entry>4(1)</entry><entry>3(1)</entry><entry>1(1)</entry></row><row rowsep="0"><entry>C(19)</entry><entry>38(2)</entry><entry>38(2)</entry><entry>14(2)</entry><entry>2(1)</entry><entry>5(1)</entry><entry>4(1)</entry></row><row rowsep="0"><entry>C(18)</entry><entry>36(2)</entry><entry>42(2)</entry><entry>14(2)</entry><entry>4(1)</entry><entry>0(1)</entry><entry>0(1)</entry></row><row rowsep="0"><entry>C(13)</entry><entry>39(2)</entry><entry>34(2)</entry><entry>16(2)</entry><entry>2(1)</entry><entry>9(1)</entry><entry>-3(1)</entry></row><row rowsep="0"><entry>C(16)</entry><entry>46(2)</entry><entry>29(2)</entry><entry>19(2)</entry><entry>1(1)</entry><entry>10(1)</entry><entry>-3(1)</entry></row><row rowsep="0"><entry>C(17)</entry><entry>43(2)</entry><entry>33(2)</entry><entry>14(2)</entry><entry>-2(1)</entry><entry>7(1)</entry><entry>-6(1)</entry></row><row rowsep="0"><entry>C(14)</entry><entry>38(2)</entry><entry>38(2)</entry><entry>11(2)</entry><entry>1(1)</entry><entry>3(1)</entry><entry>-3(2)</entry></row><row rowsep="0"><entry>C(23)</entry><entry>46(2)</entry><entry>40(2)</entry><entry>20(2)</entry><entry>2(1)</entry><entry>13(1)</entry><entry>3(2)</entry></row><row rowsep="0"><entry>C(21)</entry><entry>51(2)</entry><entry>45(2)</entry><entry>8(2)</entry><entry>2(1)</entry><entry>7(1)</entry><entry>0(2)</entry></row><row rowsep="0"><entry>C(22)</entry><entry>44(2)</entry><entry>41(2)</entry><entry>16(2)</entry><entry>-7(1)</entry><entry>1(1)</entry><entry>2(2)</entry></row><row rowsep="0"><entry>C(20)</entry><entry>40(2)</entry><entry>46(2)</entry><entry>20(2)</entry><entry>-1(2)</entry><entry>3(1)</entry><entry>1(2)</entry></row><row rowsep="0"><entry>C(24)</entry><entry>44(2)</entry><entry>49(2)</entry><entry>24(2)</entry><entry>-2(2)</entry><entry>10(1)</entry><entry>5(2)</entry></row><row rowsep="0"><entry>C(25)</entry><entry>52(2)</entry><entry>43(2)</entry><entry>24(2)</entry><entry>3(1)</entry><entry>12(2)</entry><entry>9(2)</entry></row><row rowsep="0"><entry>C(26)</entry><entry>48(2)</entry><entry>40(2)</entry><entry>24(2)</entry><entry>1(1)</entry><entry>14(1)</entry><entry>0(2)</entry></row><row rowsep="0"><entry>N(12)</entry><entry>40(1)</entry><entry>41(2)</entry><entry>12(2)</entry><entry>0(1)</entry><entry>5(1)</entry><entry>0(1)</entry></row><row rowsep="0"><entry>O(11)</entry><entry>48(1)</entry><entry>47(1)</entry><entry>13(1)</entry><entry>1(1)</entry><entry>4(1)</entry><entry>5(1)</entry></row><row rowsep="0"><entry>O(4)</entry><entry>46(1)</entry><entry>46(2)</entry><entry>12(1)</entry><entry>3(1)</entry><entry>4(1)</entry><entry>7(1)</entry></row><row rowsep="0"><entry>O(17)</entry><entry>44(1)</entry><entry>45(2)</entry><entry>18(1)</entry><entry>-6(1)</entry><entry>4(1)</entry><entry>4(1)</entry></row><row><entry>C(10)</entry><entry>37(2)</entry><entry>37(2)</entry><entry>21(2)</entry><entry>0(1)</entry><entry>8(1)</entry><entry>-2(2)</entry></row></tbody></tgroup></table></tables><tables id="tabl0008" num="0008"><table frame="topbot"><title>Table 5. Hydrogen coordinates ( x 10⁴) and isotropic displacement parameters (Å²× 10³) for Compound <b>1</b>.</title><tgroup cols="5" colsep="0"><colspec colnum="1" colname="col1" colwidth="30mm" /><colspec colnum="2" colname="col2" colwidth="30mm" /><colspec colnum="3" colname="col3" colwidth="30mm" /><colspec colnum="4" colname="col4" colwidth="30mm" /><colspec colnum="5" colname="col5" colwidth="28mm" /><thead valign="top"><row><entry /><entry align="center"><b>x</b></entry><entry align="center"><b>y</b></entry><entry align="center"><b>z</b></entry><entry><b>U(eq)</b></entry></row></thead><tbody><row rowsep="0"><entry>H(7)</entry><entry align="right">11560(30)</entry><entry align="right">-2840(70)</entry><entry align="right">5320(30)</entry><entry>35(10)</entry></row><row rowsep="0"><entry>H(9)</entry><entry align="right">12670(30)</entry><entry align="right">-3980(70)</entry><entry align="right">8120(30)</entry><entry>38(10)</entry></row><row rowsep="0"><entry>H(8)</entry><entry align="right">12680(30)</entry><entry align="right">-4860(70)</entry><entry align="right">6660(30)</entry><entry>36(10)</entry></row><row rowsep="0"><entry>H(6)</entry><entry align="right">10550(30)</entry><entry align="right">350(80)</entry><entry align="right">5580(30)</entry><entry>51(13)</entry></row><row rowsep="0"><entry>H(15)</entry><entry align="right">5770(30)</entry><entry align="right">9730(70)</entry><entry align="right">6900(30)</entry><entry>30(9)</entry></row><row rowsep="0"><entry>H(18)</entry><entry align="right">9150(20)</entry><entry align="right">8310(50)</entry><entry align="right">9160(20)</entry><entry>12(7)</entry></row><row rowsep="0"><entry>H(17)</entry><entry align="right">8620(30)</entry><entry align="right">10600(60)</entry><entry align="right">10030(30)</entry><entry>25(9)</entry></row><row rowsep="0"><entry>H(20A)</entry><entry align="right">7470(30)</entry><entry align="right">3650(70)</entry><entry align="right">6460(30)</entry><entry>32(10)</entry></row><row rowsep="0"><entry>H(20B)</entry><entry align="right">6130(30)</entry><entry align="right">4320(80)</entry><entry align="right">6280(30)</entry><entry>43(11)</entry></row><row rowsep="0"><entry>H(21A)</entry><entry align="right">8840(30)</entry><entry align="right">6840(70)</entry><entry align="right">5980(30)</entry><entry>40(11)</entry></row><row rowsep="0"><entry>H(21B)</entry><entry align="right">8160(30)</entry><entry align="right">9370(80)</entry><entry align="right">5790(30)</entry><entry>42(11)</entry></row><row rowsep="0"><entry>H(22B)</entry><entry align="right">5990(30)</entry><entry align="right">9620(70)</entry><entry align="right">5480(30)</entry><entry>31(9)</entry></row><row rowsep="0"><entry>H(22A)</entry><entry align="right">5790(30)</entry><entry align="right">7200(80)</entry><entry align="right">4820(30)</entry><entry>48(12)</entry></row><row rowsep="0"><entry>H(24A)</entry><entry align="right">3800(40)</entry><entry align="right">12810(90)</entry><entry align="right">7750(30)</entry><entry>57(13)</entry></row><row rowsep="0"><entry>H(24B)</entry><entry align="right">4210(30)</entry><entry align="right">10410(70)</entry><entry align="right">7420(30)</entry><entry>34(10)</entry></row><row rowsep="0"><entry>H(25A)</entry><entry align="right">5370(30)</entry><entry align="right">15130(60)</entry><entry align="right">8770(20)</entry><entry>24(9)</entry></row><row rowsep="0"><entry>H(25B)</entry><entry align="right">6240(30)</entry><entry align="right">15040(70)</entry><entry align="right">8150(30)</entry><entry>41(11)</entry></row><row rowsep="0"><entry>H(25C)</entry><entry align="right">6690(30)</entry><entry align="right">14100(80)</entry><entry align="right">9230(30)</entry><entry>44(11)</entry></row><row rowsep="0"><entry>H(26A)</entry><entry align="right">4600(30)</entry><entry align="right">11790(60)</entry><entry align="right">9320(20)</entry><entry>17(8)</entry></row><row rowsep="0"><entry>H(26B)</entry><entry align="right">5000(30)</entry><entry align="right">9350(70)</entry><entry align="right">9090(30)</entry><entry>28(9)</entry></row><row rowsep="0"><entry>H(1)</entry><entry align="right">12000(30)</entry><entry align="right">-1450(70)</entry><entry align="right">9140(30)</entry><entry>40(11)</entry></row><row rowsep="0"><entry>H(2)</entry><entry align="right">11050(40)</entry><entry align="right">1550(80)</entry><entry align="right">9460(40)</entry><entry>56(13)</entry></row><row rowsep="0"><entry>H(26C)</entry><entry align="right">5950(30)</entry><entry align="right">10770(80)</entry><entry align="right">9820(30)</entry><entry>51(12)</entry></row><row rowsep="0"><entry>H(24C)</entry><entry align="right">4720(40)</entry><entry align="right">12850(100)</entry><entry align="right">7170(40)</entry><entry>69(15)</entry></row><row rowsep="0"><entry>H(22C)</entry><entry align="right">5150(40)</entry><entry align="right">7470(70)</entry><entry align="right">5610(30)</entry><entry>42(11)</entry></row><row rowsep="0"><entry>H(21C)</entry><entry align="right">7820(40)</entry><entry align="right">7310(90)</entry><entry align="right">5040(40)</entry><entry>62(14)</entry></row><row rowsep="0"><entry>H(20C)</entry><entry align="right">6780(30)</entry><entry align="right">3790(70)</entry><entry align="right">5480(30)</entry><entry>48(12)</entry></row><row><entry>H(12)</entry><entry align="right">9030(40)</entry><entry align="right">5290(90)</entry><entry align="right">7280(40)</entry><entry>62(16)</entry></row></tbody></tgroup></table></tables>

<u>Preparation of Amorphous Form from Form B</u>

0201A Buchi Spray drier was used in this method under the following conditions: <ul id="ul0002" list-style="none" compact="compact"><li>Inlet temperature set point: 130 °C</li><li>Outlet Temperature (start of run) 55 °C</li><li>Outlet temperature (end of run): 58 °C</li><li>Nitrogen pressure: 120psi</li><li>Aspirator: 100%</li><li>Pump: 40%</li><li>Filter pressure 11 mbar</li><li>Condenser Temperature: 10 °C</li><li>Run Time 15 min</li><li>Yield 86.5%</li><li>Dried in 25 °C vacuum over for 24 hours.</li><li>4g of Form B was dissolved in 86.4g of acetone and 9.6 g water under the above conditions. The run time was 15 min. The product was dried under vacuum at 25 °C for over 24 hrs to produce the Amorphous Form.</li></ul>

0202The XRPD spectrum of the Amorphous Form is shown in <figref idref="f0010"><b>FIG. 11</b></figref>.

0203The TGA trace for Amorphous Form is shown in <figref idref="f0011"><b>FIG. 12</b></figref>.

0204The DSC trace for Amorphous Form is shown in <figref idref="f0012"><b>FIG. 13</b></figref>.

PK and solubility of different solid state forms of Compound <b>1</b>

0205Bioavailability of crystalline Form B, 85% amorphous Compound <b>1</b> and HPMCAS solid dispersion of Compound <b>1</b> were evaluated in rat, the results of which are provided in Table 4 below. These forms of the compound were dosed in an oral suspension with a vehicle containing 0.5% methyl cellulose/0.5% SLS/99% water. Bioavailability of various solid forms was evaluated as compared to a multicomponent IV solution of Compound 1. Bioavailability of crystalline polymorph B was 3-6%, compared to 61-95% for amorphous material and 109-111% for solid dispersion. In FaSSIF, crystalline polymorph B has a measured solubility of 1.0 µg/ml, while the 85% amorphous material has a solubility of 67.4 µg/ml. The crystalline material showed 67-74% bioavailability when dosed as a PEG solution, indicating that absorption was solubility limited. <tables id="tabl0009" num="0009"><table frame="all"><title>Table 4</title><tgroup cols="9"><colspec colnum="1" colname="col1" colwidth="29mm" /><colspec colnum="2" colname="col2" colwidth="33mm" /><colspec colnum="3" colname="col3" colwidth="26mm" /><colspec colnum="4" colname="col4" colwidth="15mm" /><colspec colnum="5" colname="col5" colwidth="15mm" /><colspec colnum="6" colname="col6" colwidth="12mm" /><colspec colnum="7" colname="col7" colwidth="12mm" /><colspec colnum="8" colname="col8" colwidth="14mm" /><colspec colnum="9" colname="col9" colwidth="12mm" /><thead valign="middle"><row><entry align="center"><b>Drug Form</b></entry><entry align="center"><b>Vehicle</b></entry><entry align="center"><b>Dose (mg/kg)</b></entry><entry namest="col4" nameend="col5" align="center"><b>AUC (ug*hr/mL)</b></entry><entry namest="col6" nameend="col7" align="center"><b>Tmax (h)</b></entry><entry namest="col8" nameend="col9" align="center"><b>% F</b></entry></row></thead><tbody valign="middle"><row><entry align="center">85% Amorphous</entry><entry align="center">0.5% MC/0.5% SLS</entry><entry align="center">50</entry><entry align="center">135.5</entry><entry align="center">27.6</entry><entry align="center">6.0</entry><entry align="center">0.0</entry><entry align="center">95.0</entry><entry align="center">20.0</entry></row><row><entry align="center">85% Amorphous</entry><entry align="center">0.5% MC/0.5% SLS</entry><entry align="center">200</entry><entry align="center">371.9</entry><entry align="center">46.1</entry><entry align="center">6.0</entry><entry align="center">0.0</entry><entry align="center">61.0</entry><entry align="center">7.0</entry></row><row><entry align="center">Crystalline</entry><entry align="center">0.5% MC/0.5% SLS</entry><entry align="center">50</entry><entry align="center">8.0</entry><entry align="center">1.2</entry><entry align="center">4.0</entry><entry align="center">0.0</entry><entry align="center">5.5</entry><entry align="center">0.8</entry></row><row><entry align="center">Crystalline</entry><entry align="center">0.5% MC/0.5% SLS</entry><entry align="center">200</entry><entry align="center">16.9</entry><entry align="center">3.0</entry><entry align="center">4.7</entry><entry align="center">1.2</entry><entry align="center">3.1</entry><entry align="center">0.3</entry></row><row><entry align="center">Crystalline</entry><entry align="center">PEG</entry><entry align="center">50</entry><entry align="center">135.1</entry><entry align="center">43.0</entry><entry align="center">5.5</entry><entry align="center">1.0</entry><entry align="center">74.0</entry><entry align="center">23.0</entry></row><row><entry align="center">Crystalline</entry><entry align="center">PEG</entry><entry align="center">200</entry><entry align="center">431.5</entry><entry align="center">101.1</entry><entry align="center">14.5</entry><entry align="center">11.0</entry><entry align="center">67.0</entry><entry align="center">16.0</entry></row><row><entry align="center">Solid Dispersion</entry><entry align="center">0.5% MC/0.5% SLS</entry><entry align="center">25</entry><entry align="center">90.1</entry><entry align="center">8.1</entry><entry align="center">6.0</entry><entry align="center">0.0</entry><entry align="center">111.0</entry><entry align="center">10.0</entry></row><row><entry align="center">Solid Dispersion</entry><entry align="center">0.5% MC/0.5% SLS</entry><entry align="center">100</entry><entry align="center">260.8</entry><entry align="center">28.4</entry><entry align="center">6.0</entry><entry align="center">0.0</entry><entry align="center">109.0</entry><entry align="center">12.0</entry></row></tbody></tgroup></table></tables>