Modulators of cystic fibrosis transmembrane conductance regulator

[0001] The present invention relates to modulators of cystic fibrosis transmembrane conductance regulator ("CFTR"), their compositions and methods related thereto. The present invention also relates to methods of treating diseases using CFTR modulators.

BACKGROUND OF THE INVENTION [0002] Transporters containing an ATP binding cassette are a family of membrane transporter proteins that regulate the transport of many different pharmacological agents, potentially toxic drugs and xenobiotics, and anions. They are homogeneous membrane proteins that bind and use cellular adenosine triphosphate (ATP) for their specific activities. Some of these transporters have been discovered as multi-drug resistance proteins (such as MDR1-P glycoprotein or multi-drug resistance protein, MRP1) that protect malignant tumor cells from chemotherapeutic agents. To date, 48 such transporters have been identified and based on sequence and activity identity they have been divided into 7 families.

[0003] One member of the transporter family containing the ATP binding cassette usually associated with the disease is the anAMP channel mediated by cAMP / ATP, CFTR. CFTR is expressed in a wide variety of cell types, including absorbing and secreting epithelial cells, where it regulates anion flow across membranes and the activity of other ion channels and proteins. In epithelial cells, the proper functioning of CFTR plays a key role in maintaining electrolyte transport in the body, including respiratory and digestive tissue. CFTR consists of about 1480 amino acids that 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, polar regulatory (R) domain having multiple phosphorylation sites that regulates channel activity and its movement in the cell.

[0004] The CFTR coding gene has been identified and sequenced (see Gregory, RJ et al. (1990) Nature 347: 382-386; Rich, DP et al. (1990) Nature 347: 358-362), (Riordan, JR and in (1989) Science 245: 1066-1073). A defect in this gene causes mutations in CFTR leading to cystic fibrosis ("CF"), the most common fatal genetic disease in humans. About one in 2,500 babies in the United States has cystic fibrosis. In the general US population, up to 10 million people carry a single copy of a defective gene with no apparent adverse effects. In contrast, people who have two copies of the CF-related gene suffer from the debilitating and fatal effects of CF, including chronic lung disease.

[0005] In patients with cystic fibrosis, mutations in CFTR endogenously expressed in the respiratory epithelium lead to reduced peak anion secretion, resulting in an imbalance in ion and fluid transport. As a result, anion transport is reduced, which contributes to increased mucus accumulation in the lung and associated microbial infections that ultimately lead to the death of CF patients. In addition to respiratory disease, CF patients typically suffer from gastrointestinal problems and pancreatic failure, which, if left untreated, are fatal. In addition, most men with cystic fibrosis are infertile and the fertility of women with cystic fibrosis is reduced. In contrast to the severe consequences of the presence of two copies of the CF-associated gene, individuals with a single copy of the CF-associated gene exhibit increased resistance to cholera and dehydration due to diarrhea - which perhaps explains the relatively high frequency of the CF gene in the population. [0006] Sequence analysis of the CFTR gene from CF chromosomes revealed the presence of various disease-causing mutations (Cutting, GR et al (1990) Nature 346: 366-369; Dean, M. et al. (1990) Cell 61: 863: 870; and Kerem, BS. et al. (1989) Science 245: 10731080; Kerem, BS et al. (1990) Proc. Natl. Acad. Sci. USA 87: 8447-8451). To date, over 1,000 mutations in the CF gene have been identified that cause disease (<a href="http://www.genet.sickkids.on.ca/cftr/">http://www.genet.sickkids.on.ca/cftr/</a>). The most common mutation is a phenylalanine deletion at position 508 of the CFTR amino acid sequence, often referred to as ΔF508-CFTR. This mutation occurs in about 70 percent of cases of cystic fibrosis and is associated with severe disease.

[0007] Deletion of residue 508 in ΔF508-CFTR prevents correct folding of the resulting protein. This makes the mutated protein unable to exit the endoplasmic reticulum and transport to the cell membrane. As a result, the number of channels present in the membrane is much smaller than that found in cells expressing wild-type CFTR. In addition to impaired cellular transport, the mutation leads to faulty channel gating. Together, the reduced number of channels in the membrane and defective gating lead to reduced transport of anions across the epithelium, leading to defective transport of ions and fluids. (Quinton, PM (1990), FASEB J. 4: 27092727). However, studies have shown that the reduced number of ΔF508-CFTR in the membrane retains its function, although it is reduced compared to wild type CFTR. (Dolmans et al. (1991), Nature Lond. 354: 526-528; Denning et al., Supra; Pasyk and Foskett (1995), J. Cell. Biochem. 270: 12347-50). In addition to ΔF508-CFTR, R117H-CFTR and G551D-CFTR, other disease causing mutations in CFTR that lead to defective intracellular transport, synthesis and / or gating of the channel can be regulated up or down, changing anion secretion and modifying progress and / or severity of the disease. [0008] Although CFTR transports a variety of molecules in addition to anions, this role (anion, chloride and bicarbonate transport) is undoubtedly one of the elements of an important mechanism for transporting ions and water across the epithelium. Other elements are the epithelial Na channel⁺, ENaC, co-transporter Na⁺/ 2Cl^-/ K⁺, pump Na⁺-K⁺-ATPaza and K channels⁺ in the basolateral membrane, which are responsible for the uptake of chloride into the cell. [0009] These elements interact with each other in directional transport across the epithelium due to their selective expression and location within the cell. Chloride absorption occurs through coordinated action of ENaC and CFTR occurring on the apical membrane and Na pump⁺-K⁺-ATPase and C1 channels expressed on the side of the basilar cell. Secondary active chloride transport from the luminal side leads to the accumulation of chloride inside the cell, and then the chloride can passively leave the cell through Cl ion channels^-, which leads to vector transport. Arranging the transporter Na⁺/ 2Cl^-/ K⁺, Na pumps⁺-K⁺-ATPazy and K channels⁺ laterol basal membrane on the laterol basal surface, and CFTR on the luminal side coordinates chloride secretion by CFTR on the luminal side. Because water alone is probably never actively transported, its flow through the epithelium depends on the tiny transepithelial osmotic gradients produced by the convective flow of sodium and chloride.

[0010] The abnormal bicarbonate transport resulting from the CFTR mutation is believed to cause defects in some secretory functions. See, e.g., "Cystic fibrosis: impaired bicarbonate secretion and mucoviscidosis," Paul M. Quinton, Lancet 2008; 372: 415-417.

[0011] Mutations in CFTR, which are associated with moderate CFTR dysfunction, are also observed in patients with conditions that have some CF disease symptoms but do not meet the CF diagnostic criteria. These include congenital bilateral vas deferens, idiopathic chronic pancreatitis, chronic bronchitis and chronic rhinitis and sinusitis. Other diseases in which the mutated CFTR is considered a risk factor, including modifying genes or environmental factors, include primary sclerosing cholangitis, allergic bronchopulmonary aspergillosis and asthma.

[0012] Cigarette smoke, hypoxia and environmental factors that trigger hypoxia signal transmission have also been shown to interfere with CFTRs and may contribute to some form of respiratory disease such as chronic bronchitis. Diseases that may result from CFTR malfunction but do not meet the CF diagnostic criteria are characterized as CFTR-related diseases.

[0013] In addition to cystic fibrosis, modulation of CFTR activity may be beneficial in other diseases that are not directly caused by mutations in CFTR, such as secretory diseases and CFTR mediated protein folding diseases. CFTR regulates chloride and bicarbonate flow through epithelials from many cells by controlling fluid flow, protein dissolution, mucus viscosity and enzymatic activity. CFTRs can cause blockage of the airways or ducts in many organs, including the liver and pancreas. Potentiators are compounds that increase the gating activity of CFTR present in the cell membrane. A candidate for treatment with tonic substances can be any disease that includes thickening of the mucus, fluid regulation disorder, mucus clearance disorder, or blockage of the ducts leading to inflammation and tissue destruction.

[0014] The above include, but are not limited to, chronic obstructive pulmonary disease (COPD), asthma, smoke-induced COPD, chronic bronchitis, rhinitis and sinusitis, constipation, dry eye disease, and Sjogren's syndrome, gastroesophageal reflux disease esophageal, gallstones, prolapse of the rectum, and inflammatory bowel disease. COPD has a limited airflow that is progressive and not fully reversible. The restriction of airflow is caused by excessive mucus secretion, emphysema and bronchiolitis. Mutant or wild type CFTR activators are potential treatments for mucus hypersecretion and impaired mucociliary clearance commonly associated with COPD. In particular, the increased anion secretion by CFTR can facilitate fluid transport to the surface fluid of the airways, hydrating mucus and optimizing the viscosity of periciliary fluid. This can lead to increased mucociliary clearance and a reduction in the symptoms associated with COPD. In addition, by preventing ongoing infection and inflammation by improving airway cleansing, CFTR modulators can counteract or slow down the destruction of airway parenchyma that characterizes emphysema and reduce or reverse the increase in the number and size of mucus secreting cells that underlies mucus hypersecretion. respiratory diseases. Dry eye disease is characterized by a decrease in the production of tear fluid and an abnormal profile of tear film lipids, proteins and mucin. There are many reasons for dry eye; some of them are: age, Lasik ophthalmic surgery, arthritis, drugs, chemical / thermal burns, allergies and diseases such as cystic fibrosis and Sjogren's syndrome. Increased anion secretion by CFTR may increase fluid transport through the corneal epithelial cells and secretory glands surrounding the eye, with increased corneal hydration. This can help alleviate the symptoms of dry eye disease. Sjogren's syndrome is an autoimmune disease in which the immune system attacks the glands that produce fluids throughout the body, including the eye, mouth, skin, respiratory tissue, liver, vagina, and intestine. Symptoms include dry eyes, mouth and vagina, and lung disease. This disease is also associated with rheumatoid arthritis, systemic lupus erythematosus, systemic sclerosis, and polymyositis / dermatomyositis. Defective intracellular protein transport is thought to cause this disease in which treatment options are limited. Modulators of CFTR activity can hydrate various affected organs and help relieve related symptoms. People with cystic fibrosis have recurrent episodes of intestinal obstruction and a higher incidence of rectal prolapse, gallstones, gastroesophageal reflux disease, GI malignancy, and inflammatory bowel disease, which indicates that CFTR may play an important role in preventing such diseases.

[0015] As discussed above, deletion of residue 508 in ΔF508-CFTR is believed to prevent proper folding of the resulting protein, rendering it unable to exit the endoplasmic reticulum and transport to the cell membrane. As a result, there is not enough mature protein in the cell membrane and chloride transport in epithelial tissue is significantly reduced. In fact, it has been shown that this cellular phenomenon of defective processing in the CFTR endoplasmic reticulum by endoplasmic reticulum machinery underlies not only CF disease, but also a wide range of other isolated and congenital diseases. Two ways in which endoplasmic mesh machinery may malfunction are loss of conjugation with export from endoplasmic mesh of proteins leading to their degradation or accumulation of these defective / poorly folded proteins [Aridor M, et al., Nature Med., 5 (7), pp. 745-751 (1999); Shastry, BS, et al., Neurochem. International, 43, pp. 1-7 (2003); Rutishauser, J., et al., Swiss Med Wkly, 132, pp. 211-222 (2002); Morello, JP et al., TIPS, 21, p. 466-469 (2000); Bross P., et al., Human Mut., 14, pp. 186-198 (1999)]. Diseases associated with the first class of endoplasmic reticulum malfunction are: cystic fibrosis (caused by poor folding of ΔF508-CFTR, as discussed above), congenital emphysema (caused by α1-antitrypsin; variant without PiZ), congenital hemochromatosis, clotting deficiencies - fibrinolysis, such as protein C deficiency, congenital angioedema type 1, lipid processing disorders, such as congenital hypercholesterolemia, chylomicronemia type 1, abetalipoproteinemia, lysosomal storage diseases, such as cellular disease pseudo Hurler, mucopolysaccharidosis (caused by lysosomal processing enzymes), Sandhof / TaySachs syndrome (caused by β-hexosaminidase), Crigler-Najjar syndrome type II (caused by UDP-glucuronyl-sialyl transferase), polyiendocrinopathy / hyperinsulinemia, diabetes mellitus (caused by insulin receptor), Laron's dwarfism (caused by growth hormone receptor), myeloperoxidase deficiency, primary hypoparathyroidism (caused by preproparathus) due to tyrosinase). Diseases associated with this second class of endoplasmic malfunction are: type 1 CDG glycanosis, congenital emphysema (caused by a1-antitrypsin; PiZ variant), congenital hyperthyroidism, congenital bone fragility (caused by type I, II, IV procollagen), congenital hypofibrinogenemia (caused by fibrinogen), ACT deficiency (caused by αΐ-antichymotrypsin), diabetes insipidus (DI), neuronal pituitary hormone (caused by vasopressin hormone) / V2 receptor), neurogenic DI (caused by aquaporin II), Charcot-Marie-Tooth syndrome (caused by peripheral myelin protein 22), Pelizaeus-Merzbacher disease, neurodegenerative diseases, such as Alzheimer's disease (caused by βΑΡΡ and presenilins), Parkinson's disease, amyotrophic lateral sclerosis, progressive supranuclear palsy, Pick's disease, various polyglutamine neurological disorders such as Huntington's disease, type I cerebellar ataxia, spinal spinal muscular atrophy, testicular atrophy toothed, red nucleus, pale knob and low-hypothalamic nucleus as well as myotonic dystrophy as well as spongiform encephalopathies, such as congenital Creutzfeldt-Jakob disease (caused by a defect in prion protein processing), Fabry disease (caused by lysosomal αgalactosidase A) and Gerstmann-Straussler-Scheinker syndrome (caused by a defect in Prp processing), infertility, pancreatitis, pancreatic insufficiency, osteoporosis penia, Gorham syndrome, chloride channel disorders, congenital myotonia (Thomson and Becker forms), Bartter type III syndrome, Dent's disease, hyperplexia, epilepsy, hyperplexia, lysosomal storage disease, Angelman's syndrome, primary ciliary dyskinesia (PCD), PCD with reverse viscera (also known as Kartagener's syndrome), PCD without reverse viscera and ciliary aplasia, and liver disease.

[0016] Other diseases in which the mutation in CFTR is involved include male infertility due to congenital bilateral vas deferens (CBAVD), benign lung disease, idiopathic pancreatitis and allergic bronchopulmonary aspergillosis (ABPA). See, "CFIR-opathies: disease phenotypes associated with cystic fibrosis transmembrane regulator gene mutations," Peader G. Noone and Michael R. Knowles, Respir. Res. 2001, 2: 328-332.

[0017] In addition to upregulation of CFTR activity, reducing anion secretion by CFTR modulators may be beneficial in the treatment of secretory diarrhea in which epithelial water transport is dramatically increased as a result of transport of activated chloride by secretion stimulants. The mechanism involves an increase in cAMP concentration and stimulation of CFTR.

[0018] Although there are numerous causes of diarrhea, the main consequences of diarrheal diseases resulting from excessive chloride transport are common and include dehydration, acidosis, impaired growth and death. Acute and chronic diarrhea are an important medical problem in many regions of the world. Diarrhea is both a significant factor in malnutrition and a leading cause of death (5,000,000 deaths per year) in children under five years of age.

[0019] Secretory diarrhea is also a serious condition in patients with acquired immunodeficiency syndrome (AIDS) and chronic inflammatory bowel disease (IBD). Sixteen million people travel to developing countries from industrialized countries with diarrhea, with the severity and number of diarrhea cases varying depending on the country and region of travel.

[0020] Accordingly, there is a need to develop potent and selective CFTR enhancers in the form of mutant and wild-type human CFTR. These mutated forms of CFTR include, but are not limited to, ΔF508del, G551D, R117H, 2789 + 5G&gt; A.

[0021] There is also a need for modulators of CFTR activity and compositions thereof that can be used to modulate CFTR activity in a mammalian cell membrane. [0022] There is a need for methods of treating diseases caused by mutations in CFTR using such modulators of CFTR activity.

[0023] There is a need for methods to modulate CFTR activity in an ex vivo mammalian cell membrane.

[0024] WO 2006/002421 A2 relates to N-phenylquinolinecarboxamides and their medical use for the treatment of inflammatory and pulmonary diseases.

SUMMARY OF THE INVENTION [0025] It has now been found that the compounds of this invention and their pharmaceutically acceptable compositions are useful as modulators of CFTR activity. The compounds have the general formula (I):

<img file="PL2349263T3_D0001.tif" />

or a pharmaceutically acceptable salt thereof, wherein R¹, R², R³ and A are described below generally and in classes and subclasses.

[0026] These compounds and pharmaceutically acceptable compositions are useful for treating or reducing the severity of various diseases, disorders or conditions associated with mutations in CFTR.

DETAILED DESCRIPTION OF THE INVENTION

General description of the compounds of the invention:

[0027] The present invention relates to compounds of Formula (I) useful as modulators of CFTR activity:

<img file="PL2349263T3_D0002.tif" />

&lt;D;

or pharmaceutically acceptable salts thereof, where:

ring A is selected from:

R¹ is -CF3, -CN or -C ^ CCH2N (CH₃)2;

R² is hydrogen, -CH3, -CF3, -OH or -CH2OH;

R³ is hydrogen, -CH3, -OCH3 or -CN;

with R² and R³ they are not hydrogen at the same time.

Relationships and definitions:

[0028] The compounds of this invention include those described generally above, and are further illustrated below by the classes, subclasses and types disclosed herein. As used herein, the following definitions apply, unless otherwise indicated.

[0029] The term "ABC transporter" as used herein means an ABC transpoter protein or fragment thereof comprising at least one binding domain, wherein the protein or fragment thereof occurs in vivo or in vitro. The term "binding domain" as used herein means a domain within an ABC transporter that can bind to a modulator. See, e.g., Hwang, TC et al., J. Gen. Physiol. (1998): 111 (3), 477-90.

[0030] The term "CFTR" as used herein means cystic fibrosis regulator of membrane conductance or its mutation capable of regulatory activity, including but not limited to, ΔF508 CFTR, R117H CFTR, and G551D CFTR (see CFTR mutations, e.g. <a href="http://www.genet.sickkids.on.ca/cftr/">http://www.genet.sickkids.on.ca/cftr/</a>).

[0031] The term "modulation" as used herein means increasing or decreasing by a measurable amount.

[0032] As used herein, the term "normal CFTR" or "normal CFTR performance" means wild type CFTR without any disturbance due to environmental factors such as smoking, environmental pollution or anything that causes inflammation in the lung.

[0033] As used herein, the term "lowered CFTR" or "lowered CFTR" means a lower level of CFTR than normal or a CFTR lower than normal.

[0034] For the purposes of the present invention, chemical elements are identified according to the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75th edition. In addition, the general principles of organic chemistry are described in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999, and "March's Advanced Organic Chemistry", 5th edition, Editions: Smith, MB and March, J., John Wiley &amp; Sons, New York: 2001.

[0035] The combinations of substituents envisaged by this invention are preferably those that result in the formation of stable or chemically possible compounds. The term "stable", as used herein, means compounds that do not change significantly when subjected to conditions that allow them to be produced, detected and preferably recovered, purified and used for one or more of the purposes disclosed herein. In some embodiments, the stable compound or chemically possible compound is one that does not change significantly when stored at 40 ° C or lower, without access of moisture or other chemically reactive conditions, for at least a week.

[0036] The term "protecting group", as used herein, refers to an agent used temporarily to block one or more desired sites in a multifunctional compound. In some embodiments, the protecting group exhibits one or more, or preferably all, of the following features: a) reacts selectively in good yield to provide a protected starting compound that is stable under reaction conditions occurring at one or more other reactive sites; and b) is selectively able to be removed in good yield by reagents that do not attack this regenerated functional group. Exemplary protecting groups have been described in detail by Greene, TW, Wuts, P. G in "Protective Groups in Organic Synthesis", 3rd Edition, John Wiley &amp; Sons, New York: 1999, and other editions of this book.

[0037] Unless otherwise stated, structures described herein are believed to include all isomeric (e.g., enantiomeric, diastereomeric and geometric (or conformational) forms of the structure; for example, R and S configurations for each asymmetric center, (Z) and (E) double bond isomers, and (Z) and (E) conformational isomers. Thus, individual stereochemical isomers as well as enantiomeric, diastereomeric and geometric (or conformational) mixtures of the present compounds are within the scope of the invention. Unless otherwise stated, all tautomeric forms of the compounds of the invention are within the scope of the invention; for example.

compounds of Formula (I) may exist as tautomers:

<img file="PL2349263T3_D0003.tif" />

The description of the structure also includes compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having structures according to the invention except that the hydrogen atom has been replaced with a deuterium or tritium atom or the carbon atom has been replaced with an enriched carbon atom¹³C- or ¹⁴C - are within the scope of the present invention. Such compounds are useful, for example, as analytical tools or samples in biological analyzes. Such compounds, in particular compounds that contain deuterium atoms, may have modified metabolic properties.

Description of example compounds:

[0039] The present invention provides a compound of Formula (I):

<img file="PL2349263T3_D0004.tif" />

or a pharmaceutically acceptable salt thereof, wherein:

ring A is selected from:

<img file="PL2349263T3_D0005.tif" />

(a) (b) (c) (d)

R¹ is -CF3, -CN or -C = CCH2N (CH3) 2;

R² is hydrogen, -CH3, -CF3, -OH or -CH2OH;

R³ is hydrogen, -CH3, -OCH3 or -CN;

both R² and R³ they are not hydrogen at the same time.

In one embodiment, ring A is

<img file="PL2349263T3_D0006.tif" />

(and).

[0041] In one embodiment, Ring A is. [0042] In another embodiment, Ring A is

<img file="PL2349263T3_D0007.tif" />

<img file="PL2349263T3_D0008.tif" />

(C).

[0043] In yet another embodiment, ring A is

<img file="PL2349263T3_D0009.tif" />

(D).

[0044] In one embodiment, R¹ means -CF3.

[0045] In another embodiment, R¹ means -CN.

[0046] In another embodiment, R¹ means -C = CCH2N (CH3) 2.

[0047] In one embodiment, R² means -CH3.

[0048] In another embodiment, R² means -CF3.

[0049] In another embodiment, R² means -OH.

[0050] In another embodiment, R² means -CH2OH.

[0051] In one embodiment, R³ means -CH3.

[0052] In one embodiment, R³ means -OCH3.

[0053] In another embodiment, R³ means -CN.

[0054] In one embodiment, R² is hydrogen; and R³ means -CH3, -OCH3 or -CN. [0055] In another embodiment, R² is -CH3, -CF3, -OH or -CH2OH; and R³ is hydrogen.

[0056] In several embodiments of the present invention, ring A is

<img file="PL2349263T3_D0010.tif" />

R¹ is -CF3, R² is hydrogen; and R³ means -CH3, -OCH3 or -CN. In other forms, R¹ means -CN. In yet further forms, R¹ means

-C = CCH2N (CH₃) 2. In one embodiment, R³ means -CH3. Alternatively, R³ means -OCH3. Alternatively, R³ means -CN.

[0057] In further embodiments of the present invention, ring A is

<img file="PL2349263T3_D0011.tif" />

R¹ is -CF3, R² is -CH3, -CF3, -OH or -CH2OH, and R³ is hydrogen. In other forms, R¹ means -CN. In yet further forms, R¹ means -C = CCH2N (CH3) 2. In one form, R² means -CH3. Alternatively, R² means -CF3. Alternatively, R² means -OH. Alternatively, R² means -CH2OH.

[0058] In several embodiments of the present invention, ring A is

<img file="PL2349263T3_D0012.tif" />

R¹ is -CF3, R² is hydrogen; and R³ means -CH3, -OCH3 or -CN. In other forms, R¹ means -CN. In yet further forms, R¹ means -C = CCH2N (CH3) 2. In one embodiment, R³ means -OCH3. Alternatively, R³ means -CH3. Alternatively, R³ means -CN.

In further embodiments of the present invention, ring A is

<img file="PL2349263T3_D0013.tif" />

R¹ is -CF3, R² is -CH3, -CF3, -OH or -CH2OH, and R³ is hydrogen. In other forms, R¹ means -CN. In yet further forms, R¹ means -C = CCH2N (CH3) 2. In one embodiment, R² means -CH3. Alternatively, R² means -CF3. Alternatively, R² means -OH. Alternatively, R² means -CH2OH.

[0060] In several embodiments of the present invention, ring A is

<img file="PL2349263T3_D0014.tif" />

R¹ is -CF3, R² is hydrogen; and R³ means -CH3, -OCH3 or -CN. In other forms, R¹ means -CN. In yet further forms, R¹ means -C = CCH2N (CH3) 2. In one embodiment, R³ means -CH3. Alternatively, R³ means -OCH3. Alternatively, R³ means -CN.

[0061] In further embodiments of the present invention, ring A is

<img file="PL2349263T3_D0015.tif" />

R¹ is -CF3, R² is -CH3, -CF3, -OH or -CH2OH, and R³ is hydrogen. In other forms, R¹ means -CN. In yet further forms, R¹ means -C = CCH2N (CH3) 2. In one embodiment, R² means -CH3. Alternatively, R² means -CF3. Alternatively, R² means -OH. Alternatively, R² means -CH2OH.

[0062] In several embodiments of the present invention, ring A is

<img file="PL2349263T3_D0016.tif" />

R¹ is -CF3, R² is hydrogen; and R³ is -CH3, -OCH3, or -CN. In other forms, R¹ means -CN. In yet further forms, R¹ means -C = CCH2N (CH3) 2. In one embodiment, R³ means -CH3. Alternatively, R³ means -OCH3. Alternatively, R³ means -CN.

[0063] In further embodiments of the present invention, ring A is

<img file="PL2349263T3_D0017.tif" />

R¹ is -CF3, R² is -CH3, -CF3, -OH or -CH2OH, and R³ is hydrogen. In other forms, R¹ means -CN. In yet further forms, R¹ means

-C = CCH2N (CH3) 2. In one embodiment, R² means -CH3. Alternatively, R² means -CF3.

Alternatively, R² means -OH. Alternatively, R² means -CH2OH.

[0064] Representative compounds of the present invention are shown below in Table 1.

Table 1

<td> 1</td><td> 2</td><td> 3</td>

<td></td><td>FU</td><td>FH 5'A2</td>

<td> 4</td><td> 5</td><td> 6</td>

<td>F i.e.</td><td>_NJ (X Ϊο * Η</td><td>H</td>

<td> 7</td><td> 8</td><td> 9</td>

<td>"IN⁵'· %</td><td></td><td>YFF</td>

<td> 10</td><td> 11</td><td> 12</td>

<td>AK Jl jfl)</td><td>FH .in %</td><td>FH .IN %</td>

<td> 13</td><td> 14</td><td></td>

<td>H Z \ JL &lt;X_ ⁰ OF — and— F</td><td>Λ _xfa.'.'</td><td></td>

General Synthesis Schemes [0065] The compounds of the present invention can be readily prepared by methods known in the art, and as outlined in Schemes 1-3.

Scheme 1. Preparation of compounds of formula (I).

<img file="PL2349263T3_D0018.tif" />

What₂R)₂CH = CH (OR), toluene, heat; b) dauterm or diphenyl ether, reflux, atmosphere N₂; c) Removal of a quantitative protecting group, if present (e.g. -Cl), Pd / C, H₂, EtOH; d) removing the protecting group R by an acid or base; e) CH₃CN, Et₃N, heat; f) Pd / C, H₂, EtOH; g) HATU, Et₃N, DMF or cyclic propylphosphonic acid anhydride (T3P®), pyridine, 2-methyltetrahydrofuran.

[0066] Scheme 1 illustrates a convergent approach for preparing compounds of Formula (I) from substituted benzene derivatives 1a and 2a. In the final transformation, amide formation by coupling carboxylic acid 1d with amine 2c to give a compound of Formula (I) can be achieved using either O- (7-azabenzotriazol-1-yl) -N, N, N ', N'- hexafluorophosphate tetramethyluronium (HATU) and triethylamine in N, N-dimethylformamide (DMF) or cyclic propylsulfonic anhydride (T3P®) and pyridine in 2-methyltetrahydrofuran. Carboxylic acid 1d is prepared from the corresponding substituted benzene derivative 1a via a reaction sequence beginning with the condensation mediated by heating 1a with the appropriate malonate (CO2R) 2CH = CH (OR), where R is an alkyl group such as methyl, ethyl or the like, with delivery 1b.

[0067] Compound 1b is converted to carboxylic acid 1d by a sequence of three steps involving intermolecular cyclization under reflux conditions in Dowtherm or diphenyl ether (step b) followed by removal (if needed) of blocking groups halogen (stage c) under palladium catalysed halogenation and acid or base catalyzed saponification (stage d). The order of the steps of deprotection and saponification can be reversed; i.e., step c may occur before or after step d as depicted in Scheme 1.

[0068] Referring again to Scheme 1, aniline derivative 2c can be prepared from nitrobenzene 2a by a three-step sequence. Thus, coupling of nitrobenzene 2a with a cyclic amine

<img file="PL2349263T3_D0019.tif" />

3, as defined herein, in the presence of triethylamine provides compound 2b.

<img file="PL2349263T3_D0020.tif" />

[0069] Scheme 2 depicts the synthesis of compounds of Formula (I) having propinylamine side chain. In this way, the coupling of nitrobenzene 2a, where Hal is a bromide, chloride or similar moiety with

<img file="PL2349263T3_D0021.tif" />

3, as defined herein, in the presence of potassium carbonate in DMSO provides compound 4. Palladium-catalyzed coupling of compound 4 with N, N-dimethylprop-2-y-1-amine followed by iron or zinc catalyzed reduction of the nitro moiety provides amine 5. Equipment cleavage of amine 5 with carboxylic acid 1d provides compound 6, which is a compound of Formula (I).

<img file="PL2349263T3_D0022.tif" />

[0070] Scheme 3 illustrates the synthesis of a compound of Formula (I) wherein * A 'is 7-azabicyclo [2.2.1] heptane, optionally having an exo or endo hydroxyl group in position 2. These hydroxyl-substituted adducts (+) - endo-7-azabicyclo [2.2.1] heptan-2-ol, (-) - endo-7-azabicyclo [2.2.1] heptan-2-ol, (+) - exo -7-azabicyclo [2.2.1] heptan-2-ol and (-) - exo-7-azabicyclo [2.2.1] heptan-2-ol can be prepared using procedures as described in Fletcher, SR, et al., "Total Synthesis and Determination of the Absolute Configuration of Epibatidine," J. Org. Chem, 59, pp. 1771-1778 (1994). 7 Azabicyclo [2.2.1] heptane as such is commercially available from Tyger Scientific Inc. 324 Stokes Avenue Ewing, NJ, 08638 USA.

[0071] Thus, as a result of a series of transformations shown in Schemes 1 and 2, coupling compound 2a with bicyclo [2.2.1] amine 7 provides compound 8. If compound 8 has a hydroxyl group, it may be necessary to protect the hydroxyl group using protecting group against further transformations. Thus, treatment of compound 8 with tert-butyldimethylsilyl chloride using known conditions provides protected compound 9 against reduction of the nitro moiety to obtain amine 10. Formation of amide with 1d (compare Scheme 3) and removal of the hydroxyl protecting group (if necessary) provides compound 11, which is a compound of Formula (I).

Applications, Preparations and Administration

Pharmaceutically Acceptable Compositions [0072] In one aspect of the present invention, pharmaceutically acceptable compositions are provided, said compositions comprising any of the compounds described herein and optionally comprising a pharmaceutically acceptable carrier, adjuvant or vehicle. In some embodiments, these compositions optionally further comprise one or more additional therapeutic agents.

[0073] It will further be appreciated that certain compounds of the present invention may exist in the free form for treatment purposes, or, where appropriate, as a pharmaceutically acceptable derivative or prodrug thereof. In accordance with the present invention, a pharmaceutically acceptable derivative or prodrug includes, but is not limited to, pharmaceutically acceptable salts, esters, salts of such esters, or any other adduct or derivative that, when administered to a patient in need thereof, is capable of directly or indirectly providing a compound as described herein description, or its metabolite or residue.

[0074] As used herein, the term "pharmaceutically acceptable salt" refers to those salts that, based on competent medical evaluation, are suitable for use in contact with human and lower animal tissues without undue toxic effects, irritation, and allergic reaction. similar, with a reasonable benefit / risk ratio. "Pharmaceutically acceptable salt" means any non-toxic salt or ester salt of a compound of this invention that, when administered to a recipient, is capable of providing, directly or indirectly, a compound of this invention or an inhibitory active metabolite or residue thereof.

[0075] Pharmaceutically acceptable salts are well known in the art. For example, SM Berge et al., Describes in detail the pharmaceutically acceptable salts in J. Pharmaceutical Sciences, 1977, 66, 1-19, incorporated herein by reference. Pharmaceutically acceptable salts of the compounds of this invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable, non-toxic acid addition salts are salts of the amino group prepared using inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid, or using 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.

[0076] Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphors, camphorsulfonate, citrate, cyclopentanopropionate, digluconate, dodecyl sulfate, edisylate, methanesulfonate, , glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxyethane sulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalene sulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, tartrate, undecanoate, valerate and the like. Salts derived from appropriate bases include alkali metal salts, alkaline earth metal salts, ammonium salts and N salts⁺(C1-4 alkyl) 4. This invention also envisages the quaternization of any groups containing basic nitrogen atoms of the compounds disclosed herein. As a result of quaternization, products soluble or dispersible in water or oils can be obtained. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium salts and the like. In addition, pharmaceutically acceptable salts contain, where appropriate, non-toxic ammonium, quaternary ammonium and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower alkyl sulfonate and aryl sulfonate.

[0077] As described above, the pharmaceutically acceptable compositions of the present invention further include a pharmaceutically acceptable carrier, excipient or vehicle which, as described herein, includes any or all solvents, diluents or other liquid substrates, dispersing or suspending agents, surface agents active, isotonic agents, thickeners or emulsifiers, preservatives, solid binders, lubricants and the like, suitable for the particular dosage form desired. Remington's Pharmaceutical Sciences, 16th Edition, EW Martin (Mack Publishing Co., Easton, Pa., 1980) discloses a variety of carriers used to formulate pharmaceutically acceptable compositions and known techniques for their preparation. Unless any conventional carrier medium is incompatible with the compounds of the invention, that is, it does not cause undesirable biological effects or otherwise does not adversely affect any other ingredient or ingredients of the pharmaceutically acceptable composition, its use is considered to be within the scope of this invention. . Some examples of materials that 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, buffering substances such as phosphates, glycine, acid sorbic or potassium sorbate, mixtures of partial glycerides 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, polyvinylpyrrolidone, 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 as 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, and other non-toxic, compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweeteners, flavors and flavors, may be present in the composition preservatives and antioxidants, according to formulating assessment. Use of compounds and pharmaceutically acceptable compositions [0078] In yet another aspect, the present invention provides a method of treating or reducing the severity of a condition, disease or disorder in which a CFTR mutation is involved. In certain embodiments, the present invention provides a method of treating a condition, disease or disorder mediated by deficiency of CFTR activity, which method comprises administering to a subject in need thereof, preferably a mammal, a composition comprising a compound of Formula (I).

[0079] In some embodiments, the present invention provides a method of treating diseases associated with reduced CFTR activity as a result of mutations in the CFTR coding gene or environmental factors (e.g., smoke). These diseases include, cystic fibrosis, chronic bronchitis, recurrent bronchitis, acute bronchitis, male infertility due to congenital bilateral vas deferens (CBAVD), female infertility due to congenital uterine and vaginal (CAUV), idiopathic chronic pancreatitis, idiopathic chronic pancreatitis recurrent pancreatitis, idiopathic acute pancreatitis, chronic rhinitis and paranasal sinuses, primary sclerosing cholangitis, allergic bronchopulmonary aspergillosis, diabetes, dry eye disease, constipation, allergic bronchopulmonary aspergillosis (ABPA), bone diseases (e.g. osteoporosis), and asthma.

[0080] In some embodiments, the present invention provides a method of treating diseases associated with the normal operation of CFTR. These diseases include chronic obstructive pulmonary disease (COPD), chronic bronchitis, recurrent bronchitis, acute bronchitis, chronic rhinitis and sinusitis, constipation, pancreatitis including chronic pancreatitis, recurrent pancreatitis and acute pancreatitis , pancreatic insufficiency, male infertility caused by congenital bilateral vas deferens (CBAVD), mild lung disease, idiopathic pancreatitis, liver disease, congenital emphysema, gallstones, gastroesophageal reflux disease, gastrointestinal malignancies, inflammatory bowel disease, constipation, diabetes, arthritis, osteoporosis and osteopenia.

[0081] In some embodiments, the present invention provides a method of treating diseases associated with the normal activity of CFTR, including congenital hemochromatosis, coagulation deficiencies - fibrinolysis, e.g. protein C deficiency, congenital angioedema type 1, lipid processing disorders, e.g. family hypercholesterolemia, chylomicronemia type 1, abetalipoproteinemia, lysosomal storage diseases, e.g. cellular inclusion / pseudo Hurler disease, mucopolysaccharidoses, Sandhof / Tay-Sachs syndrome, Crigler-Najjar syndrome type II, poliendocrinopathy / hyperinsulinemia, diabetes mellitus, Laron's dwarfism, myeloperoxidase deficiency, primary hypoparathyroidism, melanoma, CDG type 1, congenital hyperparathyroidism, hyperthyroidism ACT deficiency, diabetes insipidus (DI), neuro-pituitary DI, renal DI, Charcot-Marie-Tooth syndrome, Pelizacus-Merzbacher disease, neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, progressive supranuclear palsy, Pick's disease, various polyglutamic neurological disorders such as Huntington's disease, cerebellum ataxia type I, cerebrospinal muscular atrophy, dentin atrophy, testicular nuclei pale knob and low-hypothalamic nucleus, and myotonic dystrophy as well as spongiform encephalopathies, such as congenital Creutzfeldt-Jakob disease (caused by a defect in prion protein processing), Fabry disease and Gertsmann-Straussler-Scheinker syndrome, Gorham syndrome, chloride channel disorders, congenital myotonia (Thomson and Becker forms), Bartter syndrome type III, Dent's disease , hyperplexia, epilepsy, hyperplexia, lysosomal storage disease, Angelman syndrome, primary ciliary dyskinesia (PCD), Inverse PCD (also known as Cartagener's syndrome), PCD without invisible viscera and cilia aplasia, and Sjogren's disease, comprising the step of administering to said mammal an effective amount of a composition comprising a compound of the present invention.

[0082] According to an alternative preferred embodiment, the present invention provides a method of treating cystic fibrosis comprising the step of administering to said mammal a composition comprising the step of administering to said mammal an effective amount of a composition comprising a compound of the present invention.

[0083] According to the invention, an "effective amount" of a compound or a pharmaceutically acceptable composition is that amount which is effective in treating or reducing the severity of one or more of the diseases, disorders or conditions listed above.

[0084] The compounds and compositions according to the method of the present invention may be administered using any amount and any route of administration effective to treat or reduce the severity of one or more of the diseases, disorders or conditions mentioned above.

[0085] In certain embodiments, the compounds and compositions of the present invention are useful for treating or reducing the severity of cystic fibrosis in patients having residual CFTR activity in the apical membrane of the respiratory or non-respiratory epithelium. The presence of residual CFTR activity on the epithelial surface can easily be detected using methods known in the art, e.g., standard electrophysiological, biochemical or histochemical techniques. Such methods identify CFTR activity using in vivo or ex vivo electrophysiological techniques to measure Cl concentrations^- in sweat or saliva or ex vivo biochemical or histochemical techniques for monitoring cell surface density. Using such methods, residual CFTR activity can easily be detected in heterozygous or homozygous patients for a wide variety of mutations, including homozygous or heterozygous patients for the most common mutation, ΔF508.

[0086] In another embodiment, the compounds and compositions of the present invention are useful for treating or reducing the severity of cystic fibrosis in patients with residual CFTR activity induced or increased using pharmacological methods or gene therapy. Such methods increase the amount of CFTR present on the cell surface, thereby inducing a previously absent CFTR activity in the patient or increasing the existing level of residual CFTR activity in the patient.

[0087] In one embodiment, the compounds and compositions of the present invention are useful for treating or reducing the severity of cystic fibrosis in patients with certain genotypes having residual CFTR activity, e.g., class III mutations (disturbed regulation or gating), class IV mutations (altered conductivity) or class V mutations (reduced synthesis) (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). Other patient genotypes that show residual CFTR activity include patients who are homozygous for one of these classes or heterozygous for any of the other mutation classes, including class I mutations, class II mutations, or an unclassified mutation.

In one embodiment, the compounds and compositions of the present invention are useful for treating or reducing the severity of cystic fibrosis in patients with certain clinical phenotypes, e.g., moderate to mild clinical phenotype, which typically correlates with the level of residual CFTR activity in the apical epithelium. Such phenotypes include patients exhibiting pancreatic insufficiency or patients diagnosed with idiopathic pancreatitis or congenital bilateral defect of the vas deferens or mild lung disease.

[0089] The specific amount will vary from individual to individual and will depend on the species, age and general condition of the individual, the severity of the infection, the particular agent, the method of its administration and the like. The compounds of the invention are preferably formulated in unit dosage form to facilitate administration and uniformity of dosage. The term "unit dosage form" as used herein refers to a physically discrete unit of agent suitable for the patient undergoing treatment. However, it will be understood that the total daily dose of the compounds and compositions of the present invention will depend on the decision of the treating physician regarding competent medical judgment. The specific effective dose level for any particular patient or organism will depend on a variety of factors including the disorder being treated and the severity of the disorder, the activity of the particular compound employed; the specific composition used; 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 particular compound employed, the duration of treatment; drugs used in combination or concurrently with the specific compound employed, and similar factors well known in the medical arts. The term "patient" as used herein means a living being, preferably a mammal, and most preferably a human.

[0090] The pharmaceutically acceptable compositions of this invention can be administered to humans and other living entities orally, rectally, parenterally, intrathecal, vaginally, intraperitoneally, topically (as powders, ointments, drops or patch), buccal or as oral or nasal sprays and similar depending on the severity of the infection being treated. In some embodiments, the compounds of the invention may be administered orally or parenterally at a dosage level of about 0.01 mg / kg to about 50 mg / kg, and preferably from about 0.5 mg / kg to about 25 mg / kg, the individual's weight a day, one or more times a day to achieve the desired therapeutic effect. [0091] Liquid 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, 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 (especially cotton oil, peanut, corn oil, germ oil, olive oil, castor and sesame oil), glycerin, tetrahydrofurfuryl alcohol, polyethylene glycols and sorbitan fatty acid esters, and mixtures thereof. In addition to inert diluents, the oral compositions may also contain adjuvants such as wetting, emulsifying and suspending agents, sweetening, flavoring and flavoring agents.

[0092] Injectable preparations, for example sterile, injectable, aqueous or oily suspensions may be formulated according to the known technique using suitable dispersing or wetting agents or suspending agents. Sterile injectable preparations may also be sterile injectable solutions, suspensions or emulsions in a non-toxic parenterally-acceptable diluent or solvent, for example, a solution in 1,3-butanediol. Acceptable excipients and solvents that may be used include water, USP Ringer's solution, and isotonic sodium chloride. In addition, sterile, non-volatile vegetable oils have traditionally been used as a solvent or suspension medium. For this purpose any non-irritating, non-volatile vegetable oil may be used, including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectables.

[0093] Injectable preparations can be sterilized, for example, by filtration through a bacterial-retaining filter or by the inclusion of sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or another sterile suitable medium just before use. injection.

[0094] In order to prolong the effect of a compound of the present invention, it is often desirable to slow the absorption of the compound after subcutaneous or intramuscular injection. This can be achieved by using a liquid suspension of crystalline or amorphous material with poor water solubility. The rate of absorption of the compound will then depend on its dissolution rate, which in turn may depend on the size of the crystals or the crystal form. Alternatively, delayed absorption of the parenterally administered compound form is achieved 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 on the ratio of compound to polymer and the nature of the particular polymer used, the release rate of the compound can be controlled. Examples of other biodegradable polymers include poly (orthoesters) and poly (anhydrides).

Injectable depot preparations are also prepared by "entrapping" the compound in liposomes or microemulsions that are compatible with body tissues.

[0095] Compositions for rectal or vaginal administration are preferably suppositories which can be prepared by mixing the compounds of the present invention with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or suppository wax which are solid at ambient temperature but liquid at body temperature, and therefore melt in the rectal or vaginal cavity and release the active compound.

[0096] Solid 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 fillers such as starches, lactose, sucrose, glucose, mannitol and silicic acid, (b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and acacia, (c) humectants, such as glycerin, (d) disintegrants, such as agar, calcium carbonate, potato starch 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.

[0097] Solid compositions of a similar type may also be employed as fillers in soft and hard gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like. Solid dosage forms in the form of tablets, dragees, capsules, pills and granules can be prepared with coatings and coatings, such as enteric coatings and other coatings well known in the pharmaceutical formulation art. They may optionally contain opacifying agents and may also be formulated to release active ingredient (s) only or, preferably, in a certain portion 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 gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.

[0098] The active compounds may also be in microencapsulated form with one or more of the above-mentioned excipients. Solid dosage forms in the form of tablets, dragees, capsules, pills and granules can be prepared with coatings and coatings such as enteric coatings, release control coatings and other coatings well known in the pharmaceutical formulation art. In such solid dosage forms, the active compound can be mixed with at least one inert diluent, such as sucrose, lactose or starch. Such dosage forms may also, in accordance with common practice, additional substances other than inert diluents, for example tabletting glidants and other tableting additives such as 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 may also be formulated to release active ingredient (s) only or, preferably, in a certain portion of the intestinal tract, optionally, in a delayed manner. Examples of usable embedding compositions include polymeric substances and waxes.

[0099] Dosage 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 ingredient is mixed under sterile conditions with a pharmaceutically acceptable carrier and any needed preservatives or buffering agents, if required. Ophthalmic preparations, ear drops and eye drops are also considered within the scope of the invention. In addition, the present invention contemplates the use of transdermal patches, which has the additional advantage of providing controlled administration of the compound into the body. Such dosage forms are prepared by dissolving or placing the compound in the appropriate medium. Absorption enhancers can be used to increase the flow of the compound through the skin. The rate can be controlled either by introducing a rate controlling membrane or by dispersing the compound in a polymer matrix or gel.

[0100] The performance of the compound used in this invention as a CFTR modulator can be evaluated in accordance with methods generally described in the art and in the examples provided herein.

It should be noted that the compounds and pharmaceutically acceptable compositions of the present invention can be used in combination therapies, i.e. the compounds and pharmaceutically acceptable compositions can be administered simultaneously with, before or after the use of one or more other desired drugs or medical procedures. When choosing a specific combination of therapies (drugs or procedures) for use in combination therapy mode, consideration should be given to the compatibility of desired drugs and / or procedures and the desired therapeutic effect to be achieved. It should also be noted that the therapies used may have an effect on the disorder itself (for example, a compound of the invention may be administered concurrently with another agent used to treat the same disorder) or may produce other effects (e.g. control any side effects ). Additional therapeutic agents used herein that are normally administered to treat or prevent a particular disease or condition are known as "appropriate for the disease or condition being treated."

[0102] In one embodiment, the additional agent is selected from a mucolytic agent, bronchodilator, antibiotic, anti-infective agent, anti-inflammatory agent, CFTR modulator other than the compound of the present invention or a nutrient. In a further embodiment, this additional agent is a CFTR modulator other than the compound of the present invention.

[0103] In one embodiment, the additional agent is an antibiotic. Exemplary antibiotics useful herein include tobramycin, including tobramycin inhalation powder (TIP), azithromycin, aztreonam, including aztreonam aerosol, amikacin, including its liposome preparations, ciprofloxacin, including its preparations suitable for administration levoflaxacin, including aerosol preparations, a combination of two antibiotics, e.g. fosfomycin and tobramycin.

[0104] In another embodiment, the additional agent is a mucolytic agent. An exemplary mucolytic agent useful herein includes Pulmozyme®.

[0105] In another embodiment, the additional agent is a bronchodilator. Exemplary bronchodilators include albuterol, metaprotenerol sulfate, pirbuterol acetate, salmeterol or tetrabulin sulfate.

[0106] In another embodiment, the additional agent is effective in reconstituting liquids on the surface of the lung airways. Such agents improve the movement of salt into and out of the cells, which allows better hydration of mucus in the airways of the lungs, and thus easier cleansing. Examples of such agents include hypertonic saline, denuphosol, ([[(3S, 5R) -5- (4-amino-2-oxopyrimidin-1-yl) -3-hydroxyoxolan-2-yl] methoxyhydroxyphosphoryl] - [[[ (2R, 3S, 4R, 5R) -5- (2,4-dioxopyrimidin-121-yl) -3,4-dihydroxyoxolan-2-yl] methoxyhydroxyphosphoryl] oxyhydroxyphosphoryl] hydrogen phosphate) tetrasodium or bronchitol (preparation of mannitol for inhalation).

[0107] In another embodiment, the additional agent is an anti-inflammatory agent, i.e., an agent that can cause a reduction in inflammation in the lungs. Exemplary such agents useful herein include ibuprofen, docosahexaenoic acid (DHA), sildenafil, inhalation glutathione, pioglitazone, hydroxychloroquine or simavastatin.

[0108] In another embodiment, the additional agent reduces the activity of the sodium epithelial blocker (ENaC) either directly by blocking the channel or indirectly by modulation of proteases that lead to increased ENaC activity (e.g., serine proteases, channel activating proteases). Examples of such agents include kamostat (trypsin-like protease inhibitor), QAU145, 552-02, GS-9411, INO-4995, Aerolytic and amiloride. Additional agents that reduce the activity of the epithelial sodium channel blocker (ENaC) can be found, for example, in PCT Publication No. WO2009 / 074575. [0109] Among others, the diseases described herein, combinations of CFTR modulators such as compounds of Formula I, and agents that lower ENaC activity can be used to treat Liddle syndrome, inflammation or allergy involving cystic fibrosis, primary ciliary dyskinesia, chronic bronchitis , chronic obstructive pulmonary disease, asthma, respiratory tract infections, lung cancer, xerostomy and keratoconjunctivitis sire, respiratory tract infections (acute and chronic; viral and bacterial) and lung cancer.

[0110] Combinations of CFTR modulators, such compounds of Formula I, and agents that lower ENaC activity are also useful in the treatment of diseases involving blockage of the epithelial sodium channel, which also includes diseases other than respiratory diseases that are associated with abnormal fluid regulation by epithelium, possibly involving abnormal physiology of protective surface liquids on its surface, e.g. xerostomy (dry mouth) or keratoconjunctivitis sire (dry eye). In addition, blockade of the epithelial sodium channel in the kidney can be used to improve diuresis, and thus induce a hypotensive effect.

[0111] Asthma includes both intrinsic (non-allergic) and extrinsic (allergic) asthma, mild asthma, moderate asthma, severe asthma, bronchial asthma, exercise-induced asthma, occupational asthma, and post-bacterial asthma. Asthma treatment should also be understood to include treatment of individuals, e.g. younger than 4 or 5 years of age, exhibiting wheezing symptoms and diagnosed or diagnosed as "children with wheezing breath", an established category of patients with high medical significance and currently often identified as early or early stage asthmatics. (For convenience, this particular asthmatic condition is called "wheezing syndrome.") Prophylactic efficacy in the treatment of asthma will be demonstrated by a reduced incidence or severity of an attack of symptoms, such as an acute asthmatic attack or bronchoconstriction attack, improved lung function, or improved path hyperactivity. tract. This may be further demonstrated by the reduced requirement for other symptomatic therapy, i.e. therapy for or intended to limit or stop a symptomatic attack when it occurs, e.g. anti-inflammatory (e.g. corticosteroid) or bronchodilator. The prophylactic benefit in asthma may, in particular, be seen in patients prone to "morning decrease in peak expiratory flow". "Morning decreases in peak expiratory flow" are a recognized asthmatic syndrome, common among a significant percentage of asthmatics, and characterized by an asthma attack, e.g. between hours around 4-6 am, ie at any time point significantly distant from any previously reported symptomatic asthma therapy [0112] Chronic obstructive pulmonary disease includes chronic bronchitis or associated dyspnoea, emphysema, and exacerbation of airway hyperresponsiveness consequences of using other drugs, in particular other inhalation drugs. In certain embodiments, combinations of CFTR modulators, such as compounds of Formula I, and agents that reduce ENaC activity are useful for the treatment of bronchitis of any type or origin including, e.g., acute peanut-induced, catarrhal, croup, chronic or tuberculous inflammation bronchi.

[0113] In another embodiment, the additional agent is a CFTR modulator other than the compound of Formula I, i.e., an agent that exerts a modulation effect on CFTR activity. For example, such agents include ataluren ("PTC124®"; 3- [5- (2-fluorophenyl) -1,2,4-oxadiazol-3-yl] benzoic acid), synapultide, lanolutide, depelestate (human recombinant neutrophil elastase inhibitor ), kobiproston (7 - {(2R, 4aR, 5R, 7aR) -2 - [(3S) -1,1-difluoro3-methylpentyl] -2-hydroxy-6-oxooctahydrocyclopenta [b] pyran-5-yl} acid heptanoic acid) or (3- (6- (1- (2,2-difluorobenzo [d] [1,3] dioxol-5-yl) cyclopropanecarboxamido) -3-methylpyridin-2-yl) benzoic acid. In another embodiment, the additional agent is (3- (6 (1- (2,2-difluorobenzo [d] [1,3] dioxol-5-yl) cyclopropanecarboxamido) -3-methylpyridin-2-yl) benzoic acid.

[0114] In another embodiment, the additional agent is a nutrient. Examples of such agents include pancrelipase (pancreatic enzyme replacement), including Pancrease®, Pancreacarb®, Ultrase® or Creon®, Liprotomase® (formerly Trizytek®), Aquadeks® or inhalation glutathione. In one embodiment, the additional nutrient is pancrelipase.

[0115] The amount of additional therapeutic agent present in the compositions of this invention will not be greater than the amount that would normally be administered in a composition containing 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 containing that agent as the only therapeutically active agent.

[0116] The compounds of this invention, or pharmaceutically acceptable compositions thereof, can also be incorporated into compositions for coating implantable medical devices, such as prostheses, artificial valves, vascular implants, stents and catheters. Accordingly, the present invention, in another aspect, comprises a composition for coating an implantable device, comprising a compound of the present invention as described generally above and in sections and subsections of the present description, and a carrier suitable for coating this implantable device. In yet another aspect, the present invention includes an implantable device coated with a composition comprising a compound of the present invention as described above generally, and in items and subsections of the present description, and a carrier suitable for coating this implantable device. Suitable coatings and general method of making coated implantable devices are described in US Patent Nos. 6099562, 5886026 and 5304121. Coatings are typically biocompatible polymeric materials such as hydrogel polymer, polymethyldisiloxane, polycaprolactone, polyethylene glycol, polylactic acid, ethylene / vinyl acetate and mixtures thereof. Optionally, the coatings may be further coated with a suitable fluorosilicon topcoat, polysaccharides, polyethylene glycol, phospholipids or combinations thereof to provide controlled release characteristics in the composition.

[0117] Another aspect of the invention relates to modulation of CFTR activity in a biological sample (in vitro), the method comprising contacting said biological sample with a compound of Formula (I) or a composition comprising the compound. The term "biological sample" as used herein includes, without limitation, cell cultures or their extracts; biopsy materials obtained from a mammal or their extracts and blood, saliva, urine, feces, semen, tears or other body fluids or their extracts.

[0118] CFTR modulation in a biological sample is useful for many purposes that are known to the skilled person. Examples of such purposes include, but are not limited to, CFTR testing in biological and pathological phenomena, and comparative assessment of new CFTR modulators.

[0119] In yet another embodiment, there is provided a method of modulating anion channel activity in vivo, comprising the step of contacting said channel with a compound of Formula (I). In preferred embodiments, the anion channel is a chloride channel or a bicarbonate channel. In other preferred embodiments, the anion channel is a chloride channel.

[0120] According to an alternative embodiment, the present invention provides a method for in vitro increasing the number of functional CFTRs in a cell membrane, comprising the step of contacting that cell with a compound of Formula (I).

[0121] According to another preferred embodiment, CFTR activity is measured by measuring the transmembrane potential voltage. Methods for measuring transmembrane potential voltage in a biological sample may include any methods known in the art, such as an optical membrane potential test or other electrophysiological methods.

[0122] The optical membrane potential test uses voltage sensitive FRET sensors described by Gonzalez and Tsien (See, Gonzalez, JE and RY Tsien (1995) "Voltage sensing by fluorescence resonance energy transfer in single cells" Biophys J 69 (4) : 1272-80 and Gonzalez, JE and RY Tsien (1997) "Improved indicators of cell membrane potential that use fluorescence resonance energy transfer" Chem Biol 4 (4): 269-77) in conjunction with instrumentation for measuring fluorescence changes, such as the Voltage / Ion Probe Reader (VIPR) ( See, Gonzalez, JE, K. Oades, et al. (1999) "Cell-based assays and instrumentation for screening ion-channel targets" Drug Discov Today 4 (9): 431-439).

[0123] These voltage-sensitive tests are based on a change in resonance fluorescence excitation energy transfer (FRET) between a membrane-soluble, voltage-sensitive dye, DiSBAC2 (3), and a fluorescent phospholipid, CC2DMPE, which is attached to the outer layer of the cell membrane and acts as a FRET donor. Changes in membrane potential (Vm) cause the redistribution of negatively charged DiSBAC2 (3) in the cell membrane and the amount of energy transferred from CC2-DMPE changes accordingly. Changes in fluorescence emissions can be monitored using a VIPR device^™ II, which is an integrated liquid dispenser and fluorescence detector designed to conduct cell-based screening in 96- or 384-well microtiter plates.

[0124] In another aspect, the present invention provides a kit for use for measuring the activity of CFTR or a fragment thereof in an in vitro or in vivo biological sample, comprising (i) a composition comprising a compound of Formula (I) or any of the above forms and (ii) instructions to a) contacting this composition with a biological sample; and b) measuring the activity of this CFTR or a fragment thereof. In one embodiment, the kit further includes instructions for a) contacting the additional composition with the biological sample; b) measuring the activity of this CFTR or a fragment thereof in the presence of this additional compound, and c) comparing the CFTR activity in the presence of the additional compound with the density of CFTR in the presence of the composition of formula (I). In preferred embodiments, the kit is used to measure the density of CFTR.

[0125] For a more complete understanding of the invention described herein, the following examples are set forth. It should be understood that these examples are for illustrative purposes only and may not be construed as limiting this invention in any way.

Methods and intermediates for preparing compounds of Formula (I) [0126] Another aspect of the invention relates to a process for preparing a compound of Formula (Ic):

<img file="PL2349263T3_D0023.tif" />

or a pharmaceutically acceptable salt thereof, said method comprising:

(a) reacting an acid of formula 1d with an amine of formula 2c to form

<img file="PL2349263T3_D0024.tif" />

Ring A is selected from:

H

Ak ^ -OR⁸

V (a)

<img file="PL2349263T3_D0025.tif" />

+, or- + (d) where

R¹ is -CF3, -CN or -C = CCH2N (CH3) 2;

R² is hydrogen, -CH3, -CF3, -OH or -CH2OH;

R³ is hydrogen, -CH3, -OCH3 or -CN; both R² and R³ they are not both hydrogen and a

R^and is a hydrogen atom or a silyl protecting group selected from the group consisting of trimethylsilyl (TMS), tert-butyldiphenylsilyl (TBDPS), tert-butyldimethylsilyl (TBDMS), triisopropylsilyl (TIPS) and [2 (trimethylsilyl) ethoxy] methyl (SEM).

[0127] In one embodiment, the reaction of the acid of formula 1d with the amine of formula 2c is carried out in a solvent in the presence of O- (7-azabenzotriazol-1-yl) -N, N, N ', N'20 tetramethyluronium (HATU) hexafluorophosphate and triethylamine or in a solvent in the presence of cyclic propyl phosphonic anhydride (T3P®) and pyridine. More specifically, this solvent includes N, N-dimethylformamide, ethyl acetate or 2-methyltetrahydrofuran.

[0128] In another embodiment, R^and is hydrogen or TBDMS.

[0129] In another embodiment, R^and means TBDMS.

[0130] In another embodiment, the method comprises an additional deprotection step; for example, when ring A is or

<img file="PL2349263T3_D0026.tif" />

where R^and is a silyl protecting group to form a compound of Formula (Ic) wherein ring A is or

<img file="PL2349263T3_D0027.tif" />

Typically, removal of the silyl protecting group requires treatment with an acid such as acetic acid or diluted mineral acid or the like, although other reagents such as a fluoride ion source (e.g. tetrabutylammonium fluoride) can be used.

[0131] In this method, the amine of formula 2c is prepared from the compound of formula 2a wherein the method comprises the steps of:

(a) reacting a compound of formula 2a with an amine of formula 3 to provide

<img file="PL2349263T3_D0028.tif" />

wherein:

Hal is F, Cl, Br or I; and this amine of formula 3 is

<img file="PL2349263T3_D0029.tif" />

and (b) reducing the compound of formula 2b to the amine of formula 2c.

<img file="PL2349263T3_D0030.tif" />

[0132] In one embodiment of the method for producing the amine of formula 2c, the amine of formula 3 in step (a) is prepared in situ from a suitable quaternary ammonium salt, such as an amine hydrochloride salt, although other ammonium salts (e.g., salt may also be used trifluoroacetate).

[0133] In one embodiment of the step (a) for preparing an amine of formula 2c, when said amine of formula 3 is

<img file="PL2349263T3_D0031.tif" />

R^and is hydrogen or TBDMS. More specifically, R^and means TBDMS.

[0134] In another embodiment, step (a) is carried out in a polar aprotic solvent in the presence of a tertiary amine base. Examples of tertiary amines that can be used include triethylamine, diisopropylethylamine, 1,5-diazabicyclo [4.3.0] non-5-ene (DBN), 1,8-diazabicyclo [5.4.0] undec-7-ene (DBU) , 1,4-diazabicyclo [2.2.2] octane (DABCO) and pyridine. Examples of usable solvents include N, N-dimethylformamide, dimethyl sulfoxide or acetonitrile.

[0135] In one embodiment, the tertiary amine base is triethylamine.

[0136] In another embodiment, step (a) is carried out in acetonitrile in the presence of triethylamine.

[0137] In another embodiment, the reaction temperature of step (a) is between approximately 75 ° C and approximately 85 ° C.

[0138] In another embodiment, the reaction time of step (a) is between approximately 2 and approximately 30 hours.

[0139] In one embodiment of the process for preparing the amine of formula 2c, step (b) is carried out in a polar protic solvent or a mixture of polar protic solvents in the presence of a palladium catalyst. When the catalyst is palladium, the solvent in step (b) is usually a polar protic solvent such as an alcohol. More specifically, it includes methanol or ethanol.

[0140] In another embodiment, step (b) is carried out in a polar protic solvent such as water in the presence of Fe and FeSO4 or Zn and AcOH.

[0141] Another aspect of the invention relates to a method of producing a compound of Formula (Ic):

<img file="PL2349263T3_D0032.tif" />

or a pharmaceutically acceptable salt thereof, comprising the steps of:

(a) reacting a compound of formula 2a with an amine of formula 3 to provide

<img file="PL2349263T3_D0033.tif" />

(b) converting a compound of formula 2b to an amine of formula 2c by reduction

<img file="PL2349263T3_D0034.tif" />

(c) reacting the amine of formula 2c with an acid of formula 1d to provide

<img file="PL2349263T3_D0035.tif" />

this amine of formula 3 is

<img file="PL2349263T3_D0036.tif" />

and ring A is selected from:

<img file="PL2349263T3_D0037.tif" />

where

R¹ is -CF3, -CN or -C = CCHN (CH3) 2;

R² is hydrogen, -CH3, -CF3, -OH or -CH2OH;

R³ is hydrogen, -CH3, -OCH3 or -CN;

both R² and R³ they are not both hydrogen and a

R^and is a hydrogen atom or a silyl protecting group selected from the group consisting of trimethylsilyl (TMS), tert-butyldiphenylsilyl (TBDPS), tert-butyldimethylsilyl (TBDMS), triisopropylsilyl (TIPS) and [2- (trimethylsilyl) ethoxy] methyl (SEM).

[0142] In one embodiment, the amine of formula 3 in step (a) is prepared in situ from a suitable quaternary ammonium salt, such as an amine hydrochloride salt, although other ammonium salts (e.g., trifluoroacetate salt) can also be used.

[0143] In one embodiment of the step (a) for preparing an amine of formula 2c, when the amine of formula 3 is

<img file="PL2349263T3_D0038.tif" />

R^and is hydrogen or TBDMS. More specifically, R^and means TBDMS.

[0144] In another embodiment, step (a) is carried out in a polar aprotic solvent in the presence of a tertiary amine base. Examples of possible tertiary amines include triethylamine, diisopropylethylamine, 1,5-diazabicyclo [4.3.0] non-5-ene (DBN), 1,8-diazabicyclo [5.4.0] undec-7-ene (DBU), 1 , 4-diazabicyclo [2.2.2] octane (DABCO) and pyridine.

[0145] In one embodiment, the tertiary amine base is triethylamine. [0146] In another embodiment, step (a) is carried out in acetonitrile in the presence of triethylamine. [0147] In another embodiment, the reaction temperature of step (a) is between approximately 75 ° C and approximately 85 ° C.

[0148] In another embodiment, the reaction time for step (a) is between approximately 2 and approximately 30 hours.

[0149] In one embodiment of the process for producing the amine of formula 2c, step (b) is carried out in a polar protic solvent or a mixture of polar protic solvents in the presence of a palladium catalyst. When the catalyst is palladium, the solvent in step (b) is usually a polar protic solvent such as an alcohol.

More specifically, the solvent includes methanol or ethanol.

[0150] In another embodiment, step (b) is carried out in a polar protic solvent, such as water, in the presence of Fe and FeSO4 or Zn and AcOH.

[0151] In one embodiment of step (c), the reaction of the acid of formula 1d with the amine of formula 2c is carried out in a solvent in the presence of O- (7-azabenzotriazol-1-yl) N, N, N ', N'-tetramethyluronium hexafluorophosphate (HATU) and triethylamine or in a solvent in the presence of cyclic propylphosphonic acid anhydride (T3P®) and pyridine. More specifically, this solvent includes N, N-dimethylformamide, ethyl acetate or 2-methyltetrahydrofuran.

[0152] In another embodiment, R^and is hydrogen or TBDMS.

[0153] In another embodiment, R^and means TBDMS.

[0154] In another embodiment, the method comprises an additional deprotection step, for example when ring A is ^ra° ,,, (c) '»or

<img file="PL2349263T3_D0039.tif" />

where R^and is a silyl protecting group to form a compound of Formula (I) wherein ring A is or

<img file="PL2349263T3_D0040.tif" />

Typically, removal of the silyl protecting group requires treatment with an acid such as acetic acid or diluted mineral acid or the like, although other reagents such as a fluoride ion source (e.g. tetrabutylammonium fluoride) can be used.

[0155] Another aspect of this invention relates to a compound that is

<img file="PL2349263T3_D0041.tif" />

wherein ring A is

R^andOH

H / ^ VOR^and , or (a) (b) (c) (d) in which

R¹ is -CF3, -CN or -C = CCHN (CH3) 2, and

R^and is a silyl protecting group selected from the group consisting of trimethylsilyl (TMS), tert-butyldiphenylsilyl (TBDPS), tert-butyldimethylsilyl (TBDMS), triisopropylsilyl (TIPS) and [2- (trimethylsilyl) ethoxy] methyl (SEM).

[0156] Another aspect of this invention relates to the compound it is

<img file="PL2349263T3_D0042.tif" />

wherein ring A is

<img file="PL2349263T3_D0043.tif" />

wherein

R¹ is -CF3, -CN or -C = CCH2N (CH3) 2, and

R^and is a silyl protecting group selected from the group consisting of trimethylsilyl (TMS), tert-butyldiphenylsilyl (TBDPS), tert-butyldimethylsilyl (TBDMS), triisopropylsilyl (TIPS) and [2- (trimethylsilyl) ethoxy] methyl (SEM).

[0157] Another aspect of this invention relates to a compound of Formula (IA):

<img file="PL2349263T3_D0044.tif" />

or pharmaceutically acceptable salts thereof, where:

is selected from

<img file="PL2349263T3_D0045.tif" />

where

R¹ is -CF3, -CN or -C = CCH2N (CH3) 2;

R² is hydrogen, -CH3, -CF3, -OH or -CH2OH;

R³ is hydrogen, -CH3, -OCH3, or -CN;

both R² and R³ they are not both hydrogen and a

R^and is a silyl protecting group selected from the group consisting of trimethylsilyl (TMS), tert-butyldiphenylsilyl (TBDPS), tert-butyldimethylsilyl (TBDMS), triisopropylsilyl (TIPS) and [2- (trimethylsilyl) ethoxy] methyl (SEM).

[0158] Another aspect of this invention relates to a compound of Formula (I)

<img file="PL2349263T3_D0046.tif" />

or a pharmaceutically acceptable salt thereof, wherein: Ring A is selected from:

<img file="PL2349263T3_D0047.tif" />

f ^IWM (a) (b) (c) (d) where

R¹ is -CF3, -CN or -C = CCH2N (CH3) 2;

R² is hydrogen, -CH3, -CF3, -OH or -CH2OH; R³ is hydrogen, -CH3, -OCH3, or -CN;

both R² and R³ they are not simultaneously hydrogen; prepared by any method disclosed herein.

[0159] Another aspect of this invention relates to a compound selected from the group consisting of:

<img file="PL2349263T3_D0048.tif" />

prepared by any method disclosed herein.

EXAMPLES [0160] Intermediate 1: 4-oxo-5- (trifluoromethyl) -1,4-dihydroquinoline-3-carboxylic acid (17). The synthesis of the title compound is outlined in Scheme 4.

<img file="PL2349263T3_D0049.tif" />

[0161] Preparation of diethyl 2 - ((2-chloro-5- (trifluoromethyl) phenylamino) methylene) malonate. 2-Chloro-5- (trifluoromethyl) aniline 12 (200 g, 1.023 mol), 2- (ethoxymethylene) diethyl malonate (276 g, 1.3 mol) and toluene (100 ml) combined under nitrogen in a 1-liter atmosphere round-bottom, three-necked flask equipped with a Dean-Stark apparatus. The solution was heated to 140 ° C with stirring and the temperature maintained for 4 h. The reaction mixture was cooled to 70 ° C and hexane (600 ml) was slowly added. The resulting suspension was stirred and allowed to warm to room temperature. The solid was collected by filtration, washed with 10% ethyl acetate in hexane (2x 400 mL) and then dried in vacuo to give a white solid (350 g, 94% yield) as the desired condensation product, 2- ( Diethyl (2-chloro-5- (trifluoromethyl) phenylamino) methylene) malonate 14.¹H NMR (400 MHz, DMSO-d6) δ 11.28 (d, J = 13.0 Hz, 1H), 8.63 (d, J = 13.0 Hz, 1H), 8.10 (s, 1H ), 7.80 (d, J = 8.3 Hz, 1H), 7.50 (dd, J = 1.5, 8.4 Hz, 1H), 4.24 (q, J = 7.1 Hz , 2H), 4.17 (q, J = 7.1 Hz, 2H), 1.27 (m, 6H).

[0162] Preparation of ethyl 8-chloro-4-oxo-5- (trifluoromethyl) -1,4-dihydroquinoline-3-carboxylate (15). A 3-neck, 1-L flask was filled into Dowtherm® (200 mL, 8 mL / g), which was degassed at 200 ° C for 1 h. The solvent was heated to 260 ° C and 2 portions were added to it in 10 min. - (2-chloro-5- (trifluoromethyl) phenylamino) methylene) diethyl malonate (25 g, 0.07 mol). The resulting mixture was stirred at 260 ° C for 6.5 hours (h) and the resulting ethanol by-product was removed by distillation. The mixture was allowed to cool slowly to 80 ° C. Hexane (150 ml) was slowly added over 30 minutes (min), then an additional 200 ml hexane was added in one portion. The suspension was then stirred until room temperature was reached. The solid was filtered off, washed with hexane (3 x 150 ml) and then dried in vacuo to give ethyl 8-chloro-4-oxo-5- (trifluoromethyl) -1,4-dihydroquinoline-3-carboxylate as a light brown solid (13.9 g, 65% yield). ¹H NMR (400 MHz, DMSO-d6) δ 11.91 (s, 1H), 8.39 (s, 1H), 8.06 (d, J = 8.3 Hz, 1H), 7.81 (d , J = 8.4 Hz, 1H), 4.24 (q, J = 7.1 Hz, 2H), 1.29 (t, J = 7.1 Hz, 3H).

[0163] Preparation of ethyl 4-oxo-5- (trifluoromethyl) -1H-quinoline-3-carboxylate (16). A 3-necked, 5-liter flask was charged with ethyl 8-chloro-4-oxo-5- (trifluoromethyl) -1,4-dihydroquinoline-3-carboxylate 15 (100 g, 0.3 mol), ethanol (1250 ml, 12 , 5 ml / g) and triethylamine (220 ml, 1.6 mol). Then 10 g of 10% Pd / C (50% humidity) was placed in the dish at 5 ° C. The reaction mixture was vigorously stirred under a hydrogen atmosphere for 20 h at 5 ° C, at which time the reaction mixture was concentrated to a volume of approximately 150 ml. The product, ethyl 4-oxo-5- (trifluoromethyl) -1H-quinoline-3-carboxylate 16, in the form of a suspension with Pd / C, was used directly in the next step.

[0164] Preparation of 4-oxo-5- (trifluoromethyl) -1,4-dihydroquinoline-3-carboxylic acid (17). Ethyl 4-oxo-5- (trifluoromethyl) -1H-quinoline-3-carboxylate 16 (58 g, 0.2 mol, crude reaction suspension containing Pd / C) was suspended in NaOH (814 ml 5 M, 4, 1 mol) in a 1 L reflux flask and heated at 80 ° C for 18 h and then still heated at 100 ° C for 5 h. The hot reaction mixture was filtered through packed Celite to remove Pd / C and Celite was rinsed with 1 N NaOH. The filtrate was acidified to about pH 1 to give a thick, white precipitate. The precipitate was filtered off, then rinsed with water and cold acetonitrile. The solid was then dried under vacuum to give 4-oxo-5- (trifluoromethyl) -1,4-dihydroquinoline-3-carboxylic acid 17 as a white solid (48 g, 92% yield).¹H NMR (400.0 MHz, DMSO-d6) δ 15.26 (s, 1H), 13.66 (s, 1H), 8.98 (s, 1H), 8.13 (dd, J = 1, 6, 7.8 Hz, 1H), 8.06 - 7.99 (m, 2H).

Intermediate 2: 4- (7-azabicyclo [2.2.1] heptan-7-yl) -2- (trifluoromethyl) -

<img file="PL2349263T3_D0050.tif" />

[0166] Preparation of 7- [4-nitro-3- (trifluoromethyl) phenyl] -7-azabicyclo [2.2.1] heptane (20). To a flask containing 7-azabicyclo [2.2.1] heptane 7a hydrochloride (4.6 g, 34.43 mmol, obtained from Tyger Scientific Inc., 324 Stokes Avenue, Ewing, NJ, 08638 USA under a nitrogen atmosphere, a 4-fluoro solution was added -1-nitro-2- (trifluoromethyl) benzene 18 (6.0 g, 28.69 mmol) and triethylamine (8.7 g, 12.00 mL, 86.07 mmol) in acetonitrile (50 mL). heated at 80 ° C under nitrogen for 16 h. The reaction mixture was allowed to cool and then partitioned between water and dichloromethane. The organic layer was washed with 1 M HCl, dried over Na2SO4, filtered and concentrated to dryness. Purification by silica gel chromatography (0-10% ethyl acetate in hexanes) gave 7- [4-nitro-3- (trifluoromethyl) phenyl] -7-azabicyclo [2.2.1] heptane 19 (7.2 g, yield 88%) as a yellow solid.¹H NMR (400.0 MHz, DMSO-d6) δ 8.03 (d, J = 9.1 Hz, 1H), 7.31 (d, J = 2.4 Hz, 1H), 7.25 (dd , J = 2.6, 9.1 Hz, 1H), 4.59 (s, 2H), 1.69 - 1.67 (m, 4H), 1.50 (d, J = 7.0 Hz, 4H).

[0167] Preparation of 4- (7-azabicyclo [2.2.1] heptan-7-yl) -2- (trifluoromethyl) aniline (20). From a flask filled with 7- [4-nitro-3- (trifluoromethyl) phenyl] -7-azabicyclo [2.2.1] heptane 19 (7.07 g, 24.70 mmol) and 10% Pd / C (0.71 g , 6.64 mmol), gas was removed and then nitrogen was bubbled through it. Ethanol (22 ml) was added and a hydrogen balloon was placed on the reaction flask. After vigorous stirring for 12 h, nitrogen was bubbled through the reaction mixture and Pd / C was removed by filtration. The filtrate was concentrated to a dark oil under reduced pressure and the residue was purified by silica gel chromatography (0-15% ethyl acetate in hexanes) to give 4- (7-azabicyclo [2.2.1] heptan-7-yl) -2 - (trifluoromethyl) aniline 20 as a purple solid (5.76 g, 91% yield). ¹H NMR (400.0 MHz, DMSO-d6) δ 6.95 (dd, J = 2.3, 8.8 Hz, 1H), 6.79 (d, J = 2.6 Hz, 1H), 6 , 72 (d, J = 8.8 Hz, 1H), 4.89 (s, 2H), 4.09 (s, 2H), 1.61 - 1.59 (m, 4H) and 1.35 ( d, J = 6.8 Hz, 4H).

[0168] Intermediate 3: 2-amino-5- (7-azabicyclo [2.2.1] heptan-7-yl) benzonitrile (23). The synthesis of the title compound is outlined in Scheme 6.

<img file="PL2349263T3_D0051.tif" />

[0169] Preparation of 5- (7-azabicyclo [2.2.1] heptan-7-yl) -2-nitrobenzonitrile (22). To a solution of 5-fluoro-2-nitrobenzonitrile 21 (160 mg, 0.96 mmol) in acetonitrile (1 mL) 7-azabicyclo [2.2.1] heptane 7a hydrochloride (129 mg, 0.96 mmol) and triethylamine ( 244 mg, 335.7 gl, 2.41 mmol). The reaction mixture was stirred at 60 ° C for 4 h. The reaction mixture was quenched with water, acidified with 1 N HCl to pH 1 and extracted with dichloromethane (3x 10 mL). The combined organic layers were washed with water, dried over MgSO4, filtered and concentrated to give 5- (7-azabicyclo [2.2.1] heptan-7-yl) -2-nitrobenzonitrile 22 (205 mg, 87% yield). LC / MS m / z 244.3 [M + H]⁺, retention time 1.69 min (RP-C18, 10-99% CH3CN / 0.05% TFA for 3 min).

[0170] Preparation of 2-amino-5- (7-azabicyclo [2.2.1] heptan-7-yl) benzonitrile (23).

A flask filled with 5- (7-azabicyclo [2.2.1] heptan-7-yl) -2-nitrobenzonitrile 22 (205 mg, 0.8427 mmol) and 10% Pd / C (41 mg, 0.39 mmol) nitrogen and then gas removed under vacuum. Methanol (4 ml) was added under a nitrogen atmosphere and a hydrogen balloon was placed on the flask. After stirring for 15 min, Pd / C was removed by filtration and the solvent removed under reduced pressure to give 2-amino-5- (7-azabicyclo [2.2.1] heptan-7-yl) benzonitrile 23 (170 mg, 95% yield). ¹H NMR (400.0 MHz, DMSO-d6) δ 7.02 (dd, J = 2.8, 9.0 Hz, 1H), 6.87 (d, J = 2.7 Hz, 1H), 6 , 68 (d, J = 9.0 Hz, 1H), 5.36 (s, 2H), 4.09 (s, 2H), 1.59 (d, J = 6.8 Hz, 4H), 1 , 34 (d, J = 6.8 Hz, 4H).

[0171] Intermediate 4: 4- (7-azabicyclo [2.2.1] heptan-7-yl) -2- (3- (dimethylamino) prop-1-ynyl) aniline (27). The synthesis of the title compound is outlined in Scheme 7.

<img file="PL2349263T3_D0052.tif" />

[0172] Preparation of 7- (3-bromo-4-nitrophenyl) -7-azabicyclo [2.2.1] heptane (25). To a solution of 2-bromo-4-fluoro-1-nitrobenzene 24 (1.1 g, 4.8 mmol) and K2CO3 (2.0 g, 14.3 mmol) in DMSO (8.400 mL) was added portionwise with 7-azabicyclo [ 2.2.1] heptane 7a (765.4 mg, 5.7 mmol). The reaction mixture was stirred at 80 ° C for 24 h. The reaction mixture was diluted with water to precipitate the product. The solid was redissolved in dichloromethane, washed with 1.0 N HCl, dried over MgSO4, filtered and concentrated to give 7- (3-bromo-4-nitrophenyl) -7-azabicyclo [2.2.1] heptane 25 (1 , 1 g, 78% yield). The crude product was used directly in the next step. LC / MS m / z 299.1 [M + H]⁺, retention time 1.97 min (RP-C18, 10-99% CH3CN / 0.05% TFA for 3 min).

[0173] Preparation of 3- [5- (7-Azabicyclo [2.2.1] heptan-7-yl) -2-nitrophenyl] -N, N-dimethylprop-2-yno-1-amine (26). To 7- (3-bromo-4-nitrophenyl) -7-azabicyclo [2.2.1] heptane 25 (500 mg, 1.683 mmol), Pd (PPh3) 2Cl2 (59 mg, 0.08 mmol) and cuprous iodide (9.616 mg, 1.708 g, 0.05049 mmol), a solution of N, N-dimethylprop-2-y-1-amine (420 mg, 538 g, 5.05 mmol) in degassed DMF (5 mL) and triethylamine (5 mL) was added. . The reaction mixture was microwaved under N2 for 10 min at 100 ° C. The reaction mixture was diluted with ethyl acetate, washed with 50% saturated sodium bicarbonate solution (2x 20 mL), water and brine. The solution was dried over anhydrous Na2SO4 and filtered to give a red solid. Purification by silica gel chromatography (0-50% dichloromethane in ethyl acetate) gave 3- [5- (7-azabicyclo [2.2.1] heptan-7-yl) -2-nitrophenyl] -N, N-dimethylprop- 2-yno-1amine 26 (400 mg, 79% yield). LC / MS m / z 300.5 [M + H]⁺, retention time 1.11 min (RP-C18, 10-99% CH3CN / 0.05% TFA for 3 min).

[0174] Preparation of 4- (7-azabicyclo [2.2.1] heptan-7-yl) -2- (3-dimethylaminoprop-1-ynyl) aniline (27). 3- [5- (7-Azabicyclo [2.2.1] heptan-7-yl) -2-nitrophenyl] -N, N-dimethylprop-2-y-1-amine 26 (340 mg, 1.14 mmol) iron (634 mg, 11.36 mmol) and ferrous sulfate heptahydrate (316 mg, 1.136 mmol) were suspended in water (1 mL) and heated to reflux for 20 min. The reaction mixture was filtered and the solid washed with methanol and dichloromethane. The filtrate was concentrated and purified by silica gel chromatography using 0-5% methanol in dichloromethane to give 4- (7-azabicyclo [2.2.1] heptan-7-yl) -2- (3-dimethylaminoprop-1-ynyl) aniline 27 (148 mg, 48% yield). LC / MS m / z 270.3 [M + H]⁺, retention time 0.25 min (RP-C18, 10-99% CH3CN / 0.05% TFA for 3 min).

[0175] Intermediate 5: ex0-4- (2- (tert-butyldimethylsilyloxy) -7-azabicyclo [2.2.1] heptan-7-yl) -2- (trifluoromethyl) aniline (30). The synthesis of the title compound before

<img file="PL2349263T3_D0053.tif" />

[0176] Preparation of exo-7- [4-nitro-3- (trifluoromethyl) phenyl] -7-azabicyclo [2.2.1] heptan-5-ol (28). To a flask containing exo-7-azabicyclo [2.2.1] heptan-2-ol 7b (0.86 g, 5.74 mmol) under a nitrogen atmosphere, a solution of 4-fluoro-1-nitro-2- (trifluoromethyl) benzene was added under a nitrogen atmosphere. (1 g, 4.78 mmol) and triethylamine (1.45 g, 2.0 mL, 14.35 mmol) in acetonitrile (8 mL). The reaction mixture was heated at 84 ° C under nitrogen for 22 h. The reaction mixture was allowed to cool and then partitioned between water and ethyl acetate. The layers were separated and the aqueous layer was extracted twice with ethyl acetate. The combined organic layers were dried over Na2SO4, filtered and concentrated to dryness. Purification by silica gel chromatography (0-50% ethyl acetate in hexanes) gave exo-7- [4-nitro-3- (trifluoromethyl) phenyl] -7-azabicyclo [2.2.1] heptan-5-ol 28 in a yellow solid (0.67 g, 46% yield). LC / MS m / z 303.3 [M + H]⁺, retention time 1.51 min (RP-C18, 10-99% CH3CN / 0.05% TFA for 3 min).

[0177] Preparation of exo-tert-butyldimethyl - [[7- [4-nitro-3- (trifluoromethyl) phenyl] 7-azabicyclo [2.2.1] heptan-5-yl] oxy] silane 29. Tert-Butylchlorodimethylsilane (197 mg, 1.267 mmol) was added to a solution of 4H-imidazole (144 mg, 2.11 mmol) in DMF (0.5 mL). When no more bubbling ceased from the solution, exo-7- [4-nitro-3- (trifluoromethyl) phenyl] -7-azabicyclo [2.2.1] heptan-5-ol 28 (255 mg, 0.84 mmol) was added as solution in DMF (0.6 ml) and stirred at room temperature for 14 h. The reaction mixture was quenched with water and extracted twice with diethyl ether, dried over MgSO 4, filtered and concentrated to a colorless oil. Purification by silica gel chromatography (0-40% dichloromethane in hexanes) gave exo-tert-butyldimethyl - [[7- [4-nitro-3- (trifluoromethyl) phenyl] -7-azabicyclo [2.2.1] heptane- 5-yl] oxy] silane 29 (318 mg, 90% yield) as a yellow oil.¹H NMR (400.0 MHz, DMSO- d6) δ 8.01 (d, J = 9.2 Hz, 1H), 7.29 (d, J = 2.4 Hz, 1H), 7.19 (dd , J = 2.6, 9.2 Hz, 1H), 4.60 (t, J = 4.4 Hz, 1H), 4.47 (d, J = 5.2 Hz, 1H), 4.07 (dd, J = 2.0, 6.8 Hz, 1H), 1.94 (dd, J = 6.4, 12.8 Hz, 1H), 1.71 - 1.47 (m, 3H), 1.39 - 1.32 (m, 2H), 0.65 (s, 9H), 0.03 (s, 6H).

[0178] Preparation of exo-4- [5- [tert-butyl (dimethyl) silyl] oxy-7-azabicyclo [2.2.1] heptan-7-yl] -2- (trifluoromethyl) aniline (30). From the flask containing palladium on activated carbon (10 wt% 30 mg, 0.28 mmol), gas was removed, N2 passed through it and filled with exo-tert-butyldimethyl - [[7- [4-nitro-3- (trifluoromethyl) solution) phenyl] -7-azabicyclo [2.2.1] heptan-5-yl] oxy] silane 29 (301 mg, 0.72 mmol) in ethanol (3 mL). The gas was removed from the flask and then a H2 balloon was placed on the reaction flask and stirred for 4 h at room temperature. The mixture was filtered and concentrated to dryness to give exo-4- [5- [tert-butyl (dimethyl) silyl] oxy-7-azabicyclo [2.2.1] heptan-7-yl] -2- (trifluoromethyl) aniline ( 268 mg, 96% yield) as an off-white solid.¹H NMR (400.0 MHz, DMSO-d6) δ 6.92 (dd, J = 2.4, 8.8 Hz, 1H), 6.77 (d, J = 2.6 Hz, 1H), 6 , 70 (d, J = 8.8 Hz, 1H), 4.84 (s, 2H), 4.11 (t, J = 4.4 Hz, 1H), 3.91 - 3.89 (m, 2H), 1.82 (dd, J = 7.1, 12.3 Hz, 1H), 1.54 - 1.39 (m, 3H), 1.20 1.16 (m, 2H), 0, 79 (s, 9H), 0.02 (s, 6H).

[0179] Intermediate 6: e "Jo-4- (2- (tert-butyldimethylsilyloxy) -7-azabicyclo [2.2.1] heptan-7-yl) -2- (trifluoromethyl) aniline (34). Preparation of the title compound

<img file="PL2349263T3_D0054.tif" />

[0180] Preparation of 7-azabicyclo [2.2.1] heptan-5-one (31). To a solution of oxalyl dichloride (165 mg, 113 μΐ, 1.27 mmol) in dichloromethane (3 mL) under a nitrogen atmosphere at -78 ° C, a solution of DMSO (199 mg, 180 g, 2.54 mmol) in dichloromethane (0, 7 ml). The reaction mixture was allowed to stir for 30 min, and then a solution of exo-7- [4-nitro-3- (trifluoromethyl) phenyl] -7-azabicyclo [2.2.1] heptan-5-ol 28 (320 mg, 1 , 06 mmol) in dichloromethane (2.5 mL). The reaction mixture was stirred for an additional hour at -78 ° C, then triethylamine (536 mg, 738 g, 5.30 mmol) was added dropwise and the reaction mixture was warmed to room temperature. The reaction mixture was diluted with dichloromethane, partitioned between dichloromethane and water, and the layers were separated. The aqueous layer was extracted again with dichloromethane. The combined organic layers were dried over Na2SO4, filtered and concentrated to a yellow oil. Purification by silica gel chromatography (0-30% ethyl acetate in hexanes) gave 7- [4-nitro-3- (trifluoromethyl) phenyl] -7-azabicyclo [2.2.1] heptan-5-one 31 (266 mg , 84% yield) as a yellow solid. ¹H NMR (400.0 MHz, DMSO- d6) δ 8.06 (d, J = 9.1 Hz, 1H), 7.47 (d, J = 2.4 Hz, 1H), 7.39 (dd , J = 2.6, 9.1 Hz, 1H), 4.98 (t, J = 4.5 Hz, 1H), 4.84 (d, J = 5.4 Hz, 1H), 2.44 (d, J = 3.1 Hz, 1H), 2.23 (d, J = 16 Hz, 1H), 2.00 - 1.92 (m, 1H), 1.88 - 1.70 (m, 2H), 1.66 - 1.60 (m, 1H), [0181] Preparation of endo-7- [4-nitro-3- (trifluoromethyl) phenyl] -7-azabicyclo [2.2.1] heptan-5-ol (32). To a solution of 7- [4-nitro-3- (trifluoromethyl) phenyl] -7-azabicyclo [2.2.1] heptan-5-one 31 (261 mg, 0.87 mmol) in THF (11 mL) at -55 ° A solution of lithium tri-sec-butyl borohydride (1.04 mL 1 M, 1.04 mmol) was added dropwise under a nitrogen atmosphere. After 30 min, the reaction mixture was transferred to an ice-water bath and stirring continued. The reaction mixture was quenched with methanol (1.2 mL) at 0 ° C. The reaction mixture was partitioned between dichloromethane / water, separated and the aqueous layer was extracted twice more with dichloromethane. The organic layers were combined, dried over Na2SO4, filtered and concentrated to dryness. Purification by silica gel chromatography (0-50% ethyl acetate in hexanes) gave endo-7- [4-nitro-3- (trifluoromethyl) phenyl] -7-azabicyclo [2.2.1] heptan-5-ol 32 (222 mg, 84% yield) as a yellow solid.¹H NMR (400.0 MHz, DMSO- d6) δ 8.01 (d, J = 9.1 Hz, 1H), 7.27 (d, J = 3.0 Hz, 1H), 7.22 (dd , J = 2.6, 9.1 Hz, 1H), 5.17 (d, J = 4.4 Hz, 1H), 4.49 (t, J = 4.9 Hz, 1H), 4.44 (t, J = 4.5 Hz, 1H), 4.16 - 4.10 (m, 1H), 2.20 - 2.06 (m, 2H), 1.67 - 1.44 (m, 3H ), 1.09 (dd, J = 3.5, 12.4 Hz, 1H).

[0182] Preparation of endo-tert-butyldimethyl - [[7- [4-nitro-3- (trifluoromethyl) phenyl] 7-azabicyclo [2.2.1] heptan-5-yl] oxy] silane (33). Tert-butylchlorodimethylsilane (168 mg, 1.08 mmol) was added to a solution of 4H-imidazole (122 mg, 1.80 mmol) in DMF (425.3 μL.) When the solution stopped bubbling, endo-7- [ 4-nitro-3- (trifluoromethyl) phenyl] -7-azabicyclo [2.2.1] heptan-5-ol 32 (217 mg, 0.72 mmol) as a solution in DMF (1 mL) and stirred at room temperature for 14 h. The reaction mixture was quenched with water and extracted twice with diethyl ether, dried over MgSO 4, filtered and concentrated to a colorless oil. Purification by silica gel chromatography (0-40% dichloromethane in hexanes) gave endo-tert-butyldimethyl - [[7- [4-nitro-3- (trifluoromethyl) phenyl] -7-azabicyclo [2.2.1] heptane- 5-yl] oxy] silane 33 (251 mg, 84% yield) as a yellow oil.¹H NMR (400.0 MHz, DMSO- d6) δ 8.01 (d, J = 9.1 Hz, 1H), 7.32 (d, J = 2.3 Hz, 1H), 7.26 (dd , J = 2.5, 9.1 Hz, 1H), 4.54-4.51 (m, 2H), 4.29 - 4.26 (m, 1H), 2.20-2.11 (m , 2H), 1.67 - 1.45 (m, 3H), 1.08 (dd, J = 3.2, 12.4 Hz, 1H), 0.88 (s, 9H), 0.07 ( d, J = 2.6 Hz, 6H).

[0183] Preparation of endo-4- [5- [tert-butyl (dimethyl) silyl] oxy-7-azabicyclo [2.2.1] heptan-7-yl] -2- (trifluoromethyl) aniline (34). A gas was removed from the flask containing palladium on activated carbon (10 wt%, 24 mg, 0.23 mmol) and then flushed with nitrogen. To the mixture, a solution of endo-tert-butyldimethyl - [[7- [4-nitro-3- (trifluoromethyl) phenyl] -7-azabicyclo [2.2. 1] heptan-5-yl] oxy] silane 33 (240 mg, 0.58 mmol) in ethanol (5 mL). Gas was removed from the reaction mixture, then a H2 balloon was placed on the flask and stirred for 4 h at room temperature. The mixture was filtered and concentrated to dryness to give endo-4- [5- [tert37 butyl (dimethyl) silyl] oxy-7-azabicyclo [2.2.1] heptan-7-yl] -2- (trifluoromethyl) aniline 34 (222 mg, 100% yield) as an off-white solid.¹H NMR (400.0 MHz, DMSO- d6) δ 6.95 (dd, J = 2.4, 8.8 Hz, 1H), 6.79 (d, J = 2.6 Hz, 1H), 6 , 72 (d, J = 8.8 Hz, 1H), 4.91 (s, 2H), 4.24 - 4.19 (m, 1H), 4.06 - 4.03 (m, 2H), 2.12 - 1.99 (m, 2H), 1.55 - 1.53 (m, 1H), 1.42 - 1.36 (m, 2H), 0.96 (dd, J = 3.2 , 12.2 Hz, 1H), 0.87 (s, 9H), 0.05 (s, 6H).

Example Compound 3: N- (4- (7-azabicyclo [2.2.1] heptan-7-yl) -2- (trifluoromethyl) phenyl) -4-oxo-5- (trifluoromethyl) -1,4-dihydroquinoline-3 carboxamide.

<img file="PL2349263T3_D0055.tif" />

[0185] To a solution of 4-oxo-5- (trifluoromethyl) -1H-quinoline-3-carboxylic acid 17 (9.1 g, 35.39 mmol) and 4- (7-azabicyclo [2.2.1] heptane-7 -yl) -2- (trifluoromethyl) aniline 20 (9.2 g, 35.74 mmol) in 2-methyltetrahydrofuran (91.00 mL) cyclic propyl phosphonic anhydride (T3P, 50% solution in ethyl acetate, 52.68 ml, 88.48 mmol) and pyridine (5.6 g, 5.73 ml, 70.78 mmol) at room temperature. The reaction flask was heated at 65 ° C for 10 h under nitrogen. After cooling to room temperature, the reaction mixture was then diluted with ethyl acetate and quenched with saturated Na2CO3 solution (50 mL). The layers were separated and the aqueous layer was extracted twice with ethyl acetate. The combined organic layers were washed with water, dried over Na2SO4, filtered and concentrated to a light brown solid. The crude solid product was suspended in ethyl acetate / diethyl ether (2: 1), collected by vacuum filtration and washed twice with ethyl acetate / diethyl ether (2: 1) to give the product as a light yellow crystalline powder. The powder was dissolved in warm ethyl acetate and absorbed on Celite. Purification by silica gel chromatography (0-50% ethyl acetate in dichloromethane) gave N- (4- (7-azabicyclo [2.2.1] heptan-7-yl) -2- (trifluoromethyl) phenyl) -4-oxo5 - (trifluoromethyl) -1,4-dihydroquinoline-3-carboxamide as a white crystalline solid (13.5 g, 76% yield). LC / MS m / z 496.0 [M + H]⁺, retention time 1.48 min (RP-C18, 10-99% CH3CN / 0.05% TFA for 3 min). ¹H NMR (400.0 MHz, DMSO-d6) δ 13.08 (s, 1H), 12.16 (s, 1H), 8.88 (s, 1H), 8.04 (dd, J = 2, 1, 7.4 Hz, 1H), 7.95 - 7.88 (m, 3H), 7.22 (dd, 2.5, 8.9 Hz, 1H), 7.16 (d, J = 2 , 5 Hz, 1H), 4.33 (s, 2H), 1.67 (d, J = 6.9 Hz, 4H), 1.44 (d, J = 6.9 Hz, 4H).

[0186] Example Compound 3 Form A Salt HCl N- (4- (7-azabicyclo [2.2.1] heptan-7-yl) -2- (trifluoromethyl) phenyl) -4-oxo-5- (trifluoromethyl) 1,4 hydrochloride -dihydroquinoline-3-carboxamide (Form A-HCl).

<img file="PL2349263T3_D0056.tif" />

[0187] Preparation of 7- [4-nitro-3- (trifluoromethyl) phenyl] -7-azabicyclo [2.2.1] heptane (19). 4-fluoro-1-nitro-2- (trifluoromethyl) benzene (18) (901 g) was added to a 30 L jacketed vessel. Sodium carbonate (959.1 g) and 5 L dimethyl sulfoxide (DMSO) were added and the mixture was stirred under nitrogen. 7-Azabicyclo [2.2.1] heptane (7a) hydrochloride (633.4 g) was added portionwise to the vessel. The temperature of the mixture was gradually raised to 55 ° C and the reaction was monitored by HPLC. When less than 1% of the AUC of the starting compound remained, the reaction was considered complete. The mixture was then diluted with 10 volumes of 2-methyltetrahydrofuran and washed three times with 5.5 volumes of water until no DMSO remained in the aqueous layer as shown by HPLC analysis to give 7- [4-nitro-3- (trifluoromethyl) phenyl] -7-azabicyclo [2.2.1] heptane (19) in 2-methyltetrahydrofuran (yield about 95%).

[0188] Preparation of 4- (7-azabicyclo [2.2.1] heptan-7-yl) -2 (trifluoromethyl) aniline hydrochloride salt (20'HCl). Palladium on carbon (150 g, 5% w / w) was placed in a hydrogen generator from the company ΒϋΛί (20 l capacity) under a nitrogen atmosphere, then 7- [4-nitro-3- (trifluoromethyl) phenyl] -7 was added -azabicyclo [2.2.1] heptane (19) (1500 g) and 2-methyl tetrahydrofuran (10.5 l, 7 volumes). Hydrogen gas was introduced into the closed hydrogen generator to a pressure of 0.5 bar. Vacuum was applied for about 2 min and then hydrogen gas was introduced to a pressure of 0.5 bar. This process was repeated 2 times. Hydrogen gas was then continuously introduced into the mixture at a pressure of 0.5 bar. The mixture was then stirred at a temperature between 18 and 23 ° C using vessel jacket cooling. When hydrogen gas was no longer consumed and the reaction was no longer exothermic, a vacuum was created in the vessel. Then nitrogen gas was introduced into the vessel at 0.5 bar pressure and again vacuum was created, followed by the introduction of a second portion of nitrogen gas at 0.5 bar pressure. When HPLC analysis of the filtered samples showed that no more 7- [4-nitro3- (trifluoromethyl) phenyl] -7-azabicyclo [2.2.1] heptane (19) remained (e.g. &lt;0.5%), the reaction mixture was transferred to the receiving flask under a nitrogen atmosphere using a filter funnel using a Celite filter. The Celite filter cake was washed with 2-methyltetrahydrofuran (3 L, 2 volumes). The washed solutions and filtrates were placed in a vessel equipped with a stirrer, a temperature control device and under a nitrogen atmosphere. 4 M HCl in 1,4-dioxane (1 volume) was added continuously to the vessel for 1 h at 20 ° C. The mixture was stirred for another 10 h (or overnight), filtered and washed with 2-methyltetrahydrofuran (2 volumes) and dried to give 1519 g of salt, 4- (7-azabicyclo [2.2.1] heptan-7-yl) hydrochloride -2- (trifluoromethyl) aniline (20'HCl) as a white crystalline solid (yield about 97%).

[0189] Alternative preparation of 7- [4-amino-3- (trifluoromethyl) phenyl-7-azabicyclic 2.2.11 heptane (20-HCl) hydrochloride salt.

<img file="PL2349263T3_D0057.tif" />

i) 5% Pd / C, K₂ &lt;G). 2-MeTHF) ii) 4N HCl in 1,4-dioxane

<img file="PL2349263T3_D0058.tif" />

CF₃

20HCI [0190] In a Buchi hydrogen generator (20 L capacity) palladium on carbon (5% w / w) (150 g) was placed under a nitrogen atmosphere, then 7- [4-nitro-3 (trifluoromethyl) was added phenyl] -7-azabicyclo [2.2. 1] heptane 19 (1500 g) and 2-methyltetrahydrofuran (10.5 L, 7 volumes). Hydrogen gas was introduced into the vessel to a pressure of 0.5 bar. Vacuum was applied briefly (2 min) and then hydrogen gas was introduced to a pressure of 0.5 bar. This process was repeated two more times, then hydrogen gas was introduced into the hydrogen generator under a pressure of 0.5 bar and mixing was started. The temperature of the reaction mixture was kept at 18 to 23 ° C by cooling the jacketed vessel. When hydrogen gas was no longer consumed and the reaction was no longer exothermic, a vacuum was created in the vessel. Then nitrogen gas was introduced into the vessel and vacuum was again created, followed by a second portion of nitrogen gas under a pressure of 0.5 bar. The reaction was considered complete when HPLC analysis of the filtered sample showed that 7- [4-nitro-3- (trifluoromethyl) phenyl] -7-azabicyclo [2.2.1] heptane (&lt;0.5%) was no longer detected. The reaction mixture was then filtered through Celite. The remaining suspension was transferred to a receiving flask under a nitrogen atmosphere using a filter funnel containing a Celite filter. The Celite layer was washed with 2-methyltetrahydrofuran (3 L, 2 volumes). The filtrate and washing solutions were transferred to a vessel equipped with a mixing device, temperature control device and under a nitrogen atmosphere. 4 M HCl in 1,4-dioxane (1 volume) was added continuously to the vessel for 1 h at 20 ° C. The resulting mixture was stirred for an additional 10 h, filtered and washed with 2-methyltetrahydrofuran (2 volumes) and dried to give 1519 g of 7- [4-amino-3- (trifluoromethyl) phenyl] -7-azabicyclo [2.2.1] hydrochloride heptane (20'HCl) as a white crystalline solid.

[0191] N- (4- (7-azabicyclo [2.2.1] heptan-7-yl) -2- (trifluoromethyl) phenyl) -4-oxo-5- (trifluoromethyl) -1,4-dihydroquinoline-3 hydrochloride -carboxamide (Form AHCl). 2-Methyltetrahydrofuran (0.57 L, 1.0 volume) was placed in a 30 L jacketed reactor vessel, followed by the addition of 4- (7-azabicyclo [2.2.1] heptan-7-yl) -2- (trifluoromethyl) hydrochloride salt anilines (20'HCl) (791 g, 2.67 mol) and 4-oxo-5- (trifluoromethyl) -1,4-dihydroquinoline-3-carboxylic acid (17) (573 g, 2.23 mol) and an additional 5 , 2 L (9.0 volumes) 2-methyltetrahydrofuran. Stirring was started and T3P in 2-methyltetrahydrofuran (2.84 kg, 4.46 mol) was added to the reaction mixture over 15 min. Then, pyridine (534.0 g, 546.0 mL, 6.68 mol) was added dropwise over 30 min using a dropping funnel. The mixture was heated to 45 ° C for about 30 min and stirred for 12-15 h. HPLC analysis showed that 4-oxo-5- (trifluoromethyl) -1,4-dihydroquinoline-3-carboxylic acid was present in less than 2%. Then the mixture was cooled to room temperature. 2-methyltetrahydrofuran (4 volumes, 2.29 L) was added, followed by the addition of water (6.9 volumes, 4 L) while keeping the temperature below 30 ° C. The aqueous layer was removed and the organic layer was carefully washed twice with saturated aqueous NaHCO3 solution. The organic layer was then washed with 10% w / w citric acid (5 volumes) and finally water (7 volumes).

The mixture after the final treatment is filtered and transferred to another dry vessel. N- (4- (7-azabicyclo [2.2.1] heptan-7-yl) 2- (trifluoromethyl) phenyl) -4-oxo-5- (trifluoromethyl) -1,4-dihydroquinoline-3-carboxamide hydrochloride seed crystals were added ( Form A-HCl) (3.281 g, 5.570 mmol) from the previous batch. HCl bubbles (g) (10 eq) were bubbled through for 2 h and the mixture was stirred overnight. The resulting suspension was filtered, washed with 2-methyltetrahydrofuran (4 volumes), dried by suction and dried in an oven at 60 ° C until a constant weight of 868 g of N- (4- (7-azabicyclo [2.2.1]] heptane-7- hydrochloride was obtained. yl) -2- (trifluoromethyl) phenyl) -4-oxo-5- (trifluoromethyl) -1,4-dihydroquinoline-3-carboxamide (Form A-HCl).

[0192] Example Compound 3 Form B Salt HCl A- (4- (7-azabicyclo [2.2.1] heptan-7-yl) -2- (trifluoromethyl) phenyl) -4-oxo-5- (trifluoromethyl) hydrochloride - 1,4-dihydroquinoline-3-carboxamide (Form B-HCl).

<img file="PL2349263T3_D0059.tif" />

[0193] 2-Methyltetrahydrofuran (100 mL) was placed under a nitrogen atmosphere in a 3-neck flask equipped with a stirrer. Example Compound 3 Form A-HCl (Example 3B, 55 g, 0.103 mol) was added to the flask followed by 349 ml of 2-methyltetrahydrofuran and stirring was started. 28 ml of water was added to the flask and the flask was warmed to an internal temperature of 60 ° C and the mixture was stirred for 48 h. The flask was cooled to room temperature and the mixture was stirred for 1 h. The reaction mixture was vacuum filtered until the filter cake was dry. The solid filter cake was washed twice with 2-methyltetrahydrofuran (4 volumes). The solid filter cake was allowed to suck under vacuum for a period of about 30 minutes and transferred to a drying tray. The filter cake was dried to constant weight under vacuum at 60 ° C to give Example Compound 3 Form B-HCl as a white crystalline solid (49 g) (yield about 90%).

[0194] Example Compound 6: Preparation of N- (4- (7-azabicyclo [2.2.1] heptan-7-yl) 2-cyanophenyl) -5-methyl-4-oxo-1,4-dihydroquinoline-3-carboxamide . Wytwarza-

<img file="PL2349263T3_D0060.tif" />

[0195] For a solution of 5-methyl-4-oxo-1,4-dihydroquinoline-3-carboxylic acid (162 mg, 0.80 mmol) and 2-amino-5- (7-azabicyclo [2.2.1] heptane -7-yl) benzonitrile 23 (170 mg, 0.80 mmol) in 2-methyltetrahydrofuran (1.5 mL) cyclic propylphosphonic acid anhydride (50% solution in ethyl acetate, 949.5 gl, 1.605 mmol) and pyridine ( 126 mg, 129 g, 1.60 mmol). The reaction mixture was sealed and heated at 100 ° C for 65 min under microwave irradiation. The reaction mixture was cooled to room temperature, diluted with ethyl acetate (10 mL) and quenched with saturated Na2CO3 solution (6 mL). The organic layer was dried over Na2SO4, filtered and concentrated. Purification by silica gel chromatography (0-35% ethyl acetate in dichloromethane) gave N- (4- (7-azabicyclo [2.2.1] heptan-7-yl) -2-cyanophenyl) -5-methyl-4- oxo-1,4-dihydroquinoline-3-carboxamide (157 mg, 49% yield). LC / MS m / z 399.3 [M + H]⁺, retention time 1.47 min (RP-C18, 10-99% CH3CN / 0.05% TFA for 3 min). ¹H NMR (400.0 MHz, DMSO-d6) δ 12.77 (s, 1H), 12.75 (s, 1H), 8.77 (s, 1H), 8.11 (d, J = 9, 1 Hz, 1H), 7.64 - 7.60 (m, 1H), 7.55 (d, J = 8.0 Hz, 1H), 7.34 (d, J = 2.8 Hz, 1H) , 7.27 (dd, J = 2.8, 9.1 Hz, 1H), 7.23 (d, J = 7.2 Hz, 1H), 4.32 (s, 2H), 2.91 ( s, 3H), 1.65 (d, J = 7.2 Hz, 4H), 1.42 (d, J = 6.8 Hz, 4H).

[0196] Exemplary Compound 13: N- [4- (7-azabicyclo [2.2.1] heptan-7-yl) -2- (3-dimethylaminoprop-1-ynyl) phenyl] -4-oxo-5- (trifluoromethyl ) -1 H-quinoline-3-carboxamide. Preparation of the title compound is outlined in Scheme 12.

<img file="PL2349263T3_D0061.tif" />

[0197] To a solution of 4-oxo-5- (trifluoromethyl) -1H-quinoline-3-carboxylic acid 17 (19 mg, 0.07 mmol) and 4- (7-azabicyclo [2.2.1] heptan-7-yl ) -2- (3-dimethylaminoprop-1-ynyl) aniline 27 (20 mg, 0.07 mmol) in 2-methyltetrahydrofuran (190.9 g) added T3P (118 mg, 0.19 mmol) and pyridine (12 mg, 12 gl, 0.15 mmol). The reaction mixture was heated at 100 ° C for 30 min under microwave irradiation. The reaction mixture was diluted with EtOAc and quenched with saturated aqueous NaHCO3 (50 mL). The layers were separated and the aqueous layer was extracted twice with EtOAc. The combined organics were washed once with water, dried over Na2SO4, filtered and concentrated. The residue was purified by reverse phase HPLC (0-99% CH3CN / 0.05% TFA) to give N- [4- (7-azabicyclo [2.2.1] heptan-7-yl) 2- (3 -dimethylaminoprop-1-ynyl) phenyl] -4-oxo-5- (trifluoromethyl) -1H-quinoline-3-carboxamide (8 mg, 17% yield). LC / MS m / z 509.7 [M + H]⁺, retention time 1.06 min (RP-C18, 10-99% CH3CN / 0.05% TFA for 3 min). ¹H NMR (400.0 MHz, DMSO-d6) δ 13.23 (d, J = 6.8 Hz, 1H), 12.40 (s, 1H), 10.31 (s, 1H), 8.96 (d, J = 6.6 Hz, 1H), 8.40 (d, J = 9.0 Hz, 1H), 8.08 - 8.06 (m, H), 8.07 (dd, J = 1.5 Hz, 8.1 Hz, 1H), 8.00 - 7.95 (m, 2H), 7.15 - 7.09 (m, 2H), 4.49 (s, 2H), 4, 29 (s, 2H), 2.94 (s, 6H), 1.67 (d, J = 7.2 Hz, 4H), 1.44 (d, J = 7.0 Hz, 4H).

Example Compound 5: £ W0-N- (4 - [(5S) -5- [tert-butyl (dimethyl) silyl] oxy-7-azabicyclo [2.2.1] heptan-7-yl] -2- (trifluoromethyl phenyl] -4-oxo-5- (trifluoromethyl) 1H-quinoline-3-carboxamide. Preparation of the title compound is outlined in Scheme 13.

<img file="PL2349263T3_D0062.tif" />

[0199] Preparation of endo-N- [4 - [(5S) -5- [tert-butyl (dimethyl) silyl] oxy-7-azabicyclo [2.2.1] heptan-7-yl] -2- (trifluoromethyl) phenyl ] -4-oxo-5- (trifluoromethyl) -1 H-quinoline-3-carboxamide. To a solution of 4-oxo-5- (trifluoromethyl) -1H-quinoline-3-carboxylic acid 17 (148 mg, 0.58 mmol), O- (7-azabenzotriazol-1-yl) -N, N, N ', N'- hexafluorophosphate tetramethyluronium (HATU) (306 mg, 0.81 mmol) in 2-methyltetrahydrofuran (2.2 mL) was added endo-4- [5- [tert-butyl (dimethyl) silyl] oxy-7-azabicyclo [2.2.1] heptan-7-yl] -2- (trifluoromethyl) aniline 34 (222 mg, 0.58 mmol), followed by the addition of triethylamine (146 mg, 201 g, 1.44 mmol). The reaction mixture was heated at 62 ° C for 16 h. The reaction mixture was allowed to cool to room temperature and partitioned between 2-methyltetrahydrofuran / water, separated and the aqueous layer reextracted with 2-methyltetrahydrofuran, the organic layers were combined, dried over Na2SO4, filtered and concentrated to dryness. Purification by silica gel chromatography (0-30% ethyl acetate in dichloromethane) gave endo-N- [4 - [(5S) -5- [tert-butyl (dimethyl) silyl] oxy-7-azabicyclo [2.2.1 ] heptan-7-yl] 2- (trifluoromethyl) phenyl] -4-oxo-5- (trifluoromethyl) -1H-quinoline-3-carboxamide (285 mg, 79% yield). ¹H NMR (400.0 MHz, DMSO-d6) δ 13.07 (s, 1H), 12.16 (s, 1H), 8.88 (s, 1H), 8.04 (dd, J = 2, 2, 7.3 Hz, 1H), 7.95 - 7.89 (m, 3H), 7.22 (dd, J = 2.4, 8.9 Hz, 1H), 7.16 (d, J = 2.6 Hz, 1H), 4.29 (m, 3H), 2.16 - 2.07 (m, 2H), 1.62 - 1.43 (m, 3H), 1.05 - 1, 01 (m, 1H), 0.89 (s, 9H), 0.08 (d, J = 1.4 Hz, 6H).

[0200] Preparation of endo-N- [4 - [(5S) -5-hydroxy-7-azabicyclo [2.2.1] heptan-7-yl] -2 (trifluoromethyl) phenyl] -4-oxo-5- (trifluoromethyl ) -1 H-quinoline-3-carboxamide.

Endo-N- [4 - [(5S) -5- [tert-butyl (dimethyl) silyl] oxy-7-azabicyclo [2.2.1] heptan-7-yl] -2- (trifluoromethyl) phenyl] -4-oxo -5- (trifluoromethyl) -1H-quinoline-3-carboxamide (281 mg, 0.45 mmol) was dissolved in 1% HCl / Ethanol (2 mL 1% w / w) and allowed to stir at room temperature for 16 h, and a white precipitate was obtained. The reaction mixture was diluted with diethyl ether and filtered. The collected solid was dissolved in ethyl acetate / saturated aqueous NaHCO3 solution. The layers were separated and the aqueous layer re-extracted with ethyl acetate. The organic layers were washed twice with water, dried over Na2SO4, filtered and concentrated to dryness to give endo-N- [4 - [(5S) -5-hydroxy-7-azabicyclo [2.2.1] heptan-7-yl] -2- (trifluoromethyl) phenyl] -4-oxo-5- (trifluoromethyl) -1H-quinoline-3-carboxamide (190 mg, 83%).¹H NMR (400.0 MHz, DMSO-d6) δ 13.07 (s, 1H), 12.15 (s, 1H), 8.88 (s, 1H), 8.04 (dd, J = 2, 2, 7.4 Hz, 1H), 7.95 - 7.88 (m, 3H), 7.19 (dd, J = 2.4, 9.0 Hz, 1H), 7.12 (d, J = 2.6 Hz, 1H), 5.00 (d, J = 4.2 Hz, 1H), 4.25 - 4.13 (m, 1H), 4.21 - 4.19 (m, 1H) , 4.16 - 4.13 (m, 1H), 2.15 - 2.08 (m, 2H), 1.61 - 1.55 (m, 1H), 1.47 - 1.44 (m, 2H) and 1.03 (dd, J = 3.4, 12.3 Hz, 1H).

[0201] Analytical data for the compounds of Table 1 are shown below:

Table 2

<td>No. Sample relationship</td><td>LC / MS M + 1</td><td>LC / RT^andminutes</td><td>NMR</td>

<td> 1</td><td> 456,50</td><td> 1,76</td><td>¹H NMR (400.0 MHz, DMSO-d6) δ 12.94 (d, J = 6.1 Hz, 1H), 12.36 (s, 1H), 8.80 (d, J = 6.8 Hz , 1H), 8.13 (s, 1H), 7.98 (d, J = 8.9 Hz, 1H), 7.68-7.63 (m, 2H), 7.10 (d, J = 8.5 Hz, 1H), 7.01 (s, 1H), 4.28 (s, 2H), 2.48 (s, 3H), 2.02 - 2.00 (m, 2H), 1, 86 - 1.77 (m, 5H), 1.45 - 1.42 (m, 1H), 1.31 (d, J = 11.1 Hz, 2H).</td>

<td> 2</td><td> 458,20</td><td> 1,20</td><td>¹H NMR (400.0 MHz, DMSO-d6) δ 12.89 (s, 1H), 12.43 (s, 1H), 8.79 (s, 1H), 8.00 (d, J = 8, 9 Hz, 1H), 7.72 - 7.68 (m, 2H), 7.44 (dd, J = 2.9, 9.0 Hz, 1H), 7.22 (dd, J = 2.5 , 9.0 Hz, 1H), 7.15 (d, J = 2.6 Hz, 1H), 4.33 (s, 2H), 3.91 (s, 3H), 1.67 (d, J = 6.9 Hz, 4H), 1.43 (d, J = 6.9 Hz, 4H).</td>

<td> 3</td><td> 496,0</td><td> 1,48</td><td>¹H NMR (400.0 MHz, DMSO-d6) δ 13.08 (s, 1H), 12.16 (s, 1H), 8.88 (s, 1H), 8.04 (dd, J = 2.1, 7.4 Hz, 1H), 7.95 - 7.88 (m, 3H), 7 , 22 (dd, 2.5, 8.9 Hz, 1H), 7.16 (d, J = 2.5 Hz, 1H), 4.33 (s, 2H), 1.67 (d, J = 6.9 Hz, 4H), 1.44 (d, J = 6.9 Hz, 4H).</td>

<td> 4</td><td> 458,50</td><td> 1,22</td><td>¹H NMR (400.0 MHz, DMSO-d6) δ 13.12 (d, J = 6.7 Hz, 1H), 12.50 (s, 1H), 8.78 (d, J = 6.8 Hz , 1H), 8.10 (d, J = 9.1 Hz, 1H), 7.78 - 7.72 (m, 3H), 7.65 (d, J = 7.7 Hz, 1H), 7 , 39 (d, J = 7.3 Hz, 1H), 7.33 (s, 1H), 5.20 (s, 2H), 4.47 (s, 2H), 1.74 (d, J = 6.7 Hz, 4H), 1.51 (d, J = 7.1 Hz, 4H).</td>

<td> 5</td><td> 512,50</td><td> 1,55</td><td>¹H NMR (400.0 MHz, DMSO-d6) δ 13.07 (s, 1H), 12.15 (s, 1H), 8.88 (s, 1H), 8.04 (dd, J = 2, 2, 7.4 Hz, 1H), 7.95 - 7.88 (m, 3H), 7.19 (dd, J = 2.4, 9.0 Hz, 1H), 7.12 (d, J = 2.6 Hz, 1H), 5.00 (d, J = 4.2 Hz, 1H), 4.25 - 4.13 (m, 1H), 4.21 - 4.19 (m, 1H) .</td>

<td></td><td></td><td></td><td>4.16 - 4.13 (m, 1H), 2.15 - 2.08 (m, 2H), 1.61 - 1.55 (m, 1H), 1.47 - 1.44 (m, 2H ) and 1.03 (dd, J = 3.4, 12.3 Hz, 1H).</td>

<td> 6</td><td> 399,30</td><td> 1,47</td><td>¹H NMR (400.0 MHz, DMSO-d6) δ 12.77 (s, 1H), 12.75 (s, 1H), 8.77 (s, 1H), 8.11 (d, J = 9, 1 Hz, 1H), 7.64 - 7.60 (m, 1H), 7.55 (d, J = 8.0 Hz, 1H), 7.34 (d, J = 2.8 Hz, 1H) , 7.27 (dd, J = 2.8, 9.1 Hz, 1H), 7.23 (d, J = 7.2 Hz, 1H), 4.32 (s, 2H), 2.91 ( s, 3H), 1.65 (d, J = 7.2 Hz, 4H), 1.42 (d, J = 6.8 Hz, 4H).</td>

<td> 7</td><td> 453,0</td><td> 1,62</td><td>¹H NMR (400.0 MHz, DMSO-d6) δ 13.28 (d, J = 6.4 Hz, 1H), 12.07 (s, 1H), 8.95 (d, J = 6.5 Hz , 1H), 8.69 (s, 1H), 8.16 (dd, J = 1.5, 8.7 Hz, 1H), 8.01 (d, J = 8.8 Hz, 1H), 7 , 91 (d, J = 8.7 Hz, 1H), 7.28 (d, J = 7.8 Hz, 1H), 7.22 (s, 1H), 4.38 (s, 2H), 1 , 69 (d, J = 6.4 Hz, 4H), 1.46 (d, J = 6.9 Hz, 4H).</td>

<td>No. Sample relationship</td><td>LC / MS M + 1</td><td>LC / RT^andminutes</td><td>NMR</td>

<td> 8</td><td> 512,10</td><td> 1,35</td><td>¹H NMR (400.0 MHz, DMSO-d6) δ 13.07 (s, 1H), 12.13 (s, 1H), 8.88 (s, 1H), 8.05 - 8.02 (m, 1H), 7.95 7.86 (m, 3H), 7.18 (d, J = 9.0 Hz, 1H), 7.12 (d, J = 2.5 Hz, 1H), 4.74 (d, J = 5.2 Hz, 1H), 4.33 (m, 1H), 4.11 (m, 1H), 3.77 (m, 1H), 1.82 (dd, J = 7.3, 12.5 Hz, 1H), 1.56 - 1.48 (m, 3H) , 1.25 (m, 2H).</td>

<td> 9</td><td> 442,10</td><td> 1,20</td><td>¹H NMR (400.0 MHz, DMSO-d6) δ 12.84 (s, 1H), 12.43 (s, 1H), 8.81 (s, 1H), 8.12 (s, 1H), 8 , 00 (d, J = 8.9 Hz, 1H), 7.64 (m, 2H), 7.21 (dd, J = 2.5, 9.0 Hz, 1H), 7.15 (d, J = 2.6 Hz, 1H), 4.32 (s, 2H), 2.47 (s, 3H), 1.67 (d, J = 7.4 Hz, 4H), 1.43 (d, J = 6.9 Hz, 4H).</td>

<td> 10</td><td> 442,10</td><td> 1,40</td><td>¹H NMR (400.0 MHz, DMSO-d6) δ 12.68 (s, 1H), 12.41 (s, 1H), 8.75 (s, 1H), 7.94 (d, J = 8, 9 Hz, 1H), 7.63 - 7.60 (m, 1H), 7.54 (d, J = 7.8 Hz, 1H), 7.23 7.20 (m, 2H), 7.15 (d, J = 2.7 Hz, 1H), 4.33 (s, 2H), 2.89 (s, 3H), 1.67 (d, J = 6.7 Hz, 4H), 1.44 (d, J = 6.9 Hz, 4H).</td>

<td> 11</td><td> 444,0</td><td> 1,30</td><td>¹H NMR (400.0 MHz, DMSO-d6) δ 13.55 (s, 1H), 13.31 (d, J = 7.2 Hz, 1H), 11.58 (s, 1H), 8.86 (d, J = 6.9 Hz, 1H), 8.01 (d, J = 9.0 Hz, 1H), 7.66 (t, J = 8.2 Hz, 1H), 7.29 (d , J = 8.8 Hz, 1H), 7.23 (s, 1H), 7.17 (d, J = 7.7 Hz, 1H), 6.80 (d, J = 7.5 Hz, 1H ), 4.39 (s, 2H), 1.69 (d, J = 7.3 Hz, 4H), 1.46 (d, J = 7.0 Hz, 4H).</td>

<td> 12</td><td> 510,5</td><td> 1,95</td><td>¹H NMR (400.0 MHz, DMSO-d6) δ 13.16 (d, J = 5.7 Hz, 1H), 12.07 (s, 1H), 8.87 (d, J = 6.6 Hz , 1H), 8.05 (dd, J = 2.1, 7.3 Hz, 1H), 7.96 - 7.92 (m, 2H), 7.87 (d, J = 9.0 Hz, 1H), 7.09 (d, J = 9.1 Hz, 1H), 7.00 (s, 1H), 4.28 (s, 2H), 2.02 - 2.00 (m, 2H),</td>

<td></td><td></td><td></td><td>I, 88 - 1.73 (m, 5H), 1.45 - 1.42 (m, 1H), 1.31 (d, J = II, 6 Hz, 2H).</td>

<td> 13</td><td> 509,7</td><td> 1,06</td><td>¹H NMR (400.0 MHz, DMSO-d6) δ 13.23 (d, J = 6.8 Hz, 1H), 12.40 (s, 1H), 10.31 (s, 1H), 8.96 (d, J = 6.6 Hz, 1H), 8.40 (d, J = 9.0 Hz, 1H), 8.08 - 8.06 (m, H), 8.07 (dd, J = 1.5 Hz, 8.1 Hz, 1H), 8.00-7.95 (m, 2H), 7.15 - 7.09 (m, 2H), 4.49 (s, 2H), 4, 29 (s, 2H), 2.94 (s, 6H), 1.67 (d, J = 7.2 Hz, 4H), 1.44 (d, J = 7.0 Hz, 4H).</td>

<td> 14</td><td> 455,7</td><td> 1,04</td><td>¹H NMR (400.0 MHz, DMSO-d6) δ 12.63 (s, 2H), 8.75 (s, 1H), 8.38 (d, J = 8.8 Hz, 1H), 7.62 - 7.58 (m, 1H), 7.52 (d, J = 8.1 Hz, 1H), 7.21 (d, J = 7.2 Hz, 1H), 6.96 - 6.93 ( m, 2H), 4.24 (s, 2H), 3.70 (s, 2H), 2.92 (s, 3H), 2.28 (s, 6H), 1.65 (d, J = 7 , 0 Hz, 4H), 1.39 (d, J = 6.8 Hz, 4H).</td>

<td colspan="4">^and Retention time</td>

Tests for detection and measurement of ΔF508-CFTR amplification properties by compounds

Optical methods using membrane potential to study the modulating properties of ΔF508-CFTR by compounds [0202] Fluorescent voltage detection dyes are used in the test to measure changes in membrane potential using a fluorescent plate reader (e.g. FLIPR III, Molecular Devices, Inc.) as functional growth read ΔF508-CFTR in NIH 3T3 cells. The driving force for the response is the creation of a chloride ion gradient in combination with the activation of the channel by a single step of adding liquid after previously treating the cells with compounds and then adding a voltage detecting dye.

Identification of enhancing compounds [0203] To identify ΔF508-CFTR enhancers, a double addition HTS test format was developed. This HTS test uses fluorescent voltage sensing dyes to measure changes in membrane potential on FLIPR III as a measure of the gating (conductivity) of ΔF508 CFTR in temperature-corrected ΔF508 CFTR NIH 3T3 cells. The driving force of the answer is the Cl ion gradient^- in combination with channel activation of forskolin in a single liquid addition step using a fluorescent plate reader such as FLIPR III after treatment of cells with enhancing compounds (or DMSO medium control) followed by the addition of a redistributable dye.

Solutions [0204] Washing solution # 1: (in mM) NaCl 160, KCl 4.5, CaCl2 2, MgCl2 1, HEPES 10, pH 7.4 with NaOH.

Chloride-free washing solution: The chloride salts in washing solution # 1 are replaced with gluconate salts.

Cell culture [0205] For optical measurements of membrane potential, NIH3T3 mouse fibroblasts stably expressing ΔF508-CFTR are used. The cells are maintained at 37 ° C in 5% CO2 and 90% humidity in Dulbecco's modified Eagle medium supplemented with 2 mM glutamine, 10% fetal bovine serum, 1 X NEAA, β-ME, 1 X pen / strep and 25 mM HEPES in 175 cm² breeding bottles. For all optical tests, cells were plated at ~ 20,000 / well in 384-well matrigel-coated plates and cultured for 2 h at 37 ° C before culturing at 27 ° C for 24 h for the assay with enhancer. For the corrected tests, cells were cultured at 27 ° C or 37 ° C with or without compounds for 16-24 hours. Electrophysiological tests to study the modulating properties of ΔF508-CFTR by compounds.

Test in the Using chamber.

[0206] Ussing chamber experiments were performed on polarized airway epithelial cells expressing ΔF508-CFTR to further characterize the ΔF508-CFTR modulators identified in optical tests. Non-CF and CF airway epithelium was isolated from bronchial tissue, cultured as previously described (Galietta, LJV, Lantero, S., Gazzolo, A., Sacco, O., Romano, L., Rossi, GA, and Moran, O. (1998) In Vitro Cell. Dev. Biol. 34, 478-481) and spread on Costar® Snapwell filters^™ previously coated with NIH3T3 conditioned medium. After four days, the apical medium was removed and the cells were grown on an air-liquid interface for&gt; 14 days before use. This resulted in the formation of a single layer of completely differentiated cylindrical cells that produced cilia, which are characteristic of the airway epithelium. HBE nonCF was isolated from non-smokers who did not have any known lung disease. CF-HBE was isolated from patients homozygous for ΔF508-CFTR.

[0207] HBE grown on Coster® Snapwell ™ cell culture inserts was embedded in a Using chamber (Physiologic Instruments, Inc., San Diego, CA) and transepithelial resistance and short-circuit current were measured in the presence of side-basal to a peak Cl gradient^- (Isc) using a voltage-clamp system (Department of Bioengineering, University of Iowa, IA). Briefly, HBE was tested under voltage-clamp recording conditions (Vbold = 0 mV) at 37 ° C. The basolateral solution contained (in mM) 145 NaCl, 0.83 K2HPO4, 3.3 KH2PO4, 1.2 MgCl2, 1.2 CaCl2, 10 Glucose, 10 HEPES (pH adjusted to 7.35 using NaOH) and a peak solution contained (in mM) 145 NaG-gluconate, 1.2 MgCl2, 1.2 CaCl2, 10 glucose, 10 HEPES (pH adjusted to 7.35 using NaOH).

Identification of enhancing compounds [0208] A typical protocol used a Cl concentration gradient^- from the basolateral to the apical membrane. To establish this gradient, normal Ringer's fluid was used on the laterolateral membrane, while on the top of the NaCl membrane it was replaced with an equimolar solution of sodium gluconate (adjusted to pH 7.4 to NaOH) to produce a large Cl "concentration gradient through the epithelium. Forskolin (10 μΜ) and all test compounds were added to the apical side of the cell culture inserts. The performance of putative ΔF508-CFTR enhancers was compared with the known enhancer genistein.

Patch-clamp recording [0209] Total current Cl^- in ΔF508-NΊH3T3 cells were monitored using a perforated-patch recording configuration as previously described (Rae, J., Cooper, K., Gates, P. and Watsky, M. (1991) J. Neurosci. Methods 37, 15 -26). Voltage clamp recording was performed at 22 ° C using an Axopatch 200B patch clamp amplifier (Axon Instruments Inc., Foster City, CA). The pipette solution contained (in mM) 150 N-methyl-D-glucamine (NMDG) -Cl, 2 MgCl2, 2 CaCl2, 10 EGTA, 10 HEPES and 240 μg / ml amphotericin-B (pH adjusted to 7.35 using HCl) . The extracellular medium contained (in mM) 150 NMDG-Cl, 2 MgCl2, 2 CaCl2, 10 HEPES (pH adjusted to 7.35 using HCl). Pulse generation, data collection and analysis were performed on a computer equipped with a Digidata 1320 A / D interface in combination with Clampex 8 (Axon Instruments Inc.). To activate ΔF508-CFTR, 10 μΜ forskolin and 20 μΜ genistein were added to the bath and the voltage-voltage relationship was monitored every 30 s.

Identification of enhancers [0210] The ability of enhancers ΔF508-CFTR to increase the macroscopic current ΔF508-CFTR Cl^- (Iaf508) in NIH3T3 cells stably expressing ΔF508-CFTR were also examined using peforated-patch registration techniques. Enhancers identified in optical tests induced a dose-dependent increase in ^ F508 of similar strength and performance as observed in optical tests. In all cells tested, the reversal potential before and during use of the enhancer was about -30 mV, which is calculated EC1 (-28 mV).

Cell culture [0211] NIH3T3 mouse fibroblasts stably expressing ΔF508-CFTR are used for whole cell registration. Cells are maintained at 37 ° C in 5% CO2 and 90% humidity in Dulbecco's modified Eagle medium supplemented with 2 mM glutamine, 10% fetal bovine serum, 1 X NEAA, β-ΜΕ, 1 X pen / strep and 25 mM HEPES in 175 cm²breeding bottles. For whole-cell registration, 2500-5000 cells were plated on poly-L-lysine-coated coverslips and cultured for 24-48 h at 27 ° C before use to test for enhancer activity; and incubated with or without a correction compound at 37 ° C to measure the activity of the correction compounds. Single Channel Recording [0212] wt-CFTR gating activity and temperature-corrected ΔF508-CFTR expressed in NIH3T3 cells were observed using recording on excised and inverted membrane fragments as previously described (Dalemans, W., Barbry, P. , Champigny, G., Jallat, S., Dott, K., Dreyer, D., Crystal, RG, Pavirani, A., Lecocq, JP., Lazdunski, M. (1991) Nature 354, 526-528) using an Axopatch 200B patch clamp amplifier (Axon Instruments Inc.). The pipette contained (in mM): 150 NMDG, 150 aspartic acid, 5 CaCl2, 2 MgCl2 and 10 HEPES (pH adjusted to 7.35 with Tris base). The bath contained (in mM): 150 NMDG-Cl, 2 MgCl2, 5 EGTA, 10 TES and 14 Tris bases (pH adjusted to 7.35 using HCl). After excision, both wt- and ΔF508-CFTR were activated by the addition of 1 mM Mg-ATP, 75 nM cAMP-dependent protein kinase catalytic unit (PKA; Promega Corp. Madison, WI) and 10 mM NaF to inhibit protein phosphatases, which prevented electricity from running out. The pipette potential was maintained at 80 mV. Channel activity was analyzed for membrane fragments containing &lt;2 active channels. The maximum number of simultaneous openings determined the number of active channels during the experiment. To determine the current amplitude for a single channel, the data recorded from 120 s activity of ΔF508-CFTR was filtered off-line at 100 Hz, and then used to construct amplitude histograms with all points that were matched with multigaussian functions using Bio-Patch Analysis software ( Bio-Logic Comp. France). The total microscopic current and opening probability (Po) were determined from 120 s of channel activity. Po was determined using the Bio-Patch software or the relationship Po = I / i (N), where I = average current, i = current amplitude for a single channel, and N = the number of active channels in the fragment.

Cell culture [0213] For patch-clamp recording using dissected membranes, NIH3T3 mouse fibroblasts stably expressing ΔF508-CFTR are used. Cells are maintained at 37 ° C in 5% CO2 and 90% humidity in Dulbecco's modified Eagle medium supplemented with 2 mM glutamine, 10% fetal bovine serum, 1 X NEAA, β-ME, 1 X pen / strep and 25 mM HEPES in 175 cm² breeding bottles. For single-channel recording, 2500-5000 cells were plated on poly-L-lysine-coated coverslips and grown for 24-48 h at 27 ° C before use.

[0214] The compounds of the invention are useful as transporter modulators having an ATP binding cassette. Examples of the action and effectiveness of the compounds of Formula (I) are shown below in Table 3. The activity of the compound is shown by "+++" when the measured activity was less than 2.0 μM, "++" when the measured activity was from 2 μM to 5.0 μM, "+" when the measured activity was greater than 5.0 μM, and "-" when data was not available. Efficiency is shown with "+++" when the calculated efficiency was greater than 100%, "++" when the calculated efficiency was between 100% and 25%, "+" when the calculated efficiency was less than 25%, and " - "when data was not available. It should be noted that 100% efficacy means the maximum response obtained with 4-methyl-2- (5-phenyl-1H-pyrazol-3-yl) phenol.

Table 3

<td>Example compound no</td><td>EC50 activity (gm)</td><td>Effectiveness %</td>

<td> 1</td><td> +++</td><td> ++</td>

<td> 2</td><td> +++</td><td> ++</td>

<td> 3</td><td> +++</td><td> ++</td>

<td> 4</td><td> +++</td><td> ++</td>

<td> 5</td><td> +++</td><td> +++</td>

<td> 6</td><td> +++</td><td> +++</td>

<td> 7</td><td> +++</td><td> ++</td>

<td>Example compound no</td><td>EC50 activity (pm)</td><td>Effectiveness %</td>

<td> 8</td><td> +++</td><td> ++</td>

<td> 9</td><td> +++</td><td> ++</td>

<td> 10</td><td> +++</td><td> +++</td>

<td> 11</td><td> +++</td><td> ++</td>

<td> 12</td><td> +++</td><td> ++</td>

<td> 13</td><td> +++</td><td> ++</td>

<td> 14</td><td> +++</td><td> ++</td>

1. Relationship with Formula (I):

<img file="PL2349263T3_D0063.tif" />

JtzOH or a pharmaceutically acceptable salt thereof, where:

ring A is selected from:

where:

<img file="PL2349263T3_D0064.tif" />

^H? v^H

V 'ΛΥ (a) (b)

Μ M, or V (c) (d)

R¹ is -CF3, -CN or -C = CCH2N (CH3) 2;

R² is hydrogen, -CH3, -CF3, -OH or -CH2OH;

R³ is hydrogen, -CH3, -OCH3, or -CN;

both R² and R³ they are not hydrogen at the same time.

2. The compound according to condition 1, wherein ring A is a

3. The compound according to condition 1, wherein ring A is (a).

<img file="PL2349263T3_D0065.tif" />

AND

4. The compound according to condition 1, wherein ring A is

HO. .h (c).

5. The compound according to condition 1, wherein ring A is ^H- .OH (d).

6. A compound according to any one of conditions 2-5, wherein R¹ means -CF3.

7. A compound according to any one of conditions 2-5, wherein R¹ means -CN.

8. A compound according to any one of conditions 2-5, wherein R¹ means

-C = CCH2N (CH3) 2.

9. A compound according to any one of conditions 6-8, wherein R² is hydrogen.

10. A compound according to any one of conditions 6-8, wherein R² means -CH3.

11. A compound according to any one of conditions 6-8, wherein R² means -CF3.

12. A compound according to any one of conditions 6-8, R² means -OH.

13. A compound according to any one of conditions 6-8, wherein R² means -CH2OH.

14. A compound according to any one of conditions 10-13, wherein R³ is hydrogen.

15. A compound according to any one of conditions 9-13, wherein R³ means -CH3.

16. A compound according to any one of conditions 9-13, wherein R³ means -OCH3.

17. A compound according to any one of conditions 9-13, wherein R³ means -CN.

18. A compound selected from

<img file="PL2349263T3_D0066.tif" />

<img file="PL2349263T3_D0067.tif" />

19. A pharmaceutical composition comprising a compound according to any one of conditions 1-18 and a pharmaceutically acceptable carrier or adjuvant.

twenty. The pharmaceutical composition of clause 19, further comprising an additional agent selected from a mucolytic agent, bronchodilator, antibiotic, anti-infective agent, anti-inflammatory agent, CFTR modulator other than the compound of Formula (I) or nutrient.

21. The pharmaceutical composition according to condition 20, wherein said additional agent is a CFTR modulator other than the compound of Formula (I).

22. A method of treating or reducing the severity of a disease in a patient wherein the disease is selected from cystic fibrosis, asthma, smoke-induced COPD, chronic bronchitis, chronic rhinitis and paranasal sinuses, constipation, pancreatitis, pancreatic failure, male infertility due to congenital bilateral absence vas deferens (CBAVD), benign lung disease, idiopathic pancreatitis, allergic bronchopulmonary aspergillosis (ABPA), liver disease, congenital emphysema, congenital hemochromatosis, coagulation deficiencies - fibrinolysis, e.g. protein C deficiency, congenital angioedema type 1, lipid processing disorders, e.g. family hypercholesterolemia, type 1 chylomicronemia, abetalipoproteinemia, lysosomal diseases, e.g. cell / pseudo Hurler, mucopolysaccharidoses, Sandhof / Tay-Sachs syndrome, Crigler-Najjar syndrome type II, poliendocrinopathy / hyperinsulinemia, diabetes mellitus, Laron's dwarfism, myeloperoxidase deficiency, primary hypoparathyroidism, melanoma, type 1 CDG glycanosis, congenital hyperparathyroidism, congenital bone fragility, congenital hypofibrinogenemia, ACT deficiency, diuretic syndrome, DI Charcot-Marie-Tooth, Pelizacus-Merzbacher disease, neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, progressive supranuclear palsy, Pick's disease, various polyglutamine neurological disorders, such as Huntington's disease, type I spinal cerebellar ataxia, cerebrospinal muscular atrophy, atrophy of the toothed nucleus, red nucleus, pale knob and low-pituitary nucleus, and dystrophy as well as spongiform encephalopathies such as congenital Creutzfeldt-Jakob disease (caused by a defect in prion protein processing), Fabry disease, Straussler-Scheinker syndrome, COPD, dry eye disease, pancreatic insufficiency, osteoporosis, osteopenia, Gorham syndrome, chloride channel disorders, congenital myotonia (Thomson and Becker forms), Bartter syndrome type III, Dent's disease, hyperplexia, epilepsy, hyperplexia, lysosomal disease storage, Angelman syndrome, primary ciliary dyskinesia (PCD), PCD with reverse viscera (also known as Kartagener syndrome), PCD without reverse visceral and cilia aplasia, or Sjogren's disease, the method comprising the step of administering to said patient an effective amount of a compound according to any of conditions 1-18.

23. The method of condition 22, wherein the disease is cystic fibrosis.

24. A method of treating or reducing the severity of a disease in a patient, wherein the disease is associated with reduced CFTR activity as a result of mutation of the CFTR coding gene or environmental factors, the method comprising the step of administering to said patient an effective amount of a compound according to any one of conditions 118.

25. The method according to condition 24, wherein the disease is cystic fibrosis, chronic bronchitis, recurrent bronchitis, acute bronchitis, male infertility due to congenital bilateral vas deferens (CBAVD), female infertility due to congenital uterine and vaginal (CAUV), chronic idiopathic inflammation (ICP), idiopathic recurrent pancreatitis, idiopathic acute pancreatitis, chronic rhinosinusitis, primary sclerosing cholangitis, allergic bronchopulmonary aspergillosis, diabetes, dry eye disease, constipation, allergic bronchopulmonary aspergillosis (ABPA), bone disease, and asthma.

26. A method of treating or reducing the severity of a disease in a patient, wherein the disease is associated with the normal operation of CFTR, the method comprising the step of administering to that patient an effective amount of a compound according to any of conditions 1-18.

27. The method of condition 26, wherein the disease is chronic obstructive pulmonary disease (COPD), chronic bronchitis, recurrent bronchitis, acute bronchitis, chronic rhinitis and sinusitis, constipation, chronic pancreatitis, recurrent pancreatitis and acute inflammation pancreas, pancreatic insufficiency, male infertility caused by congenital bilateral vas deferens (CBAVD), mild lung disease, idiopathic pancreatitis, liver disease, congenital emphysema, gallstones, gastroesophageal reflux disease, gastrointestinal malignancies, inflammatory bowel disease, constipation, diabetes, arthritis, osteoporosis, and osteopenia.

28. The method according to condition 26, wherein the disease is selected from congenital hemochromatosis, coagulation deficiencies - fibrinolysis, e.g. protein C deficiency, congenital angioedema type 1, lipid processing disorders, e.g. family hypercholesterolemia, e.g. for example diseases of cellular inclusions / pseudo Hurler, mucopolysaccharidoses, Sandhof / Tay-Sachs syndrome, Crigler-Najjar type II syndrome, poliendocrinopathy / hyperinsulinemia, diabetes mellitus, Laron's dwarfism, myeloperoxidase deficiency, primary hypoparathyroidism, melanoma, CDG type 1 glycanosis, congenital hyperparathyroidism, congenital bone fragility, congenital hypofibrinogenesis, and urinary deficiency neuro-pituitary DI, renal DI, Charcot-Marie-Tooth syndrome, Pelizacus-Merzbacher disease, neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, progressive supranuclear palsy, Pick's disease, various polyglutamine neurological disorders such as Huntington's disease, cerebellum cerebellar ataxia type I, dermatocerebral muscular atrophy, atrophy of the dentate nucleus, red nucleus, pale and low brain and myotonic dystrophy as well as spongiform encephalopathies such as congenital Creutzfeldt-Jakob disease (caused by a defect in the processing of prion proteins), Fabry disease, and Straussler-Scheinker syndrome, Gorham syndrome, chloride channel disorders, congenital myotonia (Thomson and Becker forms), type III Bartter syndrome, Denta disease, hyperplexia, epilepsy, hyperplexia, lysosomal storage disease, Angelman syndrome, primary dyskinesia cilia (PCD), PCD with reverse viscera (also known as Kartagener syndrome), PCD without reverse viscera and cilia aplasia, or Sjogren's disease.

29. Kit for use for measuring the activity of CFTR or a fragment thereof in a biological sample in vitro or in vivo, comprising:

(i) a composition comprising a compound of Formula (I) according to claim 1;

(ii) instructions for:

a) contacting the composition with a biological sample;

b) measuring the activity of this CFTR or a fragment thereof.

thirty. Kit according to condition 29, additionally containing instructions for:

a) contacting the additional composition with the biological sample;

b) measuring the activity of this CFTR or a fragment thereof in the presence of this additional compound, and

c) comparing the CFTR activity in the presence of an additional compound with the density CFTR in the presence of the composition of Formula (I).

31. A method of modulating CFTR activity in a biological sample comprising the step of contacting said CFTR with a compound according to any one of conditions 1-18.

32. A method for producing a compound of Formula (I):

<img file="PL2349263T3_D0068.tif" />

or a pharmaceutically acceptable salt thereof, said method comprising:

(a) reacting an acid of formula 1d with an amine of formula 2c to provide

<img file="PL2349263T3_D0069.tif" />

ring A is selected from:

<img file="PL2349263T3_D0070.tif" />

(a) (b) (c) (d) where

R¹ is -CF3, -CN or -C = CCH2N (CH3) 2;

R² is hydrogen, -CH3, -CF3, -OH or -CH2OH;

R³ is hydrogen, -CH3, -OCH3 or -CN;

both R² and R³ they are not both hydrogen and a

R^and is a hydrogen atom or a silyl protecting group selected from the group consisting of trimethylsilyl (TMS), tert-butyldiphenylsilyl (TBDPS), tert-butyldimethylsilyl (TBDMS), triisopropylsilyl (TIPS) and [2 (trimethylsilyl) ethoxy] methyl (SEM).

33. The method of condition 32, wherein the reaction of the acid of formula 1d with the amine of formula 2c is carried out in a solvent in the presence of O- (7-azabenzotriazol-1-yl) -N, N, N ', N'-tetramethyluronium (HATU) and triethylamine hexafluorophosphate or in a solvent in the presence of cyclic propyl phosphonic anhydride (T3P®) and pyridine.

34. The method of clause 33, wherein the solvent comprises N, N-dimethylformamide, ethyl acetate or 2-methyltetrahydrofuran.

35. The method according to condition 32, in which R^and is hydrogen or TBDMS.

36. The method according to condition 32, in which R^and means TBDMS.

37. The method of condition 32 further comprising a deprotection step to remove the silyl protecting group when ring A is

<img file="PL2349263T3_D0071.tif" />

<img file="PL2349263T3_D0072.tif" />

or where R^and is a silyl protecting group to form a compound of Formula (I) wherein ring A is

HO or (c)

<img file="PL2349263T3_D0073.tif" />

38. The method of clause 32, wherein the amine of formula 2c is prepared from the compound of formula 2a comprising the steps of:

(a) reacting a compound of formula 2a with an amine of formula 3 to provide a compound of formula 2b

<img file="PL2349263T3_D0074.tif" />

wherein:

Hal is F, Cl, Br or I; and this amine of formula 3 is

<img file="PL2349263T3_D0075.tif" />

and (b) reducing the compound of formula 2b to the amine of formula 2c.

<img file="PL2349263T3_D0076.tif" />

39. The method of clause 38, wherein the amine of formula 3 in step (a) is prepared in situ from the amine hydrochloride salt.

40. The method according to condition 38, in which R^and is hydrogen or TBDMS.

41. The method according to condition 38, in which R^and means TBDMS.

42. The method of clause 38, wherein step (a) is carried out in a polar aprotic solvent in the presence of a tertiary amine base.

43. The method of clause 42, wherein step (a) is carried out in acetonitrile in the presence of triethylamine.

44. The method of clause 38, wherein the reaction temperature of step (a) is between approximately 75 ° C and approximately 85 ° C.

45. The method of clause 38, wherein the reaction time is between approximately 2 and approximately 30 hours.

46. The method of clause 38, wherein step (b) is carried out in a polar protic solvent in the presence of a palladium catalyst.

47. The method of clause 46, wherein the solvent in step (b) comprises methanol or ethanol.

48. The method of clause 38, wherein step (b) is carried out in a polar protic solvent in the presence of Fe and FeSO4 or Zn and AcOH.

49. The method of clause 48, wherein the polar protic solvent is water.

50. A method for producing a compound having Formula (Ic),

<img file="PL2349263T3_D0077.tif" />

or a pharmaceutically acceptable salt thereof, comprising the steps of:

(a) reacting a compound of formula 2a with an amine of formula 3 to provide a compound of formula 2b

<img file="PL2349263T3_D0078.tif" />

(b) converting a compound of formula 2b to an amine of formula 2c by reduction

<img file="PL2349263T3_D0079.tif" />

and (c) reacting the amine of formula 2c with an acid of formula 1d to provide

<img file="PL2349263T3_D0080.tif" />

wherein Hal is F, Cl, Br or I; this amine of formula 3 is i

ring A is selected from:

<img file="PL2349263T3_D0081.tif" />

<img file="PL2349263T3_D0082.tif" />

(a) (b) (c) (d) where

R¹ is -CF3, -CN or -C = CCHN (CH3) 2;

_R2 is hydrogen, -CH3, -CF3, -OH or -CH2OH;

_R3 is hydrogen, -CH3, -OCH3 or -CN; both R² and R³ they do not simultaneously represent a hydrogen atom, a _Ra is a hydrogen atom or a silyl protecting group selected from the group consisting of trimethylsilyl (TMS), tert-butyldiphenylsilyl (TBDPS), tert-butyldimethylsilyl (TBDMS), triisopropylsilyl (TIPS) and [2- (trimethylsilyl) ethoxy] methyl (SEM).

51. The method of condition 50, wherein the amine of formula 3 in step (a) is prepared in situ from the amine hydrochloride salt.

52. The method according to condition 51, in which R^and is hydrogen or TBDMS.

53. The method according to condition 52, in which R^and means TBDMS.

54. The method of condition 50, wherein step (a) is carried out in a polar aprotic solvent in the presence of a tertiary amine base.

55. The method of clause 54, wherein step (a) is carried out in acetonitrile in the presence of triethylamine.

56. The method of clause 50, wherein the reaction temperature of step (a) is between approximately 75 ° C and approximately 85 ° C.

57. The method of clause 50, wherein the reaction time is between approximately 2 and approximately 30 hours.

58. The method of condition 50, wherein step (b) is carried out in a polar protic solvent in the presence of a palladium catalyst.

59. The method of clause 58, wherein the solvent in step (b) comprises methanol or ethanol.

60. The method of condition 50, wherein step (b) is carried out in a polar protic solvent in the presence of Fe and FeSO4 or Zn and AcOH.

61. The method of condition 50, wherein the polar protic solvent is water.

62. The method of condition 50, wherein step (c) is carried out in a solvent in the presence of O- (7-azabenzotriazol-1-yl) -N, N, N ', N'-tetramethyl uronium hexafluorophosphate (HATU) and triethylamine or in a solvent in the presence of cyclic propylphosphonic acid anhydride (T3P®) and pyridine.

63. The method of clause 62, wherein the solvent in step (c) comprises N, N-dimethylformamide (DMF), ethyl acetate or 2-methyltetrahydrofuran.

64. The method according to condition 62, in which R^and is hydrogen or TBDMS.

65. The method according to condition 64, in which R^and means TBDMS.

66. The method of condition 50 further comprising a deprotection reaction when ring A is

<img file="PL2349263T3_D0083.tif" />

where R^and is a silyl protecting group to form a compound of Formula (I) wherein ring A is

HO H, &lt;. &lt;^H(«), Or &lt;d).

67. A compound which means where where ring A is

X (a)

<img file="PL2349263T3_D0084.tif" />

/ XOR^and (D)

R¹ is -CF3, -CN or -C = CCH2N (CH3) 2, and

R^and is a hydrogen atom or a silyl protecting group selected from the group consisting of trimethylsilyl (TMS), tert-butyldiphenylsilyl (TBDPS), tert-butyldimethylsilyl (TBDMS), triisopropylsilyl (TIPS) and [2- (trimethylsilyl) ethoxy] methyl (SEM).

68. A compound which means where ring A is (a)

<img file="PL2349263T3_D0085.tif" />

/ TL-OR "

(d) where

R¹ is -CF3, -CN or -C = CCH2N (CH3) 2, and

R^and is a hydrogen atom or a silyl protecting group selected from the group consisting of trimethylsilyl (TMS), tert-butyldiphenylsilyl (TBDPS), tert-butyldimethylsilyl (TBDMS), triisopropylsilyl (TIPS) and [2- (trimethylsilyl) ethoxy] methyl (SEM).

69. Relationship with Formula (IA):

<img file="PL2349263T3_D0086.tif" />

or a pharmaceutically acceptable salt thereof, wherein: ring A is selected from

RaO H xvh Ay, OR^and (C&gt;

where, or (d)

R¹ is -CF3, -CN or -C = CCH2N (CH3) 2;

_R2 is hydrogen, -CH3, -CF3, -OH or -CH2OH;

_R3 is hydrogen, -CH3, -OCH3 or -CN; both R² and R³ they are not both hydrogen and a _Ra is a silyl protecting group selected from the group consisting of trimethylsilyl (TMS), tert-butyldiphenylsilyl (TBDPS), tert-butyldimethylsilyl (TBDMS), triisopropylsilyl (TIPS) and [2- (trimethylsilyl) ethoxy] methyl (SEM). 70. Relationship with Formula (I)

<img file="PL2349263T3_D0087.tif" />

or a pharmaceutically acceptable salt thereof, wherein: ring A is selected from:

<img file="PL2349263T3_D0088.tif" />

(a) (b) (c) (d) where

R¹ is -CF3, -CN or -C = CCH2 (CH3) 2;

R² is hydrogen, -CH3, -CF3, -OH or -CH2OH;

R³ is hydrogen, -CH3, -OCH3 or -CN;

both R² and R³ they are not simultaneously hydrogen;

prepared by the method of any of conditions 45-61.

71. A compound which is selected from the group consisting of

<img file="PL2349263T3_D0089.tif" />

<img file="PL2349263T3_D0090.tif" />

prepared by a method according to any one of conditions 50-66.