3-carboxamide-4-oxoquinoline derivatives, useful as modulators of regulator of transmembrane cystic fibrosis conductivity

FIELD: chemistry.

SUBSTANCE: invention relates to formula compound or to its pharmaceutically acceptable salt, where R represents COOH or CH2OH. Invention also relates to pharmaceutical composition based on formula I, method of modulating CFTR activity in biological sample, based on application of formula I compound, method of treatment, based on application of formula I compound, set based on formula I compound.

EFFECT: obtained are novel derivatives of quinolin-4-one, useful as CFTR modulators.

18 cl, 1 tbl, 8 ex

 

The application for approval of priority

This application claims priority under provisional patent application U.S. with serial number 61/162130 filed March 20, 2009, which is incorporated in full in this application by reference.

The technical field to which the present invention

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

Background of invention

Cystic fibrosis (CF) is a recessive genetic disease that affects approximately 30,000 children and adults in the United States and approximately 30,000 children and adults in Europe. Despite progress in the treatment of CF in the method of its treatment none.

CF is caused by mutations in the gene regulator transmembrane conductance cystic fibrosis (CFTR), which encodes epithelial chloride ion channel responsible for assisting in the regulation of salt and water absorption and secretion in various tissues. Low molecular weight drugs, known as potentiate tools that increase the open probability of CFTR channel, represent one potential terapeuticas the th strategy for the treatment of CF.

In particular, CFTR is a cAMP/ATP-mediated anion channel that is expressed in various cell types, including absorptive and secretory epithelial cells, where it regulates the flow of anions through the membrane and the activity of other ion channels and proteins. In the epithelial cells of the normal functioning of CFTR is critical to maintain the transport of electrolytes in the body, including the respiratory and digestive tissue. CFTR consists of approximately 1480 amino acids that encode a protein consisting of a tandem repeat transmembrane domains, each of which contains six transmembrane double helices and a nucleotide binding domain. These two transmembrane domain are linked by a large polar regulatory (R)domain with multiple phosphorylation sites that regulate channel activity and cell traffic.

The gene encoding CFTR, was identified and sequenced (See Gregory, R.J. et al. (1990) Nature 347:382-386; Rich, D.P. et al. (1990) Nature 347:358-362), (Riordan, J. R. et al. (1989) Science 245:1066-1073). The defect in this gene causes mutations in CFTR, leading to cystic fibrosis ("CF"), the most common fatal genetic disease in humans. Cystic fibrosis affects approximately one out of every 2,500 children of early age in the United States. Of the U.S. population in General up to 10 mil the ions of people have one copy of the defective gene without the explicit effects of the disease. In contrast, subjects with two copies of the CF associated gene suffer from debilitating and fatal effects of CF, including chronic lung disease.

In patients with CF mutations in CFTR, endogenous expressed in respiratory epithelium, leading to reduced apical anion secretion, causing an imbalance in the transport of ions and fluids. The resulting decrease in anion transport contributes to increased accumulation of mucus in the lungs associated with microbial infections that ultimately lead to the death of CF patients. In addition to respiratory disease in CF patients typically suffer from gastrointestinal problems and insufficient pancreatic function, which if untreated leads to death. In addition, most men with cystic fibrosis are infertile, and reduced fertility in women with cystic fibrosis. In contrast to the severe effects of the two copies of the CF associated gene in subjects with one copy of the CF associated gene exhibit enhanced resistance to cholera and dehydration due to diarrhoea, possibly explaining the relatively high prevalence of CF gene among the population.

Sequence analysis of the CFTR gene CF chromosomes revealed a number of different disease-causing mutations (Cutting, G.R. et al. (1990) Nature 346:366-369; Dean, M. et al. (1990) Cel 61:863:870; and Kerem, B-S. et al. (1989) Science 245:1073-1080; Kerem, B-S, et al. (1990) Proc. Natl. Acad. Sci. USA 87:8447-8451). Currently identified more than 1000 disease causing mutations in the CF gene (http://www.genet.sickkids.on.ca/cftr/app). The most common mutation is a deletion of phenylalanine at position 508 amino acid sequence of CFTR, and it is usually indicated as ΔF508-CFTR. This mutation occurs in approximately 70% of cases of cystic fibrosis, and it is associated with severe disease.

The deletion of residue 508 in ΔF508-CFTR prevents proper installation of the nascent protein. This leads to the inability of the mutant protein to exit the ER and move to the plasma membrane. As a result, the number of channels present in the membrane, far from determined in cells expressing wild-type CFTR. In addition to the disrupted traffic this mutation leads to the defect of the gate mechanism of the channel. All together the reduced number of channels in the membrane and the defect of the gate mechanism, lead to a reduction of anion transport across the epithelium, leading to impaired transport of ions and fluids (Quinton, P.M. (1990), FASEB J. 4: 2709-2727). Research, however, showed that reduced the number of ΔF508-CFTR in the membrane are functional, albeit less than wild-type CFTR. (Dalemans 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 other sabol is a cation, causing mutations in CFTR, which leads to disrupted traffic, synthesis and/or the gate mechanism of the channel could be adjusted either by activation or down-regulation for changes in anion secretion and modification of progression and/or severity of the disease.

Although CFTR transports various molecules in addition to the anions, it is clear that this role (transport anions) represents one important element in the mechanism of transport of ions and water across the epithelium. Other elements include the epithelial Na+channel, ENaC, Na+/2C1-/K+co-Transporter, Na+-K+-ATPase pump and basolateral membrane K+channels, which are responsible for the absorption of chlorine in the cell.

These elements work together to achieve directed transport through the epithelium through their selective expression and localization in the cell. Absorption of chlorine occurs as a result of coordinated activity of ENaC and CFTR present on the apical membrane, and Na+-K+-ATPase pump and Cl ion channels expressed on basolateral the cell surface. Secondary active transport of chlorine with luminaries side leads to accumulation of intracellular chloride, which is then passively leaves the cell through Cl-channels, leading to a vector transport. The location of Na+/2C1-/K+ co-Transporter, Na+-K+-ATPase pump and basolateral membrane K+channels on basolateral surface and CFTR on luminale side coordinates the secretion of chlorine through CFTR on luminale side. Since water will probably never itself is not actively transported, its flow through the epithelium depends on very small transepithelial osmotic gradients created by the flow of sodium and chlorine.

As discussed above, it is considered that the deletion of residue 508 in ΔF5O8-CFTR prevents proper installation of the nascent protein, which leads to the inability of the mutant protein to exit the ER and move to the plasma membrane. As a result, the plasma membrane contains an insufficient number of Mature protein and transport of chlorine in epithelial tissues is significantly reduced. Indeed, it was shown that this cellular phenomenon of defective ER processing ABC transporters using ER mechanism underlies not only the CF disease, but also a wide range of other individual and hereditary diseases.

Accordingly, there is a need for modulators of CFTR activity and compositions on their basis, which can be used to modulate the activity of the CFTR in the cell membrane of a mammal.

There is a need for methods of treatment of diseases, is caused by mutation in CFTR, using such modulators of CFTR activity.

There is a need for methods of modulating CFTR activity inex vivothe cell membrane of a mammal.

BRIEF description of the INVENTION

In one aspect the present invention relates to a compound of the formula I:

or its pharmaceutically acceptable salts, where R represents a COOH or CH2OH.

In one variant embodiment, the compound has the structure:

or its pharmaceutically acceptable salt.

In another variant embodiment, the compound has the structure:

or its pharmaceutically acceptable salt.

In one aspect the present invention relates to pharmaceutical compositions comprising a compound of formula I or its pharmaceutically acceptable salt, where R has the meaning defined above, and pharmaceutically acceptable carrier or adjuvant.

In one variant embodiment of the pharmaceutical composition, the compound has the structure:

or its pharmaceutically acceptable salt.

In another variant embodiment of the pharmaceutical composition, the compound has the structure:

or its pharmaceutically acceptable salt.

Variants of this embodiment aspects which include pharmaceutical composition, containing additional means selected from the group comprising a mucolytic agent, bronchodilator agent, antibiotic, anti-infective agent, anti-inflammatory agent, a CFTR modulator, or a nutritional agent.

In one aspect of the present invention includes a method of modulating CFTR activity in a biological sample, comprising the stage of contact specified biological sample with a compound of formula I or its pharmaceutically acceptable salt, where R has the meaning defined above.

In one variant embodiment of the method of modulating CFTR activity, the compound has the structure:

or its pharmaceutically acceptable salt.

In another variant embodiment of the method of modulating CFTR activity, the compound has the structure:

or its pharmaceutically acceptable salt.

In another aspect the invention also provides a method of treating or attenuating the severity of the disease in a patient, comprising the introduction of a specified patient one of the compositions defined in the present application, and the specified disease selected from the group including cystic fibrosis, asthma, caused by Smoking COPD (chronic obstructive pulmonary disease), chronic bronchitis, rhinosinusitis, constipation, pancreatitis, insufficient the durability of the function of the pancreas, male infertility caused by congenital bilateral absence of the VAS deferens (CBAVD), uncomplicated form of pulmonary disease, idiopathic pancreatitis, allergic bronchopulmonary aspergillosis (ABPA), liver disease, hereditary emphysema, hereditary hemochromatosis, coagulation-fibrinolysis deficiencies, such as protein deficiency, hereditary angioedema type 1, lipid processing, such as familial hypercholesterolaemia, chylomicronemia type 1, abetalipoproteinemia, lysosomal storage disorders, such as disease cellular inclusions/disease Deri, mucopolysaccharidosis, disease Sandhof/Tay-Sachs disease criglernajjar Najjar syndrome type II, polyendocrinopathy/hyperinsulemia, diabetes, dwarfism of Larona, myeloperoxidase deficiency, primary hypoparathyroidism, melanoma, glycans CDG type 1, congenital hyperthyroidism, imperfect osteogenesis, hereditary hypofibrinogenemia, lack of active clotting time of blood, diabetes insipidus (ND), insipidus neurotically diabetes insipidus neurogenic diabetes, muscular atrophy, Charcot-Marie-Toot disease Pelizaeus-Merzbacher neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, progressive soprano learny paralysis, atrophy of the Peak, some polyglutamine neurological disorders such as Huntington's disease, spinal-cerebellar ataxia type I, spinal and bulbar muscular atrophy, dentato-rubro-pallido-Lewis atrophy and myotonias dystrophy, as well as spongiform encephalopathies, such as hereditary disease Creutzfeldt-Jakob disease (due to defective processing of the prion protein), Fabry disease, syndrome Straussler-Sheinker, COPD, dry eye syndrome, or sjögren's disease, osteoporosis, osteopenia, bone healing and bone growth (including the restoration of the bone, bone regeneration, reducing bone resorption and increasing bone deposition), the syndrome Goreme, chloride of kalapati, such as congenital myotonia form (Thomson and Becker), the syndrome Bartter type III, dent disease, hyperekplexia, epilepsy, hyperekplexia, lysosomal disease accumulation, Angelman syndrome, and primary ciliary dyskinesia (PCD), a term for inherited disorders of the structure and/or function resisteth structures, including PCD with transposition of internal organs (also known as syndrome Addition, PCD without transposition of internal organs and ciliary aplasia.

In another variant embodiment of the method of cure or ameliorate the severity of the disease, the disease selected from cystic fibrosis, COPD caused by Smoking is m COPD, pancreatitis and rhinosinusitis.

In some embodiments embodiment the disease is a cystic fibrosis.

In one variant embodiment of the method of cure or ameliorate the severity of the disease, the compound has the structure:

or its pharmaceutically acceptable salt.

In another variant embodiment of the method of cure or ameliorate the severity of the disease, the compound has the structure:

or its pharmaceutically acceptable salt.

In another aspect the present invention provides a kit for use in measuring the activity of CFTR or a fragment in the biological samplein vitroorin vivoincluding:

(i) a composition comprising a compound of formula I or any of the options above embodiments; and (ii) instructions for: (a) contacting the composition with the biological sample, and (b) measuring the activity of the specified CFTR or fragment.

In one variant embodiment of this aspect, the kit further includes instructions for: (a) contacting an additional composition with the biological sample; b) measuring the activity of the specified CFTR or fragment in the presence of the specified additional connections; and (C) comparing the activity of CFTR or fragment in the presence of the additional compound with the density of CFTR and the and its fragment in the presence of a composition of formula 1.

In preferred embodiments the embodiment of the set are used to measure the density of CFTR or fragment.

In another preferred variant of embodiment, the kit is used for measuring the density specified CFTR or fragment, and phase comparison activity specified CFTR or fragment provides a density specified CFTR or fragment.

In one variant embodiment of a kit for use in measuring the activity of CFTR compound has the structure:

or its pharmaceutically acceptable salt.

In another variant embodiment of a kit for use in measuring the activity of CFTR compound has the structure:

or its pharmaceutically acceptable salt.

The compounds of formula I provide useful properties, such as, but not limited to, increased polarity, good solubility in water, a smaller volume of distribution and lower penetration in the tissue.

DETAILED description of the INVENTION

DEFINITION

Compounds of the present invention include compounds generally described above, and they further illustrated by the classes, subclasses, and species disclosed in this application. Used in this application, the following definitions apply, unless otherwise specified.

p> The term "ABC-Transporter"as used in this application means ABC-Transporter protein or its fragment that contains at least one binding domain, where this protein or its fragment is presentin vivoorinvitro. The term "binding domain"as used in this application, means on the domain ABC-Transporter, which can communicate with the modulator. See, for example, Hwang, T.C. et al., J. Gen. Physiol. (1998): 111(3), 477-90.

The term "CFTR"as used in this application means a transmembrane conductance regulator cystic fibrosis or mutation that is capable of regulator activity, including, but not limited to, ΔF508 CFTR and G551D CFTR (see, for example, http://www.genet.sickkids.on.ca/cftr/, for CFTR mutations).

The term "modulating"as used in this application, means an increase or decrease in the quantity that can be measured.

For the purposes of the present invention, the chemical elements are identified in accordance with the Periodic table of the elements, CAS version, Handbook of Chemistry and Physics, 75th Ed. In addition, General principles of organic chemistry are described in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999, and "March''s Advanced Organic Chemistry", 5th Ed., Ed.: Smith, M.B. and March, J., John Wiley & Sons, New York: 2001, the full content of which is incorporated into the present application by reference.

As described in this application is soedineniya of the present invention optionally can be substituted by one or more substituents, such as in the General form illustrated above or illustrated with particular classes, subclasses and species of the present invention. It should be clear that the phrase "optionally substituted" is used interchangeably with the phrase "substituted or unsubstituted". As a rule, the term "substituted" regardless, standing before him, the word "optional" or not, refers to the replacement of hydrogen radicals in a given structure with the radical of a specified substituent. Unless otherwise specified, optionally substituted group may contain a substituent at each substitutable position of the group, and when more than one position in any particular structure can be substituted by more than one Deputy, selected from a specified group, such substituents may be either the same or different from each other in each position. The combination of the substituents contemplated by this invention, preferred are those which lead to the formation of a stable or chemically achievable connections. The term "stable"as used in this application, refers to compounds, which essentially do not change, being subject to conditions that make possible the reception, detection and, preferably, their separation, purification, and use one or skolkin appointments, disclosed in this application. In some embodiments of the incarnation stable compound or chemically achievable connection is a such that essentially does not change during curing at a temperature of 40°C or below in the absence of moisture or other chemically reactive conditions, for at least a week.

The term "protective group" (PG)as used in this application means a group designed to protect a functional group, such as, for example, alcohol, amine, carboxyl, carbonyl, and the like, against undesirable reactions during the procedure of synthesis. Traditionally used protective groups are disclosed in Greene and Wuts, Protective Groups in Organic Synthesis, 3rd Edition (John Wiley & Sons, New York, 1999), which is incorporated into the present application by reference. Examples azatadine groups include acyl, aroline or karamelnye groups, such as formyl, acetyl, propionyl, pivaloyl, t-butylacetyl, 2-chloroacetyl, 2-bromoacetyl, TRIFLUOROACETYL, trichloroacetyl, phthalyl, o-nitrophenoxyacetic, α-chlorobutyryl, benzoyl, 4-chlorobenzoyl, 4-bromobenzoyl, 4-nitrobenzoyl and the chiral auxiliary groups, such as protected or unprotected D, L or D, L-amino acids such as alanine, leucine, phenylalanine and the like; sulfonylurea groups, such as benzazolyl, p-toluensulfonyl and the like; urethane groups is, such as benzyloxycarbonyl, p-chlorobenzenesulfonyl, p-methoxybenzenesulfonyl, p-nitrobenzyloxy, 2-nitrobenzenesulfonyl, p-bromobenzyloxycarbonyl, 3,4-dimethoxyphenylacetone, 3,5-dimethoxybenzoquinone, 2,4-dimethoxybenzoquinone, 4-methoxybenzenesulfonyl, 2-nitro-4,5-dimethoxybenzonitrile, 3,4,5-trimethoxybenzylamine, 1-(p-biphenylyl)-1-methylethanolamine, α,α-dimethyl-3,5-dimethoxybenzoquinone, benzylaminocarbonyl, tert-butyloxycarbonyl, diisopropylperoxydicarbonate, isopropoxycarbonyl, etoxycarbonyl, methoxycarbonyl, allyloxycarbonyl, 2,2,2,-trichlorocyanuric, phenoxycarbonyl, 4-nitrophenoxyacetic, fluorenyl-9-methoxycarbonyl, cyclopentanecarbonyl, adamantanecarbonyl, cyclohexyloxycarbonyl, phenylthiocarbamyl and such, arylalkyl groups such as benzyl, triphenylmethyl, benzyloxyethyl and the like and silyl groups such as trimethylsilyl and the like. Preferred N-protecting group is tert-butyloxycarbonyl (Boc).

Examples of useful protective groups for acids are substituted alkalemia esters such as 9-fluorenylmethyl, methoxymethyl, methylthiomethyl, tetrahydropyranyloxy, tetrahydropyranyloxy, methoxyethoxymethyl, 2-(trimethylsilyl)ethoxymethyl, benzyloxy is methyl, pivaloyloxymethyl, phenylacetonitrile, triisopropylchlorosilane, cinematology, aretology, finally, substituted peacelove esters, 2,2,2-trichlorethylene, 2-halogenations, ω-chloralkali, 2-(trimethylsilyl)ethyl, 2-methylthioethyl, tert-butyl, 3-methyl-3-pentalogy, dicyclopentadienyl, cyclopentyloxy, cyclohexyloxy, allyl, metalloy, cinnamony, phenyl, Silovye esters, benzyl and substituted benzyl esters, 2,6-dialkylphenol esters such as pentafluorophenyl, 2,6-dialkylanilines. Other protective groups for acids represent a methyl or ethyl esters.

Methods of annexation (method, commonly referred to as "protection"), and delete (method commonly referred to as "removing protection") of such protective groups for the amine and acid are well known in the art and are available, for example, see P.J.Kocienski, Protecting Groups, Thieme, 1994, which is incorporated into the present application by reference in its entirety, and Greene and Wuts, Protective Groups in Organic Synthesis, 3rd Edition (John Wiley & Sons, New York, 1999).

Unless otherwise stated, all tautomeric forms of the compounds of the present invention is included in the scope of the present invention. That is, the compounds of formula I may exist as tautomers:

In addition, if not indicated is ANO other the patterns presented in this application include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having presented the structure, except for the substitution of hydrogen by deuterium or tritium, or the replacement of carbon carbon13C or14C included in the scope of the present invention. Such compounds are useful, for example, as analytical tools, probes in biological assays or as therapeutic agents.

Examples of suitable solvents include, but are not limited to, water, methanol, dichloromethane (DHM), acetonitrile, dimethylformamide (DMF), ethyl acetate (EtOAc), isopropyl alcohol (IPA), isopropylacetate (IPAc), tetrahydrofuran (THF), methyl ethyl ketone (MEK), tert-butanol, and N-organic (NMP).

Examples of suitable binding agents include, but are not limited to, hydrochloride, 1-(3-(dimethylamino)propyl)-3-ethylcarbodiimide (EDCI), hexaflurophosphate 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium (HBTU), 1-hydroxybenzotriazole (HOBT), hexaflurophosphate 2-(1H-7-asobancaria-1-yl)-1,1,3,3-tetramethyluronium (HATU), tetrafluoroborate 2-chloro-1,3-dimethyl-2-imidazole, 1-H-benzotriazole-1-[bis(dimethylamino)methylene]-5-changecipherspec (HCTU), 2-chloro-4,6-dimethoxy-1,3,5-triazine and 2-papapostolou anhydride (T3P®).

Por what measures are appropriate grounds include, but are not limited to, potassium carbonate (K2CO3), N-methylmorpholine (NMM), triethylamine (TEA), diisopropylethylamine (DIEA), pyridine, potassium hydroxide, sodium hydroxide and sodium methoxide.

CONNECTION

In one variant embodiment of the invention includes a Compound of the formula I:

or its pharmaceutically acceptable salt, where R is a COOH or CH2OH.

In some embodiments embodiment R represents a CH2OH.

In some embodiments embodiment R is a COOH.

In another variant embodiment of the invention includes a compound of the structure:

or its pharmaceutically acceptable salt.

In another variant embodiment of the invention includes a compound of the structure:

or its pharmaceutically acceptable salt.

In one variant embodiment of the invention includes a pharmaceutical composition comprising a compound of formula I and pharmaceutically acceptable carrier or adjuvant.

In another variant embodiment of the invention includes a pharmaceutical composition comprising a compound of the structure:

and pharmaceutically acceptable carrier or adjuvant.

In another variant embodiment of the invention includes a pharmaceutical to the position, including a connection structure:

and pharmaceutically acceptable carrier or adjuvant.

In another variant embodiment, the pharmaceutical composition further includes an additional means selected from the group comprising a mucolytic agent, a bronchodilator agent, antibiotic, anti-infective agent, anti-inflammatory agent, a CFTR modulator, or a nutritional agent.

III. TOTAL SYNTHESIS

The compounds of formula I can be synthesized according to scheme 1.

Scheme 1

Scheme 1 anilines of the formula II, where R and OH optionally and independently contain a protective group, is subjected to the interaction with carbonnanotube intermediate compounds of formula III in the reaction conditions combination with obtaining the compounds of formula IV. The compounds of formula IV, which contain one or more protective groups, can then be subjected to the procedure of removing protection from obtaining derivatives of formula I.

The reaction mix, is shown in scheme 1 can be carried out by dissolving used in the reaction of substances in a suitable solvent, processing the obtained solution suitable binding reagent, optionally in the presence of a suitable base.

Quinoline derivatives of the formula III can sintezirovat is in accordance with scheme 2.

Scheme 2

Anilines of the formula II, where R is a-CH2OH, can be synthesized in accordance with Scheme 3.

Scheme 3

Anilines of the formula II, where R represents-COOH, can be synthesized in accordance with scheme 4.

Scheme 4

APPLICATIONS AND WAYS to USE

Pharmaceutically acceptable compositions

In one aspect of the present invention are provided pharmaceutically acceptable compositions, such compositions include any of the compounds described in this application, and optionally include a pharmaceutically acceptable carrier, adjuvant or excipient. In some embodiments, embodiments of these compositions optionally further include one or more additional therapeutic agents.

Also it should be clear that some compounds of the present invention can exist in free form for treatment, or, if it is appropriate, in the form of a pharmaceutically acceptable derivative or prodrug. In accordance with the present invention pharmaceutically acceptable derivative or prodrug includes, but is not limited to, pharmaceutically acceptable salts, esters, the Oli of such esters, or any other product or accession or derivative, which when administered to a patient in need this, is able to provide, directly or indirectly, a compound that is described in this application, or its metabolite or residue.

Used in this application, the term "pharmaceutically acceptable salt" refers to those salts which, in accordance with suspended medical rating are suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, commensurate with a reasonable ratio of benefit/risk. "Pharmaceutically acceptable salt" means any non-toxic salt or salt of ester compounds of the present invention that when administered to the recipient is capable of providing, either directly or indirectly, a compound of the present invention, or possessing inhibitory activity of the metabolite, or the remainder of such compounds.

Pharmaceutically acceptable salts are well known in the prior art. For example, S.M. Berge, et al. Describe pharmaceutically acceptable salts in J. Pharmaceutical Sciences, 1977, 66, 1-19, which is incorporated into the present application by reference. Pharmaceutically acceptable salts of the compounds of the present invention include salts derived from suitable inorganic and organic acids and bases. Examples of pharmaceutical is Ki acceptable non-toxic acid additive salts include salts of an amino group, formed with inorganic acids such as hydrochloric acid, Hydrobromic acid, phosphoric acid, sulfuric acid, Perlina acid, or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, bansilalpet, benzoate, bisulfate, borate, butyrate, comfort, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulphate, edisylate (etandisulfonat), aconsultant, formate, fumarate, glucoheptonate, glycerol, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxyethanesulfonic, lactobionate, lactate, laurate, lauryl, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluensulfonate, undecanoate, valerate and the like. Salts derived from appropriate bases include alkali metal salts, alkaline earth metals, ammonium and N+(C1-4alkyl)4 . The present invention also envisions the quaternization of any basic nitrogen-containing groups of the compounds disclosed in this application. Water - or oil-soluble or dispersible products may be obtained by using such quaternization. Representative salts of alkaline or alkaline earth metals include sodium, lithium, potassium, calcium, magnesium and the like. In addition, pharmaceutically acceptable salts include, if it is appropriate, nontoxic ammonium cations, Quaternary ammonium and amine formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower alkylsulfonate and arylsulfonate.

As described above, the pharmaceutically acceptable compositions of the present invention optionally include a pharmaceutically acceptable carrier, adjuvant or excipient, which, as used in this application includes any and all solvents, diluents, or other liquid carriers, substances that contribute to the dispersion or suspendirovanie, surface active agents, isotonic agents, thickeners or emulsifiers, preservatives, solid binders, lubricants and the like, as is appropriate for a particular dosage form, which is desirable. Remington''s Pharmaceutical Sciences, Sixteenth Editin, E.W. Martin (Mack Publishing Co., Easton, Pa., 1980) discloses various carriers used in formulating pharmaceutically acceptable compositions and known methods of obtaining them. Except only in those cases where any traditional medium used as a carrier, is incompatible with the compounds of the present invention, for example, causing any undesirable biological effect or otherwise interacting in an adverse manner with any other component(components) pharmaceutically acceptable composition, its use is envisaged as covered by the scope of the present invention. Some examples of substances which can serve as pharmaceutically acceptable carriers include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid or potassium sorbate, a mixture of partial glycerides of saturated vegetable fatty acids, water, salts or electrolytes, such as preteenslut, secondary, acidic sodium phosphate, potassium phosphate, sodium chloride, zinc salts, colloidal silicon dioxide, magnesium trisilicate, polyvinylpyrrolidone, polyacrylates, waxes, polyethylene-polyoxypropylene block polymers, lanolin, sugar is, such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethylcellulose and cellulose acetate; powdered tragakant; malt; gelatin; talc; excipients such as cocoa butter and waxes for suppositories; oils such as peanut oil, cotton seed; safflower oil; sesame oil; olive oil; corn oil and soybean oil; glycols; such a propylene glycol or polyethylene glycol; esters, such as etiloleat and tillaart; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; ringer's solution; ethyl alcohol and phosphate buffer solutions, as well as other non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, and colouring agents, substances promoting the release of shapes, substances, coatings, sweeteners, fragrances and flavors, preservatives and antioxidants can also be present in the composition in accordance with the order, as it deems necessary specialist involved in formulating the composition.

Application of compounds and pharmaceutically acceptable compositions

In another aspect of the present invention PR is the way to cure or ameliorate the severity of the condition, diseases or disorders associated with CFTR mutation. In some embodiments, embodiments of the present invention provides a method of treating a condition, disease or disorder associated with deficiency of CFTR activity, the method includes introducing a composition comprising a compound of formula 1, to a subject, preferably a mammal in need of it.

In another aspect the invention also provides a method of treating or attenuating the severity of the disease in a patient, comprising the introduction of a specified patient one of the compositions defined in the present application, and the disease is selected from diseases such as cystic fibrosis, asthma, caused by Smoking COPD (chronic obstructive pulmonary disease), chronic bronchitis, rhinosinusitis, constipation, pancreatitis, pancreatic insufficiency cancer, male infertility caused by congenital bilateral absence of the VAS deferens (CBAVD), uncomplicated form of pulmonary disease, idiopathic pancreatitis, allergic bronchopulmonary aspergillosis (ABPA), liver disease, hereditary emphysema, hereditary hemochromatosis, coagulation-fibrinolysis deficiencies, such as protein deficiency, hereditary angioedema type 1, lipid processing, such as family hypercholester the nemia, chylomicronemia type 1, abetalipoproteinemia, lysosomal storage disorders, such as disease cellular inclusions/disease Deri, mucopolysaccharidosis, disease Sandhof/Tay-Sachs disease criglernajjar Najjar syndrome type II, polyendocrinopathy/hyperinsulemia, diabetes, dwarfism of Larona, myeloperoxidase deficiency, primary hypoparathyroidism, melanoma, glycans CDG (congenital diseases of glycosylation type 1, congenital hyperthyroidism, imperfect osteogenesis, hereditary hypofibrinogenemia, the lack of an activated clotting time of blood, diabetes insipidus (ND), insipidus neurotically diabetes insipidus neurogenic diabetes, muscular atrophy, Charcot-Marie-Toot, disease Pelizaeus-Merzbacher neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, progressive supranuclear paralysis, atrophy of the Peak, some polyglutamine neurological disorders such as Huntington's disease, spinal-cerebellar ataxia type I, spinal and bulbar muscular atrophy, dentato-rubro-pallido-Lisova atrophy and myotonica dystrophy, as well as spongiform encephalopathies, such as hereditary disease Creutzfeldt-Jakob disease (due to defective processing of the prion protein), Fabry disease, syndrome Straussler-Necks is Kera, COPD (chronic obstructive pulmonary disease), dry eye syndrome, or sjögren's disease, osteoporosis, osteopenia, bone healing and bone growth (including the restoration of the bone, bone regeneration, reducing bone resorption and increasing bone deposition), the syndrome Goreme, chloride of kalapati, such as congenital myotonia form (Thomson and Becker), the syndrome Bartter type III, dent disease, hyperekplexia, epilepsy, hyperekplexia, lysosomal disease accumulation, Angelman syndrome, and primary ciliary dyskinesia (PCD), a term for inherited disorders of the structure and/or function resisteth structures, including PCD with transposition of internal organs (also known as syndrome Addition, PCD without transposition of internal organs and ciliary aplasia.

In some embodiments embodiment the method comprises the treatment or mitigation of the severity of cystic fibrosis in a patient, comprising the introduction of a specified patient one of the compositions defined in the present application. In some embodiments embodiment the patient has a mutant form of human CFTR. In other variants of the embodiment, the patient has one or more of the following mutations ΔF508, R117H, and G551D human CFTR. In one variant embodiment, the method includes the treatment or mitigation of the severity of cystic fibrosis patients with the F508 mutation of human CFTR, includes introduction to the specified patient one of the compositions defined in the present application. In one variant embodiment, the method includes the treatment or mitigation of the severity of cystic fibrosis in a patient possessing the G551D mutation of human CFTR comprising the introduction of a specified patient one of the compositions defined in the present application. In one variant embodiment, the method includes the treatment or mitigation of the severity of cystic fibrosis in a patient possessing the ΔF508 mutation of human CFTR on at least one allele, including the introduction of a specified patient one of the compositions defined in the present application. In one variant embodiment, the method includes the treatment or mitigation of the severity of cystic fibrosis in a patient possessing the ΔF508 mutation of human CFTR on both alleles, including the introduction of a specified patient one of the compositions defined in the present application. In one variant embodiment, the method includes the treatment or mitigation of the severity of cystic fibrosis in a patient possessing the G551D mutation of human CFTR on at least one allele, including the introduction of a specified patient one of the compositions defined in the present application. In one variant embodiment, the method includes the treatment or mitigation of the severity of cystic fibrosis in a patient possessing the G551D mutation of human CFTR on both alleles, kiuchumi introduction to the specified patient one of the compositions, defined in this application.

In some embodiments the embodiment of the method includes the attenuation of the severity of cystic fibrosis in a patient, comprising the introduction of a specified patient one of the compositions defined in the present application. In some embodiments embodiment the patient has a mutant form of human CFTR. In other variants of the embodiment, the patient has one or more of the following mutations ΔF508, R117H, and G551D human CFTR. In one variant embodiment, the method includes the attenuation of the severity of cystic fibrosis in a patient possessing the ΔF508 mutation of human CFTR comprising the introduction of a specified patient one of the compositions defined in the present application. In one variant embodiment, the method includes the attenuation of the severity of cystic fibrosis in a patient possessing the G551D mutation of human CFTR comprising the introduction of a specified patient one of the compositions defined in the present application. In one variant embodiment, the method includes the attenuation of the severity of cystic fibrosis in a patient possessing the ΔF508 mutation of human CFTR on at least one allele, including the introduction of a specified patient one of the compositions defined in the present application. In one variant embodiment, the method includes the attenuation of the severity of cystic fibrosis in a patient possessing the ΔF508 mutation of human CFTR on both and lelah, includes introduction to the specified patient one of the compositions defined in the present application. In one variant embodiment, the method includes the attenuation of the severity of cystic fibrosis in a patient possessing the G551D mutation of human CFTR on at least one allele, including the introduction of a specified patient one of the compositions defined in the present application. In one variant embodiment, the method includes the attenuation of the severity of cystic fibrosis in a patient possessing the G551D mutation of human CFTR on both alleles, including the introduction of a specified patient one of the compositions defined in the present application.

In some aspects, the invention provides a method of treating or attenuating the severity of osteoporosis in a patient, comprising the introduction of a given patient, the compounds of Formula 1 or its pharmaceutically acceptable salt.

In some embodiments, embodiments a method of treating or attenuating the severity of osteoporosis in a patient includes an introduction to the specified patient the pharmaceutical composition described in this application.

In some aspects, the invention provides a method of treating or attenuating the severity of osteopenia in a patient, comprising the introduction of a given patient, the compounds of formula 1 or its pharmaceutically acceptable salt.

In some embodiments, embodiments a method of treating or attenuating t the tin osteopenia in a patient includes the introduction of the indicated patient a pharmaceutical composition, described in this application.

In some aspects, the invention provides a method of bone healing and/or bone repair in a patient, comprising the introduction of a given patient, the compounds of formula 1 or its pharmaceutically acceptable salt.

In some embodiments, embodiments of a method of bone healing and/or bone repair in a patient includes an introduction to the specified patient the pharmaceutical composition described in this application.

In some aspects, the invention provides a method of reducing bone resorption in a patient, comprising the introduction of a given patient, the compounds of formula 1 or its pharmaceutically acceptable salt.

In some aspects, the invention provides a method of increasing bone deposits in a patient, comprising the introduction of a given patient, the compounds of formula 1 or its pharmaceutically acceptable salt.

In some embodiments, embodiments of the method of increasing bone deposits in a patient includes an introduction to the specified patient the pharmaceutical composition described in this application.

In some aspects, the invention provides a method of treating or attenuating the severity of COPD in a patient, comprising the introduction of a given patient, the compounds of formula 1 or its pharmaceutically acceptable salt.

In some embodiments, embodiments of the method of treatment and the loosening of severity of COPD in a patient includes the introduction of the indicated patient a pharmaceutical composition, described in this application.

In some aspects, the invention provides a method of treating or attenuating the severity caused by Smoking COPD patients, including the introduction of a given patient, the compounds of formula 1 or its pharmaceutically acceptable salt.

In some embodiments, embodiments a method of treating or attenuating the severity of COPD caused by Smoking in a patient includes an introduction to the specified patient the pharmaceutical composition described in this application.

In some aspects, the invention provides a method of treating or attenuating the severity of chronic bronchitis in a patient, comprising the introduction of a given patient, the compounds of formula 1 or its pharmaceutically acceptable salt.

In some embodiments, embodiments a method of treating or attenuating the severity of chronic bronchitis in a patient includes an introduction to the specified patient the pharmaceutical composition described in this application.

In accordance with an alternative embodiment of the present invention provides a method of treating cystic fibrosis, comprising the stage of introduction of a given mammal an effective amount of a composition comprising the compound of the present invention.

In accordance with the present invention an "effective amount" of a compound or pharmaceutically acceptable composition ol dstanley a such number, which is effective to treat or ameliorate the severity of one or more diseases, disorders or conditions mentioned above.

Another aspect of the present invention provides a method for introducing a pharmaceutical composition by oral administration to a patient at least once a day, compositions comprising the compound of formula 1. In one variant embodiment, the method includes the introduction of a pharmaceutical composition comprising a compound of formula 1, every 24 hours. In another variant embodiment of the method includes the introduction of a pharmaceutical composition comprising a compound of formula 1, every 12 hours. In the following variant embodiment of the method includes the introduction of a pharmaceutical composition comprising a compound of formula 1, three times per day. In another variant embodiment of the method includes the introduction of a pharmaceutical composition comprising a compound of formula 1, every 4 hours.

Compounds and compositions in accordance with the method of the present invention can be administered using any amount and any route of administration effective for treating or attenuating the severity of one or more of the diseases, disorders or conditions mentioned above.

In some embodiments the embodiment of the compounds and compositions of the present invention are reached what passed for a cure or ameliorate the severity of cystic fibrosis in patients which exhibit residual CFTR activity in the apical membrane of respiratory tissue and nerespectarea epithelium. The presence of residual CFTR activity on the surface of the epithelium can be easily determined using methods known from the prior art, such as standard electrophysiological, biochemical or histochemical methods. Such methods determine CFTR activity usingin vivoorex vivoelectrophysiological methods, measurement of Cl-concentrations in the discharge sweaty or salivary glands or usingex vivobiochemical or histochemical methods for controlling the density of the cell surface. Using these methods you can easily determine the residual CFTR activity in patients heterozygous or homozygous for different mutations, including patients homozygous or heterozygous for the most common mutation, ΔF508.

In another variant embodiment of the compounds and compositions of the present invention are useful to treat or ameliorate the severity of cystic fibrosis in patients who have residual CFTR activity, induced or amplified using pharmacological methods or gene therapy. Such methods increase the amount of CFTR present on the cell surface, inducyrutmi way missing before CFTR activity in a patient or increasing the existing level of residual CFTR activity in a patient.

In one variant embodiment of the compounds and compositions of the present invention are useful to treat or ameliorate the severity of cystic fibrosis in patients within certain genotypes, demonstrating residual CFTR activity, e.g mutation class III (violation of regulation or gate mechanism), the mutation class IV (changing conductivity) or mutations of class V (reduced synthesis) (Lee R. Choo-Kang, Pamela L., Zeitlin, Type I, II, III, IV, and V cystic fibrosis Transmembrane Conductance Regulator Defects and Opportunities of Therapy; Current Opinion in Pulmonary Medicine 6:521-529, 2000). Other genotypes in patients who exhibit residual CFTR activity, include patients homozygous for one of these classes or heterozygous with any other class of mutations, including mutations of class I mutation class II or mutations that are not classified.

In one variant embodiment of the compounds and compositions of the present invention are useful to treat or ameliorate the severity of cystic fibrosis in patients within certain clinical phenotypes, e.g., from moderate to mild clinical phenotype that typically correlates with the amount of residual CFTR activity in the apical membrane of epithelial tissues. Such phenotypes include patients, demonstrating the insufficiency of the pancreas, or patients cat who where diagnosed with idiopathic pancreatitis and congenital bilateral absence of the VAS deferens or mild lung disease.

The exact amount that is needed will be different for different subjects, depending on the specific species, age and General condition of the subject, the severity of infection, specific means, method of its introduction, etc. of the Compounds of the present invention are preferably formulated in dosage form containing standard dosing unit, for ease of introduction and uniform dosing. The expression "standard unit dosing", as used in this application, refers to a physically discrete unit of money that is appropriate for the patient to be treated. However, it should be clear that the total daily intake of the compounds and compositions of the present invention is determined by the physician in accordance with a weighted medical assessment. The specific effective dose for any particular patient or organism will depend upon a variety of factors including the disorder to be treated, and the severity of the disorder; activity of the specific compound; the specific composition; the age, body weight, General health, sex and diet of the patient; the time of administration, route of administration, and rate of excretion of the specific compound; the duration of the treatment; drugs used in combination or cases, is but the specific connection and similar factors, well known in medicine. The term "patient"as used in this application, means an animal, preferably a mammal, and most preferably human.

Pharmaceutically acceptable compositions of the present invention can enter humans and other animals orally, rectally, parenterally, intracisternally, intrawaginalno, IPR, local path (for example, in the form of powders, ointments, drops or patch), buccal, in the form of oral or nasal spray, or the like, depending on the severity of the infection to be treated. In some embodiments the embodiment of the compounds of the present invention can be administered orally or parenterally at dosages between about 0.01 mg/kg to about 50 mg/kg and preferably from about 0.5 mg/kg to about 25 mg/kg of body weight of the subject per day, once or several times a day to obtain the desired therapeutic effect.

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, the liquid dosage forms may contain inert diluents conventionally used in this field, such as, for example, water or other solvents, solubilizing agents and emulsion is atory, such as ethyl alcohol, isopropyl alcohol, ethylcarbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyleneglycol, dimethylformamide, oils (in particular, oil seeds, cotton, peanut oil, corn oil, oil of seedlings seeds, olive oil, castor oil and sesame oil), glycerin, tetrahydrofurfuryl alcohol, polyethylene glycols and esters of fatty acids sorbitan, and mixtures thereof. Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifiers and suspendresume agents, sweeteners, fragrances and flavorings.

Injectable preparations, for example a sterile aqueous or oily suspension for injection can be formulated according to known prior art methods using suitable dispersing or wetting agents and suspendida substances. A sterile preparation for injection may also be a sterile solution, suspension or emulsion for injection in a non-toxic parenterally acceptable diluent or solvent, for example, in the form of a solution in 1,3-butanediol. Acceptable fillers and solvents that can be used, you can specify the water, ringer's solution U.S.P. and isotonic sodium chloride solution. In addition, article is realnye non-volatile oil is traditionally used as a solvent or medium for suspension. For these purposes you can use any light non-volatile oils, including synthetic mono - or diglycerides. In addition, in preparations for injection use fatty acids such as oleic acid.

Compositions for injection can be sterilized, for example, by filtration through a retaining bacteria filter or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersing in sterile water or other sterile environment for injection before use.

To prolong the effect of the compounds according to the present invention it is often desirable to slow the absorption of the compound from subcutaneous or intralesional injection. This can be done using a liquid suspension of crystalline or amorphous material with poor water-solubility. The absorption rate of the connection in this case depends on its rate of dissolution which, in turn, may depend upon crystal size and crystalline form. Alternative a slower absorption of parenteral input form compounds obtained by dissolution or suspension of the compounds in the oil filler. Depot forms for injection is produced by the formation of matrices for microencapsulation compounds in biodegradable polymers such as polylactide-polyglycol the D. Depending on the ratio of the compounds to the polymer and the nature of the particular polymer used, you can control the rate of release of connection. Examples of other biodegradable polymers include poly(orthoevra) and poly(anhydrides). Composition depot injections also produced by the conclusion of the compounds in liposomes or microemulsions that are compatible with body tissues.

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

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 secondary acidic calcium phosphate and/or a) fillers or bulk substances such as cu is hmily, lactose, sucrose, glucose, mannitol and silicic acid, b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and the Arabian gum, (c) humectants, such as glycerol, d) disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates and sodium carbonate, e) slow dissolving substances such as paraffin, f) absorption accelerators such as Quaternary ammonium compounds, g) wetting agents such as, for example, cetyl alcohol and glycerylmonostearate, h) absorbents such as kaolin and bentonite clay, and i) lubricating agents such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures of such substances. In the case of capsules, tablets and pills, the dosage form may also include a buffering agent.

Solid compositions of a similar type can also be used as fillers, prisoners in soft and hard gelatin capsules using such excipients as lactose or milk sugar and high molecular weight polyethylene glycols and the like. Solid dosage forms such as tablets, pills, capsules, pills and granules can be obtained with coatings and shells, such is how intersolubility coatings and other coatings, well known in the field of pharmaceutical formulation. They do not necessarily contain opaque agents and also may have such a composition that makes possible the release of the active ingredient(ingredients)only, or preferentially, in a certain part of the digestive tract, it is not necessarily slow. Examples of compositions for encapsulating substances, which can be used include polymeric substances and waxes. Solid compositions of a similar type can also be used as fillers, prisoners in soft and hard gelatin capsules using such excipients as lactose or milk sugar and high molecular weight polyethylene glycols and the like.

The active compounds can also be microencapsulating form with one or more excipients as noted above. Solid dosage forms such as tablets, coated tablets, capsules, pills and granules can be obtained with coatings and shells, such as intersolubility coating that controls the release coatings and other coatings well known in the field of pharmaceutical formulation. In such solid dosage forms the active compound may be mixed with at least one inert diluent such as sucrose, lactose or starch. the such dosage forms can also include, as in normal practice, additional substances other than inert diluents, such as lubricant for tableting and other excipients for tableting, such stearate and microcrystalline cellulose. In the case of capsules, tablets and pills such dosage forms can also include buffering agents. They do not necessarily contain opaque agents and also may have such a composition that makes possible the release of the active ingredient(ingredients)only, or preferentially, in a certain part of the digestive tract, it is not necessarily slow. Examples of compositions for encapsulating substances, which can be used include polymeric substances and waxes.

Dosage forms for local administration or by percutaneous connection of the present invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalations or patches. The active ingredient is mixed under sterile conditions with a pharmaceutically acceptable carrier and any needed preservatives or buffers, which may be involved. Eye medications, ear drops and eye drops are also provided as covered by the scope of the present invention. In addition, the present invention p is educative using percutaneous patches, which have the added advantage of providing controlled delivery of compounds into the body. Such dosage forms are obtained by dissolving or dispersing the compound in a suitable medium. You can also use amplifiers absorption, used to enhance the penetration of compounds through the skin. Speed can be controlled either by software controlling the speed of the membrane or by dispersing the compound in a polymer matrix or gel.

Analysis of the activity of the compounds used in the present invention as a modulator of CFTR can be carried out in accordance with methods described in General in the prior art and in the Examples presented in this application.

Also it should be clear that the compounds and pharmaceutically acceptable compositions of the present invention can be used in combination therapies, that is, the compounds and pharmaceutically acceptable compositions can be administered simultaneously with introduction, before or after administration of one or more other desired therapeutics or medical procedures. Specific combined therapy (therapeutic agent or procedure) for use in a combined regimen should take into account compatibility of the desired therapeutics and/or proced the R and the desired therapeutic effect, the achievement you want. Also it should be clear that your therapy can achieve desired effect for the same disorder (for example, the compound of the present invention can be administered simultaneously with another agent used to treat the same disorder), or they can be designed to achieve different effects (e.g., control of any adverse effects). As used in this application, additional therapeutic agents that are normally administered to treat or prevent a specific disease or condition, known as "suitable for the disease or condition to be treated."

In one variant embodiment, the additional agent selected from a mucolytic agents, bronhodilatator, antibiotic, anti-infective funds protivovospalitelnoe means of CFTR modulator other than the compounds of the present invention, or nutrients.

In one variant embodiment, the additional agent is an antibiotic. Examples of antibiotics that are useful in the present invention include tobramycin, including tobramycin powder for inhalation (TIP), azithromycin, aztreonam, including aerosol form aztreonam, amikacin, including liposomal composition, ciprofloxacin, including eg the composition, suitable for administration by inhalati, levofloxacin, including aerosol composition, and combinations of two antibiotics, for example fosfomicin and tobramycin.

In another variant embodiment, the additional agent is a mucolytic agent. Examples mucolytic tools that are useful in the present invention include Pulmozyme®.

In another variant embodiment, the additional agent is a bronchodilator. An example of bronchodilators include albuterol, metaproterenol sulfate, pirbuterol acetate, salmeterol or tetralin sulfate.

In another variant embodiment, additional means is effective to restore the surface of the liquid in the Airways in the lungs. Such tools improve the movement of salt in the cells and from the cells, making the mucus in the Airways in the lungs more hydrated, so they are more easily excreted. Examples of such tools include hypertonic saline, denufosol tetranitro ([[(3S,5R)-5-(4-amino-2-oxopyrimidine-1-yl)-3-hydroxyanisole-2-yl]methoxyhydroquinone][[[(2R,3S,4R,5R)-5-(2,4-dioxopyrimidine-1-yl)-3,4-dihydroquinoxaline-2-yl]methoxyhydroquinone]occipitotemporal]phosphate) or bronchitol (drug for inhalation based on mannitol).

In another variant embodiment, additional means to depict is to place an anti-inflammatory agent, i.e. a tool that can reduce inflammation in the lungs. Examples of such tools that are useful in the present invention include ibuprofen, docosahexaenoyl acid (DHA), sildenafil, glutathione for inhalation, pioglitazone, hydroxychloroquine or simvastatin.

In another variant embodiment, the additional agent is a CFTR modulator other than compounds 1, i.e. a tool that has the effect of modulating CFTR activity. Examples of such funds include ataluren (PTC 124®"; 3-[5-(2-forfinal)-1,2,4-oxadiazol-3-yl]benzoic acid), sinapultide, lancemate, depeleted (human recombinant inhibitor of neutrophil elastase), cobiprostone (7-{(2R,4aR,5R,7aR)-2-[(3S)-1,1-debtor-3-methylpentyl]-2-hydroxy-6-accountpaydayloan[b]Piran-5-yl}heptane acid) or (3-(6-(1-(2,2-debtorrent[d][1,3]dioxol-5-yl)cyclopropanecarboxamide)-3-methylpyridin-2-yl)benzoic acid. In another variant embodiment, the additional agent is a (3-(6-(1-(2,2-debtorrent[d][1,3]dioxol-5-yl)cyclopropanecarboxamide)-3-methylpyridin-2-yl)benzoic acid.

In another variant embodiment, the additional agent is a nutrient. Examples of such substances include pancrelipase (Deputy pancreatic enzymes), including Pancrease®, Pancreacarb®, Ultrase® or Creon®, Liprotomase® (formerly Trizytek®), Aquadeks® or glutathione for inhalation. In one variant embodiment, additional nutrient is pancrelipase.

The amount of additional therapeutic agent present in the compositions of the present invention, should not exceed the amount, which is normally possible to introduce in the composition comprising therapeutic agent as the only active substance. Preferably the amount of additional therapeutic agent in the compositions disclosed in the present invention must be in the range of from about 50% to 100% of the amount, which is normally possible to introduce in the composition comprising therapeutic agent as the only active substance.

Compounds of the present invention or containing pharmaceutically acceptable compositions can also be incorporated into compositions for coating implantable medical devices such as prostheses, artificial valves, vascular grafts, stents and catheters. Accordingly, the present invention in another aspect includes a composition for coating an implantable device comprising the compound of the present invention described generally above and in classes and subclasses described in this application, and a carrier suitable for coating specified and plateruena device. In another aspect of the present invention includes an implantable device coated with a composition including the compound of the present invention described generally above and in classes and subclasses described in this application, and a carrier suitable for coating of specified implanted device. Suitable coatings and the General description of the receipt with the coating implantable devices can be found in U.S. patent No. 6099562; 5886026 and 5304121. Coverage typically represent a biocompatible polymer material, such as polymeric hydrogels, polymethylsiloxane, polycaprolactone, polyethylene glycol, polylactic acid, ethylene vinyl acetate and mixtures thereof. The coating optionally may be additionally covered with a suitable top layer of Versiliana, polysaccharides, polyethylene glycol, phospholipids or combinations thereof to impart to the composition of the characteristics of a controlled release.

Another aspect of the present invention relates to modelirovaniya CFTR activity in a biological sample or in a patient (e.g.,in vitroorin vivo), and the method comprises the administration to the patient, or contacting the specified biological sample with a compound of formula 1 or a composition comprising the specified connection. The term "biological sample", as used in this application, VK is uchet, without limitation, cell cultures or extracts; biopsy material obtained from a mammal or extracts; and blood, saliva, urine, feces, semen, tears, or other body fluids or extracts.

Modulation of CFTR in a biological sample is useful for a variety of purposes that are known to specialists in this field. Examples of such purposes include, but are not limited to, the study of CFTR in biological and pathological phenomena and comparative evaluation of new modulators of CFTR.

In the following variant embodiment provides a method of modulating activity of anionic channelin vitroorin vivoincluding the stage of contacting the specified channel with a compound of formula 1. In variants of the embodiment of the anion channel is a chloride channel or a bicarbonate channel. In other embodiments, the embodiment of the anion channel is a chloride channel.

In accordance with an alternative embodiment of the present invention provides a method of increasing the number of functional CFTR in the cell membrane, including the state of engagement of the specified cell with the compound of the formula 1.

In accordance with another preferred embodiment of the CFTR activity is determined by measuring the transmembrane potential. Means for measuring the potential of the and through the membrane in a biological sample can include any of the ways known from the prior art, such as optical analysis of membrane potential or other electrophysiological methods.

Optical analysis of the membrane potential involves the use of a potential-sensitive FRET sensors described by Gonzalez and Tsien (see Gonzalez, J.E. and R.Y. Tsien (1995) "Voltage sensing by fluorescence resonance energy transfer in single cells." Biophys J 69(4): 1272-80; Gonzalez, J.E. and R.Y. Tsien (1997); "Improved indicators of cell membrane potential that use fluorescence resonance energy transfer" Chem Biol 4(4): 269-77)in combination with devices for measuring fluorescence changes such as the Voltage/Ion Probe Reader (VIPR) (see Gonzalez, J.E., K. Oades, et al. (1999) "Cell-based assays and instrumentation for screening ion-channel targets" Drug Discov Today 4(9): 431-439).

These potential-sensitive analyses based on the change of the transmission resonance energy fluorescence (FRET) between membranectomy potential-sensitive dye, DiSBAC2(3) and fluorescent phospholipid, CC2-DMPE, which is attached to the outer leaf of the plasma membrane and acts as a FRET donor. Changes in membrane potential (Vm) causes a redistribution of the negatively charged DiSBAC2(3) through the plasma membrane, and the amount of transferred energy from the CC2-DMPE changes accordingly. Changes in the fluorescence emission can be monitored using VIPR™ II, which is an integrated device fluid supply and the fluorescence detector, designed the TES for the implementation of cellular screening assays in 96 - or 384-well microtiter plates.

In one variant embodiment of the present invention provides a method of modulating CFTR activity in a biological sample, comprising the stage of contact specified biological sample with a compound of formula 1 or its pharmaceutically acceptable salt, where R has the meaning defined above.

In one variant embodiment of the present invention provides a method of modulating CFTR activity in a biological sample, comprising the stage of contact specified biological sample with a compound having the structure:

or its pharmaceutically acceptable salt.

In one variant embodiment of the present invention provides a method of modulating CFTR activity in a biological sample, comprising the stage of contact specified biological sample with a compound having the structure:

or its pharmaceutically acceptable salt.

In one variant embodiment of the present invention provides a method of treating or attenuating the severity of the disease in a patient, comprising the introduction of a specified patient an effective amount of the compounds of formula 1 or its pharmaceutically acceptable salts, where R has the meaning defined above, and the disease is selected from the following: cystic fibrosis, asthma, call the data Smoking COPD (chronic obstructive pulmonary disease), chronic bronchitis, rhinosinusitis, constipation, pancreatitis, pancreatic insufficiency cancer, male infertility caused by congenital bilateral absence of the VAS deferens (CBAVD), uncomplicated form of pulmonary disease, idiopathic pancreatitis, allergic bronchopulmonary aspergillosis (ABPA), liver disease, hereditary emphysema, hereditary hemochromatosis, coagulation-fibrinolysis deficiencies, such as protein deficiency, hereditary angioedema type 1, lipid processing, such as familial hypercholesterolaemia, chylomicronemia type 1, abetalipoproteinemia, lysosomal storage disorders, such as disease cellular inclusions/disease Deri, mucopolysaccharidosis, disease Sandhoff/Tay-Sachs disease criglernajjar Najjar syndrome type II, polyendocrinopathy/hyperinsulemia, diabetes, dwarfism of Larona, myeloperoxidase deficiency, primary hypoparathyroidism, melanoma, glycans CDG (congenital diseases of glycosylation type 1, congenital hyperthyroidism, imperfect osteogenesis, hereditary hypofibrinogenemia, the lack of an activated clotting time of blood, diabetes insipidus (ND), insipidus neurotically diabetes insipidus neurogenic diabetes, muscular atrophy, Charcot-Marie-Toot disease Pelizaeus-Merzbacher aeroderivative disease, such as Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, progressive supranuclear paralysis, atrophy of the Peak, some polyglutamine neurological disorders as Huntington's disease, spinal-cerebellar ataxia type I, spinal and bulbar muscular atrophy, dentato-rubro-pallido-Lisova atrophy and myotonica dystrophy, as well as spongiform encephalopathies, such as hereditary disease Creutzfeldt-Jakob disease (due to defective processing of the prion protein), Fabry disease, syndrome Straussler-Sheinker, COPD (chronic obstructive pulmonary disease), dry eye syndrome or disease Sjogren's syndrome.

In one variant embodiment, the method includes treating or attenuating the severity of the disease in a patient by introducing a specified patient an effective amount of a compound having the structure:

or its pharmaceutically acceptable salt.

In one variant embodiment, the method includes treating or attenuating the severity of the disease in a patient by introducing a specified patient an effective amount of a compound having the structure:

or its pharmaceutically acceptable salt.

In one the following variant embodiment the disease is a cystic fibrosis.

In another TSA is the present invention provides a kit for use in measuring the activity of CFTR or a fragment in the biological sample in vitroorin vivoincluding (i) a composition comprising a compound of formula 1 or any of the options above embodiment and (ii) instructions for a) contacting the composition with the biological sample and b) measuring activity of the specified CFTR or fragment.

In one variant embodiment, the kit further includes instructions for a) contacting an additional composition with the biological sample; b) measuring activity of the specified CFTR or fragment in the presence of the specified additional compound, and c) comparing the activity of the CFTR in the presence of the additional compound with the density of CFTR in the presence of a composition of formula 1.

In preferred embodiments, the embodiments set used to measure the density of CFTR.

In one variant embodiment, the kit includes a composition including a compound having the structure:

or its pharmaceutically acceptable salt.

In one variant embodiment, the kit includes a composition including a compound having the structure:

or its pharmaceutically acceptable salt.

For a more complete disclosure of the invention described in this application, hereinafter presents the following examples. It should be clear that these examples are intended for illustrative purposes only and should not races is to metrovacesa as limiting in any way the present invention.

EXAMPLES

Getting 1: General synthesis of 4-oxo-1,4-dihydroquinoline-3-carboxylic acid (26)

The procedure to obtain ethyl 4-oxo-1,4-dihydroquinoline-3-carboxylate (25)

Compound 23 (4.77 g, while 47.7 mmol) was added dropwise to the compound 22 (10 g, and 46.3 mmol) with subsurface flow N2to displace ethanol at a temperature below 30°C for 0.5 hours. The solution is then heated to a temperature of 100-110°C and was stirred for 2.5 hours. After cooling the mixture to a temperature below 60°C was added diphenyl ether. The resulting solution was added dropwise to diphenyl ether, which was heated to 228-232°C for 1.5 hours with subsurface flow N2to displace ethanol. The mixture was stirred at a temperature of 228-232°C for an additional 2 hours, cooled to a temperature below 100°C and then added heptane to precipitate the product. The resulting suspension was stirred at 30°C for 0.5 hours. The solids were then filtered and the precipitate washed with heptane and dried in vacuum to obtain compound 25 as a brown solid.

1H-NMR (DMSO-d6; 400 MHz) δ 12,25 (c), δ 8,49 (d), δ 8,10 (m), δ to 7.64 (m, δ of 7.55 (m), δ 7,34 (m), δ 4,16 (square), δ 1,23 (t).

The procedure for obtaining 4-ACS who -1,4-dihydroquinoline-3-carboxylic acid (26)

Method 1

Compound 25 (1.0 EQ.) suspended in a solution of HCl (10.0 EQ.) and H2O (11,6 about.). The suspension was heated to a temperature of 85-90°C, although alternative temperature are also suitable for this hydrolysis step. For example, hydrolysis of an alternative can be performed at a temperature of from about 75 to about 100°C. In some cases, the hydrolysis is carried out at a temperature of from about 80 to about 95°C. In other cases, the stage of the hydrolysis is carried out at a temperature of from about 82 to about 93°C (for example, from about 82.5 to about 92,5°C or from about 86 to about 89°C). After stirring at a temperature of 85-90°C for approximately 6.5 hours took a sample of the reaction mixture to complete the reaction. Mixing can be carried out at any temperature suitable for the reaction of hydrolysis. The solution is then cooled to a temperature of 20-25°C and filtered. Reactor/the precipitate was washed using H2O (2 vol. ×2). The precipitate is then washed with about 2. H2O up until the pH became ≥3,0. The precipitate was then dried under vacuum at 60°C to obtain compound 26.

Method 2

Compound 25 (11.3 g, 52 mmol) was added to a mixture of 10% NaOH (aqueous solution) (10 ml) and ethanol (100 ml). The solution was heated to boiling point under reflux for 16 hours, cooled to a temperature of 20-25°C and then ur the level of pH is brought up to 2-3 using the 8% solution of HCl. The mixture then was stirred for 0.5 hours and filtered. The precipitate was washed with water (50 ml) and then dried in vacuum to obtain compound 26 as a brown solid.

1H-NMR (DMSO-d6; 400 MHz) δ 15,33 (c), δ 13,39 (c), δ 8,87 (c), δ compared to 8.26 (m), δ 7,87 (m), δ 7,80 (m), δ 7,56 (m).

Example 1:General synthesis of N-(2-tert-butyl-5-hydroxy-4-(1-hydroxy-2-methylpropan-2-yl)phenyl)-4-oxo-1,4-dihydroquinoline-3-carboxamide (27)

The General scheme of the synthesis of compound 27 is presented below, followed by the synthesis procedure of each synthetic intermediate substance.

The procedure for obtaining 2-hydroxy-5-tert-butylbenzaldehyde (2)

To a stirred solution of compound 1 (700 g of 4.66 mol) in CH3CN (7.0 l) was added MgCl2(887 g to 9.32 mol), para-formaldehyde (1190 g) and TEA (2.5 l, 17,9 mol) in an atmosphere of N2. The mixture was heated to boiling point under reflux for 5 hours. After cooling to room temperature, to the mixture was added 2 l of ice water and then adding 6 l of 3M HCl solution (aqueous solution). The suspension was left to mix until until the solution became transparent. The organic layer was separated and the aqueous layer was extracted using MTBE (3 l ×3). The organic layers were combined and co is centered dryness. The residue was dissolved in MTBE (4000 ml), washed with water (1000 ml ×2) and saturated saline (1000 ml), dried over anhydrous Na2SO4, was filtered, and then concentrated to obtain compound 2 as a pale yellow solid, which was used in the next stage of the reaction without further drying or purification.

1H-NMR (CDCl3; 400 MHz) δ 10,86 (c), δ 9,89 (c), δ to 7.59 (m), δ 7,51 (d), δ 6,94 (d), δ 10,61 (c).

The procedure for obtaining 2-(benzyloxy)-5-tert-butylbenzaldehyde (3)

To a stirred solution of compound 2 (614,5 g of 3.33 mol) in DMF (3.5 l) was added K2CO3(953 g of 6.90 mol) and benzylchloride (480 g of 3.80 mol). The mixture was heated to 90°C and kept under stirring for 3 hours. The suspension was cooled to room temperature, then was added MTBE (2 l) followed by addition of water (12 l). The mixture then was stirred for 10 minutes and the aqueous layer was separated and was extracted using MTBE (2 l ×3). The organic layers were combined and washed with water (2 l ×2) and saturated brine (1.5 l ×1) and then concentrated to obtain compound 3 as a pale yellow solid.

1H-NMR (DMSO-d6; 400 MHz) δ 10,42 (c), δ 7,71 (m), δ 7,51 (m), δ 7,43 (m), δ 7,35 (m), δ 7.24 to (m), δ 5,27 (c), δ 1.26 in (c).

The procedure for obtaining 2-(benzyloxy)-5-tert-butylbenzyl alcohol (4)

To a stirred suspension of compound 3 (974 g, 3.63 mol) in MeOH (4000 ml) was slowly added NaBH4(121 g, 3,20 mol) at a temperature of 0-20°C. the Solution was left under stirring at the temperature of 15°C for 3 hours and then was cooled to 0°C. was added dropwise 2n. a solution of HCl (aqueous solution) (1300 ml) at a temperature below 20°C. the Solution is then filtered and evaporated to dryness and the residue was dissolved in MTBE (5 l). The solution is then washed with water (2 l ×2) and saturated brine (1.5 l ×1). Evaporation of the solvent gave compound 4 as a pale yellow solid, which was used in the next stage of the reaction without additional purification.

1H-NMR (DMSO-d6; 400 MHz) δ 7,40 (m), δ 7,32 (m), δ 7,17 (m), δ 6,91 (m), δ 5,09 (c), δ 5,00 (t), δ 4,56 (d), δ 1.26 in (c).

The procedure for obtaining 2-(benzyloxy)-5-tert-butylbenzylamine (5)

To a stirred solution of compound 4 (963 g of 3.56 mol) in anhydrous DHM (2000 ml) was slowly added SOCl2(535 g, 4.5 mol) at 0°C. the Mixture was stirred at 20°C for 2 hours, then concentrated in vacuum to obtain compound 5 in the form of oil, which was used in the next stage of the reaction without further drying or purification.

The procedure for obtaining 2-(benzyloxy)-5-tert-butylbenzonitrile (6)

mix a solution of compound 5 (1045 g, 3.54 mol) in anhydrous DMF (1000 ml) was added KCN (733 g, 11.3 mol). The mixture was stirred at 35°C for 24 hours, then poured into water (10 l). Added ethyl acetate (4 l) and the mixture was stirred for 30 minutes. The organic layer was then separated and the aqueous layer was extracted with ethyl acetate (3000 ml ×2). The organic layers were combined and washed with water (4 l ×2) and saturated saline solution (3 l ×1), then concentrated in vacuum to obtain compound 6 as a yellow solid.

1H-NMR (DMSO-d6; 400 MHz) δ 7,51 (m), δ 7,37 (m), 7,02 (d), δ 5,17 (c), δ 3,88 (c)of 1.26 (c).

The procedure for obtaining 2-(2-(benzyloxy)-5-tert-butylphenyl)-2-methylpropionitrile (7)

To a stirred suspension of NaH (86 g of 2.15 mol, 60% in mineral oil) in DMF (1000 ml) was added dropwise a solution of compound 6 (100.0 g, 0,358 mol) in DMF (500 ml) at 20°C. After stirring for 30 minutes was added dropwise MeI (205 g, 1.44 mol) in DMF (500 ml) below 30°C for 2 hours. The suspension was stirred for 1.5 hours at a temperature of 25-30°C, then added slowly to ice (100 g) until the gas evolution stops. The pH was brought to about 7 by the slow addition of 2n. HCl solution. The mixture was diluted with water (4 l) and MTBE (2 l). The organic layer was separated and the aqueous layer was extracted using MTBE (500 ml ×2). The combined organization is organic layers were washed with water and saturated saline solution, dried over Na2SO4, filtered and then concentrated in vacuum to obtain compound 7 as a white solid.

1H-NMR (DMSO-d6; 400 MHz) δ 7,56 (m), δ 7,40 (m), δ 7,34 (m), δ 7,10 (d), δ to 5.21 (c), δ 1,73 (c), δ 1,27 (c).

The procedure for obtaining 2-(2-(benzyloxy)-5-tert-butylphenyl)-2-methylpropanal (8)

To a stirred solution of compound 7 (20 g, 0,065 mol) in toluene (300 ml) was added dropwise DIBAH (80 ml, 1M solution in toluene) at a temperature of from about -60 to -50°C. After stirring for 2 hours to the reaction mixture was added 6N. HCl (300 ml) and stirring was continued for 30 minutes. The organic layer was then separated, washed using a 2H. HCl solution followed by rinsing with a solution of NaHCO3then with a saturated saline solution, dried over Na2SO4and concentrated in vacuum to obtain compound 8 in the form of oil. The product was used for next reaction stage without further purification.

1H-NMR (CDCl3; 400 MHz) δ being 9.61 (c), δ was 7.36 (m), δ 7,25 (m), δ 6.87 in (m), δ is 5.06 (m), δ 1,43 (c), δ is 1.33(c).

The procedure for obtaining 2-(2-(benzyloxy)-5-tert-butylphenyl)-2-methylpropan-1-ol (9)

To a stirred solution of compound 8 (of 9.21 g, being 0.030 mol) in MeOH (150 ml) was slowly added NaBH4(2.3 g, 0.061 mol) at 0°C. Then mesh was stirred at 20°C for 3 hours, added 12 ml of 6N. HCl solution and the mixture was stirred for an additional 30 minutes. The solution is then concentrated to approximately one-fourth the original volume and extracted using EtOAc. The organic layer was separated and washed with water and saturated saline solution, dried using Na2SO4, filtered and then concentrated in vacuum to obtain compound 9 as a white solid.

1H-NMR (DMSO-d6; 400 MHz) δ 7,47 (m), δ 7,42 (m), δ 7,34 (m), δ 7,28 (m), δ 7,16 (m), δ 6,94 (m), δ 5,08 (c), δ 4,45 (t), δ 3,64 (d), δ 1.28 (in c), δ 1,25 (c).

The procedure for obtaining 2-(2-hydroxy-5-tert-butylphenyl)-2-methylpropan-1-ol (10)

Pd(OH)2(1 g) and compound 9 (9,26 g, being 0.030 mol) in MeOH (200 ml) was stirred in hydrogen atmosphere at a pressure of 20-30 psi (1,406-2,109 kg/cm2) for 16-18 hours. The mixture was then filtered through Celite® and the filtrate was concentrated to obtain compound 10 as a white solid.

1H-NMR (DMSO-d6; 400 MHz) δ 9,16 (c), δ 7,16 (d), δ 7,00 (m), δ 6,65 (m), δ 4,71 (t), δ 3,62 (d), δ 1,27 (c), δ 1,22 (c).

The procedure for obtaining 1-((methylcarbamic)oxy)-2-(1-((methylcarbamic)oxy)-2-methylpropan-2-yl)-4-tert-butylbenzene (11)

To a stirred solution of compound 10 (23,2 g, 0.10 mol), DMAP (1.44 g) and DIEA (72,8 g of 0.56 mol) in anhydrous DHM (720 ml) was added the dropwise methylchloroform (43,5 g, 0.46 mol) in DHM (160 ml) at 0°C. the mixture is Then stirred at a temperature of 20°C for 16 hours, washed with water, 1N. HCl solution and saturated saline solution, dried using MgSO4and concentrated in vacuum. The residue was purified using column chromatography on silica gel (mixture 1:20 EtOAc:petroleum ether) to give compound 11 as a white solid.

1H-NMR (DMSO-d6; 400 MHz) δ 7,32 (m), δ 7,10 (d), δ 4.26 deaths (c), δ 3,84 (c), δ 3,64 (c), δ 1,31 (c), δ 1.28 (in c).

The procedure for obtaining 1-((methylcarbamic)oxy)-2-(1-((methylcarbamic)oxy)-2-methylpropan-2-yl)-4-tert-butyl-5-nitrobenzene (12)

To a stirred solution of compound (11) (32 g, 0,095 mol) in DHM (550 ml) was added dropwise 98% solution of H2SO4(43 g, 0.43 mol) at 0°C. After stirring for 20 minutes at a temperature of 0°C. to the mixture was added dropwise 65% solution of HNO3(16.2 g, to 0.17 mol) at 0°C. the Mixture was then stirred at a temperature of 1-10°C for 4 hours and then added to a mixture of ice water (200 ml). The aqueous layer was separated and was extracted using DHM (200 ml ×3) and the combined organic layers were washed with water (aqueous solution), NaHCO3and saturated saline, then was dried using MgSO4and concentrated in vacuum. The residue was purified using column chromatography the raffia on silica gel (mixture 1:20 EtOAc:petroleum ether) to give the crude compound 12 in the form of oil.

The procedure for obtaining 2-tert-butyl-5-((methylcarbamic)oxy)-4-(1-((methylcarbamic)oxy)-2-methylpropan-2-yl)aniline (13)

Pd/C (2.6 g) and compound 12 (14 g, crude) was stirred in MeOH (420 ml) at room temperature in a hydrogen atmosphere at a pressure of 20-30 psi (1,406-2,109 kg/cm2) for 16-18 hours. The mixture is then filtered through kieselguhr® and the filtrate was concentrated in vacuum. The residue was purified using column chromatography on silica gel (mixture of 1:10 EtOAc:petroleum ether) to give compound 13 as a gray solid.

1H-NMR (CDCl3; 400 MHz) δ 7,26 (c), δ 7,19 (c), δ 4.26 deaths (c), δ 3,89 (c), δ 3,74 (c), δ 1,40 (c), δ 1,35 (c).

The procedure for obtaining N-(2-tert-butyl-5-((methylcarbamic)oxy)-4-(1-((methylcarbamic)oxy)-2-methylpropan-2-yl)phenyl)-4-oxo-1,4-dihydroquinoline-3-carboxamide (14)

To a stirred solution of compound 26 (5.0 g, was 0.026 mol) in anhydrous DMF (120 ml) was added EDCI (5.6 g, 0,029 mol), HOBT (3.8 g, 0,028 mol) and DIEA (6.6 g, 0,051 mol) at 0°C. After stirring for 1 hour the mixture was added dropwise a solution of compound 13 (3.0 g, 0,008 mol) in DHM (30 ml) at 0°C. the Mixture was stirred at 25°C for 72 hours and then concentrated in vacuum. The residue was dissolved in EtOAc (225 ml) and washed with water (120 ml ×1), 1H. HCl solution (10 ml) and saturated saline solution, dried using Na2SO4and concentrated in vacuum. The residue was purified using column chromatography on silica gel (mixture of 1:1 EtOAc:petroleum ether) to give compound 14 as a white solid.

1H-NMR (400 MHz, CDCl3) δ 12,34 (c, 1H), 11,58 (c, 1H), 9,07 (c, 1H), 8,42 (d, 1H), 7,66 (c, 1H), 7,51 (c, 1H), 7,47 (c, 1H), 7,39 (c, 1H), 6,72 (c, 1H), 4,34 (c, 2H), 3,82 (c, 3H), 3,74 (c, 3H), 1.41 to (c, 9H), 1,40 (c, 6H).

The procedure for obtaining N-(2-tert-butyl-5-hydroxy-4-(1-hydroxy-2-methylpropan-2-yl)phenyl)-4-oxo-1,4-dihydroquinoline-3-carboxamide (27)

To a stirred solution of KOH (1.2 g, 0.02 mol) in MeOH (80 ml) was added compound 14 (1.9 grams, 0,0036 mol) at 0°C. After stirring for 2-3 hours at a temperature of 5-15°C and the mixture was concentrated to dryness. The residue was then treated with water (10 ml), was filtered, washed using DHM and dried in vacuum for 24 hours to obtain compound 27 as a white solid.

1H-NMR (DMSO-d6; 400 MHz) δ 12,77 (c), δ 8,86 (c), δ 8,20 (d), δ 7,55 (d), δ 7,42 (t), δ 7,16 (square), δ 7,02 (c), δ 6,85 (m), δ 3,55 (c), δ 1.55V (c), δ 1,35 (c), δ 1,27 (c). MS found (M+H) 409,2.

Example 2:An alternative General synthesis of N-(2-tert-butyl-5-hydroxy-4-(1-hydroxy-2-methylpropan-2-yl)phenyl)-4-oxo-1,4-dihydroquinoline-3-carboxamide (27)

The procedure to obtain methyl 2-(5-tert-butyl-2-hydrox the-4-nitrophenyl)-2-methylpropanoate (38):

A mixture of 2-bromo-4-tert-butyl-5-NITROPHENOL (15,00 g, 54,72 mmol), bis(tri-tert-butylphosphine)palladium(0) (1,422 g 2,783 mmol), zinc fluoride (2,82 g, 27,27 mmol), methyltrimethoxysilane (MTDA) (19,35 g, 111,0 mmol) and dimethylformamide (150 ml) was heated at 70°C for 18 hours. The mixture was cooled to room temperature and diluted with water. After stirring for one hour, the aqueous phase was extracted using MTBE. The organic phase was dried in vacuum to obtain the crude product as a brown solid. Purification of the product was carried out by grinding the powder in n-heptane.

1H-NMR (400 MHz, DMSO-d6)δ 10,38 (c, 1H); 7,37 (c, 1H); 6,79 (c, 1H); 3,54 (c, 3H); 1,45 (c, 6H); 1.32 to(c, 9H).

The procedure for obtaining 4-tert-butyl-2-(1-hydroxy-2-methylpropan-2-yl)-5-NITROPHENOL (39):

1M solution of lithium aluminum hydride in THF (11,80 ml, RS 11.80 mmol) was added to a solution of methyl 2-(5-tert-butyl-2-hydroxy-4-nitrophenyl)-2-methylpropanoate (are 5.36 g, 18,15 mmol) in THF (50 ml). The mixture was stirred at ambient temperature for 3 hours and then was diluted with methanol. The mixture was acidified using 1N. HCl solution (pH 1-2) and the aqueous phase was extracted using MTBE. The organic phase was dried in vacuum to obtain 4-tert-butyl-2-(1-hydroxy-2-methylpropan-2-yl)-5-nitro is Nola, which was used without further purification in the next stage.

1H-NMR (400 MHz, DMSO-d6) δ 10,12 (c, 1H); 7,37 (c, 1H); 6,80 (c, 1H); 4,77 (c, 1H); 3,69-the 3.65 (m, 2H); 1.30 on (c, 9H); 1,29 (c, 6H).

The procedure for obtaining 4-tert-butyl-2-(2-methoxycarbonylamino-1,1-dimethylethyl)-5-nitrophenyl]methylcarbamate (12)

To a solution of 4-tert-butyl-2-(1-hydroxy-2-methylpropan-2-yl)-5-NITROPHENOL (1.92 g, 7,18 mmol), triethylamine (1,745 g, 17,24 mmol) and dimethylaminopyridine (87,74 mg, 0,718 mmol) in dichloromethane (30 ml) at 0°C was slowly loaded methylchloroform (2,376 g, 25,14 mmol), keeping the temperature below 5°C. After addition the mixture was allowed to warm to ambient temperature and was stirred until then, until the results of the HPLC did not show complete conversion of starting material (2-8 hours). The reaction mixture was diluted with water and acidified using 1N. HCl solution (pH 1-2). The aqueous phase was extracted using DHM and the combined organic substance was dried in vacuum. The crude semi-solid substance amber recrystallized from methanol and dichloromethane to obtain specified in the title compound as a yellow crystalline solid.

1H-NMR (400 MHz, DMSO-d6) to 7.67 δ (c, 1H); 7,52 (c, 1H); 4,30 (c, 2H); 3,86 (c, 3H); 3,64 (c, 3H); 1,35 (c, 9H); 1,35 (c, 6H).

The procedure for obtaining 5-amino-4-tert-butyl-2-(-methoxycarbonylamino-1,1-dimethylethyl)phenyl]methylcarbamate (13):

A mixture of [4-tert-butyl-2-(2-methoxycarbonylamino-1,1-dimethylethyl)-5-nitrophenyl]methylcarbamate (1.27 g, 3,313 mmol) and Pd/C (75 mg, 0.035 mmol) in methanol (50 ml) was purged with nitrogen. After purging the flask with hydrogen, the mixture was first made for 18 hours at a temperature and ambient pressure. The solution was filtered through Celite® and was dried in vacuum to obtain the product as a solid.

1H-NMR (400 MHz, DMSO-d6) δ 6,99 (c, 1H); 6,39 (c, 1H); 4.92 in(c, 2H); 4,13 (c, 2H); 3,82 (c, 3H); 3,65 (c, 3H); 1.32 to (c,9H); 1,23 (c, 6H).

The procedure for obtaining N-(2-tert-butyl-5-hydroxy-4-(1-hydroxy-2-methylpropan-2-yl)phenyl)-4-oxo-1,4-dihydroquinoline-3-carboxamide (27):

In a mixture of [5-amino-4-tert-butyl-2-(2-methoxycarbonylamino-1,1-dimethylethyl)phenyl]methylcarbamate (103 mg, 0.29 mmol), 4-oxo-1,4-dihydroquinoline-3-carboxylic acid (50 mg, 0.26 mmol) and pyridine (42 mg, of 0.53 mmol) in 2-Methf (3.0 ml) was loaded T3P in the form of a 50 wt%. solution 2-Methf (286 mg, 0.45 mmol). The mixture was heated to 50°C for 18 hours. After cooling to ambient temperature the mixture was diluted with water. The organic phase was separated and again washed with water. The sodium methoxide (39 mg, to 0.72 mmol) were loaded into the organic phase and the solution was stirred for 2 hours. The reaction mixture was extinguished with the help of 1H. HCl solution and after the partition is of the phases the organic phase is washed using 0.1 n. HCl solution. The organic phase is then dried in vacuum to obtain Compound 27 in the form of solids. Data1H-NMR spectrum was consistent with the above.

Example 3:General synthesis of 2-(5-tert-butyl-2-hydroxy-4-(4-oxo-1,4-dihydroquinoline-3-carboxamido)phenyl)-2-methylpropanoic acid (28):

The procedure for obtaining 2-(5-tert-butyl-2-hydroxyphenyl)-2 - methylpropionitrile (15)

Pd(OH)2/C (2.0 g) and compound 7 (20,0 g, 0.104 g mol) was stirred in MeOH (150 ml) at room temperature in a hydrogen atmosphere at a pressure of 10 psi(0,703 kg/cm2) for 16-18 hours. The mixture is then filtered through a layer of Celite® and the filtrate was concentrated to obtain compound 15, which was used in the next stage of the reaction without additional purification.

1H-NMR (DMSO-d6; 400 MHz) δ 9,83 (c), δ 7.24 to (c), δ 7.18 in (m), δ to 6.80 (m), δ 1,71 (c), δ 1,24 (c).

The procedure for obtaining 4-tert-butyl-2-(2-cyanoprop-2-yl)phenylmercuriborate (16)

To a stirred mixture of compound 15 (126,6 g, 0,564 mol), DMAP (6.0 g) and DIEA (188 g of 1.46 mol) in anhydrous DHM (1500 ml) was added dropwise methylchloroform (110 g at 1.17 mol) in anhydrous DHM (300 ml) at 0°C for 2 hours. After stirring for 12 hours at 0°C was added the mixture is IDA with water (1.5 l) and the mixture was stirred at 0°C for 30 minutes. The organic layer was separated and washed with 1N. HCl solution, water and saturated salt solution. The solution DHM was dried over MgSO4and concentrated in vacuum to obtain compound 16 as a yellow solid.

1H-NMR (DMSO-d6; 400 MHz) δ 7,47 (m), δ 7,39 (d), δ 7.24 to (d), δ 3,84 (c), δ 1,71 (c), δ 1,30 (c).

The procedure for obtaining 2-(1-amino-2-methyl-1-oxoprop-2-yl)-4-tert-butyl-5-nitrophenylacetate (17)

To a stirred mixture of compound 16 (10.0 g, 36,3 mmol) and KNO3(5,51 g of 54.5 mmol) in DHM (1000 ml) was added dropwise 98% solution of H2SO4(145,4 g of 1.45 mol) at 0°C. the Mixture was stirred at 30°C for 4 days. Layer H2SO4then was separated and poured into a mixture of ice water (50 g) and then was extracted using DHM (100 ml ×3). The combined organic layers were washed with water, aqueous solution of NaHCO3and saturated saline, then was dried over MgSO4and concentrated in vacuum. The residue was purified using column chromatography on silica gel (mixture of petroleum ether/EtOAc 20:1→10:1→5:1→3:1) obtaining of compound 17 as a yellow solid.

1H-NMR (CDCl3; 400 MHz) δ 8,05 (c), δ 7,74 (c), δ to 7.61 (c), δ 7,32 (c), δ 5,32 (c), δ 3,91 (c), to 3.92 δ (c)δ 1,62 (c), δ 1,59 (c), δ 1,42 (c), δ 1,38 (c).

The procedure for obtaining 2-(5-tert-butyl-2-hydroxy-4-nitrophenyl)-2-m is thisproposal acid (18)

To a mixture of compound 17 (7,3 g, 21.6 mmol) in methanol (180 ml) was added water (18 ml) and NaOH (8,64 g, 216 mmol). The solution was heated and maintained at the boiling point under reflux for 3 days. The solvent is evaporated in vacuum and the residue was dissolved in 140 ml of water. Then the solution was acidified to pH 2 by adding 2n. HCl solution. The aqueous phase was extracted with ethyl acetate (100 ml ×3) and the combined organic phases are washed with water and saturated saline, dried over anhydrous Na2SO4and then concentrated to obtain compound 18 as a yellow solid, which was used in the next stage of the reaction without additional purification.

The procedure for obtaining 5-tert-butyl-3,3-dimethyl-6-nitrobenzofurazan-2(3H)-she (19)

To a solution of compound 18 (7,10 g of 25.2 mmol) in 710 ml of anhydrous THF was added EDCI (14.5 g, to 75.6 mmol). The resulting suspension was left under stirring at a temperature of 30°C during the night. The precipitate was filtered and thoroughly washed using DHM. The filtrate was concentrated to dryness and the residue was dissolved in DHM (100 ml). The solution was washed with water (50 ml ×2) and saturated saline (50 ml ×1). Layer DHM then dried over anhydrous Na2SO4and concentrated to obtain the crude product, which PTS is attended with the help of column chromatography on silica gel (mixture of petroleum ether/EtOAc 200:1→100:1→50:1) obtaining compound 19 as a white solid.

1H-NMR (CDCl3; 400 MHz) δ of 7.36 (c), δ 7,10 (c), δ 1,53 (c), δ 1.41 to (c).

The procedure to obtain 6-amino-5-tert-butyl-3,3-dimethylbenzofuran-2(3H)-it (20)

Pd/C (1.50 g) and compound 19 (3.00 g, to 1.14 mmol) suspended in THF (1500 ml) at 25°C in an atmosphere of hydrogen at a pressure of 30 psi(2,109 kg/cm2) for 4 hours. The mixture is then filtered through a layer of Celite® and the filtrate was concentrated in vacuum to obtain compound 20 as a white solid.

1H-NMR (DMSO-d6; 400 MHz) δ 7,05 (c)of 6.49 δ (c)δ 5,01 (c), δ 1,35 (c), δ is 1.33 (c).

The procedure for obtaining N-(5-tert-butyl-3,3-dimethyl-2-oxo-2,3-dihydrobenzofuran-6-yl)-4-oxo-1,4-dihydroquinoline-3-carboxamide (21)

Suspension HATU (17.6 g, 46,3 mol) and compound 26 (at 8.36 g, a 44.2 mmol) in anhydrous acetonitrile (1 l) was stirred at room temperature for 1 hour. To the suspension was added Compound 20 (3,40 g, 14.6 mmol) and then dropwise added DIEA (11.5g, 89,0 mmol). The mixture was stirred at 45°C for 4 days. The precipitate was filtered and thoroughly washed using DHM. The filtrate was concentrated to dryness and the residue was dissolved in DHM (200 ml) and washed with 1N. HCl solution (200 ml ×2) followed by washing with 5% aqueous solution of NaHCO3(200 ml ×3) and then with saturated saline (200 ml ×1). Mixture C is the dried over Na 2SO4and concentrated in vacuum. The residue was purified using column chromatography on silica gel (mixture of CH2Cl2/MeOH 100:1→50:1) to give compound 21 as a pale yellow solid.

1H-NMR (400 MHz, DMSO-d6) δ 12,96 (d, J=6,4 Hz, 1H); 12,1 (c, 1H); 8,9 (d, 76,4 Hz, 1H); with 8.33 (d, 7 8 Hz, 1H); 7,84 to 7.75 (m, 2H); 7,55-of 7.48 (m, 3H); 1,47 (c, 6H); 1,45 (c, 9H).

The procedure for obtaining 2-(5-tert-butyl-2-hydroxy-4-(4-oxo-1,4-dihydroquinoline-3-carboxamido)phenyl)-2-methylpropanoic acid (28)

To a stirred solution of compound 21 (0.9 g, 2.45 mmol) in MeOH (50 ml) was added NaOH (1.5 g, 37.5 mmol) at 0°C. After stirring for 16 hours at 40°C the solvent is evaporated in vacuo, then the residue was dissolved in H2O (50 ml). The precipitate was filtered and the filtrate was washed using DHM (100 ml ×1) and ethyl acetate (100 ml ×1). The aqueous layer was acidified using 2n. HCl solution to pH 1-2. The precipitate was filtered and washed using H2O (80 ml) and heptane (50 ml). This precipitate was dried in vacuum to obtain compound 28 as a white solid.

1H-NMR (DMSO-d6; 400 MHz) δ is 12.85 (c), δ 11,84 (c), δ 11,77 (c), δ 9,39 (c), δ 8,86 (c), δ 8,33 (c), δ 7,79 (m), δ 7,52 (m), δ 7.18 in (c), δ 7,09 (c), δ 1,44 (c), δ 1,40 (c). MS found (M+H) 423,08.

Example 4:The second alternative synthesis of N-(2-tert-butyl-5-hydroxy-4-(1-hydroxy-2-methyl shall ropan-2-yl)phenyl)-4-oxo-1,4-dihydroquinoline-3-carboxamide (27)

3-Necked round bottom flask of 50 ml volume was supplied with a magnetic stirrer, bubbling device for nitrogen and thermocouple. Into the flask was loaded connection 21 (514 mg, of 1.27 mmol) and 2-Methf (4 ml). The reaction mixture was stirred at room temperature. Added alumoweld lithium (204 mg, 6.6 mmol) in the form of a solid substance to achieve 100% conversion, which was monitored by means of HPLC. Potassium sodium 2,3-dihydroxybutanedioate salt tetrahydrate (50 ml 400 g/l) and MTBE (50 ml) was added to the reaction mixture. The resulting solution was stirred for 15 minutes and then allowed to stand for 15 minutes. The organic layer was separated and the pH of the aqueous layer was brought to approximately 6-7 by adding tartaric acid. The aqueous layer was extracted using MTBE. The organic layer was concentrated and dried under high vacuum to obtain specified in the connection header in the form of not-quite-white powder. Data1H-NMR spectrum was consistent with the above.

Example 5:An alternative General synthesis of 2-(5-tert-butyl-2-hydroxy-4-(4-oxo-1,4-dihydroquinoline-3-carboxamido)phenyl)-2-methylpropanoic acid (28):

The procedure for obtaining 2-bromide carbonic acid, 4-tert-butil delovogo ester methyl ester (35)

3-Necked round bottom flask provided with magnetic stirrer, bubbling device for nitrogen and thermocouple. Was added 2-bromo-4-tert-butylphenol (50 g, 211,7 mmol) followed by the addition DHM (1.75 l), DMAP (1.29 g, of 10.58 mmol) and Et3N (44,3 ml, 317,6 mmol). The reaction mixture was cooled to 0°C. To the reaction mixture was added dropwise methylchloroform (19,62 ml, 254 mmol). The mixture was left to warm to room temperature with stirring overnight. After completion of the reaction the mixture was filtered through a funnel from the sintered glass. The filtrate was transferred into a separating funnel with a volume of 1 L. For quenching the reaction mixture to the filtrate was added 1N. HCl (300 ml) and the organic layer was separated. The organic layer is then washed with a mixture of 291 ml of a saturated solution of NaHCO3and 100 ml of water. The layers were separated and determined that the water layer has a pH of about 8. The organic layer was concentrated and dried under high vacuum for about 16 hours to obtain specified in the connection header in the form of a clear yellow oil, which was used in the next stage without additional purification.

1H-NMR (400 MHz, DMSO-d6) 7,66 (d, J=2.0 Hz, 1H), 7,46 (DD, J=8,4, 2.0 Hz, 1H), 7,32 (d, J=8,4 Hz, 1H), 3,86 (c, 3H), 1.28 (in c, 9H).

The procedure for obtaining (2-bromo-4-tert-butyl-5-nitrophenyl)methylcarbamate (36)

3-Necked round bottom flask with a volume of 2 l was supplied with a magnetic stirrer, bubbling device for nitrogen and thermocouple. Into the flask was loaded connection 35 (176 g, 612,9 mmol) and concentrated sulfuric acid (264 ml). The reaction mixture was cooled to a temperature of -5°C-0°C. was added dropwise nitric acid (28,6 ml, 612,9 mmol) and the reaction mixture was stirred at 0°C for 2 hours. After stirring was added water (264 ml) followed by addition of MTBE (264 ml). The solution was stirred for 15 minutes, then allowed to stand for 15 minutes. The organic layer was separated, concentrated and dried under high vacuum to obtain specified in the title compound as a dark brown oil, which was used in the next stage without additional purification.

1H-NMR (400 MHz, DMSO-d6) of 7.96 (c, 1H), 7,92 (c, 1H), 3,89 (c, 3H), 1,34 (c, 9H).

The procedure to obtain 2-bromo-4-tert-butyl-5-NITROPHENOL (37)

(2-Bromo-4-tert-butyl-5-nitrophenyl)methylcarbamate with 72.9 g, to 219.5 mmol) were loaded into the reactor and added DHM (291,6 ml). The yellow reaction solution was cooled, using a bath of ice. Portions was added sodium methoxide (67,04 g, 69,11 ml of a 5.4 M solution, 373,2 mmol) at a temperature of 2,2-6,9°C. After complete addition, the reaction mixture was slowly heated up to the temperature environment is the first environment. After heating, the reaction mixture was cooled to 0°C and extinguished using 1M HCl solution (373,2 ml, 373,2 mmol). A two-phase mixture was stirred for 20 minutes and transferred into a separating funnel. The organic layer was separated and washed with water (300 ml) followed by washing with saturated saline (300 ml). The organic layer was concentrated and the crude product was dried under high vacuum. The product was further purified using supercritical fluid chromatography (SFC) on the device Berger MultiGram III (Mettler Toledo AutoChem, Newark DE). The conditions for this method were as follows: 20% methanol at a flow rate of 250 ml/min column on the PPU (30*150) from Princeton Chromatography, 100 bar, 35°C, 220 nm. Introduced in 3.5 ml 55-70 mg/ml Data were collected using the SFC program ProNTo. The purified product obtained using SFC purification was methanol MES. Removal of the methanol was carried out by azeotropic distillation. In a round bottom flask with a volume of 1 l was downloaded dark yellow solid, 2-bromo, 4-tert-butyl, 5-nitrophenetole MES, (111,3 g to 59.9 mmol) followed by loading of heptane (500 ml). The suspension was heated to 64°C to obtain a clear solution. The solvent is kept under reduced pressure (649 mbar) for 30 minutes and then was evaporated to dryness. This procedure was repeated three times until, until he identified the absence of MeOH using 1H-NMR. The product was dried under high vacuum for 16 hours to obtain the product as a dark yellow semi-solid substance.

1H-NMR (400 MHz, DMSO-d6) δ 11,2 (users, OH), 7,69 (c, 1H); 7.03 is (c, 1H); 1.30 on (c, 9H).

The procedure for obtaining 5-tert-butyl-3,3-dimethyl-6-nitrobenzofurazan-2(3H)-she (19)

Divorcing (6,093 g, 58,92 mmol) was added in a round bottom flask, which was purged with nitrogen. Under the flow of nitrogen was then added Pd(tBu3P)2(2 g 3,835 mmol). 2-Bromo-4-tert-butyl-5-NITROPHENOL (16,15 g, 58,92 mmol)dissolved in DMF (80,75 ml), then added to the flask. The reaction mixture was a orange suspension. To the mixture was added 1-methoxy-2-methyl-prop-1-enocsi)trimethylsilane (21,61 g, 25,13 ml, 117,8 mmol) and the resulting mixture was heated up to 80°C and was stirred for 16 hours. After completion, the reaction mixture was cooled to ambient temperature and filtered through Celite®. The filter residue was washed using MTBE (536,0 ml) and to the filtrate was added water (893,3 ml). The mixture was stirred for 15 minutes and advocated for an additional 15 minutes. The layers were separated and the organic phase was added 0,5M aqueous HCl (500 ml, 250,0 mmol). The layers were separated and the organic layer was washed with water (500 ml). The layers were separated and the organic layer was washed using NaCl (500 ml; 8% wt.). The organic layer from elali and the solvent was removed in vacuum. Received the crude product as a brown crystalline solid and was then purified through a plug of silica using as eluent a mixture of hexane:MTBE 20:1-10:1. The fractions containing the product were combined and the solvent was removed in vacuum to obtain the pure product as a white crystalline solid.

1H-NMR (400 MHz, DMSO-d6) δ 7,80 (c, 1H); 7,62 (c, 1H); 1,49 (c, 6H); 1,34 (c, 9H).

The procedure to obtain 6-amino-5-tert-butyl-3,3-dimethylbenzofuran-2(3H)-it (20)

Palladium on carbon (wet; 5% of the mass) were placed in a round bottom flask under a stream of nitrogen. The vessel was then added 5-tert-butyl-3,3-dimethyl-6-nitro-benzofuran-2-he (4.7 g, 17,85 mmol). The vessel was then carefully loaded methanol (120 ml) under nitrogen atmosphere. The vessel was then purged with the help of N2that was pumped out the air, then filled with gaseous hydrogen. From the vessel was pumped out the air and re-filled with gaseous hydrogen, and then typed a continuous stream of gaseous hydrogen. After the introduction of gas, the reaction mixture was filtered through Celite® and the residue was washed using MeOH (300 ml). The solvent was removed in vacuum and the product was dried under high vacuum to obtain a white crystalline solid.

1H-NMR (400 MHz, DMSO-d6) δ 7,05 (c, 1H); 6.48 in (c, 1H); 5,02 (c, 2H, NH2); 1,34 (c, 6H; 1,30 (c, 9H).

The procedure for obtaining N-(5-tert-butyl-3,3-dimethyl-2-oxo-2,3-dihydrobenzofuran-6-yl)-4-oxo-1,4-dihydroquinoline-3-carboxamide (21)

Into a reaction vessel were loaded connection 26 (2,926 g, 15,43 mmol), compound 20 (4,32 g holds 18.52 mmol), 2-Methf (35,99 ml) and then 50% T3P 2-Methf (made 13.36 g, 21.00 mmol). Added pyridine (2,441 g 2,496 ml, 30,86 mmol) and the suspension was heated at a temperature of 47.5°C±5°C for 18 hours. After heating, the reaction mixture was cooled to ambient temperature and was added 2-Methf (36) and water (30 ml). The layers were separated and the organic layer was washed for 10% of the mass. citric acid solution (30 ml), water (30 ml) and twice with the help of NaHCO3(20 ml). The organic layer was washed with saturated saline (50 ml), separated and the solvent was removed in vacuum. The crude product was dissolved in MTBE (100 ml) and was added hexane (200 ml) as antibacterial. The solid was besieged, and the resulting suspension was stirred for two hours. The solid was collected by vacuum filtration and the precipitate was washed with hexane. The obtained product was dried in a vacuum oven at 55°C with a selection of nitrogen with obtaining specified in the title compound as a beige solid.

1H-NMR (400 MHz, DMSO-d6) δ 12,96 (d, J=6,4 Hz, 1H); 12,1 (c, 1H); 8,9 (d, J=6,4 Hz, 1H); with 8.33 (d, J=8gts, 1H); 7,84 to 7.75 (m, 2H); 7,55-of 7.48 (m, 3); 1,47 (c, 6H); 1,45 (c, 9H).

The procedure for obtaining 2-(5-tert-butyl-2-hydroxy-4-(4-oxo-1,4-dihydroquinoline-3-carboxamido)phenyl)-2-methylpropanoic acid (28)

Compound 26 (81,30 mg, 0,4288 mmol) and compound 20 (110 mg, 0,4715 mmol) were loaded into a round bottom flask. Then added 2-Methf (1 ml) followed by addition of 50% T3P 2-Methf (371,4 mg, 0,5836 mmol) and pyridine (67,84 mg, 69,37 μl, 0,8576 mmol) in 2-Methf. The suspension was heated at 47,5°C±5°C during the night. After heating, the reaction mixture was cooled to ambient temperature. Was added 2-Methf (1,014 registered ml) and water (811,2 µl). The layers were separated and the organic layer was washed with water (811,2 ml) and twice with the help of NaHCO3(2 ml). The organic layer was transferred into a round bottom flask. Added LiOH (38,6 mg, 0.9 mmol)dissolved in water (2 ml) and the reaction mixture was heated to 45°C. After heating the layers were separated and the organic layer is discarded. The aqueous layer was cooled in a bath of ice and to the solution was added hydrochloric acid (of 10.72 ml of 1.0 M solution, 10.72 mmol) up until the pH reached a value of about 3-4. The aqueous layer was extracted twice using 2-Methf (5 ml) and the organic layers were combined and washed with saturated saline (5 ml). The organic layer was separated and the solvent was removed in vacuum. P the obtained solid substance was dried in a vacuum oven with a selection of nitrogen at 50°C with obtaining specified in the connection header.

1H-NMR (400 MHz, DMSO-d6) δ 12,89 (d, 7 6,8 Hz, 1H); 11,84 (c, 1H); 11,74 (c, 1H); 9,36 (c, 1H); 8,87-8,61 (d, 76,4 Hz, 1H); 8.34 per-8,32 (d, 79,1 Hz, 1H); 7,83-7,745 (m, 2H); 7,17-to 7.09 (m, 1H); 7,17 (c, 1H); 7,09 (c, 1H); 1,43 (c, 6H); 1,40 (c, 9H).

Example 6:The procedure of the biosynthesis of N-(2-tert-butyl-5-hydroxy-4-(1-hydroxy-2-methylpropan-2-yl)phenyl)-4-oxo-1,4-dihydroquinoline-3-carboxamide (27) and 2-(5-tert-butyl-2-hydroxy-4-(4-oxo-1,4-dihydroquinoline-3-carboxamido)phenyl)-2-methylpropanoic acid (28)

Streptomyces rimosus(DSM 40260) was purchased from DSMZ as a frozen culture. This culture was used for inoculation beveled Ugarov who supported and kept at 4°C. Yeast extract-malt extract-peptone (YMP) medium containing yeast extract (4 g/l), malt extract (10 g/l) and soy flour (5 g/l), were obtained and were sterilized at 130°C for 60 minutes. Five flasks containing 1 l YMP environment, directly inoculableS. rimosusfrom the beveled Ugarov. The culture was allowed to grow for 2 to 3 days at 30°C with gentle agitation at approximately 100 rpm under these conditions, observed two types of growth: either a cloudy solution or spheroidal particles that have accumulated on the bottom of the flask. It has been shown that this latter type of growth leads to higher conversions in connection 27. Cells are then centrifuged, sobi is Ali and resuspendable in two flasks, containing 1 l of 0,1M potassium phosphate buffer, pH 7.0. To the flask was added 5.0 g of compound 34 in 50 ml of N,N-dimethylformamide (DMF). The reaction was carried out for 24 hours at 30°C with gentle agitation at about 100 rpm, at this point HPLC analysis showed conversion to 7.59% connection 27 and 1,17% connection 28.

The contents of both flasks were pooled, centrifuged at 3500 rpm for 10 minutes and resuspendable in 500 ml of methanol. This suspension was intensively stirred for 30 minutes and then centrifuged again at 6000 rpm for 10 minutes. The organic layer was collected and the process repeated twice. The methanol extracts were concentrated in vacuum to obtain 2.50 g, of 1.57 g of 1.11 g of solids, respectively. It was shown that the solids from these extracts contain 74,78-91,96% connection 34, 7,66-19,73% connection 27 and 0.39-5,49% connection 28. For the selection of the connection portion 34 of the products of the oxidation process, the solids of the first two extraction were combined, suspended in 250 ml of methanol, was intensively stirred for 1 hour and filtered under vacuum. Although compounds 27 and 28 were enriched in the filtrate (22,09 and 6.14%, respectively), the solids still contain the compound 27 (8,96%) and compound 28 (0,50%).

The methanol filtrate, containing approximately 2.2 g of dissolved solids, adsorbing 4.5 is silicon dioxide, and purified flash chromatography using a gradient of 100% dichloromethane→88:12 dichloromethane/methanol. The fractions containing the compound 27 was concentrated in vacuo and further dried by freeze drying to obtain 130 mg of the compound 27 (of 98.5% purity according to HPLC). The fraction containing the crude compound 28 was also concentrated in vacuum to obtain less than 10 mg of solid substances.

Cellular precipitate after centrifugation resuspendable in 500 ml of methanol and homogenized in a BeadBeater for detachment of cells and allocate any remaining product. The organic layer was obtained by centrifugation of the homogenized suspension at 6000 rpm for 10 minutes. It was added to the solid substance, derived from the third extraction and filtered solids-rich slurry of the first two extraction, and suspended at the boiling point under reflux overnight. The suspension is then cooled and subjected to vacuum filtration to obtain 1,99 g solids. The solid was again dissolved in 300 ml of methanol, then adsorbing approximately 5 g of silicon dioxide, and purified flash chromatography using a gradient of 100% dichloromethane→94:6 dichloromethane/methanol to obtain 820 mg of a solid product containing the connection 34 and the connection 27, as well as other impurities. The resulting material was again subjected to column purification using more than the high gradient of solvent (100% DHM→a mixture of 6% MeOH/94% DHM) additional 89 mg of compound 27. Range1H-NMR was consistent with the above.

Example 7:General procedure for testing the solubility at a pH of 7.4

Performance analysis in shake flask was used to determine the solubility of compounds in a pH of 7.4 buffer. To calculate the concentration of compounds in solution experiments were performed using two conditions for each connection: 300 μm in 100% DMSO and 200 μm at a pH of 7.4 phosphate buffer in the presence of 2% DMSO. Each sample was left to shake overnight, then was injected into the column HPLC-UV for the determination of the peak area using the following conditions: column Phenomenex 00A-4251-B0 - 30×2.00 mm Luna 3 μm C18(2) 100A; flow rate 0.8 ml/min; volume of injected sample 20 μl; mobile phase: standard water for HPLC with 0.1% formic acid and standard acetonitrile for HPLC with 0.1% formic acid; the peak area was determined at 254 nm. Solubility in µm was calculated using the following equation: conc.=(the peak area pH 7,4)/(peak area of 300 μm DMSO standard conditions)×300 μm, the concentration of the standard conditions. Interest peaks was determined in buffer conditions on the basis of retention time (RT) of the largest peak area 300 μm DMSO standard conditions.

ANALYSES of ACTIVITY

Example 8:General procedure for analysis of activity

Assays for detecting and measuring ΔF508-CFTR-potentiating properties of compounds

Optical methods for the determination of membrane potential for the analysis of ΔF508-CFTR-modulating properties of compounds

In the analysis using sensitive fluorescent potential dyes to measure changes in membrane potential using the tablet reader for reading fluorescence (e.g., FLIPR III, Molecular Devices, Inc.) to obtain data showing the increased level of functional ΔF508-CFTR in cells NIH 3T3. The driving force for the answer is the creation of a gradient of chloride ions due to activation of the channel by a single addition of liquid, after cells were pretreated with compounds with subsequent load potential-sensitive dye.

Identification potentiating compounds

To identify ΔF508-CFTR-potentiate tools was developed HTS analysis in the format of double add. In this HTS analysis using sensitive fluorescent potential dyes to measure changes in membrane potential on FLIPR III as the gain of the gate mechanism (conductivity) ΔF508 CFTR in ΔF508 CFTR NIH 3T3 cells with a correction for temperature. The driving force for the answer is the gradient of Cl-ions due to activation of the channel by Forskolin when onocr the nom adding fluid using a tablet reader for reading fluorescence, such as FLIPR III, after cells were pretreated potentiate compounds (or DMSO carrier control) followed by loading dye redistribution.

Solutions

The solution in the bath #1: (in mm) NaCl 160, KCl 4.5 Is, CaCl22, MgCl21, HEPES 10, pH to 7.4 using NaOH.

The solution in the bath without chloride: chloride salts in solution in the bath #1 substituted gluconate salts.

Cell cultureMurine NIH3T3 fibroblasts stably expressing ΔF508-CFTR was used for optical measurements of membrane potential. Cells were maintained at 37°C in 5% CO2and 90% humidity in the modified Dulbecco environment, Needle, supplemented with 2 mm glutamine, 10% fetal bovine serum, ×1 NEAA, β-ME, ×1 penicillin/streptomycin, and 25 mm HEPES in 175 cm2the culture flasks. For all optical analysis cells were sown at a density of ~20000/well in 384-well Matrigel-coated tablets and were cultured for 2 hours at 37°C, then were cultured at 27°C for 24 hours for analysis potentiate funds. For corrective analyses cells were cultured at 27°C or 37°C with and without a connection for 16-24 hours.

Electrophysiological analyses to determine ΔF508-CFTR-modulating properties of connections

1. Analysis using the Ussing chamber

Experiments using the m Ussing chamber was performed on polarized epithelial cells of the respiratory tract, expressing ΔF508-CFTR to further characterize modulators ΔF508-CFTR identified in the optical assays. Non-CF and CF epithelium of the respiratory tract was isolated from bronchial tissue were cultured as described previously (Galietta, L.J.V., Lantero, S., Gazzolo, A., Sacco, A., Romano, L., Rossi, G.A., & Zegarra-Moran, O. (1998) In Vitro Cell. Dev. Biol. 34, 478-481), and were sown on Costar® Snapwell™ filters that were pre-coated with NIH3T3-conditioned medium. Four days later, the apical medium was removed and cells were grown at the interface of the air-liquid for >14 days before using them. This led to the formation of a monolayer of fully differentiated cylindrical cells, which were ciliated, a feature that is characteristic of the epithelium of the respiratory tract. Non-CF HBE were isolated from non-Smoking subjects who did not suffer from any known lung disease. CF-HBE were identified in patients homozygous for the ΔF508-CFTR.

Enable HBE grown on Costar® Snapwell™ cell culture was placed in an Ussing chamber (Physiologic Instruments, Inc., San Diego, CA) and measured the transepithelial resistance and short circuit current in the presence of basolateral → apical gradient of Cl-(IScusing a system of fixed potential (Department of Bioengineering, University of Iowa, IA). Briefly, HBE investigated in terms of registration of fixed potential (Vhold=0 mV) at 37°C. Basolateral solution contained (in mm) 145 NaCl, 0,83 K2HPO4, 3,3 KH2PO4that 1.2 MgCl2, 1,2 CaCl2, 10 Glucose, 10 HEPES (pH brought up to 7.35 using NaOH), and the apical solution contained (in mm) 145 Na, 1,2 MgCl2, 1,2 CaCl2, 10 glucose, 10 HEPES (pH brought up to 7.35 using NaOH).

Identification potentiating compounds

A typical Protocol included the use of a gradient of concentration of Cl-on basolateral→apical membrane. To establish this gradient on basolateral membrane used normal solutions ringer, whereas apical NaCl was replaced with equimolar amounts of sodium gluconate (titrated to pH to 7.4 using NaOH) to obtain the high concentration gradient of Cl-through the epithelium. Forskolin (10 μm) and all test compounds were added to the apical side of the inclusions in cell culture. The efficiency of the alleged ΔF508-CFTR-potentiate means were compared with known efficiency potentiate means of genistein.

2. The patch-clamp registration

General Cl-the current in ΔF508-NIH3T3 cells was monitored using the perforated patch-clamp method, as described previously (Rae, J., Cooper, K., Gates, P., & Watsky, M. (1991) J. Neurosci. Methods 37, 15-26). Determination of potential fixation was carried out at 22°C using Axopatch 200B the patch-clamp amplifier (Axon Instrument Inc., Foster City, CA). The solution in the pipette 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 brought up to 7.35 using HCl). Extracellular medium contained (in mm) 150 NMDG-Cl, 2 MgCl2, 2 CaCl2, 10 HEPES (pH brought up to 7.35 using HCl). Generating pulses, data collection and analysis was performed using a PC equipped with a Digidata 1320 AJD interface in conjunction with Clampex 8 (Axon Instruments Inc.). To activate ΔF508-CFTR 10 μm of Forskolin and 20 μm genistein was added to the bath and the ratio of the current-voltage controlled every 30 seconds.

Identification potentiating compounds

The ability of ΔF508-CFTR-potentiate funds to increase the macroscopic ΔF508-CFTR Cl-current (IΔF508) in NIH3T3 cells stably expressing ΔF508-CFTR was also investigated using the perforated patch-clamp method. Potentiate the funds identified in the optical analyses, caused a dose-dependent increase of IΔF5O8with the same activity and efficiency, as observed in the optical assays. In all investigated cell potential reversion before and during application potentiate funds was around -30 mV, which represents a calculated value of ECl(-28 mV).

Cell culture

Murine NIH3T3 fibroblasts stably expressing ΔF508-CFTR used is to register in the configuration of the “whole cell”. Cells were maintained at 37°C in 5% CO2and 90% humidity in the modified Dulbecco environment, Needle, supplemented with 2 mm glutamine, 10% fetal bovine serum, ×1 NEAA, β-ME, ×1 penicillin/streptomycin, and 25 mm HEPES in 175 cm2flasks for cultivation. To register in the configuration of the “whole cell” 2500-5000 cells were sown on poly-L-lysine-coated cover glasses and were cultured for 24-48 hours at 27°C before using them for testing activity potentiate funds and incubated with or without corrective compounds at 37°C for measuring the activity of correction compounds.

3. Analyses of single channels

The portal activity of wt-CFTR and temperature-corrected ΔF508-CFTR expressed in NIH3T3 cells was observed using the patch-registrations in isolated areas of the membranes in the configuration of the “inside out”, as described previously (Dalemans, W., Barbry, P., Champigny, G., Jallat, S., Dott, K., Dreyer, D., Crystal, R.G., Pavirani, A., Lecocq, J-P., Lazdunski, M. (1991) Nature 354, 526 - 528), using Axopatch 200B the patch-clamp amplifier (Axon Instruments Inc.). The solution in the pipette contained (in mm): 150 NMDG, 150 aspartic acid, 5 CaCl2, 2 MgCl2and 10 HEPES (pH brought up to 7.35 using Tris base). The solution in the bath contained (in mm): 150 NMDG-Cl, 2 MgCl2, 5 EGTA, 10 TES and 14 Tris base (pH brought up to 7.35 using HCl). After the excision of both wt - and ΔF508-CFTR, an act which was verovali by adding 1 mm Mg-ATP, 75 nm catalytic subunit of cAMP-dependent protein kinase (PKA; Promega Corp. Madison, WI) and 10 mm NaF to inhibit proteinopathies, which prevented the passage of current. The potential in the pipette maintained at 80 mV. The activity of the channels analyzed in small, isolated areas of membranes containing ≤2 active channel. The maximum number of simultaneous discoveries were determined by the number of active channels in the course of the experiment. To determine the current amplitude of the single channel data recorded from 120 sec ΔF508-CFTR activity was filtered off-line at 100 Hz and then used to construct the amplitude histogram of all pixels that are customized with many features Gauss using the Bio-Patch Analysis (Bio-Logic Comp. France). General microscopic current and the probability of disclosure (P0) was determined from 120 sec for channel activity. P0was determined using the Bio-Patch or of the dependence of P0=I/i(N), where I = mean current, i = the current amplitude of the single channel, and N = number of active channels in the patch.

Cell culture

Murine NIH3T3 fibroblasts stably expressing ΔF508-CFTR, used for patch-clamp registrations in isolated areas of the membranes. Cells were maintained at 37°C in 5% CO2and 90% humidity in the modified Dulbecco environment the Needle, the stage is Lannoy 2 mm glutamine, 10% fetal bovine serum, ×1 NEAA, β-ME, ×1 penicillin/streptomycin, and 25 mm HEPES in 175 cm2flasks for cultivation. For registration of single channels 2500-5000 cells were sown on poly-L-lysine-coated cover glasses and were cultured for 24-48 hours at 27°C before use.

Compounds 27 and 28 were tested for solubility using the procedure outlined in example 3, and the activity using the procedures in the tests of example 4. The results are presented in table 1. The potency of the compound is shown as “+++”if the measured value EU50(or IR50) less than 2.0 μm, “++”, if the measured activity is from 2 μm to 5.0 μm, “+”, if the measured activity of more than 5.0 µm, and “-”if no data is available. The efficiency is shown as “+++”, if the calculated efficiency more than 100%, “++”, if the calculated efficiency ranges from 100% to 25%, “+”, if the calculated efficiency is less than 25%, and “-”if no data.


The connection 28
Table 1
Examples of activities and efficacies of the compounds of formula I
Structure
Connection 27
HTSF solubility at pH 7.487 mcm>200 μm
Optical analysis of the EU50+++++
ΔF508-HBE
EU50
++++++
ΔF508/G551D-HBE
EU50
+++++
HLM/RLM (% residual. @ 30 min)+++++
SPS 1.2 IR50+-

OTHER VARIANTS of the INCARNATION

All publications and patents referenced in the present disclosure, is incorporated into the present application by reference to the extent as if each such publication or patent application was specifically and individually indicated as incorporated by reference. If the meaning of the terms in any of the patents or publications, incorporated by reference, contrary to the meaning of the terms used in this disclosure, it is assumed that the values of terms in the present disclosure are identifying the mi. In addition, the above discussion discloses and describes only illustrative variants of the embodiment of the present invention. Specialists in this field can easily be calculated from such discussion and from the accompanying drawings and claims that various changes, modifications and variations without departure from the essence and scope of the present invention, as set forth in the following claims.

1. The compound of formula I:

or its pharmaceutically acceptable salt, in which R represents a COOH or CH2OH.

2. The compound according to claim 1, in which R represents a CH2OH.

3. The compound according to claim 1, in which R is a COOH.

4. Pharmaceutical composition for treating or attenuating the severity of the disease, mediated by CFTR, including:
the compound of formula I according to claim 1; and
pharmaceutically acceptable carrier or adjuvant.

5. Pharmaceutical composition for treating or attenuating the severity of the disease, mediated by CFTR, including a connection structure:

and pharmaceutically acceptable carrier or adjuvant.

6. Pharmaceutical composition for treating or attenuating the severity of the disease, mediated by CFTR, including a connection structure:
and pharmaceutically acceptable carrier or adjuvant.

7. Method of modulating CFTR activity in a biological sample, comprising the stage of contact specified biological sample with the compound of the formula I according to claim 1.

8. Method of modulating CFTR activity in a biological sample, comprising the stage of contact specified biological sample with the compound of the structure:

or its pharmaceutically acceptable salt.

9. Method of modulating CFTR activity in a biological sample, comprising the stage of contact specified biological sample with the compound of the structure:

or its pharmaceutically acceptable salt.

10. A method of treating or attenuating the severity of the disease in a patient, comprising the introduction of a specified patient an effective amount of the compounds of formula I according to claim 1, where the disease is selected from cystic fibrosis, COPD caused by Smoking, pancreatitis, insufficiency of pancreatic cancer, hereditary emphysema, COPD and dry eye syndrome.

11. The method according to claim 10, in which the compound of formula I has the structure:

12. The method according to claim 10, in which the compound of formula I has the structure:

13. The method according to any of p-12, which is supplied Zabol the existence represents cystic fibrosis.

14. Kit for use in measuring the activity of CFTR or a fragment in a biological sample in vitro or in vivo, including:
i. a composition including a compound of formula I according to claim 1; and
ii. instructions:
a. contacting the composition with the biological sample; and
b. measurement of the activity specified CFTR or fragment.

15. Set on 14, further comprising instructions for:
i. contacting an additional compound with the biological sample;
ii. measurement of the activity specified CFTR or fragment in the presence of the specified additional connections; and
iii. comparison of the activity of CFTR or fragment in the presence of the additional compound with the activity of CFTR or fragment in the presence of a composition of formula 1.

16. Set on 15, where phase comparison activity specified CFTR or fragment provides a density specified CFTR or fragment.

17. Set according to any one of p-16, in which the compound of formula I has the structure:

18. Set PP-16, in which the compound of formula I has the structure:



 

Same patents:

FIELD: chemistry.

SUBSTANCE: claimed invention relates to field of organic chemistry, namely to novel compound of formula (I), where Y and Z, each independently, are selected from group, consisting of: a) phenyl, if necessary substituted with 1 or 2 R6; b) pyridine, imidazole, thiazole, furan, triazole, quinoline or imidazopyridine, if necessary substituted with 1 R6; and c) substituent, independently selected from group, consisting of hydrogen, C1-C6alkyl or pyperidine; R1, R2 and R3, each independently selected from group, consisting of hydrogen and halogen; A and B is each independently selected from hydrogen, OH and C1-C6alkyl; RA and RB are independently selected from group, consisting of hydrogen, C1-C6alkyl and C3-C8cycloalkyl; or RA and RB together with atom, to which they are attached, form 4-6-membered heterocycle, if necessary having additionally one heteroatom or functional heterogrpoup, selected from group, consisting of -O-, -NH, -N(C1-C6-alkyl)- and -NCO(C1-C6-alkyl)-, and 6-membered heterocycle can be additionally substituted with one or two C1-C6-alkyl groups; R4 and R5, each stands for hydrogen; and each R6 is selected from Br, Cl, F, I, C1-C6-alkyl, pyrrolidine, if necessary substituted with one C1-C6-alkyl, C1-C6alkoxy, halogen-C1-C6alkyl, hydroxyl-C1-C6alkylene, -(NRARB)C1-C6alkylene and (NRARB)carbonyl; or to its individual isomer, stereoisomer or enantiomer, or their mixture, if necessary pharmaceutically acceptable salt. Invention also relates to compound of formula (II), particular compounds of formula (I) and (II), pharmaceutical composition and industrial product based on compound of formula (I) and (II), method of treating said pathological conditions, method of obtaining formula (I) compound and to intermediate compound of formula 3.

EFFECT: novel compounds, useful as inhibitors of poly(ADP-ribose)polymerase, are obtained.

50 cl, 1 tbl, 159 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to naphthalene carboxamide derivatives of general formula I which possess the properties of protein kinase or histone deacetylase inhibitors. The compounds can find application for preparing a drug for treating inflammatory diseases, autoimmune diseases, oncological disease, diseases of the nervous system and neurodegenerative diseases, allergies, asthma, cardiovascular diseases and metabolic diseases or disease related to hormonal diseases. In general formula I: , Z represents CH or N; each of the groups R1, R2 and R3 represents hydrogen, halogen, alkyl, alkoxy or trifluoromethyl; R4 represents or X represents a benzene ring or a pyridine ring; R5 represents one or more substitutes specified in a group consisting of hydrogen, halogen, alkyl, alkoxy or trifluoromethyl. The invention also refers to a method for preparing the above compounds, a pharmaceutical preparation and using them.

EFFECT: preparing the compounds which possess the properties of protein kinase or histone deacetylase inhibitors.

13 cl, 10 tbl, 6 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to new quinolone derivatives of general formula (1) or a pharmaceutically acceptable salts thereof, wherein R1 represents a hydrogen atom, a lower alkyl group, cyclo C3-8 alkyl, a lower alkyl group or a lower alkoxy, a lower alkyl group; R2 represents a hydrogen, a lower alkyl group or a halogen-substituted lower alkyl group; R3 represents a phenyl group, a difurylglyoxal group, a thienyl group or pyridyl group with each group of the above is optionally substituted by one or two groups specified in a group consisting of the following (1) to (16) in an aromatic or heterocyclic ring, presented by the above R3: (1) lower alkyl groups, (2) lower alkoxy groups, (3) halogen-substituted lower alkoxy groups; (4) a phenoxy group, (5) lower alkylthio groups, (6) a hydroxy group, (7) hydroxy lower alkyl groups, (8) halogen atoms, (9) lower alkanoyl groups, (10) lower alkoxycarbonyl groups, (11) amino groups optionally substituted by one or two lower alkyl groups, (12) carbamoyl groups optionally substituted by one or two lower alkyl groups, (13) cyclo C3-8 alkyl lower alkoxy groups, (14) pyrrolidinyl carbonyl groups, (15) morpholinyl carbonyl groups and (16) a carboxyl group; R1 represents a halogen atom; R5 represents a hydrogen atom or a halogen atom; R6 represents a hydrogen atom; and R7 represents any of the above groups (1) to (15): (1) a hydroxyl group, (2) a halogen atom, (3) a lower alkoxy group, (4) a halogen-substituted lower alkoxy group, (5) a hydroxy lower alkoxy group, (6) a lower alkoxy lower alkoxy group, (7) an amino group optionally substituted by one or two members specified in a group consisting of lower alkyl groups, lower alkoxy lower alkyl groups and cyclo C3-8 alkyl groups, (8) an amino lower alkoxy group optionally substituted in an amino group by one or two members specified in a group consisting of lower alkyl groups, lower alkanoyl group, lower alkyl sulphonyl groups and carbamoyl groups optionally substituted by one or two lower alkyl groups, (9) a cyclo C3-8 alkoxy group, (10) a cyclo C3-8 alkyl lower alkoxy group, (11) a tetrahydrofuryl lower alkoxy group, (12) a lower alkylthio group, (13) a heterocyclic group specified in a group consisting of morpholinyl groups, pyrrolidinyl groups, difurylglyoxal groups, thienyl groups and benzothienyl groups, (14) a phenyl lower alkoxy lower alkoxy group and (15) a pyrrolidinyl carbonyl group. Also, the invention refers to a pharmaceutical composition, and a preventive and/or therapeutic agent based on the compound of formula (1), using the compound of formula (1), a method of treating or preventing the above diseases, to a method of preparing the compound of formula (1).

EFFECT: there are prepared new quinolone derivatives effective for treating and/or preventing the neurodegenerative diseases, diseases caused by neurological dysfunction, or diseases induced by deterioration of mitochondrial function.

11 cl, 1 tbl, 104 ex

FIELD: chemistry.

SUBSTANCE: invention relates to derivatives with anticancer activity of formulae:

, , , , ,

R2', R3', R4', R5' and R6' are selected from H, Y(CH2)nCH3, X and (CH2)nNR8R9; Y is selected from O and S; X is selected from F, Cl and Br; R8 and R9 are selected from (CH2)nCH3; R2, R3, R4 and R5 are selected from H, Y(CH2)nCH3, X and (CH2)nNR8R9, or R3 and R4 together form -Y(CH2)nY-; R1 and R1' are selected from H, Li+, Na+, K+, N+R8R9R10R11 or benzyl, where R10 and R11 are selected from H, (CH2)nYH, (CH2)nN(CnH2n+1)(CmH2m+1) or (CH2)nCH3, where n and m are integers from 0 to 4, q is an integer from 1 to 4.

EFFECT: obtaining novel compounds with anticancer activity.

37 cl, 3 dwg, 10 ex, 2 tbl

The invention relates to the technology of known derivatives hinolincarbonova acid, in particular to a method for producing derivatives of 3-hinolincarbonova acid

The invention relates to new derivatives of 5-amino-8-methyl-7-pyrrolidineethanol-3-carboxylic acids and their stereoisomers and their pharmacologically acceptable salts, have excellent antibacterial activity, and to methods for their preparation

FIELD: chemistry.

SUBSTANCE: invention relates to form A of N-(4-(7-azabicyclo[2.2.1]heptan-7-yl-)-2-(trifluoromethyl)phenyl)-4-oxo-5-(trifluoromethyl)-1,4-dihydroquinoline-3-carboxamide, where said form A is characterised by peak at approximately 7.9 degree, peak at approximately 11.9 degree, peak at approximately 14.4 degree and peak at approximately 15.8 degree in powder X-ray. Invention also relates to pharmaceutical composition and set based on said form A, application of form A, method of CFTR modulation.

EFFECT: obtained is novel form of quinoline derivative, which is modulator of CFTR activity.

12 cl, 3 dwg, 2 tbl, 3 ex

FIELD: biotechnologies.

SUBSTANCE: in general formula (I), R1 and R2 that have been taken jointly are a group of formula (II), R3 represents atom of hydrogen or F and R4 represents F; R1, R2, R3 and R4 are connected to 17, 16, 6 and 9 carbon atoms of steroid structure in position α or β; R represents (III) or (IV), and the rest values of radicals are given in the description.

EFFECT: improving use efficiency of compositions for curing of eye diseases, such as diabetic macular edema, diabetic retinopathy, macular degeneration, age-related macular degeneration and other macular and retina diseases.

11 cl, 4 dwg, 5 tbl, 11 ex

FIELD: medicine.

SUBSTANCE: invention relates to medicine, in particular opthalmology, and may be used for treating the patients with a "dry" form of age-related macular degeneration. That is combined with using lutein-containing antioxidants and carotenoids for one year every 2-3 months. Fenofibrate (Tricor 145) is additionally used for a period of time long enough to normalise a lipid profile and to maintain the above values at the attained level. Vitrum vision forte and nutrof total are used as the above lutein-containing antioxidants and carotenoids.

EFFECT: invention provides arresting the progression of the "dry" form of age-related macular degeneration.

2 cl, 1 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to biotechnology, specifically to preparations inhibiting angiogenesis, and may be used in medicine. The method involves administering an EGFL8 antagonist to a subject suffering a pathological condition associated with angiogenesis.

EFFECT: invention enables inhibiting the vascular growth in pathological, eg tumour tissues.

14 cl, 4 dwg, 2 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to ophthalmology, endocrinology and restorative medicine, and may be used to treat the patients with non-proliferative diabetic retinopathy. For this purpose, with underlying background individual antidiabetic therapy, endonasal electrophoresis with 0.1% Semax in a daily dose of 700 mcg is applied in a patient lying on his/her back. A bifurcated positive electrode is coated with cotton swabs each of which is moistened with 7 drops of Semax. These electrodes are introduced into the middle nasal passages. At the same time, a cathode spacer of the second negative electrode 8×10 cm is moistened with 2% aminophylline and placed in the back of the neck. The current intensity is 1.2 mA. The exposure length is 20-22 minutes. The therapeutic course is 8-10 sessions, daily or every second day.

EFFECT: provided therapeutic effect in the early stages of the disease, including in the patients with severe comorbidities, as well as slow progression of the retinal pathological process, and stabilised long-term result up to 6 months.

2 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, in particular to neuroophthalmology, and may be used for treating the visual pathway disorders in the patients with multiple sclerosis. For this purpose, the preparation cortexin is introduced retrobulbar or parabulbar. Then 30 minutes after the introduction of cortexin, upper cervical and stellate ganglia of the sympathetic nervous system are percutaneously exposed to a rotating electric pulse field. This field is generated in a space between the left and right ganglia using two multiple electrodes. The electrodes consist of a number of partial galvanic isolated conductive elements functioning as cathodes and two anodes which are placed in a projection of the ganglia. While forming the electric current pulses, the partial elements of the multiple electrodes are switched; the exposure is paused. Thereafter, a zone of ganglia activity block is switched. It is followed by electrical stimulation of the visual pathway via an electrode applied on the eyelids. For this purpose, the rotating electric pulse field is generated in the space between this electrode which is as the anode, and the partial elements of the multiple electrodes.

EFFECT: method provides relieved demyeliniation, neurodegenerative processes within the visual pathway, improved visual function, including visual acuity and functional characteristics.

5 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely neurology, otorhinolaryngology and orthopaedics, and may be used for treating cochleovestibular disorders in the patients with cervical osteochondrosis. That is ensured by daily injections of the drugs improving cerebral circulation; those are followed by three-stage exposure of head-to-lower extremities centrifugal force on the patient. At the first stage, the patient is exposed to one session a day of centrifugal force at 28 rpm for 5 minutes. The second stage involves 3 days of the exposure to centrifugal force at 30 rpm for 8 minutes, one session a day. The third stage involves 4 days of the daily exposure to centrifugal force at 30 rpm for 10 minutes.

EFFECT: implementing the method provides the higher therapeutic effect ensured by the traction exposure on the cervical spine, passive training of vestibular analyser stability, improved vertebrobasilar circulation ensured by vertebral artery spasmolysis under craniocaudal hypergravitation.

1 dwg, 2 ex

FIELD: medicine.

SUBSTANCE: group of inventions relates to medicine, namely to ophthalmology and can be used for treatment of glaucoma or higher intraocular pressure (IOP) in patient. For this purpose efficient amount of composition, containing preparation inhibiting PAI-1 binding with vitronectin, is introduced to patient. Also claimed is method of treating PAI-1-associated eye disease.

EFFECT: group of inventions ensures treatment of glaucoma and IOP due to reduction of separation of trabecular meshwork tissue and reduction of increased resistance to intraocular fluid outflow, and also due to increase of TM tissue cellularity and preservation of phagocytosis.

14 cl, 8 dwg, 8 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely ophthalmology and physiotherapy, and may be used for treating vascular and dystrophic retinal and visual nerve diseases. That is ensured by the intravenous drop-by-drop infusion of the 2/3 amount of the medical solution made of cytoflavin 10 ml and 0.9% sodium chloride 200 ml. It is followed by 15-minite transverse halvanisation in the Potok-1 apparatus at current force making 0.5 mA. For this purpose 2 oval anode electrodes through a pad impregnated in distilled water are placed on skin of closed eyelids of both eyes and connected by a Y-wire with the anode of the apparatus. The third cathode electrode is placed transversally in relation to the two first anode electrodes on the area of upper cervical spines S2-S4 and connected with the cathode of the apparatus. The therapeutic course makes 10 daily procedures.

EFFECT: method provides high clinical effectiveness ensured by high functional effectiveness of cytoflavin electric accumulation in eye tissue with improving visual acuity by 0,15-0,3, prolonged action of the ensured therapeutic effect up to 5-6 months, eliminated risk of damaging an external eyeball capsule.

1 tbl, 2 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to suspension for visualisation of transparent tissue of eye, which contains finely dispersed particles of biodegradable macromolecular compound and salt, selected from salt of divalent metal in amount 0.1-1 wt/vol.% or salts of trivalent metal in amount 0.01-1 wt/vol.%. Said suspension is intended for pouring into eye to come in contact with transparent eye tissues with the purpose to improve their distinguishability. Invention also relates to ophthalmologic preparation which contains hard particles of biodegradable macromolecular compound and water medium, which contains said salts of divalent or trivalent metal, solid and water media being placed separately and without allowing contact with each other. Invention also relates to methods of obtaining suspension for eye visualisation, which include obtaining solid composition, containing finely dispersed particles of biodegradable macromolecular compound or finely dispersed particles of biodegradable macromolecular compound and 0.1-1 wt/vol.% of salt of divalent or 0.01-1 wt/vol.% of salt of trivalent metal, and suspending said solid composition in water solution.

EFFECT: invention ensures obtaining suspension for visualisation of transparent eye tissue, which can be applied as convenient and safe means for improvement of distinguishability of transparent eye tissues when performing surgical operation on them.

21 cl, 1 tbl, 7 ex

FIELD: chemistry.

SUBSTANCE: invention relates to form A of N-(4-(7-azabicyclo[2.2.1]heptan-7-yl-)-2-(trifluoromethyl)phenyl)-4-oxo-5-(trifluoromethyl)-1,4-dihydroquinoline-3-carboxamide, where said form A is characterised by peak at approximately 7.9 degree, peak at approximately 11.9 degree, peak at approximately 14.4 degree and peak at approximately 15.8 degree in powder X-ray. Invention also relates to pharmaceutical composition and set based on said form A, application of form A, method of CFTR modulation.

EFFECT: obtained is novel form of quinoline derivative, which is modulator of CFTR activity.

12 cl, 3 dwg, 2 tbl, 3 ex

Up!