HK1129581B - Compositions of n-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide - Google Patents

Description

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

Priority declaration

Priority of U.S. patent application serial No. 60/799,795, filed 2006, 5, 12, is claimed herein according to USC § 119(e), the entire disclosure of which is incorporated herein by reference.

Technical Field

The present invention relates to pharmaceutical compositions of N- [2, 4-bis (1, 1-dimethylethyl) -5-hydroxyphenyl ] -1, 4-dihydro-4-oxoquinoline-3-carboxamide and methods thereof.

Background

CFTR is a cAMP/ATP-mediated anion channel that is expressed in a variety of cell types, including absorptive and secretory epithelial cells, where CFTR regulates anion flux across membranes, as well as the activity of other ion channels and proteins. In epithelial cells, the maintenance of normal function of CFTR is critical to maintain electrolyte transport throughout the body, including respiratory and digestive tissues. CFTR consists of approximately 1480 amino acids, which encode a protein consisting of tandem repeats of transmembrane domains, each containing six transmembrane helices and a nucleotide binding domain. The two transmembrane domains are connected by a large polarity-regulating (R) -domain with multiple phosphorylation sites that regulate channel activity and cellular trafficking.

The gene encoding CFTR has been identified and sequenced (see Gregory, R, J. et al, (1990) Nature 347: 382. sup. 386; Rich, D.P. et al, (1990) Nature 347; 358. sup. 362), (Riordan, J.R. et al, (1989) Science 245; 1066. sup. 1073). Defects in this gene cause CFFR mutations, which lead to cystic fibrosis ("CF"), the most common fatal genetic disease in humans. In the united states, approximately one in every 2,500 newborns is affected by cystic fibrosis. Up to 1 million people in the entire us population carry a single copy of a defective gene, but have no apparent disease effects. In contrast, individuals with two copies of the CF-associated gene can be affected by the debilitating and fatal effects of CF, including chronic lung disease.

In patients with cystic fibrosis, mutations in endogenously expressed CFTR in respiratory epithelial cells result in reduced apical anion secretion, which can lead to an imbalance in ionic and humoral transport. The resulting decrease in anion transport promotes increased lung mucus accumulation and concomitant increased microbial infection, which ultimately leads to death in CF patients. In addition to respiratory diseases, CF patients typically suffer from gastrointestinal problems and pancreatic insufficiency, which if left untreated, can lead to death. In addition, most cystic fibrosis men are infertile and the fertility of women with cystic fibrosis is reduced. In contrast to the severe effects of having two copies of the CF-associated gene, individuals with a single copy of the CF-associated gene show increased tolerance to cholera and to dehydration due to diarrhea, which may explain the relatively high frequency of CF genes in the population.

Sequencing analysis of the CFTR gene of the CF chromosome revealed a number of disease-causing mutations (Cutting, G.R. et al, (1990) Nature 346: 366-. To date, has identified>1000 pathogenic CF Gene mutations (ii) ((iii))http://www.genet.sickkids.on.ca/cftr/). The most common mutation is a deletion of phenylalanine at amino acid sequence 508 of CFTR, commonly referred to as Δ F508-CFTR. This mutation occurs in approximately 70% of cases of cystic fibrosis and is associated with severe disease.

Deletion of residue 508 in Δ F508-CFTR prevents the nascent protein from folding correctly. This results in the inability of the mutein to exit the ER and to be transported to the plasma membrane. As a result, the number of channels present in the membrane is much smaller than that observed in cells expressing pristine CFTR. In addition to impaired trafficking, mutations also lead to defective channel gating. In summary, the reduced number of channels in the membrane and defective channel gating lead to reduced anion transport through epithelial cells, resulting in defective ion and fluid transport (Quinton, P.M, (1990), FASEB J.4: 2709-. However, studies have shown that the reduction in the number of AF 508-CFTR in the membrane is functional, albeit smaller than the pristine CFTR (Dalemans et al, (1991), NatureLond.354: 526-. In addition to af 508-CFTR, other pathogenic CFTR mutations that result in defective trafficking, synthesis, and/or channel gating may be up-or down-regulated to alter anion secretion and modify disease progression and/or severity.

Although CFTR transports a variety of molecules in addition to anions, it is clear that this effect (anion transport) represents an important element in the transport of ions and water through epithelial cells. Other elements include epithelial Na⁺Channels, ENaC, Na⁺/2Cl^-/K⁺Cotransporter, Na⁺-K⁺Adenosine triphosphate pump and basolateral membrane (basemembranmembrane) K⁺Channels, which are responsible for the uptake of chlorine into cells.

These elements work together by their selective expression and localization within the cell to obtain directional transport across epithelial cells. The uptake of chlorine is via ENaC and CFTR present in the apical membrane and Na expressed on the basolateral surface of the cell⁺-K⁺Adenosine triphosphate pump and Cl^-Coordinated activity of the channels occurs. Secondary active transport of chlorine from the luminal side results in the accumulation of intracellular chlorine, which can then pass through Cl^-The channel passively leaves the cell, resulting in vector transport. Na (Na)⁺/2Cl^-/K⁺Cotransporter, Na⁺-K⁺Adenosine triphosphate pump and substrate outer membrane K on substrate outer surface⁺The arrangement of the CFTR on the channel and luminal side coordinates the secretion of chlorine through the CFTR on the luminal side. Because water may never actively transport itself, its flow across epithelial cells relies on a slight trans-epithelial osmotic gradient created by the bulk flow of sodium and chlorine.

In addition to cystic fibrosis, modulation of CFTR activity may be beneficial in other diseases not directly caused by mutations in CFTR, such as secretory diseases and other protein folding diseases mediated by CFTR. These diseases include, but are not limited to, Chronic Obstructive Pulmonary Disease (COPD), dry eye disease, and sjogren's syndrome. COPD is characterized by airflow limitation, which is progressive and not fully reversible. The airflow limitation is due to mucus hypersecretion, emphysema and bronchiolitis. Activators of mutant or primitive CFTR offer a potential treatment for mucus hypersecretion and decreased mucociliary clearance common in COPD. In particular, increasing anion secretion across CFTR may facilitate fluid transport into the liquid of the airway surface, thereby hydrating the mucus and optimizing the viscosity of the fluid around the cilia. This will result in increased mucociliary clearance and a reduction in symptoms associated with COPD. Dry eye is characterized by a decrease in tear 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 sjogren's syndrome. Increasing anion secretion via CFTR will increase transport of body fluids from corneal endothelial cells and secretory glands surrounding the eye, thereby increasing corneal hydration. This will help alleviate symptoms associated with dry eye. Sjogren's syndrome is an autoimmune disease in which the immune system attacks the whole body of moisture-producing glands, including the eye, mouth, skin, respiratory tissues, liver, vagina and intestine. Symptoms include dryness of the eye, mouth and vagina, and lung disease. The disease is also associated with rheumatoid arthritis, systemic lupus, systemic sclerosis and polymyositis/dermatomyositis. Defective protein trafficking is thought to lead to the disease, and therefore treatment options are limited. Modulators of CFTR activity can hydrate a variety of disease-affected organs and help ameliorate the associated symptoms.

As discussed above, it is believed that the deletion of residue 508 in af 508-CFTR prevents the nascent protein from folding correctly, resulting in the inability of this mutein to exit the ER and be transported to the plasma membrane. As a result, the amount of mature protein present in the plasma membrane is insufficient and chloride transport in epithelial tissues is significantly reduced. In fact, it has been shown that this cellular phenomenon of defective ER processing of ABC transporters by the ER mechanism is not only the basis of CF disease, but is also a potential basis for a variety of other isolated or genetic diseases. Two ways in which the ER system may malfunction are either failure of the protein to bind to ER export leading to degradation, or ER accumulation of these defective/misfolded proteins [ Aridror M., et al, Nature Med., 5(7), pp 745-751 (1999); sharry, b.s., et al, Neurochem, International,43pp 1-7 (2003); rutishauser, j, et al, SwissMed WkIy,132pp 211-; morelio, JP et al, TIPS,21pp.466-469 (2000); bross P., et al, Human mut,14，pp.186-198(1999)]. And the first kindDiseases associated with ER disorders are cystic fibrosis (caused by the misfolded AF 508-CFTR described above), hereditary emphysema (caused by alpha 1-antitrypsin; non-Piz variants), hereditary hemochromatosis, coagulation-fibrinolysis defects, e.g.protein C defects, hereditary angioedema type 1, lipid processing defects, e.g.familial hypercholesterolemia, chylomicronemia type 1, betalipoproteinemia, lysosomal storage diseases, e.g.I-cell disease/pseudoherlerian disease, mucopolysaccharidosis (caused by lysosomal processing enzymes), Morhoff/tay-Saccharopathy (caused by beta-hexosaminidase), Creutzfeldt-Jacob syndrome type II (caused by UDP-glucuronic-sialic acid (sialyc) -transferase), polyendocrinopathy/hyperinsulinemia, diabetes (caused by the insulin receptor), larval dwarfism (caused by the growth hormone receptor), myeloperoxidase deficiency, primary hypoparathyroidism (caused by the prepro-parathyroid hormone), melanoma (caused by tyrosinase). Diseases associated with disturbances of the latter ER are the glycan disease CDG 1 type, hereditary emphysema (caused by alpha 1-insulin resistance (PiZ variant)), congenital hyperthyroidism, osteogenesis imperfecta (caused by procollagen types I, II, IV), hereditary hypofibrinogenemia (caused by fibrinogen), ACT deficiency (caused by alpha 1-antichymotrypsin), Diabetes Insipidus (DI), posterior leaflet hormone transporter DI (caused by vasopressin/V2-receptor), renal DI (caused by aquaporin II), Charcot-Marie-Tooth syndrome (caused by peripheral myelin protein 22), Per-Merzbacher disease, neurodegenerative diseases such as Alzheimer's disease (caused by beta APP and presenilin), Parkinson's disease, amyotrophic lateral sclerosis, progressive supranuclear palsy, pick's disease, several polyglutamine nervous system disorders such as huntington's disease, spinocerebellar ataxia type I, spinal and bulbar muscular atrophy, dentatorubral pallidoluysian atrophy, and myotonic atrophy, and spongiform encephalopathies such as hereditary Creutzfeldt-Jakob disease (caused by prion protein processing defects), Fabry disease (caused by lysosomal α -galactosidase a) and Straussler-Scheinker syndrome (caused by Prp processing defects).

In addition to upregulating CFTR activity, reduction of anion secretion by CFTR modulators may also be beneficial in the treatment of secretory diarrhoea, in which epithelial water transport is dramatically increased due to secretagogue activation of chloride transport. This mechanism involves elevation of cAMP and stimulation of CFTR.

Although there are many causes of diarrhea, the major consequences of diarrheal disease caused by excessive chloride transport are common, including dehydration, acidosis, reduced growth and death.

Acute and chronic diarrhea are major medical problems occurring in many areas of the world. Diarrhea is both a significant cause of malnutrition and a leading cause of death (5,000,000 deaths per year) in children less than five years of age.

Secretory diarrhea is also a risk symptom in patients with acquired immunodeficiency syndrome (AIDS) and chronic Inflammatory Bowel Disease (IBD). Every year, 1 thousand 6 million people among travelers from industrialized countries to developing countries develop diarrhea, and the severity and number of cases of diarrhea varies depending on the country and region of travel.

Diarrhoea, also known as scours, in livestock and pets, such as cattle, pigs and horses, sheep, goats, cats and dogs, is a major cause of death in these animals. Diarrhea can be caused by any major cause, such as weaning or physical movement, as well as a response to various bacterial or viral infections and usually occurs within the first few hours of the animal's life.

The most common diarrhea causing bacteria is enterotoxigenic escherichia coli (ETEC) with K99 cilia antigen. Common viruses that cause diarrhea include rotaviruses and coronaviruses. Other infectious agents include, inter alia, cryptosporidium (cryptosporidium), giardia lamblia (giardia1amblia) and salmonella.

Symptoms of rotavirus infection include excretion of aqueous feces, dehydration and weakness. Coronaviruses cause more severe disease in newborn animals and have a higher mortality rate than rotavirus infection. However, often young animals may be infected with more than one virus at a time or with a combination of viral and bacterial microorganisms. This strongly increases the severity of the disease.

Accordingly, there is a need for pharmaceutical compositions of modulators of CFTR activity that can be used to modulate CFTR activity in the cell membrane of a mammal.

There is a need for methods of treating CFTR mediated diseases using such pharmaceutical compositions.

Disclosure of Invention

The present invention relates to pharmaceutical compositions of N- [2, 4-bis (1, 1-dimethylethyl) -5-hydroxyphenyl ] -1, 4-dihydro-4-oxoquinoline-3-carboxamide (hereinafter referred to as "Compound 1"), which has the following structure:

the pharmaceutical composition of compound 1 can be effectively used for treating or reducing the severity of various diseases mediated by CFTR.

Drawings

FIG. 1 is an X-ray powder diffraction pattern of Compound 1.

FIG. 2 is a drawing of Compound 1¹H NMR spectrum chart.

FIG. 3 is a DSC of Compound 1.

Detailed Description

According to one embodiment, the present invention provides a pharmaceutical composition comprising:

(i) n- [2, 4-bis (1, 1-dimethylethyl) -5-hydroxyphenyl ] -1, 4-dihydro-4-oxoquinoline-3-carboxamide (Compound 1) or a pharmaceutically acceptable salt thereof;

(ii) suitable liquid PEG; and

(iii) optionally a suitable viscosity enhancing agent.

As used herein, the phrase "suitable liquid PEG" means a polyethylene glycol polymer that is in liquid form at ambient temperature and is suitable for use in pharmaceutical compositions. Such suitable polyethylene glycols are well known in the art; see, for example, http:// www.medicinescomplete.com/mc/excipients/current, which is incorporated herein by reference. Exemplary PEGs include low molecular weight PEGs such as PEG 200, PEG 300, PEG400, and the like. The numbers following the term "PEG" refer to the average molecular weight of a particular polymer, e.g., PEG400 is a polyethylene glycol polymer having a polymer average molecular weight of about 400.

In one embodiment, the suitable liquid PEG has an average molecular weight of about 200 to about 600. In another embodiment, the suitable liquid PEG is PEG400 (e.g., PEG having a molecular weight of about 380 to about 420 g/mol).

In another embodiment, the present invention provides a pharmaceutical composition comprising compound 1 or a pharmaceutically acceptable salt thereof; polyethylene glycol; and optionally a suitable viscosity enhancing agent.

In another embodiment, the pharmaceutical composition of the present invention comprises a suitable viscosity enhancing agent. In one embodiment, a suitable viscosity increasing agent is a polymer soluble in PEG. Such suitable viscosity enhancing agents are well known in the art, for example, polyvinylpyrrolidine (hereinafter "PVP"). PVP is characterized by its viscosity in aqueous solution, expressed as K-value (hereinafter conjugate, e.g., PVP K20), relative to water, ranging from about 10 to about 120. See, e.g., http:// www.medicinescomplete.com/mc/excipients/current. Embodiments of the PVP used in the present invention have a K-value of about 90 or less. An exemplary embodiment is PVP K30.

In one embodiment, the present invention provides a pharmaceutical composition comprising:

(i) n- (5-hydroxy-2, 4-di-tert-butyl-phenyl) -4-oxo-1H-quinoline-3-carboxamide (Compound 1) or a pharmaceutically acceptable salt thereof;

(ii) PEG 400; and

(iii)PVP K30。

in another embodiment, the present invention provides a pharmaceutical composition wherein said N- (5-hydroxy-2, 4-di-tert-butyl-phenyl) -4-oxo-1H-quinoline-3-carboxamide is present in an amount of about 0.01% w/w to about 6.5% w/w.

In another embodiment, the present invention provides a pharmaceutical composition wherein the PEG is present in an amount of about 87.5% w/w to about 99.99% w/w.

In another embodiment, the present invention provides a pharmaceutical composition wherein the PVP K30 is present in an amount between 0% w/w to about 6% w/w.

In another embodiment, the invention provides a pharmaceutical composition, wherein the composition comprises PEG400 (e.g., about 97.8 to about 98.0% w/w, e.g., about 97.88% w/w), PVP K30 (e.g., about 1.9 to about 2.1% w/w, e.g., about 2.0% w/w), and compound 1 (e.g., about 0.10 to about 0.15% w/w, e.g., about 0.13% w/w).

In another embodiment, the invention provides a pharmaceutical composition, wherein the composition comprises PEG400 (e.g., about 97.5 to about 98.0% w/w, e.g., about 97.75% w/w), PVP K30 (e.g., about 1.8 to about 2.2% w/w, e.g., about 2.0% w/w), and compound 1 (e.g., about 0.2 to about 0.3% w/w, e.g., about 0.25% w/w).

In another embodiment, the invention provides a pharmaceutical composition, wherein the composition comprises PEG400 (e.g., about 97.2 to about 97.8% w/w, e.g., about 97.50% w/w), PVP K30 (e.g., about 1.8 to about 2.2% w/w, e.g., about 2.0% w/w), and compound 1 (e.g., about 0.4 to about 0.6% w/w, e.g., about 0.50% w/w).

In another embodiment, the invention provides a pharmaceutical composition, wherein the composition comprises PEG400 (e.g., about 96.5 to about 97.5% w/w, e.g., about 97.0% w/w), PVP K30 (e.g., about 1.8 to about 2.2% w/w, e.g., about 2.0% w/w), and compound 1 (e.g., about 0.9 to about 1.1% w/w, e.g., about 1.0% w/w).

In another embodiment, the invention provides a pharmaceutical composition, wherein the composition comprises PEG400 (e.g., about 96.60 to about 96.65% w/w, e.g., about 96.63% w/w), PVP K30 (e.g., about 1.8 to about 2.2% w/w, e.g., about 2.0% w/w), and compound 1 (e.g., about 1.30 to about 1.45% w/w, e.g., about 1.38% w/w).

In another embodiment, the invention provides a pharmaceutical composition, wherein the composition comprises PEG400 (e.g., about 96.0 to about 96.3% w/w, e.g., about 96.12% w/w), PVP K30 (e.g., about 1.8 to about 2.0% w/w, e.g., about 2.0% w/w), and compound 1 (e.g., about 1.8 to about 2.2% w/w, e.g., about 1.88% w/w).

In another embodiment, the invention provides a pharmaceutical composition, wherein the composition comprises PEG400 (e.g., about 95.5 to about 96.0% w/w, e.g., about 95.75% w/w), PVP K30 (e.g., about 1.8 to about 2.2% w/w, e.g., about 2.0% w/w), and compound 1 (e.g., about 2.0 to about 2.5% w/w, e.g., about 2.25% w/w).

In another embodiment, the invention provides a pharmaceutical composition, wherein the composition comprises PEG400 (e.g., about 95 to about 96% w/w, e.g., about 95.5% w/w), PVP K30 (e.g., about 1.8 to about 2.2% w/w, e.g., about 2.0% w/w), and compound 1 (e.g., about 2.3 to about 2.7% w/w, e.g., about 2.50% w/w).

In another embodiment, the invention provides a pharmaceutical composition, wherein the composition comprises PEG400 (e.g., about 94.5 to about 94.8% w/w, e.g., about 94.63% w/w), PVP K30 (e.g., about 1.8 to about 2.2% w/w, e.g., about 2.0% w/w), and compound 1 (e.g., about 3.5 to about 4.0% w/w, e.g., about 3.38% w/w).

In another embodiment, the invention provides a pharmaceutical composition, wherein the composition comprises PEG400 (e.g., about 93.5 to about 94.5% w/w, e.g., about 94.0% w/w), PVP K30 (e.g., about 1.8 to about 2.2% w/w, e.g., about 2.0% w/w), and compound 1 (e.g., about 3.7 to about 4.3% w/w, e.g., about 4.0% w/w).

In one embodiment, the present invention provides a pharmaceutical composition comprising:

(i) n- (5-hydroxy-2, 4-di-tert-butyl-phenyl) -4-oxo-1H-quinoline-3-carboxamide (Compound 1) or a pharmaceutically acceptable salt thereof;

(ii) suitable PEG lipids; and

(iii)PVP。

in some embodiments, the PEG lipid has an average molecular weight of about 400 to about 600, such as PEG 400. In some embodiments, the PVP is PVP K30.

According to another embodiment, the pharmaceutical composition of the present invention comprises a therapeutically effective amount of compound 1. The phrase "therapeutically effective amount" refers to an amount effective to treat or alleviate any of the diseases, symptoms, or disorders described below.

Unless otherwise indicated, a structure described herein is also meant to include all isomeric forms of the structure (e.g., enantiomers, diastereomers, and geometric isomers (or conformers)); for example, the R and S configurations of each asymmetric center, (Z) and (E) double bond isomers, and (Z) and (E) conformational isomers. Thus, single stereochemical isomers as well as mixtures of enantiomers, diastereomers and geometric isomers (or conformers) of the compounds of the present invention are also within the scope of the present invention. Unless otherwise indicated, all tautomeric forms of the compounds of the invention are within the scope of the invention, e.g., compound 1 may exist as a tautomer:

unless otherwise indicated, structures described 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 (1) wherein one or more hydrogen atoms are replaced by deuterium or tritium, or one or more carbon atoms are replaced by 13C-or 14C-enriched carbon atoms, are within the scope of the present invention. Such compounds may be useful, for example, as analytical tools, probes in bioassays, or compounds with improved therapeutic properties.

Use, formulation and administration

Pharmaceutically acceptable compositions

In another aspect of the invention, pharmaceutically acceptable compositions are provided, wherein the compositions include additional pharmaceutically acceptable carriers, adjuvants or vehicles. In certain embodiments, these compositions optionally further comprise one or more additional therapeutic agents.

It is also envisioned that certain compounds of the present invention may exist in free form for therapeutic use or, where appropriate, in the form of a pharmaceutically acceptable derivative or prodrug thereof. According to the present invention, a pharmaceutically acceptable derivative or prodrug includes, but is not limited to, a pharmaceutically acceptable salt, ester, salt of such an ester, or any other adduct or derivative that, upon administration to a patient in need thereof, is capable of providing the compound, or a metabolite or residue thereof, directly or indirectly in other forms described herein.

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

Pharmaceutically acceptable salts of the compounds of the present invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable non-toxic acid addition salts are amino salts 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 using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include adipates, alginates, ascorbates, aspartates, benzenesulfonates, benzoates, bisulfates, borates, butyrates, camphorates, camphorsulfonates, citrates, cyclopentanepropionates, digluconates, dodecylsulfates, ethanesulfonates, formates, fumarates, glucoheptanoates, glycerophosphates, gluconates, hemisulfates, heptanoates, hexanoates, hydroiodides, 2-hydroxy-ethanesulfonates, lactobionates, lactates, laurates, malates, maleates, malonates, methanesulfonates, 2-naphthalenesulfonates, nicotinates, nitrates, oleates, oxalates, palmitates, pamoates, pectates, persulfates, 3-phenylpropionates, persulfates, benzoates, bisulfates, salts of acids, salts, Phosphates, picrates, pivalates, propionates, stearates, succinates, sulfates, tartrates, thiocyanates, p-toluenesulfonates, undecanoates, valerates, and the like. Salts derived from suitable bases including alkali metals, alkaline earth metals, ammonium and N⁺(C_1-4Alkyl radical)₄And (3) salt. The present invention also encompasses the quaternization of any basic nitrogen-containing group of the compounds as disclosed herein. By means of such quaternization, products which are soluble or dispersible in water or oil can be obtained. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Other pharmaceutically acceptable salts include, where appropriate, non-toxic ammonium salts, quaternary ammonium salts and amine cations, which are formed using counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkyl sulfonates and aryl sulfonates.

In one embodiment, the present invention provides a method of treating a CFTR mediated disease, symptom or disorder in a patient, the method comprising the step of administering to the patient a pharmaceutical composition according to the present invention.

As used herein, a "CFTR mediated disease" is a disease selected from the group consisting of: cystic fibrosis, hereditary emphysema, hereditary hemochromatosis, coagulation-fibrinolysis defects, e.g., protein C deficiency, type 1 hereditary angioedema, lipid processing defects, e.g., familial hypercholesterolemia, type 1 chylomicronemia, non-beta lipoproteinemia, lysosomal storage diseases, e.g., I-cell disease/pseudohelosis, mucopolysaccharidosis, Sanhoff/Tay-Saccharitis, type II Creutzfeldt-Jakob syndrome, polyendocrinopathy/hyperinsulinemia, diabetes, Radwarfism, myeloperoxidase deficiency, primary hypoparathyroidism, melanoma, CDG 1 type glycan, hereditary emphysema, congenital hyperthyroidism, osteogenesis imperfecta, hereditary hypofibrinogenemia, ACT deficiency, Diabetes Insipidus (DI), posterior leaflet hormonally-carried DI, renal DI, charcot-marie-tooth syndrome, peimeri's disease, neurodegenerative diseases such as alzheimer's disease, parkinson's disease, amyotrophic lateral sclerosis, progressive supranuclear palsy, pick's disease, several polyglutamine nervous system disorders such as huntington's disease, spinocerebellar ataxia type I, spinal and bulbar muscular atrophy, dentatorubral pallidoluysian atrophy, and myotonic atrophy, and spongiform encephalopathies such as hereditary creutzfeldt-jakob disease, Fabry disease, swiss-schneider syndrome, COPD, dry eye disease, and Sjogren's disease.

According to an alternative embodiment, the present invention provides a method of treating cystic fibrosis comprising the step of administering to said mammal a pharmaceutical composition according to the present invention.

In certain embodiments, the pharmaceutical compositions of the invention may be used to treat or reduce the severity of cystic fibrosis in patients who exhibit residual CFTR activity in the apical membrane of respiratory and non-respiratory epithelial cells. The presence of residual CFTR activity on the epithelial cell surface can be readily detected using methods known in the art, such as standard electrophysiological, biochemical or histochemical techniques. Such methods use in vivo or ex vivo electrophysiological techniques, measurement of Cl "concentration in sweat or saliva, or ex vivo biochemical or histochemical techniques to monitor the density of cell surfaces to identify CFTR activity. Using such methods, residual CFTR activity can be readily detected in patients who are heterozygous or not heterozygous for a number of different variants, including patients who are not heterozygous or heterozygous for the most common variant, af 508.

In one embodiment, the pharmaceutically acceptable compositions of the invention may be used to treat or reduce the severity of cystic fibrosis in a patient having a certain gene Type that shows residual CFTR activity, such as a class III variant (impaired regulation or gating), a class IV variant (altered Conductance), or a class V variant (reduced synthesis) (LeeR. Choo-Kang, Pamela L., Zeitinin, Type I, II, III, IV, and Vcstic fibrosis tarsomemembrane conductor Regulator Defects and opportunities of Therapy 6: 521-. Other patient gene types that exhibit residual CFTR activity include patients who are not heterozygous for one of these types, or who are heterozygous for any other type of variant, including class I variants, class II variants, or unclassified variants.

In one embodiment, the pharmaceutically acceptable compositions of the invention can be effectively used in methods of treating or reducing the severity of cystic fibrosis in a patient having a clinical phenotype, such as a moderate to mild clinical phenotype typically associated with the amount of residual CFTR activity in the apical membrane of epithelial cells. Such phenotypes include patients exhibiting pancreatic defects or diagnosed with congenital pancreatitis and congenital bilateral vasectomy, or mild pulmonary disease.

The exact amount of compound 1 required in the pharmaceutical composition of the present invention will vary depending on the subject, depending on the species, age and general condition of the subject, the severity of the infection, the particular drug, its administration form and the like. The compounds of the present invention are preferably formulated in dosage unit form for ease of administration and uniformity of dosage. The expression "dosage unit form" as used herein denotes a physically discrete pharmaceutical unit, which is adapted to the patient to be treated. It will be understood, however, that the total daily amount of the compounds and compositions of the present invention will be determined 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 particular compound used; the specific composition used; the age, weight, health, sex, and diet of the patient; the time of administration, the route of administration, and the rate of excretion of the particular compound employed; the duration of the treatment; drugs used in combination or concomitantly with the specific compound employed; and other factors well known in the medical arts. The term "patient" as used herein means an animal, preferably a mammal, most preferably a human.

The pharmaceutically acceptable compositions of the present invention may be administered orally at a dosage level of from about 0.01mg/kg to about 50mg/kg, preferably from about 1mg/kg to about 25mg/kg, once or more a day, based on the weight of the subject, to achieve the desired therapeutic effect.

The pharmaceutical compositions of the present invention may additionally 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. In addition to inert diluents, oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

It is also envisioned that the pharmaceutical compositions of the present invention may be used in combination therapy, that is, they may be administered simultaneously, prior to, or after one or more other desired therapeutic agents or medical procedures. The particular combination of therapies (therapies or procedures) used in a combination regimen will take into account the compatibility of the desired therapeutic agent and/or procedure with the desired therapeutic effect to be achieved. It is contemplated that the therapies used may achieve the desired effect on the same disorder (e.g., the pharmaceutical compositions of the present invention may be administered simultaneously with other agents used to treat the same disorder), or they may achieve different effects (e.g., control of any side effects). As used herein, other therapeutic agents that are typically administered to treat or prevent a particular disease or condition are said to be "appropriate for the disease or condition being treated.

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

The amount of the other therapeutic agent present in the compositions of the present invention does not exceed that which would normally be administered in a composition containing the therapeutic agent as the only active agent. Preferred amounts of the other therapeutic agent in the compositions disclosed herein are from 50% to 100% of the amount typically present in a composition comprising said agent as the sole therapeutically active agent.

In order that the invention described herein may be more fully understood, the following examples are now disclosed. It is to be understood that these examples are for illustrative purposes only and are not to be construed as limiting the invention in any way.

Examples

Method and material

Differential Scanning Calorimetry (DSC)

DSC data were collected on a TA apparatus Q1000 equipped with a 50-bit autosampler. The energy and temperature calibration standard is indium. The sample was heated at a rate of 10 c/min between 20 and 350 c. A 30 ml/min nitrogen purge was maintained on the sample.

Between 0.5 and 4mg of sample was used and all samples were run in a small hole aluminum pan.

NMR

All spectra were collected at Bruker 400MHz with an autosampler. Unless otherwise indicated, in d₆Samples were prepared in DMSO.

XRPD (X-ray powder diffraction)

Bruker AXS C2 GADDS diffractometer

The X-ray powder diffractogram of the sample was obtained on a Bruker AXS C2 GADDS diffractometer using Cu ka radiation (40kV, 40mA) in the autorun XYZ stage, a laser video microscope for determining the autoinjection position and a HiStar 2-dimensional area detector. The X-ray optical part is a single unit connected with a pinhole sight of 0.3mmA multilayer mirror.

The beam divergence, i.e. the effective size of the X-ray beam on the sample, is about 4 mm. Using a theta-theta continuous scan mode, the sample to detector distance is 20cm, which gives an effective 2 theta range of 3.2-29.8 deg.. The typical exposure time for the sample was 120 s.

The powder was prepared as a flat plate sample as a sample for ambient handling without grinding after the sample was received. Approximately 1-2mg of the sample was gently pressed onto a glass slide to obtain a smooth surface. Samples operating under non-ambient conditions were placed on a silicon wafer with a thermally conductive compound. The sample was then heated to the appropriate temperature at a rate of approximately 20 deg.C/min, then held isothermal for approximately 1 minute before data collection was initiated.

Synthesis of N- (5-hydroxy-2, 4-di-tert-butyl-phenyl) -4-oxo-1H-quinoline-3-carboxamide (Compound 1):

2-Phenylaminomethylene malonic acid diethyl ester

A mixture of aniline (25.6g, 0.275mol) and diethyl 2- (ethoxymethylene) malonate (62.4g, 0.288mol) was heated at 140 ℃ for 2 hours. The mixture was cooled to room temperature and dried under reduced pressure to obtain 2-phenylaminomethylene-malonic acid diethyl ester as a solid, which was used in the next step without further purification.¹HNMR(DMSO-d₆)δ 11.00(d，1H)，8.54(d，J＝13.6Hz，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

A1L three-necked flask with a mechanical stirrer was charged with diethyl 2-phenylaminomethylene-malonate (26.3g, 0.100mol), polyphosphoric acid (270g), and phosphorus oxychloride (750 g). The mixture was heated to 70 ℃ and stirred for 4 h. The mixture was cooled to room temperature and filtered. The residue is taken up with Na₂CO₃The aqueous solution was treated, filtered, washed with water and dried to obtain ethyl 4-hydroxyquinoline-3-carboxylate (15.2g, 70%) as a pale brown solid. The crude product can be used in the next step without further purification.

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

Ethyl 4-hydroxyquinoline-3-carboxylate (15g, 69mmol) was suspended in sodium hydroxide solution (2N, 150mL) and stirred at reflux for 2 h. After cooling, the mixture was filtered and the filtrate was acidified to pH4 with 2N HCl. The resulting precipitate was collected by filtration, washed with water, and dried in vacuo to obtain 4-oxo-1, 4-dihydroquinoline-3-carboxylic acid (10.5g, 92%) as an off-white solid.¹H NMR(DMSO-d₆)δ 15.34(s，1H)，13.42(s，1H)，8.89(s，1H)，8.28(d，J＝8.0Hz，1H)，7.88(m，1H)，7.81(d，J＝8.4Hz，1H)，7.60(m，1H)。

2, 4-di-tert-butylphenyl carbonate methyl ester

To 2, 4-di-tert-butylphenol (103.2g, 500mmol), Et, cooled to 0 ℃ in an ice-water bath was added₃Methyl chloroformate (58mL, 750mmol) was added dropwise to a solution of N (139mL, 1000mmol) and DMAP (3.05g, 25mmol) in dichloromethane (400 mL). The mixture was warmed to room temperature, kept stirring overnight, and then filtered through silica gel (approximately 1L) using 10% ethyl acetate-hexane (-4L) as eluent. The combined filtrates were concentrated to obtain 2, 4-di-tert-butyl-phenyl carbonate methyl ester as a yellow oil (132g, equiv. reaction).¹H NMR(400MHz，DMSO-d₆)δ 7.35(d，J＝2.4Hz，1H)，7.29(dd，J＝8.5，2.4Hz，1H)，7.06(d，J＝8.4Hz，1H)，3.85(s，3H)，1.30(s，9H)，1.29(s，9H)。

2, 4-di-tert-butyl-5-nitro-phenyl carbonate methyl ester and 2, 4-di-tert-butyl-6-nitro-phenyl carbonate methyl ester

To a stirred mixture of 2, 4-di-tert-butyl-phenyl methyl carbonate (4.76g, 180mmol) in concentrated sulfuric acid (2mL) cooled in an ice-water bath was added a cooled mixture of sulfuric acid (2mL) and nitric acid (2 mL). The addition is carried out slowly so that the reaction temperature does not exceed 50 ℃. The reaction was stirred for 2 hours while warming to room temperature. The reaction mixture was then added to ice water and extracted into diethyl ether. Dry Ether layer (MgSO)₄) Concentrated and purified by column chromatography (0-10% ethyl acetate-hexane) to obtain a mixture of 2, 4-di-tert-butyl-5-nitro-phenyl methyl carbonate and 2, 4-di-tert-butyl-6-nitro-phenyl methyl carbonate (4.28g) as a pale yellow solid, which was used directly in the next step.

2, 4-di-tert-butyl-5-nitro-phenol and 2, 4-di-tert-butyl-6-nitro-phenol

A mixture of 2, 4-di-tert-butyl-5-nitro-phenyl methyl carbonate and 2, 4-di-tert-butyl-6-nitro-phenyl methyl carbonate (4.2g, 14.0mmol) was dissolved in MeOH (65mL) and KOH (2.0g, 36mmol) was added. The mixture was stirred at room temperature for 2 hours. The reaction mixture was then made acidic (pH 2-3) by addition of concentrated HCl and partitioned between water and diethyl ether. Dry Ether layer (MgSO)₄) Concentrated and purified by column chromatography (0-5% ethyl acetate-hexane) to obtain 2, 4-di-tert-butyl-5-nitro-phenol (1.31g, 29% yield in two steps) and 2, 4-di-tert-butyl-6-nitro-phenol. 2, 4-di-tert-butyl-5-nitro-phenol:¹H NMR(400MHz，DMSO-d₆) 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-tert-butyl-6-nitro-phenol:¹H NMR(400MHz，CDCl₃)δ11.48(s，1H)，7.98(d，J＝2.5Hz，1H)，7.66(d，J＝2.4Hz，1H)，1.47(s，9H)，1.34(s，9H)。

5-amino-2, 4-di-tert-butyl-phenol

To a mixture of 2, 4-di-tert-butyl-5-nitro-phenol (1.86g, 7.40mmol) and formamide (1.86g) in ethanol (75 m)L) to the reflux solution was added Pd-5% wt on activated carbon (900 mg). The reaction mixture was stirred at reflux for 2 hours, cooled to room temperature, and filtered through celite. Celite was washed with methanol and the combined filtrates were concentrated to give 5-amino-2, 4-di-tert-butyl-phenol (1.66g, equiv. reaction) as a grey solid.¹H NMR(400MHz，DMSO-d₆)δ 8.64(s，1H，OH)，6.84(s，1H)，6.08(s，1H)，4.39(s，2H，NH₂) 1.27(m, 18H); HPLC retention time 2.72 min, 10-99% CH₃CN, running for 5 minutes; ESI-MS 222.4M/z [ M + H ]]⁺。

N- (5-hydroxy-2, 4-di-tert-butyl-phenyl) -4-oxo-1H-quinoline-3-carboxamide

To a suspension of 4-oxo-1, 4-dihydroquinoline-3-carboxylic acid (35.5g, 188mmol) and HBTU (85.7g, 226mmol) in DMF (280mL) at ambient temperature was added Et₃N (63.0mL, 451 mmol). The mixture became homogeneous and was stirred for 10 minutes, then 5-amino-2, 4-di-tert-butyl-phenol (50.0g, 226mmol) was added in small portions. The mixture was stirred at ambient temperature overnight. The mixture becomes heterogeneous during the reaction. After complete consumption of the acid (LC-MS analysis, MH +190, 1.71 min), the solvent was removed under vacuum. EtOH was added to the orange solid material, resulting in a slurry. The mixture was stirred for 15 minutes on a rotary evaporator (bath temperature 65 ℃) without placing the system under vacuum. The mixture was filtered, and the resulting solid was washed with hexane to obtain a white solid, which was EtOH crystalline. Et was added to the material obtained above₂O, until a slurry is formed. The mixture was stirred for 15 minutes on a rotary evaporator (bath temperature 25 ℃) without placing the system under vacuum. The mixture was filtered and the solid collected. The procedure was performed five times in total. The solid obtained after the fifth precipitation was placed under vacuum overnight to obtain 8N- (5-hydroxy-2, 4-dibutyl-phenyl) -4-oxo-1H-quinoline as a white powdery solidQuinoline-3-carboxamide (38g, 52%).

HPLC retention time 3.45 min, 10-99% CH₃CN, running for 5 minutes;¹H NMR(400MHz，DMSO-d₆)δ12.88(s，1H)，11.83(s，1H)，9.20(s，1H)，8.87(s，1H)，8.33(dd，J＝8.2，1.0Hz，1H)，7.83-7.79(m，1H)，7.76(d，J＝7.7Hz，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 393.3m/z[M+H]⁺。

disclosed below are characteristic data for compound 1:

TABLE 2

Compound numbering	LC-MS M+1	LC-RT min
			1	393.2	3.71

The XRPD spectrum of compound 1 is shown in figure 1.

Process for preparation of Compound 1¹The H NMR data are shown in FIG. 2.

The DSC chart of compound 1 is shown in FIG. 3.

Preparation of pharmaceutical compositions

Materials:

● glass bottle for preparation (250 cc dark brown glass bottle with Teflon lining cover)

● glass bottle for confirming sample dosage (30 cc dark brown glass bottle with Teflon lining cover)

● mixing disk with temperature probe (ensure the probe has been cleaned)

● novel magnetic stirring rod

● spatula to dispense excipients and active ingredients.

Step 1: to a clean 250cc dark brown glass bottle was added a stir bar and the tare weight of the bottle, stir bar, label and lid was recorded. Weigh the bottle with label and stir bar.

Step 2: a target amount of PEG400 was dispensed in the vial and accurately weighed. The vial was placed on a stir plate and stirred to form a small vortex (300 and 500rpm or as needed) on the surface of the liquid. A clean temperature probe was inserted into a liquid depth of-1 cm and the heater set point was raised to 40 ℃. The mouth of the bottle was covered with aluminum foil. PEG400 was stabilized at 40+/-5 ℃.

And step 3: the required amount of PVP K30 was dispensed and added to the stirred PEG 400. PVP was added slowly (over 2-3 minutes) and the particles were dispersed. If the particles agglomerate, dissolution will take longer. The bottle mouth was covered with foil and the mixture was stirred further at 40+/-5 ℃. The mixture was sampled at 10 minutes using a small pipette to determine if the PVP was completely dissolved. Large undissolved lumps in the stirred solution should also be checked. If the solution is clear, the next step is performed. If undissolved polymer is present, stirring is continued. The dissolution was checked every 10 minutes, the maximum stirring time being 30 minutes in total. When complete dissolution was observed, the next step was followed, and if complete dissolution was not observed within 30 minutes after PVP addition, the preparation was terminated, the material was discarded and the preparation was restarted from scratch.

And 4, step 4: the required amount of compound 1 was dispersed and slowly added to the stirred PEG/PVP solution. The bottle mouth was covered with foil and the mixture was stirred further at 40+/-5 ℃. The mixture was sampled after 30 minutes using a small pipette to determine if compound 1 had completely dissolved. After 30 minutes if the solution is clear, the next step is carried out. If undissolved compound 1 is present, stirring is continued. Dissolution was checked every 30 minutes, with a maximum stirring time of 300 minutes (5 hours) after addition of compound 1. If complete dissolution is not observed within 300 minutes (5 hours) after addition of compound 1, the preparation is terminated, the material is discarded and the preparation is restarted from scratch.

After complete dissolution of compound 1, it was removed from the stir plate and capped. The formulation should be kept at room temperature until dosing is performed, but must be dosed within 24 hours after preparation. If precipitation of VX-770 is observed, quantification of the solution is not performed.

Using the above method, the following ten pharmaceutical compositions shown in table a were prepared:

TABLE A

Composition #	％PEG400w/w	％PVP K30w/w	% compound 1w/w	Amount of Compound 1 per 20g of formulation (mg)
					1	97.875	2.0	0.125	25
2	97.750	2.0	0.250	50
					3	97.500	2.0	0.500	100
4	97.000	2.0	1.000	200
					5	96.625	2.0	1.375	275
6	96.125	2.0	1.875	375
					7	95.750	2.0	2.25	450
8	95.500	2.0	2.500	500
					9	94.625	2.0	3.375	675
10	94.000	2.0	4.000	800

Claims (21)

1. A pharmaceutical composition comprising:

(i) n- (5-hydroxy-2, 4-di-tert-butyl-phenyl) -4-oxo-1H-quinoline-3-carboxamide or a pharmaceutically acceptable salt thereof;

(ii) liquid PEG with an average molecular weight of 200-600; and

(iii) optionally PVP.

2. The pharmaceutical composition according to claim 1, wherein the composition comprises PVP.

3. The pharmaceutical composition according to claim 2, wherein the PVP has a K-value of 30 or less.

4. A pharmaceutical composition according to claim 3, wherein the PVP has a K-value of 30.

5. The pharmaceutical composition according to claim 2, wherein the suitable liquid PEG has an average molecular weight of 400.

6. The pharmaceutical composition according to claim 1, comprising:

(i) n- (5-hydroxy-2, 4-di-tert-butyl-phenyl) -4-oxo-1H-quinoline-3-carboxamide;

(ii) PEG 400; and

(iii)PVP K 30。

7. the pharmaceutical composition according to claim 1, wherein the N- (5-hydroxy-2, 4-di-tert-butyl-phenyl) -4-oxo-1H-quinoline-3-carboxamide is present in an amount between 0.01% w/w and 6.5% w/w.

8. The pharmaceutical composition according to claim 1, wherein the PEG is present in an amount between 87.5% w/w to 99.99% w/w.

9. The pharmaceutical composition according to claim 1, wherein the polyvinylpyrrolidone is present in an amount between 0% w/w and 6% w/w.

10. The pharmaceutical composition according to claim 1, wherein the composition comprises PEG 40097.875% w/w, PVP K302.0% w/w, and N- (5-hydroxy-2, 4-di-tert-butyl-phenyl) -4-oxo-1H-quinoline-3-carboxamide 0.125% w/w,

or wherein the composition comprises PEG 40097.75% w/w, PVP K302.0% w/w, and N- (5-hydroxy-2, 4-di-tert-butyl-phenyl) -4-oxo-1H-quinoline-3-carboxamide 0.25% w/w,

or wherein the composition comprises PEG 40097.5% w/w, PVP K302.0% w/w, and N- (5-hydroxy-2, 4-di-tert-butyl-phenyl) -4-oxo-1H-quinoline-3-carboxamide 0.5% w/w,

or wherein the composition comprises PEG 40097.0% w/w, PVP K302.0% w/w, and N- (5-hydroxy-2, 4-di-tert-butyl-phenyl) -4-oxo-1H-quinoline-3-carboxamide 1.0% w/w,

or wherein the composition comprises PEG 40096.625% w/w, PVP K302.0% w/w, and N- (5-hydroxy-2, 4-di-tert-butyl-phenyl) -4-oxo-1H-quinoline-3-carboxamide 1.375% w/w,

or wherein the composition comprises PEG 40096.12% w/w, PVP K302.0% w/w, and N- (5-hydroxy-2, 4-di-tert-butyl-phenyl) -4-oxo-1H-quinoline-3-carboxamide 1.88% w/w,

or wherein the composition comprises PEG 40095.75% w/w, PVP K302.0% w/w, and N- (5-hydroxy-2, 4-di-tert-butyl-phenyl) -4-oxo-1H-quinoline-3-carboxamide 2.25% w/w,

or wherein the composition comprises PEG 40095.5% w/w, PVP K302.0% w/w, and N- (5-hydroxy-2, 4-di-tert-butyl-phenyl) -4-oxo-1H-quinoline-3-carboxamide 2.5% w/w,

or wherein the composition comprises PEG 40094.625% w/w, PVP K302.0% w/w, and N- (5-hydroxy-2, 4-di-tert-butyl-phenyl) -4-oxo-1H-quinoline-3-carboxamide 3.375% w/w,

or wherein the composition comprises PEG 40094.0% w/w, PVP K302.0% w/w, and N- (5-hydroxy-2, 4-di-tert-butyl-phenyl) -4-oxo-1H-quinoline-3-carboxamide 4.0% w/w).

11. Use of a pharmaceutical composition according to claim 1 in the manufacture of a medicament for treating a CFTR mediated disease in a patient.

12. Use according to claim 11, wherein the disease is selected from the group consisting of cystic fibrosis, hereditary emphysema, hereditary hemochromatosis, coagulation-fibrinolysis defects, lipid processing defects, lysosomal storage diseases, neurodegenerative diseases, polyglutamate nervous system disorders and spongiform encephalopathy.

13. The use according to claim 12, wherein the coagulation-fibrinolysis defect is selected from the group consisting of a protein C defect and hereditary angioedema type 1.

14. Use according to claim 12, wherein the lipid processing deficiency is selected from the group consisting of familial hypercholesterolemia, chylomicronemia type 1 and betalipoproteinemia.

15. The use according to claim 12, wherein the lysosomal storage disease is selected from the group consisting of I-cell disease/pseudoheller's disease, mucopolysaccharidosis, sandhoff disease/tay-sachs disease, crenella syndrome type II, polyendocrinosis/hyperinsulinemia, diabetes, ralon dwarfism, myeloperoxidase deficiency, primary hypoparathyroidism, melanoma, CDG type 1 glycan, hereditary emphysema, congenital hyperthyroidism, osteogenesis imperfecta, hereditary hypofibrinogenemia, ACT deficiency, diabetes insipidus, charcot-marie-tooth syndrome and peliza-mei disease.

16. The use according to claim 15, wherein said diabetes insipidus is a posterior lobe hormone transporter diabetes insipidus or a renal diabetes insipidus.

17. The use according to claim 12, wherein the neurodegenerative disease is selected from the group consisting of alzheimer's disease, parkinson's disease, amyotrophic lateral sclerosis, progressive supranuclear palsy and pick's disease.

18. Use according to claim 12, wherein the polyglutamate nervous system disorder is selected from huntington's disease, spinocerebellar ataxia type I, spinal and bulbar muscular atrophy, dentatorubral globus thalamic atrophy and myotonic atrophy.

19. The use according to claim 12, wherein the spongiform encephalopathy is selected from hereditary Creutzfeldt-Jakob disease, Fabry disease, Sjogren's syndrome, COPD, dry eye disease and Sjogren's disease.

20. Use according to claim 12, wherein the disease is cystic fibrosis.

21. The use according to claim 11, wherein the pharmaceutical composition is administered once daily to a patient in need thereof.