Inhibitors of protease of hepatitis c virus and their application

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to compounds, which can be used as inhibitors of protease of hepatitis C virus, pharmaceutical compositions, containing the said compounds, and methods of their application.

EFFECT: obtaining compounds which can be used as inhibitors of protease of hepatitis C virus.

41 cl, 10 dwg, 7 tbl, 26 ex

 

Cross references to related applications

[0001] the Present invention claims the priority under provisional patent application U.S. serial No. 61/016110, filed December 21, 2007, provisional application for U.S. patent, serial No. 61/016,473, filed on December 23, 2007, provisional application for U.S. patent, serial No. 61/075,001, filed on June 23, 2008, and provisional patent application U.S. serial No. 61/098,675, filed September 19, 2008, the contents of which are fully incorporated herein by reference.

The technical field

[0002] the Present invention relates to compounds that may be useful as protease inhibitors of hepatitis C. the Present invention also provides pharmaceutically acceptable compositions containing the compounds according to the present invention, and methods of using these compositions to treat various diseases.

The level of technology

[0003] researchers estimate that about 170 million people worldwide are affected by the hepatitis C virus (HCV). HCV is a dominant serotype of 3% worldwide and is the primary cause of most cases of hepatitis other than hepatitis a and b (Alberti, A. et al., J. Hepatology 31., (Suppl. I): 17-24, 1999). Although in some patients the symptoms of acute hepatitis weaken, at least in 85% of cases the infection, caused by HCV becomes chronic and 20% of those infected will develop cirrhosis. The survival rates of four years after the diagnosis of cirrhosis is less than 50%. Chronic HCV infection is also associated with increased cases of disease hepatocellular carcinoma.

[0004] HCV refers to viruses that contain "±chain" RNA genome which encodes polyprotein, consisting of approximately 3000 amino acid residues. After processing of this protein-precursor is formed at least 10 viral structural and non-structural proteins C, E1, E2, P7, NS2, NS3, NS4A, NS4B, NS5A, and NS5B (Blight, K.J. et al., AntiviralTher. 3, Suppl. 3: 71-81, 1998). Non-structural (NS) proteins of HCV are formed as a result of proteolytic cleavage of polyprotein, and suggest that they provide basic proteins that catalyze the replication of the virus.

[0005] NS3 is a protein of approximately 68 Kda, contains in its N-terminal domain serine protease, domain and RNA-dependent ATP-ASE at the end. It was shown that protein NS4A performs the functions of a co-factor activity of NS3 as a serine protease. NS3 functions as a proteolytic enzyme, releasing other non-structural proteins necessary for replication of HCV, and is a useful therapeutic target for antiviral chemotherapy.

[0006] At present is it time there is no vaccine against HCV, and developed a therapy involving treatment with interferon is effective only in 15-20% of cases (Weiland, O., FEMSMicrobiol. Rev. 14: 279-88, 1994) and has significant side effects (Walker, M.A., et al., DDT4: 518-29, 1999; Moradpour, D., et al., Eur. J. Gastroenterol. Hepatol. 11: 1199-1202, 1999). Although the current standard of care, which is a therapy paglierani interferon-α in combination with ribavirin is more effective and, apparently, reduces the likelihood of developing hepatocellular carcinoma in patients associated with HCV cirrhosis (Hung, C.H. et al., JViral Hepatitis 13(6): 409-414, 2006), this method of treatment also showed side effects such as the development of thyroid dysfunction (Huang, J.F., et al., J Viral Hepatitis 13(6): 396-401, 2006).

[0007] a poor prognosis for patients suffering from HCV infection, and the current lack of effective approved therapies indicate an existing need for new inhibitors of the HCV protease NS3.

Brief description of the invention

[0008] it Was found that the compounds according to the present invention and pharmaceutically acceptable compositions that contain these compounds are effective as inhibitors of HCV protease. These compounds are compounds of General formula I:

or pharmaceutically pickup is acceptable salts of these compounds, where the values of R1, R1', R2a, R3, R4, Rw, Rx, Rx'and Rydefined in the present description.

[0009] the Compounds according to the present invention, and pharmaceutically acceptable compositions containing such compounds that are effective for the treatment of many diseases, disorders and conditions associated with HCV. Such diseases, disorders and conditions include described in the text of this application.

[0010] the Compounds according to the present invention are also useful in the study of the role of HCV protease in the biological and pathological processes; in the study of pathways of intracellular signal transduction mediated by the HCV protease; and for the comparative evaluation of new HCV protease inhibitors.

Brief description of drawings

[0011] Figure 1 shows the results of mass spectral analysis of the HCV protease NS3/4A wild type in the presence of test compound I-1.

[0012] Figure 2 shows the results of mass spectral analysis of the HCV protease NS3/4A wild type in the presence of test compound I-25.

[0013] Figure 3 shows the results of mass spectral analysis of the HCV protease NS3/4A.

[0014] Figure 4 shows the results of mass spectral analysis of mutant HCV protease NS3/4A D168V in the presence of test compound I-11.

[0015] Figure 5 shows resulthi mass-spectral the analysis of mutant HCV protease NS3/4A A156S in the presence of test compound I-11.

[0016] Figure 6 shows the results of mass spectral analysis of mutant HCV protease NS3/4A R155K in the presence of test compound I-11.

[0017] Figure 7 shows the results of mass spectral analysis of mutant HCV protease NS3/4A AT in the presence of test compound I-11.

[0018] Figure 8 illustrates that the inner products autoproteolysis NS3 protease inhibited when exposed to cells carrying the replicon, Compound 1-47 for 16 hours.

[0019] Figure 9 shows that irreversible covalent inhibitor of the NS3 protease (compound I-11) provides long-lasting inhibition of the activity of the NS3 protease in wild-type cells carrying the replicon, which was determined through the value of autoproteolysis after removal of the compounds.

[0020] Figure 10 shows that irreversible covalent inhibitor of the NS3 protease (compound I-25) provides long-lasting inhibition of the activity of the NS3 protease in wild-type cells carrying the replicon, which was determined through the value of autoproteolysis, after removal of the compounds.

Detailed description of some implementation options

1. A General description of the compounds according to the present invention

[0021] According to some variants of implementation of the present invention proposed a compound of the formula I:

or pharmaceutically acceptable salt of the decree of the aqueous compounds, where:

R1and R1'independently represent hydrogen or may contain substituents With1-6aliphatic hydrocarbon, or R1and R1'joined together with formation may contain substituents 3-7-membered carbocyclic ring;

R2arepresents-IT or-NHSO2R2;

R2represents-N(R)2or possibly containing substituents group selected from the following:3-7cycloalkyl associated bridge Bicycle, 6-10-membered aryl, 5-10-membered heteroaryl containing 1-4 heteroatoms that are independently selected from nitrogen, oxygen, or sulfur, or 4-7-membered heterocycle containing 1-2 heteroatoms that are independently selected from nitrogen, oxygen, or sulfur;

each R independently represents hydrogen, possibly containing substituents With1-6aliphatic group, or:

two R together with the nitrogen atom to which they are attached, form a 4-7-membered heterocyclic ring containing 1-2 heteroatoms that are independently selected from nitrogen, oxygen, or sulfur;

R3is a head group, or:

R3and R1together with in between the atoms form may contain substituents saturated or unsaturated 10-12-membered ring containing 2-6 heteroatoms selected from: nitrogen, is of ikorodu or sulfur, moreover, the specified educated these substituents, the ring includes a head group; or

R3and the ring formed by R1and R1'together with the atoms located between them form may contain substituents saturated or unsaturated 12-18 membered ring containing 2 to 6 heteroatoms, which are independently selected from nitrogen, oxygen or sulfur, with the specified educated ring includes a head group;

R4represents H, -NHC(O)R5, -NHC(O)OR6,or the side group of a natural or unnatural amino acids;

each R5independently represents-N(R)2or possibly containing substituents group selected from the following:1-6aliphatic hydrocarbons associated with bridge Bicycle, 6-10-membered aryl, 5-10-membered heteroaryl containing 1-4 heteroatoms that are independently selected from nitrogen, oxygen, or sulfur, or 4-7-membered heterocyclyl containing 1-2 heteroatoms that are independently selected from nitrogen, oxygen, or sulfur;

R6represents possibly containing substituents group selected from the following:1-6aliphatic hydrocarbons associated with bridge Bicycle, 6-10-membered aryl, 5-10-membered heteroaryl containing 1-4 heteroatoms that are independently selected from nitrogen, oxygen, or sulfur, or 4-7-clenn the th heterocyclyl, containing 1-2 heteroatoms that are independently selected from nitrogen, oxygen, or sulfur; and

R7represents possibly containing substituents group selected from the following:1-6aliphatic hydrocarbon, connected by bridge Bicycle, 6-10-membered aryl, 5-10-membered heteroaryl containing 1-4 heteroatoms that are independently selected from nitrogen, oxygen, or sulfur, or 4-7-membered heterocyclyl containing 1-2 heteroatoms that are independently selected from nitrogen, oxygen or sulfur.

RZrepresents; or R4and RZtogether with in between the atoms form may contain substituents saturated or unsaturated 16-22 membered ring containing 2 to 6 heteroatoms, which are independently selected from nitrogen, oxygen, or sulfur;

each of Ryindependently selected from the following: halogen, -ORo, -CN, -NO2, -N(Ro)2or possibly containing substituents With1-4aliphatic group; and

m is an integer from 0 to 4 inclusive;

s is an integer from 0 to 4 inclusive;

t is an integer from 0 to 4 inclusive;

moreover, the sum of s and t is not equal to zero.

2. Substances and definitions

[0022] the Compounds according to the present invention include compounds broadly described the above, and the following as examples in this application classes, subclasses and species. In the present description will be used the following definitions except as otherwise specified. In the context of the present invention the determination of the chemical elements are given in accordance with the periodic system of the elements, CAS version, Hand book of Chemistry and Physics, 75thEd. 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, 5thEd., Ed.: Smith, M.B. and March, J., John Wiley&Sons, New York: 2001, the description of which is fully incorporated herein by reference.

[0023] the Term "aliphatic hydrocarbon" or "aliphatic group", as used in the present description, refers to a straight (i.e. unbranched) or branched hydrocarbon chain containing or not containing substituents, fully saturated or containing one or more unsaturated sites, and monocyclic or bicyclic hydrocarbon that is completely saturated or contain one or more unsaturated sites, but which is not aromatic (these definitions also apply to used in the present description the terms "carbocycle", "cycloaliphatic hydrocarbon or cycloalkyl"), having one point of linkage with the rest of the molecule. C is an exception otherwise specified aliphatic group contains from 1 to 6 aliphatic carbon atoms. In some embodiments of the aliphatic group includes 1-5 aliphatic carbon atoms. In other embodiments of the aliphatic group contains 1-4 aliphatic carbon atoms. In other embodiments of the aliphatic group includes 1-3 aliphatic carbon atoms, there are also options for implementation according to which the aliphatic group include 1-2 aliphatic carbon atoms. In some embodiments, the implementation of "cycloaliphatic hydrocarbon" (or "carbocycle", or "cycloalkyl") refers to a monocyclic hydrocarbon With3-C6fully saturated or containing one or more unsaturated sites, but which is not aromatic, having a single point of linkage with the rest of the molecule. Suitable aliphatic groups include, but are not limited to the following examples: a linear or branched, containing or not containing substituents alkyl, alkeline, alkyline groups and their combinations, such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl or (cycloalkyl)alkenyl.

[0024] the Term "bicyclic hydrocarbon (Bicycle), connected (bound) bridge"used in the present description, refers to any system consisting of two rings, i.e carbocyclic or heterocyclic, saturated or containing unsaturated sites containing what about the at least one bridge. As defined in the IUPAC nomenclature, the "bridge" is an unbranched chain of atoms, or atom, or a valence bond connecting two atoms in the head of the bridge, where the atom in the head of the bridge" is any atom of the hydrocarbon skeleton of the cyclic system, associated with three or more atoms of the skeleton (excluding hydrogen). In some embodiments of bicyclic group associated with the bridge contains 7-12 atoms comprising the ring and 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur. Such bicyclic groups connected by a bridge, well known in this field and include groups below, where each of the groups attached to the rest of the molecule at any carbon atom or nitrogen, which can be appended to the Deputy. Except as otherwise specified bicyclic group, connected by a bridge, may contain one or more substituents as described for aliphatic groups. Additionally or alternatively, any nitrogen atom, to which may be attached Deputy bicyclic group, linked by a bridge, can be replaced. The examples are linked by a bridge of bicyclo include:

[0025] the Term "lower alkyl" refers to straight or branched alkyl group With1-4. Examples of the lower alkyl groups are m the Tyl, ethyl, propyl, isopropyl, butyl, isobutyl and tert-butyl.

[0026] the Term "lower haloalkyl" refers to a straight or branched alkyl group With1-4containing as substituents one or more halogen atoms.

[0027] the Term "heteroatom" means one or more of the following: oxygen, sulphur, nitrogen, phosphorus, or silicon (including, any oxidized form of nitrogen, sulfur, phosphorus, or silicon; the Quaternary form of any nitrogenous base or; the nitrogen to which can be attached Deputy, heterocyclic ring, for example N (as in the structure of 3,4-dihydro-2H-pyrrolyl), NH (as in the composition of pyrrolidinyl) or NR+(as in the N-C-pyrrolidinyl)).

[0028] the Term "unsaturated" in the text of this application refers to a group containing in their structure one or more unsaturated regions.

[0029] the Term "divalent saturated or unsaturated, branched or not branched hydrocarbon chain C1-8(or1-6)" in the text of the present application refers to a straight or branched as defined in the present description, the divalent alkilinity, alkenylamine and alkynylaryl circuits.

[0030] the Term "alkylene" refers to divalent alkyl group. "Allenova chain" is a polymethene group, ie -(CH2)n-, where n is a positive value, predpochtite is correctly located in the interval from 1 to 6, 1 to 4, from 1 to 3, 1 to 2 or from 2 to 3. Allenova chain containing substituents, represents polymethene group, in which instead of one or more atoms of hydrogen in the methylene group contained Deputy. Suitable substituents include the substituents described below for aliphatic group.

[0031] the Term "albaniles" refers to divalent alkenylphenol group. Alcanena chain containing substituents, represents polymethene group containing at least one double bond, in which instead of one or more hydrogen atoms in the molecule is contained Deputy. Suitable substituents include the substituents described below for aliphatic group.

[0032] the Term "cyclopropylethyl" in the present description refers to a divalent cyclopropene group having the structure described by the following schema:.

[0033] the Term "halogen" refers to atoms of F, Cl, Br or I.

[0034] the Term "aryl"used alone or as part of the title of the larger groups, such as aralkyl", "arakaki", or "aryloxyalkyl", refers to monocyclic or bicyclic system, the total number of atoms in which there are from five to fourteen, in which at least one ring is aromatic and each ring consists of 3 to 7 and the Ohm. The terms "aryl" and "aryl ring" may be used interchangeably.

[0035] the Term "aryl"used alone or as part of the title of the larger groups, such as aralkyl," "arakaki" or "aryloxyalkyl", refers to monocyclic or bicyclic system, the total number of atoms which ranges from five to fourteen, in which at least one ring is aromatic and each ring consists of 3 to 7 atoms. The terms "aryl" and "aryl ring" may be used interchangeably. In some embodiments of the present invention, "aryl" refers to aromatic closed in a ring system, including, but not limited to the following examples: phenyl, diphenyl, naphthyl, antracol and similar compounds, which may contain one or more substituents. Also in the present description, the term "aryl" means a group in which an aromatic ring condensed with one or more non-aromatic rings, such as indanyl, phthalimide, naftopidil, phenanthridine or tetrahydronaphthyl and the like.

[0036] the Terms "heteroaryl" and "heteroa-", used alone or as part of the title of the larger groups, such as heteroalkyl" or "heteroaromatic"are groups consisting of 5-10 atoms contained in the ring, prefer the LNA 5, 6 or 9 atoms contained in the ring; having 6, 10 or 14 electrons distributed in a circular system; and containing, besides carbon atoms, from one to five heteroatoms. The term "heteroatom" refers to nitrogen, oxygen, or sulfur, and any oxidized form of nitrogen or sulfur and to any Quaternary form a nitrogenous base. Heteroaryl groups include, but are not limited to the following examples: thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolin, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, pyrenyl, naphthyridines and pteridinyl. The meaning of the terms "heteroaryl" and "heteroa-"used in the present description also includes groups containing heteroaromatic ring fused with one or more aryl, cycloaliphatic or heterocyclic rings such that the radical or point of connection is on the heteroaromatic ring. Examples of such compounds is not limited to the following: indolyl, isoindolyl, benzothiazyl, benzofuranyl, dibenzofurans, indazoles, benzimidazoles, benzothiazoles, quinolyl, isoquinolinyl, cyclolepis, phthalazines, quipazine, quinoxalinyl, 4H-cipolini, carbazolyl, acridines, phenazines, phenothiazines, phenoxazines, tetrahedr quinolinyl, tetrahydroisoquinolines and pyrido-[2,3-b]-1,4-oxazin-3(4H)-he. Heteroaryl group may be mono - or bicyclic. The terms "heteroaryl", "heteroaryl ring", "heteroaryl group" or "heteroaromatic hydrocarbon" may be used interchangeably, and the value of any of these terms includes the cyclic system may contain substituents. The term "heteroalkyl" refers to an alkyl group containing as a substituent heteroaryl, where alkyl and heteroaryl areas may contain substituents.

[0037] the Terms "heterocycle", "heterocyclyl", "heterocyclic radical", and "heterocyclic ring"used in the present description, are used interchangeably and refer to a stable 5-7-membered monocyclic or 7-10-membered bicyclic heterocyclic molecules, which can be saturated or contain unsaturated sites, and containing, besides carbon atoms, one or more, preferably one to four, heteroatoms, as described above. The meaning of the term "nitrogen", used in relation to the atom ring heterocycle that includes the nitrogen containing substituents. As an example, in a saturated or unsaturated parcels ring containing 0-3 heteroatoms selected from oxygen, sulfur or nitrogen, the nitrogen may be in the footprint of the affected forms: N (as part of 3,4-dihydro-2H-pyrrolyl), NH (as part of pyrrolidinyl) or+NR (as part of N-substituted pyrrolidinyl).

[0038] the Heterocyclic ring may be connected with its end group at any heteroatom or carbon atom that provides a stable structure, and any of the atoms of the ring may contain substituents. Examples of such saturated or unsaturated areas of heterocyclic radicals include, but are not limited to, the following: tetrahydrofuranyl, tetrahydrothiophene pyrrolidinyl, piperidinyl, pyrrolyl, tetrahydroquinoline, tetrahydroisoquinoline, decahydroquinoline, oxazolidinyl, piperazinil, dioxane, dioxolane, diazepines, oxazepines, thiazepines, morpholinyl and quinuclidinyl. The terms "heterocycle", "heterocyclyl", "heterocyclyl ring", "heterocyclic group", "heterocyclic molecule" and "heterocyclic radical" is used in the present description are interchangeable and also include groups in which heterocyclyl ring condensed with one or more aryl, heteroaryl or cycloaliphatic rings, such as indolyl, 3H-indolyl, bromanil, or tetrahydroquinoline, where the radical or point of connection is located on heterocyclyl ring. The term "geterotsiklicheskikh" refers to an alkyl group containing as a substituent heterotic the sludge, in which alkyl and heterocyclyl areas independently may contain substituents.

[0039] the Term "partially unsaturated"as used in the present description, refers to a circular molecule containing at least one double or triple bond. The term "partially unsaturated" is used to define rings containing multiple unsaturated sites, but its value does not include aryl or heteroaryl molecules, as defined in the present description.

The phrase "group of the side chain of natural amino acids", as used in the present description, refers to a group of the side chain of any of the 20 amino acids naturally occurring in proteins. Such natural amino acids include the non-polar or hydrophobic amino acids, glycine, alanine, valine, leucine, isoleucine, methionine, phenylalanine, tryptophan and Proline. Cysteine is in some cases referred to as non-polar or hydrophobic amino acids, in other cases - polar. Natural amino acids also include polar or hydrophilic amino acids such as tyrosine, series, threonine, aspartic acid (also known as aspartate, in the charged state), glutamic acid (also called glutamate, in the charged state), asparagine and glutamine. Some polar or hydrophilic amino acids, contain charged side C is PI. These charged amino acids include lysine, arginine and histidine. An ordinary person skilled in the art it is clear that the protection of the side chain polar or hydrophilic amino acids may transfer such non-polar amino acid in the form. For example, a suitably protected hydroxyl group of tyrosine can translate this tyrosine in nonpolar and hydrophobic form by protecting a hydroxyl group.

[0040] the Phrase "group of the side chain unnatural amino acids", as used in the present description, refers to a group of the side chain amino acids are not included in the list of 20 amino acids naturally occurring in proteins, above. Such amino acids include the D-isomers of any of the 20 amino acids occurring in nature. Unnatural amino acids may include homoserine, ornithine, norleucine and thyroxine. Side chains other unnatural amino acids are widely known to experts in this field and include non-natural aliphatic side chains. Also to non-natural amino acids include modified amino acids, including N-alkylated, cyklinowanie, phosphorylated, acetylated, amidarone, azetilirovanie, amino acids, made labels, etc. In some embodiments of the unnatural amino acid is a D-isomer. In some vari is ntah the implementation of the unnatural amino acid is an L-isomer.

[0041] As described in the text of this application, the substances according to the present invention can contain groups, "possibly containing substituents". In General, the term "containing substituents, as preceded by the term "possible"or not, means that one or more hydrogen atoms of the specified group is replaced by a suitable substitute. Except otherwise specified, the group, possibly containing substituents"may include a suitable substituent in any position in which you can enter a Deputy, and in cases where one or more substituents selected from the respective groups, you can enter in one or more of the provisions in any given structure, the substituents may be the same in all positions, and different. Combinations of substituents, provided by the present invention, preferably are contributing to the formation of stable and chemically acceptable substances. The term "stable", as used in the present description, refers to substances which do not significantly change under the influence of the conditions required for their production, definition, and in some embodiments of their separation, purification, and use for one or more purposes described in the present description.

[0042] Suitable monovalent substituents at the appropriate atom of carbon is in the group, "possibly containing substituents"independently represent halogen; -(CH2)0-4Ro; -(CH2)0-4ORo; -O(CH2)0-4Ro, -O-(CH2)0-4C(O)ORo; -(CH2)0-4CH(ORo)2; -(CH2)0-4SRo; -(CH2)0-4Ph, which may contain as substituents Ro; -(CH2)0-4O(CH2)0-1Ph, which may contain as substituents Ro; -CH=CHPh, which may contain as substituents Ro; -(CH2)0-4O(CH2)0-1-pyridyl, which may contain as substituents Ro; -NO2; -CN; -N3; -(CH2)0-4N(Ro)2; -(CH2)0-4N(Ro)C(O)Ro; -N(Ro)C(S)Ro; -(CH2)0-4N(Ro)C(O)NRo2; -N(Ro)C(S)NRo2; -(CH2)0-4N(Ro)C(O)ORo; -N(Ro)N(Ro)C(O)Ro; -N(Ro)N(Ro)C(O)NRo2; -N(Ro)N(Ro)C(O)ORo; -(CH2)0-4C(O)Ro; -C(S)Ro; -(CH2)0-4C(O)ORo; -(CH2)0-4C(O)SRo; -(CH2)0-4C(O)OSiRo3; -(CH2)0-4OC(O)Ro; -OC(O)(CH2)0-4SR-SC(S)SRo; -(CH2)0-4SC(O)Ro; -(CH2)0-4C(O)NRo2; -C(S)NRo2; -C(S)SRo; -SC(S)SRo, -(CH2) 0-4OC(O)NRo2; -C(O)N(ORoRo; -C(O)C(O)Ro; -C(O)CH2C(O)Ro; C(NORoRo; -(CH2)0-4SSRo; -(CH2)0-4S(O)2Ro; -(CH2)0-4S(O)2ORo; -(CH2)0-4OS(O)2Ro; -S(O)2NRo2; -(CH2)0-4S(O)Ro; -N(Ro)S(O)2NRo2; -N(Ro)S(O)2Ro; -N(ORoRo; -C(NH)NRo2; -P(O)2Ro; -P(O)Ro2; -OP(O)Ro2; -OP(O)(ORo)2; SiRo3; - (straight or branched alkylen C1-4)O-N(Ro)2; or - (straight or branched alkylen C1-4)C(O)O-N(Ro)2where each Romay contain substituents as described above and independently represents hydrogen, C1-6aliphatic hydrocarbon, -CH2Ph, -O(CH2)0-1Ph, -CH2-(5-6-membered heteroaryl ring), or a 5-6-membered saturated, partially unsaturated, or aryl ring containing 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or, regardless of the specific above, two independent Rotogether with the intermediate atoms, form a 3-12-membered saturated, partially unsaturated, or aryl mono - or bicyclic ring containing 0-4 heteroatoms independently selected from nitrogen is, oxygen or sulfur, which may contain substituents, as defined below.

[0043] Suitable monovalent substituents at Ro(or the ring formed by two independent Rotogether with the intermediate atoms), independently represent hydrogen, -(CH2)0-2R, -(R), -(CH2)0-2HE, -(CH2)0-2OR, -(CH2)0-2CH(OR)2; -O(R), -CN, -N3, -(CH2)0-2C(O)R, -(CH2)0-2C(O)OH, - (CH2)0-2C(O)OR, -(CH2)0-2SR, -(CH2)0-2SH, -(CH2)0-2NH2, -(CH2)0-2Other, -(CH2)0-2NR2, -NO2, -SiR3, -OSiR3, -C(O)SR, -(C1-4unbranched or branched alkylene)C(O)ORor-SSRwhere each Rcontains no substituents, or in cases where his name has the prefix "halo", contains as substituents one or more halogen atoms, and are independently selected from the following groups:1-4aliphatic hydrocarbon, -CH2Ph, -O(CH2)0-1Ph, or 5-6-membered saturated, partially unsaturated, or aryl ring containing as substituents 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur. Suitable LW is valentie the substituents at the saturated carbon atom, R oinclude =O and =s

[0044] Suitable divalent substituents at the carbon atom of containing substituents" include the following: =O, =S, =NNR*2, =NNHC(O)R*, =NNHC(O)OR*, =NNHS(O)2R*, =NR*, =NOR*, -O(C(R*2))2-3O-, or-S(C(R*2))2-3S-, where each independent variant implementation of R*selected from hydrogen, C1-6aliphatic hydrocarbon that may contain substituents as described below, or not containing substituents 5-6-membered saturated, partially unsaturated, or aryl ring containing 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur. Suitable divalent substituents that are associated with the adjacent carbon atoms to which you can attach the substituents in the group, "perhaps containing substituents include-O(CR*2)2-3O-, where each independent variant implementation of R*selected from hydrogen, C1-6aliphatic hydrocarbon that may contain substituents as described below, or not containing substituents 5-6-membered saturated, partially unsaturated, or aryl ring containing 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

[0045] Suitable substituents in the aliphatic group of R*include halogen, -R,-(R), -OH, -OR, -O(R), -CN, -C(O)OH, -C(O)OR, -NH2-The other, -NR2or NO2where each Rcontains no substituents, or in cases where his name has the prefix "halo", contains as substituents one or more halogen atoms, and are independently selected from the following groups:1-4aliphatic hydrocarbon, -CH2Ph, -O(CH2)0-1Ph, or 5-6-membered saturated, partially unsaturated, or aryl ring containing 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

[0046] the Respective substituents at nitrogen in the composition of the groups may contain substituents include-R, -NR2-C(O)R, -C(O)OR, -C(O)C(O)R, -C(O)CH2C(O)R, -S(O)2R, -S(O)2NR2, -C(S)NR2, -C(NH)NR2or-N(R)S(O)2R; where each Rindependently represents hydrogen or C1-6aliphatic hydrocarbon that may contain substituents as described below, not containing substituents-OPh, or not containing substituents 5-6-membered saturated, partially unsaturated, or aryl ring containing 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or, regardless of the specific above, two independent the x option, Rtogether with in between the atoms form not containing substituents 3-12-membered saturated, partially unsaturated, or aryl mono - or bicyclic ring containing 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

[0047] Suitable substituents in the aliphatic group of Rindependently represent halogen, -R, -(R), -OH, -OR, -O(R), -CN, -C(O)OH, -C(O)OR, -NH2-The other, -NR2or NO2where each Rcontains no substituents, or, whenin its name is the prefix "halo", contains as substituents only one or more halogen atoms, and independently represents an aliphatic hydrocarbon, C1-4, -CH2Ph, -O(CH2)0-1Ph, or 5-6-membered saturated, partially unsaturated, or aryl ring containing 0-4 heteroatoms that are independently selected from nitrogen, oxygen or sulfur.

[0048] the Term "pharmaceutically acceptable salt"used in the present description, refers to salts that rational limits from a medical point of view, suitable for use in contact with the tissues of humans and animals, non-toxic, does not cause irritation and allergic responses, etc. and have the appropriate acceptable risk ratio. F. rmaceuticals acceptable salts are widely known in this field. For example, S.M.Berge et al., describe pharmaceutically acceptable salts in J. Pharmaceutical Sciences, 1977, 66, 1-19, incorporated herein by reference. Pharmaceutically acceptable salts of the substances according to the present invention include salts derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable, nontoxic acid additive salts are salts of amino groups, obtained by adding inorganic acids such as hydrochloric acid, Hydrobromic acid, phosphoric acid, sulphuric acid, perchloric acid, or by adding 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 this field, such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, bansilalpet, benzoate, bisulfate, borate, butyrate, comfort, camphorsulfonate, citrate, cyclopentane propionate, digluconate, dodecyl sulphate, aconsultant, formate, fumarate, glucoheptonate, glycerol, gluconate, hemisulfate, heptanoate, hexanoate, hydroidea, 2-hydroxyethanesulfonic, lactobionate, lactate, laurate, lauryl, malate, maleate, malonate, mutans lifenet, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluene sulfonate, undecanoate, salt valerianic acid and the like.

[0049] the Salt obtained by reacting with the appropriate bases include alkali metal salts, alkaline earth metals, ammonium salts and N+(C1-4alkyl)4. Examples of salts of alkaline or alkaline earth metals include sodium, lithium, potassium, calcium, magnesium and the like. Further pharmaceutically acceptable salts include, when appropriate, non-toxic salts of ammonia, Quaternary ammonium salts and amines cations formed using such protivoionov as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, sulfonates lower Akilov and aryl sulfonate.

[0050] except otherwise specified chemical structure shown in the present description, also mean all isomeric (e.g., enantiomeric, diastereomeric and geometric (or conformational) shape; for example, R and S configurations for each center of asymmetry, Z and E isomers around the double bond, and Z and E) conformational isomers. Thus, the scope of the present invention covers stereochemical isomers, and enantiomers, diastereomeric and geometric (and conformational) of a mixture of compounds according to the invention. Except otherwise specified, the scope of the present invention covers all tautomeric forms of the compounds. Also except otherwise specified chemical structure shown in the present description, is also used to refer to compounds that differ only by the presence of one or more as a result of isotopically enriched atoms. For example, the scope of the present invention encompasses compounds having given in this description of the structure, including the replacement of hydrogen by deuterium or tritium, or the replacement of carbon on13With or14C-enriched carbon. Such compounds may be useful as an analytical tool, as probes in biological assays or as therapeutic agents according to the present invention. In some embodiments of the parent group, R3, or connection according to the present invention comprises one or more deuterium atoms.

[0051] the Term "irreversible" or "irreversible inhibitor", as used in the present invention refers to an inhibitor (i.e. connection)capable of covalently to contact the HCV protease essentially irreversible manner. While reversible ing the inhibitor is able to join the HCV protease (but usually are not able to form with her covalent bond) and thus, it may dissociate from the HCV protease, an irreversible inhibitor after the formation of the covalent bond remains associated with the HCV protease. Irreversible inhibitors usually have a time dependence, in which the degree of inhibition increases depending on the time during which the inhibitor is in contact with the enzyme. In some embodiments of the irreversible inhibitor remains essentially associated with protease after the formation of the covalent bond and remains associated with it for a time period longer than the lifetime of the protein.

[0052] methods for determining the ability of a substance to act as an irreversible inhibitor known to the ordinary person skilled in the art. Such methods include, but are not limited to kinetic analysis of the profile of inhibition of the compounds of the HCV protease enzyme reactions, application of mass spectrometry of protein drug target modified in the presence of substances inhibitor, experiments with periodic effect, also known as "perfusion" (shaded) and making labels, for example by introducing a radioactive label in the composition of the inhibitor, for the study of covalent modification of the enzyme, as well as other methods known to the expert in this field.

[0053] an Ordinary specialist in this field is t, it will be clear some reactive functional groups can act as a "head group". The term "parent group"used in the present description, refers to a functional group included in the compounds according to the present invention with the ability to covalently to contact amino acid residue (e.g., cysteine, lysine, histidine or other amino acid residues susceptible to covalent modification), located in the region of the binding pocket" of the protein target, and thereby irreversibly inhibiting protein. It should be understood that under the group-L-Y, as defined and described in the text of this application, refers to these head groups, providing covalent and irreversible inhibition of protein.

[0054] the Term "inhibitor" in the present description is defined as a connection, substantially communicates with the HCV protease and/or inhibiting it. In some embodiments of the values of the IC50and/or constants of the binding of the inhibitor is less than about 50 nm, less than about 1 nm, less than about 500 nm, less than about 100 nm, less than about 10 nm, or less than about 1 nm.

[0055] the Compound according to the present invention can be associated with detektivami substance. Specialists in this region, the STI will be clear, what detective substance can be attached to the connection according to the present invention by means of a suitable replacement group. The term "suitable replacement group"used in the present description, refers to a functional group capable of covalently to join detectivemisa. Such functional groups are well known to ordinary experts in this field and include contains, for example, carboxylate group, amino group, thiol group or hydroxyl group, and others. It should be understood that such groups may be attached directly to the connection according to the present invention, or by using a linking group, for example, divalent saturated or unsaturated hydrocarbon chain. According to some variants of realization of these groups can be attached via Click reactions-chemistry. According to other variants of realization of these groups can be attached using the 1,3-cycloaddition of azide with Alcina, for example in the presence of a copper catalyst. How to apply Click reactions-chemistry known in the field and include the methods described in Rostovtsev et al., Angew. Chem. Int. Ed. 2002, 41, 2596-99 and Sun et al., Bioconjugate Chem., 2006, 17, 52-57.

[0056] the Term "contrast agent", as used in the present description, is used interchangeably with the term "label" and apply the group, that it is possible to detect, for example to the primary tags and secondary marks. Primary label such as a radioisotope (for example, tritium,32P,33R35S or14C), mass labels, and fluorescent labels are transmitting the signal Renasterii" groups, which can be detected without further modifications. Detected groups also include fluorescent and phosphorescent group.

[0057] the Term "secondary label"used in the present description, refers to groups such as Biotin and various antigens protein nature, requiring the presence of secondary intermediates to generate a detected signal. For Biotin secondary intermediates may include conjugates streptavidin-enzyme. For antigenic labels secondary intermediates may include the conjugates of enzyme-antibody. Some fluorescent groups act as secondary labels, as in the process of resonance energy transfer fluorescence (FRET, CHEF) they carry the charge to another functional group, and the detected signal is generated by the second group.

[0058] the Term "fluorescent label", "fluorescent dye" and "fluorophore"as used in the present description, refers to groups, absorbing the energy of the radiation at a certain wavelength excitation and radiant light energy on each the second wavelength. Examples of fluorescent labels include, but are not limited to, the following: Alexa Fluor dyes (Alexa Fluor 350, Alexa Fluor 488, Alexa Fluor 532, Alexa Fluor 546, Alexa Fluor 568, Alexa Fluor 594, Alexa Fluor 633, Alexa Fluor 660 and Alexa Fluor 680), AMCA, AMCA-S, BODIPY dyes (BODIPY FL, BODIPY R6G, BODIPY TMR, BODIPY TR, BODIPY 530/550, BODIPY 558/568, BODIPY 564/570, BODIPY 576/589, BODIPY 581/591, BODIPY 630/650, BODIPY 650/665), carboxyrhodamine 6G, carboxy-X-rhodamine (ROX), Cascade Blue, Cascade Yellow, coumarin 343, cyanine dyes (SS3, So, So, So), dansyl, doxil, dialkylaminoalkyl, 4',5'-dichloro-2',7'-dimethoxy-fluorescein, DM-NERF, eosin, erythrosine, fluorescein, FAM, hydroxycoumarin, IR Dyes (IRD 40, IRD 700 and IRD 800), JOE, lissamine-rhodamine, MarinaBlue, methoxycoumarin, naftiliaki, Oregon Green 488, Oregon Green 500, Oregon Green 514, Pacific Blue, PyMPO, pyrene, rhodamine B, rhodamine 6G, adaminaby green, adaminaby red, green rodol, 2',4',5',7'-Tetra-Braselton-fluorescein, tetramethyl-rhodamine (TMR), carboxytetramethyl (TAMRA, Texas Red, Texas Red-X.

[0059] the Term "mass tag"used in the present description, refers to any group that makes it possible to unambiguously detect on the basis of its mass using mass spectrometric (MS) detection methods. Examples of mass labels include labels with electroform release, such as N-[3-[4'-[(p-methoxycarbonylbenzyl)oxy] phenyl]-3-methylglycerol]isonicotinoyl acid, 4'-[2,3,5,6-titrator-4-(Pantothenate)]methyl acetophenone and derivatives thereof. Synthesis and application of such mass m the current is described in U.S. patent No. 4,650,750, 4,709,016, 5,360,8191, 5,516,931, 5,602,273, 5,604,104, 5,610,020 and 5,650,270. Other variants of mass labels include, but are not limited to the following examples: nucleotides, dideoxynucleotide, oligonucleotides of different length and nucleotide composition, oligopeptides, oligosaccharides and other synthetic polymers of different lengths and Monomeric composition. Also as mass labels, you can use a large variety of organic molecules, as it is not charged, and carrying charge (biomolecules or synthetic substances), the weight of which is in the range (100-2000 Yes).

[0060] the Terms "significant capacity" and "inhibits substantially"used in the present description, means a change, subject to measurement, the activity of HCV protease in the sample containing the compound according to the present invention or a composition of such compounds and the HCV protease, relative to the activity of HCV protease comprising an equivalent shape, do not contain these compounds or their compositions.

3. Examples of compounds according to the present invention:

[0061] According to some variants of the proposed compound of the formula I:

or pharmaceutically acceptable salt of the compounds where:

R1and R1'independently represent hydrogen or may contain substituents C1- aliphatic hydrocarbon, or R1and R1'together form may contain substituents 3-7-membered carbocyclic ring;

R2arepresents-IT or-NHSO2R2;

R2represents-N(R)2or possibly containing substituents group selected from the following:3-7cycloalkyl connected by a bridge Bicycle, 6-10-membered aryl, 5-10-membered heteroaryl containing 1-4 heteroatoms that are independently selected from nitrogen, oxygen, or sulfur, or 4-7-membered heterocycle containing 1-2 heteroatoms that are independently selected from nitrogen, oxygen, or sulfur;

each R independently represents hydrogen, possibly containing substituents With1-6aliphatic hydrocarbon, or:

two R together with the nitrogen atom to which they are attached, form a 4-7-membered heterocyclic ring containing 1-2 heteroatoms that are independently selected from nitrogen, oxygen, or sulfur;

R3represents-L-Y, where:

L is a covalent bond or a bivalent, saturated or unsaturated, unbranched or branched C1-8hydrocarbon chain, one, two, or three methylene unit of L can be independently substituted with cyclopropyl, -NR-, -N(R)C(O)-, -C(O)N(R)-, -N(R)SO2-, -SO2N(R)-, -O-, -C(O)-, -OC(O)-, -C(O)O-, -S-, -SO-, -SO2-, -C(=S)-, -C(=R), -N=N-, or-C(=N2)-;

Y represents hydrogen, C1-6the aliphatic hydrocarbon may contain as substituents oxoprop, halogen, NO2or CN; or 3-10-membered monocyclic or bicyclic, saturated, partially unsaturated, or aryl ring containing 0-3 heteroatoms that are independently selected from nitrogen, oxygen or sulfur, and where the specified ring contains as Vice-1-4 groups Re; and

each Reindependently selected from the following: -Q-Z, oxoprop, NO2, halogen, CN, an appropriate leaving group or a C1-6the aliphatic hydrocarbon may contain as substituents oxoprop, halogen, NO2or CN, where:

Q is a bivalent, saturated or unsaturated, unbranched or branched C1-6hydrocarbon chain, one, two, or three methylene unit of Q may be independently substituted with: -N(R)-, -S-, -O-, -C(O)-, -OC(O)-, -C(O)O-, -SO-, or-SO2-, -N(R)C(O)-, -C(O)N(R)-, -N(R)SO2- or-SO2N(R)-; and

Z represents hydrogen or C1-6the aliphatic hydrocarbon may contain as substituents oxoprop, halogen, NO2or CN; or

R3and the ring formed by R1and R1'together with in between the atoms form may contain a Vice is eating saturated or unsaturated 12-18 membered ring, containing 2-6 - heteroatoms, which are independently selected from nitrogen, oxygen or sulfur, and formed a ring contains a head group;

R4represents H, -NHC(O)R5, -NHC(O)OR6,or the side group of a natural or unnatural amino acids;

each R5independently represents-N(R)2or possibly containing substituents group selected from the following:1-6aliphatic hydrocarbon, connected by bridge Bicycle, 6-10-membered aryl, 5-10-membered heteroaryl containing 1-4 heteroatoms that are independently selected from nitrogen, oxygen, or sulfur, or 4-7-membered heterocyclyl containing 1-2 heteroatoms that are independently selected from nitrogen, oxygen, or sulfur;

R6represents possibly containing substituents group selected from the following:1-6aliphatic hydrocarbon, connected by bridge Bicycle, 6-10-membered aryl, 5-10-membered heteroaryl containing 1-4 heteroatoms that are independently selected from nitrogen, oxygen, or sulfur, or 4-7-membered heterocyclyl containing 1-2 heteroatoms that are independently selected from nitrogen, oxygen, or sulfur; and

R7represents possibly containing substituents group selected from the following examples:1-6aliphatic hydrocarbon, connected by bridge Bicycle, 6-10-member of the first aryl, 5-10-membered heteroaryl containing 1-4 heteroatoms that are independently selected from nitrogen, oxygen, or sulfur, or 4-7-membered heterocyclyl containing 1-2 heteroatoms that are independently selected from nitrogen, oxygen or sulfur.

RZrepresents; or R4and RZtogether with in between the atoms form may contain substituents saturated or unsaturated 16-22 membered ring containing 2 to 6 heteroatoms, which are independently selected from nitrogen, oxygen, or sulfur;

each of Ryindependently selected from the following: halogen, -ORo, -CN, -NO2, -N(Ro)2or possibly containing substituents With1-4aliphatic group; and

m is an integer from 0 to 4 inclusive;

s is an integer from 0 to 4 inclusive;

t is an integer from 0 to 4 inclusive;

moreover, the sum of s and t is not equal to zero.

[0062] In some embodiments of L is a covalent bond.

[0063] In some embodiments of L is a bivalent, saturated or unsaturated, straight or branched C1-8hydrocarbon chain, one, two, or three methylene units in structure L may be independently replaced by: cyclopropyl, -NR-, -N(R)C(O)-, -C(O)N(R)-, -N(R)SO2-, -SO2N(R)-, O-, -C(O)-, -OC(O)-, -C(O)O-, -S-, -SO-, -SO2-, -C(=S)-, -C(=NR)-, -N=N - or-C(=N2)-.

[0064] In some embodiments of L is a bivalent saturated or unsaturated, unbranched or branched C1-8the hydrocarbon chain. In some embodiments of L represents-CH2-.

[0065] In some embodiments of L is a covalent bond, -CH2-, -NH-, -CH2NH-, -NHCH2-, -NHC(O)-, -NHC(O)CH2OC(O)-, -CH2NHC(O)-, -NHSO2-, -NHSO2CH2-, -NHC(O)CH2OC(O)-, or-SO2NH-.

[0066] In some embodiments of L represents a divalent unbranched or branched C2-8hydrocarbon chain, and L contains at least one double bond and one or two additional methylene unit of L can be independently replaced by-NRC(O)-, -C(O)NR-, -N(R)SO2-, -SO2N(R)-, -S-, -S(O)-, -SO2-, -OC(O)-, -C(O)O-, -O-, -N(R)- or-C(O)-.

[0067] In some embodiments of L represents a divalent unbranched or branched C2-8hydrocarbon chain, and L contains at least one double bond, and at least one methylene unit of L are replaced by-C(O)-, -NRC(O)-, -C(O)NR-, -N(R)SO2-, -SO2N(R)-, -S-, -S(O)-, -SO2-, -OC(O)-, or-C(O)O-, and one or two additional methyl is a new structural unit L may be independently substituted with cyclopropyl, -O-, -N(R)- or-C(O)-.

[0068] In some embodiments of L represents a divalent unbranched or branched C2-8hydrocarbon chain, and L contains at least one double bond, and at least one methylene unit of L are replaced by-C(O)-, and one or two additional methylene unit of L may be independently substituted with cyclopropyl, -O-, -N(R)-, or-C(O)-.

[0069] as described above, in some embodiments of L represents a divalent unbranched or branched C2-8hydrocarbon chain, and L contains at least one double bond. An ordinary person skilled in the art it will be clear that this double bond may be located within the hydrocarbon chain, and can be "Exo-relationship with respect to the hydrocarbon skeleton, forming, thus, alkylidene group. As an example of such a group L containing alkylidene branched chain comprising-CH2C(=CH2)CH2-. Thus, in some embodiments of L represents a divalent unbranched or branched hydrocarbon chain C2-8where L contains at least one alkylidene double bond. Examples of groups L on the ut-NHC(O)C(CH 2)CH2-.

[0070] In some embodiments of L represents a divalent unbranched or branched hydrocarbon chain With2-8where at least one methylene unit within L replaced by-C(O)-. In some embodiments of L represents-C(O)CH=CH(CH3)-, C(O)CH=CHCH2NH(CH3)-, -C(O)CH=CH(CH3)-, -C(O)CH=CH-, -CH2C(O)CH=CH-, -CH2C(O)CH=CH(CH3)-, -CH2CH2C(O)CH=CH-, -CH2CH2C(O)CH=SNSN2-, CH2CH2C(O)CH=CHCH2NH(CH3)-, or-CH2-CH2C(O)CH=CH(CH3)-, or-CH(CH3)OS(O)CH=CH-.

[0071] In some embodiments of L represents a divalent unbranched or branched C2-8hydrocarbon chain, and L contains at least one double bond, and at least one methylene unit within L replaced by-OC(O)-.

[0072] In some embodiments of L represents a divalent unbranched or branched C2-8hydrocarbon chain, and L contains at least one double bond, and at least one methylene unit within L replaced by-NRC(O)-, -C(O)NR-, -N(R)SO2-, -SO2N(R)-, -S-, -S(O)-, -SO2-, -OC(O)-, or-C(O)O-, and one or two additional methylene units in structure L may not be avisio substituted cyclopropyl, -O-, -N(R)- or-C(O)-. In some embodiments of L represents-CH2OC(O)CH=SNSN2-, -CH2-OC(O)CH=CH - or-CH(CH=CH2)OS(O)CH=CH-.

[0073] In some embodiments of L is a-NRC(O)CH-CH-, -NRC(O)CH=CHCH2N(CH3)-, -NRC(O)CH=CHCH2O-, -CH2NRC(O)CH=CH-, -NRSO2CH=CH-, -NRSO2CH=CHCH2-, -NRC(O)(C=N2)C(O)-, -NRC(O)CH=CHCH2N(CH3)-, -NRSO2CH=CH-, -NRSO2CH-CHCH2-, -NRC(O)CH=CHCH2O-, -NRC(O)C(=CH2)CH2-, -CH2NRC(O)-, -CH2NRC(O)CH=CH-, -CH2CH2NRC(O)- or-CH2NRC(O)cyclopropyl-, where each R independently represents hydrogen or may contain substituents With1-6aliphatic group.

[0074] In some embodiments of L represents-NHC(O)CH=CH-, -NHC(O)CH=CHCH2N(CH3)-, -NHC(O)CH=CHCH2O-, -CH2NHC(O)CH=CH-, -NHSO2CH=CH-, -NHSO2CH=CHCH2-, -NHC(O)(C=N2)C(O)-, -NHC(O)CH=CHCH2N(CH3)-, -NHSO2CH-CH-, -NHSO2CH=CHCH2-, -NHC(O)CH=CHCH2O-, -NHC(O)C(=CH2)CH2-, -CH2NHC(O)-, -CH2NHC(O)CH=CH-, -CH2CH2NHC(O)- or-CH2NHC(O)cyclopropyl-.

[0075] In some embodiments of L represents a divalent unbranched or branched C2-8hydrocarbon chain, and L contains at least one triple bond. In some embodiments of L is a TLD is the valence unbranched or branched C 2-8hydrocarbon chain, and L contains at least one triple bond and one or two additional methylene unit of L can be independently replaced by-NRC(O)-, -C(O)NR-, -S-, -S(O)-, -SO2-, -C(=S)-, -C(=NR)-, -O-, -N(R)- or-C(O)-. In some embodiments of L contains at least one triple bond, and at least one methylene unit of L is substituted by-N(R)-, -N(R)C(O)-, -C(O)-, -C(O)O-, or-OC(O)-, or-O-.

[0076] Examples of groups L include-C≡C-, -C≡CLO2N(isopropyl)-, -NHC(O)C≡CCH2CH2-, -CH2-C≡C-CH2-, -C≡CCH2O-, -CH2C(O)≡C-, -C(O)≡C - or-CH2OC(=O)≡C-.

[0077] In some embodiments of L represents a divalent unbranched or branched C2-8hydrocarbon chain, and one methylene unit of L is replaced by cyclopropyl, bottom and one or more methylene unit of L is independently replaced by-C(O)-, -NRC(O)-, -C(O)NR-, -N(R)SO2- or-SO2N(R)-. Examples of groups L include-NHC(O)-cyclopropyl-SO2- , and-NHC(O)-cyclopropyl-.

[0078] As broadly defined above, Y represents hydrogen, C1-6the aliphatic hydrocarbon may contain as substituents oxoprop, halogen, NO2or CN, or a 3-10 membered monocyclic or bicyclic, saturated, h is ichno unsaturated or aryl ring, containing 0-3 heteroatoms that are independently selected from nitrogen, oxygen or sulfur, and where the specified ring contains as Vice-1-4 groups Rewhere each Reindependently selected from the following: -Q-Z, oxoprop, NO2, halogen, CN, an appropriate leaving group, or With1-6aliphatic group, where Q is a covalent bond or a bivalent saturated or unsaturated, unbranched or branched C1-6hydrocarbon chain, and one or two methylene unit of Q may be independently replaced by-N(R)-, -S-, -O-, -C(O)-, -OC(O)-, -C(O)O-, -SO - or-SO2-, -N(R)C(O)-, -C(O)N(R)-, -N(R)SO2- or-SO2N(R)-; and Z represents hydrogen or a hydrocarbon chain With1-6may contain as substituents oxoprop, halogen, NO2or CN.

[0079] In some embodiments of Y represents hydrogen.

[0080] In some embodiments of Y represents a C1-6the aliphatic group may contain as substituents oxoprop, halogen, NO2or CN. In some embodiments of Y represents a C2-6alkenyl may contain as substituents oxoprop, halogen, NO2or CN. In other embodiments of Y represents a C2-6quinil may contain as the Deputy is the Fort worth oxoprop, halogen, NO2or CN. In some embodiments of Y represents a C2-6alkenyl. In other embodiments of Y represents a C2-4quinil.

[0081] In other embodiments of Y represents a C1-6alkyl containing as substituents oxoprop, halogen, NO2or CN. Such groups Y include-CH2F, -CH2Cl, -CH2CN and CH2NO2.

[0082] In some embodiments of Y is a saturated 3-6 membered ring containing 0-3 heteroatoms that are independently selected from nitrogen, oxygen or sulfur, and Y contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application.

[0083] In some embodiments of Y is a saturated 3-4 membered heterocyclic ring containing 1 heteroatom selected from oxygen or nitrogen, with the specified ring contains as Vice-1-2 groups Rewhere each group Redefined above and described in the text of this application. Examples of such rings are epoxy and oxetanone rings, where each ring contains as Vice-1-2 groups Rewhere each group Redefined above and described in the text of this application.

[0084] In other embodiments of Y is the Wallpaper saturated 5-6-membered heterocyclic ring, containing 1-2 heteroatoms selected from oxygen and nitrogen, with the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application. Examples of such rings include piperidine and pyrolidine ring, where each ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application. In some embodiments of Y isorwhere each of R, Q, Z and Rematch the specific above and described in the text of this application.

[0085] In some embodiments of Y is a saturated 3-6 membered carbocyclic ring, with the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application. In some embodiments of Y is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, and each ring contains as Vice-1-4 groups Rewhere each Redefined above and described in the text of this application. In some embodiments of Y iswhere Redefined above and described in exce of this application. In some embodiments of Y is cyclopropyl may contain as substituents halogen, CN or NO2.

[0086] In some embodiments of Y represents a partially unsaturated 3-6 membered monocyclic ring containing 0-3 heteroatoms that are independently selected from nitrogen, oxygen or sulfur, with the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application.

[0087] In some embodiments of Y represents a partially unsaturated 3-6 membered carbocyclic ring, with the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this "proposal. In some embodiments of Y is cyclopropyl, cyclobutyl, cyclopentenyl, or cyclohexenyl, and each ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application. In some embodiments of Y iswhere each group Redefined above and described in the text of this application.

[0088] In some embodiments of Y represents a partially unsaturated 4-6 membered gets aziklicescoe ring, containing 1-2 heteroatoms that are independently selected from nitrogen, oxygen or sulfur, with the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application. In some embodiments of Y is selected from the following examples:

where each R and Redefined above and described in the text of this application.

[0089] In some embodiments of Y is a 6-membered aromatic ring containing 0-2 nitrogen atoms, with the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application. In some embodiments of Y is phenyl, pyridyl or pyrimidinyl, where each ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application.

[0090] In some embodiments of Y is selected from the following examples:

where each Redefined above and described in the text of this application.

[0091] In other embodiments of Y represents a 5-membered heteroaryl ring containing 1-3 heteroatoms which are independently selected from nitrogen, oxygen or sulfur, moreover, the specified ring contains as Vice-1-3 group, R ewhere each Redefined above and described in the text of this application. In some embodiments of Y represents a 5-membered partially unsaturated or aryl ring, containing 1-3 heteroatoms which are independently selected from nitrogen, oxygen or sulfur, with the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application. Examples of such rings include isoxazolyl, oxadiazol, thiazolyl, imidazolyl, pyrazolyl, pyrrolyl, furanyl, thienyl, triazole, thiadiazole and oxadiazole, and each ring contains as Vice-1-3 group, Rewhere each group Redefined above and described in the text of this application. In some embodiments of Y is selected from the following examples:

where each R and Redefined above and described in the text of this application.

[0092] In some embodiments of Y is an 8-10-membered bicyclic, saturated, partially unsaturated, or aryl ring containing 0-3 heteroatoms that are independently selected from nitrogen, oxygen or sulfur, with the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application. According to another what aspect of the present invention Y is a 9-10-membered bicyclic, partially unsaturated, or aryl ring containing 1-3 heteroatoms which are independently selected from nitrogen, oxygen or sulfur, where the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application. Examples of such bicyclic rings include 2,3-dihydrobenzo[d]isothiazol where the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application.

[0093] As broadly defined above, each Reindependently selected from the following examples: -Q-Z, oxoprop, NO2, halogen, CN, an appropriate leaving group, or With1-6the aliphatic group may contain as substituents oxoprop, halogen, NO2or CN, where Q is a covalent bond or a bivalent saturated or unsaturated, unbranched or branched C1-6hydrocarbon chain, and one or two methylene structural unit within Q may be independently replaced by-N(R)-, -S-, -O-, -C(O)-, -OC(O)-, -C(O)O-, -SO-, or-SO2-, -N(R)C(O)-, -C(O)N(R)-, -N(R)SO2- or-SO2N(R)-; and Z represents hydrogen or an aliphatic group C1-6may contain as substituents oxoprop, halogen, NO2or CN.

[0094]In some embodiments of R erepresents a C1-6the aliphatic group may contain as substituents oxoprop, halogen, NO2or CN. In other embodiments of Reis oxoprop, NO2, halogen or CN.

[0095] In some embodiments of Rerepresents-Q-Z, where Q is a covalent bond and Z is hydrogen (i.e. Reis hydrogen). In other embodiments of Rerepresents-Q-Z, where Q is a divalent saturated or unsaturated, unbranched or branched C1-6hydrocarbon chain, and one or two methylene unit within Q may be independently replaced by-NR-, -NRC(O)-, -C(O)NR-, -S-, -O-, -C(O)-, -SO - or-SO2-. There are also options for implementation according to which Q represents a divalent unbranched or branched C2-6hydrocarbon chain containing at least one double bond, one or two methylene unit within Q may be independently replaced by-NR-, -NRC(O)-, -C(O)NR-, -S-, -O-, -C(O)-, -SO - or-SO2-. In some embodiments of Z in the group Rerepresents hydrogen. In some embodiments of Q-Z-represents-NHC(O)CH=CH2or-C(O)CH=CH2.

[0096] In some embodiments of each Reindependently selected from the following is th: oxoprop, NO2, CN, fluorine, chlorine, -NHC(O)CH=CH2, -C(O)CH=CH2, -CH2CH=CH2, -C≡CH, -C(O)OCH2Cl, -C(O)OCH2F, -C(O)OCH2CN, -C(O)CH2Cl, -C(O)CH2F, -C(O)CH2CN or-CH2C(O)CH3.

[0097] In some embodiments of Rerepresents a suitable leaving group, i.e. a group which is subjected to nucleophilic substitution. "Suitable leaving group" is a chemical group which is easily possible to substitute coming on the desired chemical functional group, for example, thiol group or a cysteine according to the present invention. Appropriate leaving groups are well known in this field, for example, see "Advanced Organic Chemistry," Jerry March, 5thEd., pp.351-357, John Wiley and Sons, N.Y. Such leaving groups include, but are not limited to the following examples: halogen, alkoxy, sulfonyloxy-; possibly containing substituents of alkylsulfonate-; possibly containing substituents of alkanesulphonic-; may contain substituents arylsulfonate, acyl and Diaconia functional groups. Examples of appropriate leaving groups include the following groups: chlorine-, iodine-, bromine-, fluorine-, acetoxy-, methanesulfonate (mesilate-), tosyloxy, triplelux, Petro-phenolsulfonate (nonyloxy-and bromine-phenolsulfonate (brasilense-group).

[0098] In some implementations use the following implementation options and combinations-L-Y:

(a) L is a divalent unbranched or branched C2-8hydrocarbon chain, where L contains at least one double bond and one or two additional methylene unit in the structure L may be independently replaced by-NRC(O)-, -C(O)NR-, -N(R)SO2-, -SO2N(R)-, -S-, -S(O)-, -SO2-, -OC(O)-, -C(O)O-, cyclopropyl, -O-, -N(R)- or-C(O)-; and Y represents hydrogen or an aliphatic group With1-6may contain as substituents oxoprop, halogen, NO2or CN; or

(b) L is a divalent unbranched or branched C2-8hydrocarbon chain, and L contains at least one double bond, and at least one methylene unit in the composition of L are replaced by-C(O)-, -NRC(O)-, -C(O)NR-, -N(R)SO2-, -SO2N(R)-, -S-, -S(O)-, -SO2-, -OC(O)-, or-C(O)O-, and one or two additional methylene unit in the structure L may be independently substituted with cyclopropyl, -O-, -N(R)- or-C(O)-; and Y represents hydrogen or C1-6the aliphatic group may contain as substituents oxoprop, halogen, NO2or CN; or

(c) L is a divalent unbranched or branched C2-81-6the aliphatic group may contain as substituents oxoprop, halogen, NO2or CN; or

(d) L is a divalent unbranched or branched C2-8hydrocarbon chain, and L contains at least one double bond, and at least one methylene unit in the composition of L are replaced by-C(O)-; and Y represents hydrogen or C1-6the aliphatic group may contain as substituents oxoprop, halogen, NO2or CN; or

(e) L is a divalent unbranched or branched C2-8hydrocarbon chain, and L contains at least one double bond, and at least one methylene unit in the composition of the L substituted with-OC(O)-; and Y represents hydrogen or C1-6the aliphatic group may contain as substituents oxoprop, halogen, NO2or CN; or

(f) L is a-NRC(O)CH=CH-, -NRC(O)CH=CHCH2N(CH3)-, -NRC(O)CH=CHCH2O-, -CH2/sub> NRC(O)CH=CH-, -NRSO2CH=CH-, -NRSO2CH=CHCH2-, -NRC(O)(C=N2)-, -NRC(O)(C=N2)C(O)-, -NRC(O)CH=CHCH2N(CH3)-, -NRSO2CH=CH-, -NRSO2CH=CHCH2-, -NRC(O)CH=CHCH2O-, -NRC(O)C(=CH2)CH2-, -CH2NRC(O)-, -CH2NRC(O)CH=CH-, -CH2CH2NRC(O)- or-CH2NRC(O)cyclopropyl-; where R represents H or possibly containing substituents C1-6aliphatic group; and Y represents hydrogen or C1-6the aliphatic group may contain as substituents oxoprop, halogen, NO2or CN; or

(g), L represents-NHC(O)CH=CH-, -NHC(O)CH=CHCH2N(CH3)-, -NHC(O)CH=CHCH2O-, -CH2NHC(O)CH=CH-, -NHSO2CH=CH-, -NHSO2CH=CHCH2-, -NHC(O)(C=N2)-, -NHC(O)(C=N2)C(O)-, -NHC(O)CH=CHCH2N(CH3)-, -NHSO2CH=CH-, -NHSO2CH=CHCH2-, -NHC(O)CH=CHCH2O-, -NHC(O)C(=CH2)CH2, -CH2NHC(O)-, -CH2NHC(O)CH=CH-, -CH2CH2NHC(O)- or-CH2NHC(O)cyclopropyl-; and Y represents hydrogen or C1-6the aliphatic group may contain as substituents oxoprop, halogen, NO2or CN; or

(h) L is a divalent unbranched or branched C2-8hydrocarbon chain, and L contains at least one alkylidene double bond, and at least one methylene unit in the composition of L are replaced by-C(O)-, -NRC(O)-, -C(O)NR-, -(R)SO 2-, -SO2N(R)-, -S-, -S(O)-, -SO2-, -OC(O)-or-C(O)O-, and one or two additional methylene unit in the structure L may be independently substituted with cyclopropyl, -O-, -N(R)- or-C(O)-; and Y represents hydrogen or C1-6the aliphatic group may contain as substituents oxoprop, halogen, NO2or CN; or

(i) L is a divalent unbranched or branched C2-8hydrocarbon chain, and L contains at least one triple bond and one or two additional methylene unit in the structure L may be independently replaced by-NRC(O)-, -C(O)NR-, -N(R)SO2-, -SO2N(R)-, -S-, -S(O)-, -SO2-, -OC(O)- or-C(O)O-, and Y represents hydrogen or C1-6the aliphatic group may contain as substituents oxoprop, halogen, NO2or CN; or

(j) L is a-C≡C-, -C≡CCH2N(isopropyl)-, -NHC(O)C≡CCH2CH2-, -CH2-C≡C-CH2-, -C≡CCH2O-, -CH2C(O)≡C-, -C(O)≡C-, or-CH2OC(=O)≡C-; and Y represents hydrogen or C1-6the aliphatic group may contain as substituents oxoprop, halogen, NO2or CN; or

(k) (L represents a divalent unbranched or branched C2-8hydrocarbon chain, and one methylene unit in the composition of the e L replaced by cyclopropyl, and one or two additional methylene unit in the structure of L is independently replaced by-NRC(O)-, -C(O)NR-, -N(R)SO2-, -SO2N(R)-, -S-, -S(O)-, -SO2-, -OC(O)- or-C(O)O-; and Y represents hydrogen or C1-6the aliphatic group may contain as substituents oxoprop, halogen, NO2or CN; or

(l) L is a covalent bond, and Y is selected from the following examples:

(i)1-6alkyl containing as substituents oxoprop, halogen, NO2or CN;

(ii)2-6alkenyl may contain as substituents oxoprop, halogen, NO2or CN; or

(iii)2-6quinil may contain as substituents oxoprop, halogen, NO2or CN; or

(iv) a saturated 3-4 membered heterocyclic ring containing 1 heteroatom selected from oxygen and nitrogen, with the specified ring may contain as substituents 1-2 groups Rewhere each group Redefined above and described in the text of this application; or

(v) a saturated 5-6-membered heterocyclic ring containing 1-2 heteroatoms selected from oxygen and nitrogen, with the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application; or

(vi) orwhere each of R, Q, Z and Ryas defined above and described in the text of this application; or

(vii) a saturated 3-6 membered carbocyclic ring, with the specified ring contains as Vice-1-4 groups Reand each group Redefined above and described in the text of this application; or

(viii) a partially unsaturated 3-6 membered monocyclic ring containing 0-3 heteroatoms that are independently selected from nitrogen, oxygen or sulfur, with the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application; or

(ix) a partially unsaturated 3-6 membered carbocyclic ring, with the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application; or

(x)where each group Redefined above and described in the text of this application; or

(xi) a partially unsaturated 4-6 membered heterocyclic ring containing 1-2 heteroatoms that are independently selected from nitrogen, oxygen or sulfur, with the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application; or

xii) or

where each R and Rymatch the specific above and described in the text of this application; or

(xiii) a 6-membered aromatic ring containing 0-2 nitrogen atoms, with the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application;

(xiv)

where each Redefined above and described in the text of this application; or

(xv) a 5-membered heteroaryl ring containing 1-3 heteroatoms which are independently selected from nitrogen, oxygen or sulfur, with the specified ring contains as Vice-1-3 group, Rewhere each group Redefined above and described in the text of this application; or

where each Redefined above and described in the text of this application; or

(xvii) an 8-10 membered bicyclic, saturated, partially unsaturated, or aryl ring containing 0-3 heteroatoms that are independently selected from nitrogen, oxygen or sulfur, where the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application;

(m) L represents-C(O)-, and Y is selected from the following examples:

() 1-6alkyl containing as substituents oxoprop, halogen, NO2or CN; or

(ii)2-6alkenyl may contain as substituents oxoprop, halogen, NO2or CN; or

(iii)2-6quinil may contain as substituents oxoprop, halogen, NO2or GN; or

(iv) a saturated 3-4 membered heterocyclic ring containing 1 heteroatom selected from oxygen and nitrogen, with the specified ring contains as Vice-1-2 groups Rewhere each group Redefined above and described in the text of this application; or

(v) a saturated 5-6-membered heterocyclic ring containing 1-2 heteroatoms selected from oxygen and nitrogen, where the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application; or

(vi)orwhere each group R, Q, Z and Redefined above and described in the text of this application; or

(vii) a saturated 3-6 membered carbocyclic ring, with the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application; or

(viii) a partially unsaturated 3-6 membered monocyclic ring, sod is rashee 0-3 heteroatoms, are independently selected from nitrogen, oxygen or sulfur, with the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application; or

(ix) a partially unsaturated 3-6 membered carbocyclic ring, with the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application; or

(x)where each group Redefined above and described in the text of this application; or

(xi) a partially unsaturated 4-6 membered heterocyclic ring containing 1-2 heteroatoms that are independently selected from nitrogen, oxygen or sulfur, with the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application; or

(xii)or

where each of the groups R and Redefined above and described in the text of this application; or

(xiii) a 6-membered aromatic ring content in its composition 0-2 nitrogen atoms, with the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application; or

(xiv)

where each Redefined above and described in the text of this application; or

(xv) a 5-membered heteroaryl ring containing 1-3 heteroatoms which are independently selected from nitrogen, oxygen or sulfur, with the specified ring contains as Vice-1-3 group, Rewhere each group Redefined above and described in the text of this application; or

(xvi)

where each Redefined above and described in the text of this application; or

(xvii) an 8-10 membered bicyclic saturated, partially unsaturated, or aryl ring, with the specified ring contains as Vice-1-4 groups Rewhere the group Redefined above and described in the text of this application;

(n) L represents-N(R)C(O)-, and Y is selected from the following examples:

(i) alkyl With1-6containing as substituents oxoprop, halogen, NO2or CN; or

(ii)2-6alkenyl may contain as substituents oxoprop, halogen, NO2or CN; or

(iii) C2-6quinil may contain as substituents oxoprop, halogen, NO2or CN; or

(iv) a saturated 3-4 membered heterocyclic ring containing 1 heteroatom selected from oxygen or nitrogen, with the specified ring contains as Vice-1-2 gr is PI R ewhere each group Redefined above and described in the text of this application; or

(v) a saturated 5-6-membered heterocyclic ring containing 1-2 heteroatoms selected from oxygen and nitrogen, with the specified ring contains as Vice-1-4 groups R0where each group R0defined above and described in the text of this application; or

(vi)orwhere each group R, Q, Z and Reor defined above and described in the text of this application; or

(vii) a saturated 3-6 membered carbocyclic ring, with the specified ring contains as Vice-1-4 groups Reand each group Redefined above and described in the text of this application; or

(viii) a partially unsaturated 3-6 membered monocyclic ring containing 0-3 heteroatoms that are independently selected from nitrogen, oxygen or sulfur, with the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application; or

(ix) a partially unsaturated 3-6 membered carbocyclic ring, with the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application; or

(x) where each group Redefined above and described in the text of this application; or

(xi) a partially unsaturated 4-6 membered heterocyclic ring containing 1-2 heteroatoms that are independently selected from nitrogen, oxygen or sulfur, with the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application; or

(xii)or

where each of the groups R and Redefined above and described in the text of this application; or

(xiii) a 6-membered aromatic ring containing 0-2 nitrogen atoms, with the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application; or

(xiv)

where each Redefined above and described in the text of this application; or

(xv) a 5-membered heteroaryl ring containing 1-3 heteroatoms which are independently selected from nitrogen, oxygen or sulfur, with the specified ring contains as Vice-1-3 group, Rewhere each group Redefined above and described in the text of this application; or

(xvi)

where each of the groups R and Redefined above and described in the text nastoyasheva; or

(xvii) an 8-10 membered bicyclic saturated, partially unsaturated, or aryl ring containing 0-3 heteroatoms that are independently selected from nitrogen, oxygen or sulfur, with the specified ring contains as Vice-1-4 groups Rewhere the group Redefined above and described in the text of this application;

(o) L is a bivalent saturated or unsaturated, unbranched or branched C1-8hydrocarbon chain; and Y is selected from the following examples:

(i)1-6alkyl; containing as substituents oxoprop, halogen, NO2or CN;

(ii)2-6alkenyl may contain as substituents oxoprop, halogen, NO2or CN; or

(iii)2-6quinil may contain as substituents oxoprop, halogen, NO2or CN; or

(iv) a saturated 3-4 membered heterocyclic ring containing 1 heteroatom selected from oxygen and nitrogen, with the specified ring contains as Vice-1-2 groups Rewhere each group Redefined above and described in the text of this application; or

(v) a saturated 5-6-membered heterocyclic ring containing 1-2 heteroatoms selected from oxygen and nitrogen, with the specified ring contains as Vice-1-4 groups Rewhere each of the groups which R edefined above and described in the text of this application; or

(vi)orwhere each group R, Q, Z and Redefined above and described in the text of this application; or

(vii) a saturated 3-6 membered carbocyclic ring, with the specified ring contains as Vice-1-4 groups Rewhere each Redefined above and described in the text of this application; or

(viii) a partially unsaturated 3-6 membered monocyclic ring containing 0-3 heteroatoms that are independently selected from nitrogen, oxygen or sulfur, with the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application; or

(ix) a partially unsaturated 3-6 membered carbocyclic ring, with the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application; or

(x)where each group Redefined above and described in the text of this application; or

(xi) a partially unsaturated 4-6 membered heterocyclic ring containing 1-2 heteroatoms that are independently selected from nitrogen, oxygen or sulfur, with the specified ring contains as Vice-1-4 groups ewhere each group Redefined above and described in the text of this application; or

(xii)

where each of the groups R and Redefined above and described in the text of this application; or

(xiii) a 6-membered aromatic ring containing 0-2 nitrogen atoms, with the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application; or

(xiv)

where each Redefined above and described in the text of this application; or

(xv) a 5-membered heteroaryl ring containing 1-3 heteroatoms which are independently selected from nitrogen, oxygen or sulfur, with the specified ring contains as Vice-1-3 group, Rewhere each group Redefined above and described in the text of this application; or

(xvi)

where each of the groups R and Redefined above and described in the text of this application; or

(xvii) an 8-10 membered bicyclic saturated, partially unsaturated, or aryl ring containing 0-3 heteroatoms that are independently selected from nitrogen, oxygen or sulfur, with the specified ring contains as Vice-1-4 groups Rewhere the group Redefined above and described in the text of the crust is of the present application;

(p) L is a covalent bond, -CH2-, -NH-, -C(O)-, -CH2NH-, -NHCH2-, -NHC(O)-, -NHC(O)CH2OC(O)-, -CH2NHC(O)-, -NHSO2-, -NHSO2CH2-, -NHC(O)CH2OC(O)- or-SO2NH-; and Y is selected from the following examples:

(i)1-6alkyl containing as substituents oxoprop, halogen, NO2or CN; or

(ii)2-6alkenyl may contain as substituents oxoprop, halogen, NO2or CN; or

(iii)2-6quinil may contain as substituents oxoprop, halogen, NO2or CN; or

(iv) a saturated 3-4 membered heterocyclic ring containing 1 heteroatom selected from oxygen and nitrogen, with the specified ring contains as Vice-1-2 groups Rewhere each group Redefined above and described in the text of this application; or

(v) a saturated 5-6-membered heterocyclic ring containing 1-2 heteroatoms selected from oxygen and nitrogen, with the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application; or

(vi)orwhere each group R, Q, Z and Redefined above and described in the text of this application; or

(vii) a saturated 3-6 membered carbocyclization, moreover, the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application; or

(viii) a partially unsaturated 3-6 membered monocyclic ring, 0-3 heteroatoms which are independently selected from nitrogen, oxygen or sulfur, where the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application; or

(ix) a partially unsaturated 3-6 membered carbocyclic ring, with the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application; or

(x)where each group Redefined above and described in the text of this application; or

(xi) a partially unsaturated 4-6 membered heterocyclic ring containing 1-2 heteroatoms that are independently selected from nitrogen, oxygen or sulfur, with the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application; or

(xii)or

where each of the groups R and Redefined above and described in the text of this application; or

(xiii) a 6-membered aromatics the second ring, containing 0-2 nitrogen atoms, with the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application; or

(xiv)

where each Redefined above and described in the text of this application; or

(xv) a 5-membered heteroaryl ring containing 1-3 heteroatoms which are independently selected from nitrogen, oxygen or sulfur, where the specified ring contains as Vice-1-3 group, Rewhere each group Redefined above and described in the text of this application; or

(xvi)

where each of the groups R and Redefined above and described in the text of this application; or

(xvii) an 8-10 membered bicyclic saturated, partially unsaturated, or aryl ring containing 0-3 heteroatoms that are independently selected from nitrogen, oxygen or sulfur, with the specified ring contains as Vice-1-4 groups Rewhere each group Redefined above and described in the text of this application.

[0099] In certain embodiments of the group Y of formula I is selected from the groups represented in Table 1 below, where each wavy line indicates the attachment point to the rest of the molecule.

Table 1. Examples of Groups Formula I:

where each Reindependently represents an appropriate leaving group, NO2CN or oxoprop.

[00100] In some embodiments of the group R3according to the formula I selected from the groups shown in Table 2, below, where each wavy line indicates the attachment point to the rest of the molecule.

Table 2. Examples of Groups R3:

where each Reindependently represents an appropriate leaving group, NO2CN or oxoprop.

[00101] In some embodiments of group R1and R1'formula I independently represent hydrogen or may contain substituents With1-6aliphatic group. In some embodiments of R1represents hydrogen and R1'represents a C1-4aliphatic group. In other embodiments of R1represent hydrogen and R1'represents n-propyl.

[00102] In some embodiments of group R1and R1'formula I together form may contain substituents 3-7-membered carbocycle the mini-ring. In some embodiments of group R1and R1'formula I together form may contain substituents cyclopropene ring. In some embodiments of group R1and R1'formula I together form cyclopropyl ring containing as substituents ethyl or vinyl.

[00103] In some embodiments of R4represents H, -NHC(O)R5, -NHC(O)OR6,; or R4and RZtogether with in between the atoms form may contain substituents, saturated or unsaturated 16-22 membered ring containing 2 to 6 heteroatoms, which are independently selected from nitrogen, oxygen or sulfur.

[00104] In certain embodiments of the group R4formula I represents-NHC(O)R5. In some embodiments of the group R4formula I represents-NHC(O)OR6. In other embodiments of the group R4formula I represents. In certain embodiments of the group R4formula I is hydrogen.

[00105] There are options for implementation according to which, when R4represents-NHC(O)R5, R5represents a C1-6aliphatic group, or may contain substituents group selected from the following examples: connect the config bridge Bicycle, 5-10-membered heteroaryl containing 1-4 heteroatoms that are independently selected from nitrogen, oxygen, or sulfur, or 4-7-membered heterocycle containing 1-2 heteroatoms that are independently selected from nitrogen, oxygen or sulfur.

[00106] There are options for implementation according to which, if R4represents-NHC(O)R6, R6represents a C1-6aliphatic group, or may contain substituents group selected from the following examples: connected the bridge Bicycle, 5-10-membered heteroaryl containing 1-4 heteroatoms that are independently selected from nitrogen, oxygen, or sulfur, or 4-7-membered heterocycle containing 1-2 heteroatoms that are independently selected from nitrogen, oxygen or sulfur.

[00107] In some embodiments of the group R4formula I represents a side group of the amino acids. In some embodiments of the group R4formula I represents a side group of non-natural amino acids. In some embodiments of the group R4formula I is an aliphatic side group of non-natural amino acids. In some embodiments of the group R4formula I represents a side group unnatural alanine containing as substituents one, two or three groups of Rowhere each Rocorresponds to the definition is built up. In some embodiments of the group R4formula I represents a side group of a natural amino acid.

[00108] In certain embodiments of the group R4formula I represents a side group of a natural alanine (i.e. R4represents methyl). In some embodiments of the group R4formula I represents a side group of a natural D-alanine. There are options for implementation under which the group R4formula I represents a side group of a natural L-alanine.

[00109] In other embodiments of the group R4formula I represents a side group of a natural valine. In some embodiments of the group R4formula I represents a side group of a natural balance of D-valine. In some embodiments of the group R4formula I represents a side group of a natural L-valine.

[00110] In some embodiments of the group R4formula I consists of a mixture of side groups of the amino acids in the D-and L-configurations. Such groups R4referred to in the present description D,L-mixed side groups of amino acids". In some embodiments of the ratio of the D-configuration of the side groups of amino acids to L-configuration selected from the following relations: 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4; 1:5 and 1:6. Thus, in some is that embodiments of the group R 4formula I is D,L-mixed side group of alanine. In other embodiments of the group R4formula I is D,L-mixed side group of valine.

[00111] Without limitation to any specific theory, believe that for compounds of formula I of the side groups of amino acids are in the D-configuration, it is advisable to prevent the adoption of the substance of spatial orientation, facilitating the binding of HCV protease.

[00112] In some embodiments of group R5and R7formula I independently represent possibly containing substituents of group selected from the following examples: possibly containing substituents, C1-6aliphatic group, 6-10-membered aryl, 5-10-membered heteroaryl containing 1-4 heteroatoms that are independently selected from nitrogen, oxygen, or sulfur, or 4-7-membered heterocyclyl containing 1-2 heteroatoms that are independently selected from nitrogen, oxygen or sulfur. In some embodiments of R5represents possibly containing substituents 5-10-membered heteroaryl containing 1-4 heteroatoms that are independently selected from nitrogen, oxygen or sulfur, and R7represents possibly containing substituents With1-6aliphatic group. There are implementations in which R5represents and R7is cyclohexyl.

[00113] In some embodiments of R4represents-NHC(O)R5where R5independently represents-N(R)2or possibly containing substituents group selected from C1-6aliphatic groups. In some embodiments of R5represents-N(R)2and each R independently represents hydrogen may contain substituents With1-6aliphatic group, or two Ry together with the nitrogen atom to which they are attached, form a 4-7-membered heterocyclic ring containing 1-2 heteroatoms that are independently selected from nitrogen, oxygen or sulfur. In some embodiments of R5represents-N(R)2and each R independently represents hydrogen or t-butyl.

[00114] In certain embodiments of the group R5formula I represents a possible containing substituents 5-10-membered heteroaryl ring containing 1-4 heteroatoms that are independently selected from nitrogen, oxygen or sulfur. In some embodiments of R5represents possibly containing substituents 6-membered heteroaryl, contain 1 to 2 nitrogen atoms. In some embodiments of R5represents piperazinil.

[00115] In some embodiments of the groups which R 7formula I represents a possible containing substituents C1-6aliphatic group. In some embodiments of R7represents a branched alkyl group With1-5. In other embodiments of R7is cyclopentyl or cyclohexyl.

[00116] In some embodiments of RZrepresents

In some embodiments of RZrepresents.

In some embodiments of RZrepresents.

In some embodiments of RZrepresents.

In some embodiments of RZrepresents.

In some embodiments of RZrepresents.

In some embodiments of RZrepresents.

In some embodiments of RZrepresents.

In some embodiments of RZrepresents.

[00117] In some embodiments of Ryrepresents a halogen. In other embodiments of Ryrepresents a C1-4alifaticescoe. In some embodiments of Ryrepresents fluorine. In some embodiments of Ryrepresents chlorine. In some embodiments of R5yrepresents bromine. In some embodiments of Ryrepresents iodine. In other embodiments of Ryrepresents vinyl.

[00118] In some embodiments of m is an integer from 1 to 3 inclusive. In some implementations, the value of m is 1. In some implementations, the value of m is 2. In some implementations, the value of m is 3.

[00119] In some embodiments of s is an integer from 1 to 3 inclusive. In some implementations, the value of s is 0. In some implementations, the value of s is 1. There are implementations in which the value of s is 2. In some implementations, the value of s is 3. In some embodiments of the s value is 4.

[00120] In some embodiments of t represents an integer between 1 and 3 inclusive. In some implementations, the value of t is 0. In some implementations, the value of t is 1. In some implementations, the value of t is 2. In some implementations, the value of t which is 3. In some embodiments of the t-value is 4.

[00121] In certain embodiments of the group R2aformula I is a HE. In some embodiments of the group R2aformula I is-NHSO2R2where R2corresponds to above and described in the text of this application. Thus, according to the present invention proposed a compound according to formula I-a or I-b:

or pharmaceutically acceptable salt of the compounds, where each of the groups R1, R1', R2, R3, R4and RZcorresponds above for formula I, and also the description of the classes and subclasses described in the text of this application.

[00122] In certain embodiments of the group R2formula I-b is-N(R)2. In other embodiments of the group R2formula I-b is a possibly containing substituents group selected from the following examples:3-7cycloalkyl, 6-10-membered aryl, 5-10-membered heteroaryl containing 1-4 heteroatoms that are independently selected from nitrogen, oxygen, or sulfur, or 4-7-membered heterocyclyl containing 1-2 heteroatoms that are independently selected from nitrogen, oxygen or sulfur. In some embodiments of R2represents a cycloalkyl or 6-10-membered aryl. In some embodiments of R2represents possibly containing substituents 6-10-membered aryl. In some embodiments of R2represents phenyl. There are options for implementation according to which R2is cyclopropyl.

[00123] In some embodiments of R2selected from the following examples:3-7cycloalkyl connected by a bridge Bicycle, 6-10-membered aryl, 5-10-membered heteroaryl containing 1-4 heteroatoms that are independently selected from nitrogen, oxygen, or sulfur, or 4-7-membered heterocyclyl containing 1-2 heteroatoms that are independently selected from nitrogen, oxygen or sulfur.

[00124] In certain embodiments of the group R3formula I is a head group. There are implementations of the present invention, according to which the group R3and R1the compounds of formula I together with in between the atoms form may contain substituents saturated or unsaturated 12-18 membered ring containing 2 to 6 heteroatoms, which are independently selected from nitrogen, oxygen or sulfur, with the specified ring contains head group. In some embodiments of R3and the ring formed by R1and R1'together with in between the atoms that form the possibly containing substituents saturated or unsaturated 12-18 membered ring, containing 2 to 6 heteroatoms, which are independently selected from nitrogen, oxygen or sulfur, and the formed ring contains head group.

[00125] As defined generally above, the ring formed by the groups R3and R1in formula I contains the head group. The phrase "contains head group"used in the present description, may mean either that the ring formed by R3and R1contains head group as a substituent, and the fact that this group is part of the ring. For example, the ring formed by R3and R1can contain as Deputy head group-L-Y, where group-L-Y are as described above. Alternatively, the ring formed by R3and R1has the properties of the head group, which is part of the ring. For example, the ring formed by R3and R1may contain one or more unsaturated sites and possible substituents and/or heteroatoms, which in combination leads to the formation of groups capable of covalent binding of the HCV protease according to the present invention. In some embodiments of the ring formed by R3and R1may contain substituents in α-, β-, γ - or δ-positions relative to the carbon atom that is attached to the group R4.

[00126] it Should be understood, that when R3and R1bind together with the formation may contain substituents saturated or unsaturated 12-18 membered ring containing 2 to 6 heteroatoms, which are independently selected from nitrogen, oxygen or sulfur, such compounds include compounds formed by linking R3and the ring formed by R1and R1'.

[00127] Examples of compounds of formula I, where R3and the ring formed by R1and R1'tied together, include compounds according to formulas I-c-1, I-c-2, I-c-3, I-c-4, I-5 and I-6:

or pharmaceutically acceptable salts of these compounds, where each of R2a, R4, Rw, Rx, Rx'and Rycorresponds above, and the description of the classes and subclasses described in the text of this application. It should be understood that, although the formulas I-c-1, I-c-2, I-c-3, I-c-4, I-c-5, and I-c-6 shows cyclopropylidene the ring formed by R1and R1'this group is given as an example, and thus, according to the present description provides for the other group, R1and R1'.

[00128] Examples of such compounds include the compounds shown below in Table 3.

[00129] Although the compounds according to formulas I-c-1, I-c-2, I-c-3, I-c-4, I-c-5 and I-6 are depicted in the form of a Z-double bond stereoisomers in macrocyclic the ring, it should be understood that in some embodiments of compounds according to formulas I-c-1, I-c-2, I-c-3, I-c-4, I-c-5 and I-c-6 can be found in E-stereoisomeric state to the double bond in the macrocyclic ring. There are also options for implementation according to which the proposed mixture of both stereoisomers. In other embodiments of compounds according to formulas I-c-1, I-c-2, I-c-3, I-c-4, I-c-5 and I-c-6 may result in appropriate conditions with the saturation of double bonds.

[00130] In some embodiments of R1and R1'associated with education may contain substituents 3-7-membered carbocyclic ring. In certain embodiments of such compounds have the structure shown in formula I-d:

or pharmaceutically acceptable salts of these compounds, where each of R2a, R3, R4, RZand Rocorresponds to formula I and the descriptions of the classes and subclasses described in the text of this application.

[00131] In some embodiments of Rorepresents possibly containing substituents group selected among those With1-6aliphatic groups. In some embodiments of Rorepresents ethyl. In other embodiments of Rorepresents vinyl.

[00132] Examples of the groups R3/sup> formula I-d include groups described above and shown in the text of this application, and are shown in Table 3 below.

[00133] In certain embodiments of the group R4and Rxtogether with in between the atoms form may contain substituents, saturated or unsaturated 16-22 membered ring containing 2 to 6 heteroatoms, which are independently selected from nitrogen, oxygen or sulfur. In some embodiments of R4and Rxtogether with in between the atoms form may contain substituents, unsaturated 18-22-membered ring containing 3-5 heteroatoms that are independently selected from nitrogen, oxygen or sulfur. In some embodiments of the ring formed by R4and Rxcontains as substituents one or more groups Rmwhere each option Rmindependently represents halogen, -ORo; -CN; -SCN; -SRo; -SORo; -SO2Ro; -NO2; -N(Ro)2; -NHC(O)Roor possibly containing substituents group selected among those With1-6aliphatic groups and3-7cycloalkyl groups. In some implementations according to the present invention proposed the compounds of formula I-e or I-f:

or pharmaceutically acceptable salts of these compounds, where R , R3and Romeet defined for formula I and described in classes and subclasses described in the text of this application;

p can take integer values from 1 to 6, inclusive; and

each of Rmindependently represents halogen, -ORo; -CN; -N(Ro)2; or may contain substituents group selected among those With1-6aliphatic groups and3-7cycloalkyl groups.

[00134] In some embodiments of p is 1. In some embodiments of p is 2.

[00135] In some embodiments of Rmrepresents a C1-6aliphatic group. In some embodiments of Rmrepresents methyl.

[00136] In some embodiments of Rois possibly containing substituents group selected from C1-6aliphatic groups. There are options for implementation according to which Rorepresents ethyl. In other embodiments of Rorepresents vinyl.

[00137] Examples of the groups R3according to the formula I-e and I-f include groups described in this text and are listed below in Table 3.

[00138] in spite of the fact that the compounds according to formula I-e and I-f are depicted in Z-and E-conformations of the double bond in the macrocyclic ring, it should be understood that in some embodiments is realizatsii compounds according to formula I-e and I-f can be E-stereoisomeric state to the double bond in the macrocyclic ring. In some embodiments of compounds according to formula I-e and I-f may be in Z-stereoisomeric state to the double bond in the macrocyclic ring. According to some variants of the proposed mixture of both stereoisomers. In other embodiments of compounds according to formula I-e and I-f can be exposed to appropriate conditions to form compounds according to formulas I-g or I-h:

or pharmaceutically acceptable salts of such compounds.

[00139] In some embodiments of R4and Rxconnected together as described above, and R3and the ring formed by R1and R1'connected together as described above, with the formation of new imacrazymaniacchic substances. In some embodiments of the ring formed by R4and Rxcontains as substituents one or more groups Rmas described above for formula I-e and I-f. In some embodiments of the macrocyclic ring formed R3and the ring formed by R1and R1'contains as Deputy head group-L-Y to form compounds of the formula I-j or I-k:

or pharmaceutically acceptable salts of these compounds; where eachindependently researched the Simo denotes one double bond. Methods for such compounds, in addition to methods of synthesis of macrocycles and substances containing head group, described in the text of this application, include also described by McCauley, J.A. et al., Angew. Chem. Int. Ed., 2008, 47, p.9104-7.

[00140] In some embodiments of the methylene unit within the ring formed by R3and the ring formed by R1and R1'replaced by the group L-Y with obtaining compounds of formula I-m or I-n:

or pharmaceutically acceptable salts of these compounds; where eachindependently denotes one double bond.

[00141] As was briefly described in the text of this application, the group R4in the composition of the compounds of formula I represents hydrogen. According to separate variants of implementation of the present invention proposed the compound (formula II-a or II-b:

where each of the groups R1, R1', R2, R3and RZcorresponds above for formula I, and also the description of the classes and subclasses described in the text of this application.

[00142] Examples of compounds of formula I include compounds shown below in Table 3

Table 3. Examples of compounds of the formula formula I

[00143] According to some variants of implementation of the present invention proposed any of the above in Table 3, compounds, or pharmaceutically acceptable salt of the compounds.

[00144] As was as broadly defined above, R3is a head group. Without being bound to any particular theory, believe that the group R3i.e. head groups, are particularly suitable for covalent binding of key cysteine residue in binding of HCV protease. An ordinary person skilled in the art it is known that HCV protease and its mutants contain the region comprises binding a cysteine residue. In some embodiments of compounds according to the present invention contain a head group, due to the properties which these connections can be aimed at residue cysteine S HCV protease. In some embodiments of compounds according to the present invention contain a head group, due to the properties which these compounds aimed at residue cysteine C16 HCV protease.

[00145] Thus, in some embodiments of R3characterized by the fact that the group-L-Y able kovalan is but to contact the cysteine residue, and thus irreversibly inhibit the specified enzyme. In some embodiments of the cysteine residue is Cys159 of HCV protease, or a mutant form, where the numbering of residues is according to the Uniprot (code Q91RS4).

[00146] the person skilled in the art it is clear that for such covalent binding fit different head groups, as defined in the present description. Such groups R3include, but are not limited to the groups described in the text of this application and shown above in Table 3. The specified process was performed by conducting mass spectrometric experiments according to the Protocol described in detail below in Examples 17-21.

[00147] According to another aspect of the present invention proposed a conjugate containing the HCV protease, or a mutant, covalently linked to the inhibitor in the field of Cys159. In some embodiments of the inhibitor covalently linked using the linker section.

[00148] According to some variants of implementation of the present invention proposed a conjugate composition Cys159-linker-inhibitor. The person skilled in the art it is clear that the "linker" group (linker patch) corresponds to the head group-L-Y as described in the present text. Accordingly, in some embodiments of the description of the linker section with testvol the above description for the group-L-Y. However, it is clear that the linker section is divalent and, thus, the corresponding group-L-Y must also have a valence equal to two, resulting from the reaction of the parent group with Cys16 HCV protease, or a mutant form of this enzyme.

[00149] In some embodiments of the group of the inhibitor is a compound of the formula:

where each of the groups R1, R1', R2a, R4, Rxand RZformula And corresponds to the particular above for formula I and described in classes and subclasses described herein. Thus, according to some variants of implementation of the present invention proposed a conjugate of the formula:

where each of the groups R1, R1', R2a, R4and RZin the composition of the conjugate corresponds to the above description for the formula I, as well as the description of the classes and subclasses described in the text of this application.

[00150] In some embodiments of R3characterized by the fact that the group-L-Y is capable of covalently to contact the cysteine residue and thereby irreversibly inhibiting the enzyme. In some embodiments of the cysteine residue is Cys16 HCV protease, or a mutant, where the numbering of residues is according to the on Uniprot (code Q91RS4).

[00151] According to another aspect of the present invention proposed a conjugate containing a HCV protease or its mutant, covalently associated with the inhibitor in the field of Cys16. In some embodiments of covalent binding of the inhibitor is carried out with the help of the linker section.

[00152] According to some variants of implementation of the present invention proposed a conjugate composition Cys16-linker-inhibitor. The person skilled in the art it is clear that the "linker" section corresponds head group-L-Y as described in the present text. Accordingly, in some embodiments of the linker description of the plot corresponds to the above description for the group-L-Y. However, it is clear that the linker section is divalent and, thus, the corresponding group-L-Y must also have a valence equal to two, resulting from the reaction of the parent group with Cys16 HCV protease or its mutant.

[00153] In some embodiments of the inhibitor has the structure of formula a-1:

where each of the groups R1, R1', R2a, R4and RZin the composition of the conjugate corresponds to the particular above for formula I, and also the description of the classes and subclasses described in the text of this application. Accordingly, according to the present invention pre is false conjugate of the formula:

where each of the groups R1, R1', R2a, R4and RZin the composition of the conjugate corresponds to the particular above for formula I, and also the description of the classes and subclasses described in the text of this application.

General methods for producing compounds according to the present invention

[00154] According to some variants of implementation of the compounds according to the present invention in General receive in accordance with Scheme 1 below:

Scheme 1

[00155] According to one aspect of the present invention the methods of obtaining compounds of formula I in accordance with the steps mentioned above in Scheme 1, which identifies and describes each variable and each PG is a suitable protecting group. At stage S-1 N-protected (e.g., Boc (tert-butoxycarbonyl)) derived Proline of the formula And condense with alpha aminoethanol formula B in terms of the conjugation of peptides with getting the dipeptide of formula C. the Appropriate conditions for the conjugation of peptides is well known in this area and include the terms and conditions described in detail in PCT No. WO2002094822 (US6825347)whose contents are fully incorporated into the present description by reference. Except for the cases specifically mentioned in the text of this proposal, these conditions will be midrange is to seek the relevant conditions of conjugation of peptides.

[00156] In step S-2 ester group is subjected to hydrolysis using a suitable base and then neutralized to obtain the dipeptide of formula D. the Appropriate bases include, but are not limited to alkali metals, hydroxides of alkaline earth metals and their combinations. In some embodiments of the base is lithium hydroxide.

[00157] In step S-3 carry out the pairing of donated formula D with a sulfonamide of formula E in the corresponding conditions of conjugation of peptides with getting arylsulfonamides formula F.

[00158] In step S-4 by removal of the protective group (for example, removal of the Boc) from the dipeptide of formula F receive amine of the formula G. In some embodiments of the cleavage of the group Vos reach by interacting substances of the formula F with an inorganic or organic acid in a halogenated hydrocarbon solvent. In some embodiments of the acid used triperoxonane acid, and the solvent is dichloromethane.

[00159] In step S-5 is carried out mate amine of formula G with a carboxylic acid of the formula H under appropriate conditions of conjugation of peptides to obtain the intermediate compounds of formula I-0.

[00160] the Intermediate compound of formula I-0 is converted into the compounds of formula I in the course of the stages described in the quality of the ve examples to this application.

[00161] As defined generally above, the group PG formulas a, C, D, and F is a suitable protective amino group. Appropriate protective amino groups are well known in this field and include groups described in detail in Protecting Groups in Organic Synthesis, T. W. Greene and P. G. M. Wuts, 3rdedition, John Wiley & Sons, 1999, the description of which is fully included in the text of this application by reference. Protected amines are well known in this field and include groups described in Greene (1999). Next, the appropriate monoamine amines include, but are not limited to aralkylamines, carbamates, allylamine, inorganic salts and the like. Examples of relevant monogamistic amino group include the following compounds: t-butyloxycarbonyl-(-NHBOC), ethoxycarbonylethyl, methoxycarbonylamino, trichlorocarbanilide, allyloxycarbonyl-(-NHAlloc), benzyloxycarbonylamino- (-NHCBZ), allylamino, benzylamino- (-NHBn), floridalottery- (-NHFmoc), formamido-, acetamido-, chloracetamide, dichloroacetamide, trichloroacetamide, phenylacetamido, triptorelin, benzamido-, t-butyldiphenylsilyl and the like. Appropriate tizamidine amines include amines containing two substituent independently selected from the substituents described above for monogamistic amines, and, further, include cyclic imides, such as phthalimide, maleinimide, succinimide and the like.

[00162] According to other variants of implementation of the compounds according to the present invention receive in accordance with Scheme 2, below.

Scheme 2

[00163] According to one aspect of the present invention methods of preparing compounds according to formula I in accordance with the steps described above in Scheme 1. At step S-6, make a removal of the Boc group from the structure of the dipeptide of the formula under conditions of acid catalysis with getting dipeptide ester of formula J.

[00164] In step S-7, dipeptide ester of formula J condense with functionalized amino acid of the formula H with appropriate conditions of conjugation of peptides with getting Tripeptide formula K, which then convert (convert) in tripeptidyl ether of the formula L in the course of the stages described in the examples in the text of this application.

[00165] In step S-8 ester group of the compounds of formula L is subjected to hydrolysis with an appropriate base and then neutralized with obtaining Tripeptide formula M Corresponding bases include, but are not limited to alkali metals, hydroxides of alkaline earth metals, and their combinations. In some embodiments of the base is lithium hydroxide.

[00166] In the step S-9 Tripeptide of formula Contensious with a sulfonamide of formula E with appropriate conditions for the conjugation of peptides with obtaining compounds of the formula I.

[00167] the Group PG on the formula C, H, and K represents the corresponding aminosidine group as described above.

4. Use, dosage forms and routes of administration according to the present invention

Pharmaceutically acceptable compositions according to the present invention

[00168] According to another implementation variant of the present invention proposed a composition containing the compound according to the present invention or a pharmaceutically acceptable derivative of such compounds, and pharmaceutically acceptable carrier, adjuvant or diluent. The quantitative content of the compounds in the compositions according to the invention sufficient to effectively measured the inhibition of HCV protease or its mutant forms in a biological sample or in a patient's body. In some embodiments of the quantitative content of the compounds in the compositions according to the present invention corresponds to a significant inhibition of HCV protease or its mutant in a biological sample or in a patient's body. According to some variants of implementation of the present invention mentioned composition was prepared in the form of a dosage form intended for administration to a patient in need of such compositions. In some embodiments of compositions is s according to the present invention is decorated in a dosage form, intended for oral administration to a patient.

[00169] the Term "patient"used in the present description, refers to an animal, preferably to a mammal, most preferably to humans.

[00170] the Term "pharmaceutically acceptable carrier, adjuvant or solvent" refers to non-toxic carrier, adjuvant or solvent that does not violate the pharmacological activity of the substance contained in the composition of the dosage form with him. Pharmaceutically acceptable carriers, adjuvants or diluents suitable for use in compositions according to the present invention, include, but are not limited to the following examples: ion exchangers, alumina, start aluminum, lecithin, serum proteins blood, such as albumin human serum, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, mixtures of partial glycerides of saturated vegetable fatty acids, water, salts, electrolytes, such as preteenslut, sodium dihydrophosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, substance on the basis of cellulose, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and lanolin.

[00171] the Term "pharmaceutical preparations is automatic acceptable derivative" refers to any non-toxic salt, complex ether, salts of ester or other derivative compounds according to the present invention, in the injection the patient is capable of forming, directly or indirectly, a compound according to the present invention, or possessing inhibitory activity metabolite of such a compound, or its residue.

[00172] the Term "possessing inhibitory activity of the metabolite or residue"used in this description, means that a metabolite or residue metabolite is an inhibitor of HCV protease or its mutant.

[00173] the introduction of the compositions according to the present invention can be performed oral, parenteral, by inhalation, topically, rectally, nazalnam by, buccal, vaginally or via an implantable capacity. The meaning of the term "parenteral"as used in the present description includes a method of intravenous, subcutaneous, intramuscular, intra-articular, intrasynovial, intrasternally, intrathecal, intrahepatic, intracranial injections and infusions and injections or iefusi in the area of the affected tissues. Preferably the compositions according to the present invention is injected oral, intraperitoneal or intravenous route. Sterile dosage forms of the compositions according to the present invention can be in the form of water or oil is Noah suspension. Such suspensions can be made into dosage forms according to methods known in this field, including the use of appropriate dispersing agents or wetting agents and suspendida agents. A sterile preparation intended for injection, may also be a sterile solution for injection or suspension in a nontoxic, acceptable for injecting diluent or solvent, for example, in the form of a solution of 1,3-butanediol. As the acceptable vehicles and solvents that are suitable for the application, you can use water, ringer's solution, and isotonic sodium chloride solution. Also as a solvent or suspendida environment, you can apply sterile fatty oil.

[00174] In the above purposes, you can use any soft fatty oils, including synthetic mono - or diglycerides. Fatty acids such as butyric acid and its glyceride derivatives, useful in the manufacture of dosage forms intended for injection, as well as natural olive or castor oil, especially in polyoxyethylenic forms. These oil solutions or suspensions may also contain a solvent or dispersing agent based on long-chain alcohol, such as carboxymethyl cellulose or similar dispersing agents, the usual used in the manufacture of pharmaceutically acceptable dosage forms, including emulsions and suspensions. Also in the manufacture of the dosage form can be applied to other commonly used surfactants, such as Tweens, Spans and other emulsifying agents or substances that increase the bioavailability, which is usually used in the manufacture of pharmaceutically acceptable soluble, liquid and other medicines.

[00175] the Introduction of pharmaceutically acceptable compositions according to the present invention can be performed oral route in any dosage form that is acceptable for oral administration, including but not limited to, capsules, tablets, aqueous suspensions or solutions. When choosing tablets for oral administration commonly used carriers include lactose and corn starch. Also usually add lubricating agents such as magnesium stearate. Useful diluents for oral introduction capsules include lactose and dried corn starch. In the case when for oral administration requires water suspension, the active ingredient is combined with emulsifying and suspendresume agents. Optionally, you can also add certain sweetening, flavoring or coloring agents.

[00176] pharmaceutically acceptable compositions according to the present invention can be applied in the form of suppositories (medical candle is), intended for rectal administration. Such dosage forms can be manufactured by mixing the agent with the appropriate not causing irritation filler, which is in a solid form at room temperature and in liquid form at rectal temperature and therefore soluble in the rectum to release the drug. Such substances include cocoa butter, beeswax and polyethylene glycols.

[00177] the Pharmaceutically acceptable composition according to the present invention it is also possible to enter a local manner, especially in cases when the object of treatment includes areas or organs, which are readily available in the local application, including diseases of the eye, skin, or the lower digestive tract. For each of the above areas or organs are easy to manufacture dosage forms for local use.

[00178] the Local introduction in the lower gastrointestinal tract can be realized in the form of rectal suppositories (see above) or in an appropriate dosage form for injection with an enema. You can also use transdermal patches topical application.

[00179] In the case of local administration of a pharmaceutically acceptable composition according to the present invention can be produced in the form of relevant ointments, containing the active substance, suspended or dissolved in one or more carriers. Media, applicable when the local introduction of the substances according to the invention include, but are not limited to the following examples: mineral oil, liquid paraffin, white (medical) petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene, emulsifying wax and water. Also pharmaceutically acceptable composition according to the present invention can be produced in the form of relevant lotions or creams containing the active substance suspended or dissolved in one or more carriers. Appropriate carriers include, but are not limited to the following examples: mineral oil, sorbitan monostearate, Polysorbate 60, citylove esters wax, Cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.

[00180] For ophthalmic use, the pharmaceutically acceptable composition according to the present invention can be produced in the form of micronized suspensions in isotonic saline solution, sterile saline solution with adjusted pH or, preferably, in isotonic saline solution, sterile saline solution with adjusted pH value, as a preservative, for example with benzalconi chloride, and without it. Also pharmaceutically acceptable composition according to the present and the finding, designed for ophthalmic applications, can be produced in the form of ointments such as petroleum jelly.

[00181] the Pharmaceutically acceptable composition according to the present invention can also be entered in the form of a nasal aerosol or inhalation. These compositions are made in accordance with techniques well known in the field of dosage forms in the pharmaceutical industry, by dilution in saline solution, using benzyl alcohol or other suitable preservatives, substances that promote absorption, to enhance bioavailability, fluorocarbons, and/or other relevant agents that increase the solubility and dispersing agents.

[00182] In the most preferred embodiment, the implementation of the pharmaceutically acceptable composition according to the present invention is prepared in a dosage form intended for oral administration. Such dosage forms can be entered at the same time as the meal, and regardless of this. In some embodiments of pharmaceutically acceptable compositions according to the present invention is not injected simultaneously with the meal. In other embodiments of pharmaceutically acceptable compositions according to the present invention is administered simultaneously with the meal.

[00183] the Amount of compound according to N. the present invention, you can connect with substances native to obtaining a composition for a single dose varies depending on the patient being treated, the particular mode of administration. Preferably the compositions according to the present invention should be made so that the patient received the dose of the inhibitor, comprising 0.01-100 mg/kg of body weight.

[00184] it should Also be understood that a specific dosage and treatment regimen for each individual patient depends on many factors, including the specific activity of the applied compound, the patient's age, body weight, General state of health, sex, diet, which adheres to the patient, time of administration, the degree of development of the disease under treatment. Also the quantitative content of the compounds according to the present invention in the composition will vary for each specific connection.

Methods of using compounds and pharmaceutically acceptable compositions according to the present invention

[00185] the Compounds and compositions described in the text of this application, are typically useful for inhibiting the activity of HCV protease and/or its mutant. Thus, the compounds according to the present invention are useful for the treatment of hepatitis other than hepatitis a and b, including hepatitis.

[00186] HCV is a highly variable virus, forming a polymorphic group of options in the host organism. To date, identified six different genotypes worldwide (Simmonds et al., Hepatology, Vol.42, No. 4, 2005). Subsequently, these genotypes are classified more closely related, genetically distinct subtypes. Parts of the sequence used for comparison, known as consensus sequences shown below in Table 3A. Genotypes and subtypes of HCV have different rates of spread in different parts of the world, and in some areas dominated by certain genotypes. Genotypes 1-3 widely distributed around the world. Subtype 1A predominates in North and South America, Europe and Australia. Subtype 1b distributed in North America and Europe and is also found in some areas of Asia. Genotype 2 is presented in the most developed countries, but is less common than genotype 1 (http://www.hcvadvocate.org/hepatitis/factsheets_pdf/genotype_FS.pdf). Other genotypes predominate in populations of the former residents of the U.S. and therefore represent an important target.

[00187] the fact that the cysteine in the 159 amino acid position in the genotype 1b is conservative for all genotypes and subtypes of HCV NS3 sequenced to date, although in other genotypes and subtypes in this position may find the Xia another amino acid. Directed accession inhibitors on this cysteine residue gives the opportunity for the development of compounds with efficacy against multiple HCV genotypes.

[00188] As described in the proposal, according to the present invention proposed an irreversible protease inhibitors of one or more HCV genotypes and their variants. Such compounds containing head group, designated as R3include the compounds of formulas I, I-a, I-b, I-c-1, I-c-2, I-c-3, I-c-4, I-c-5, I-c-6, I-d, I-e, I-f, I-g, I-h, II-a, II-b, III-a, III-b, IV-a, IV-b, IV-c, IV-d, V-a, V-b, V-c, V-d, VI-a, VI-b, VII-a, VII-b, VIII-a and VIII-b, as described in the present text. In some embodiments of R is characterized by the fact that the group-L-Y is capable of covalently to contact the cysteine residue and thereby irreversibly inhibiting the enzyme. Without being bound to any particular theory, it is believed that such groups R3i.e. head groups, are particularly suitable for covalent binding to a key cysteine residue in binding protease of one or more genotypes of HCV or its variants. In some embodiments of one or more genotypes, protease which is subject to inhibition by interaction with the compounds according to the invention, includes 1a, 1b, 2A, and 3A. In some embodiments of one or more of these options include AC, A156S, D168V, D168A and R155K.

[00189] Usually the one skilled in the art will understand, the protease of various genotypes of HCV and its variants containing one or more cysteine residues near the binding region. Without being bound to any particular theory, it is believed that the proximity of the head group with the cysteine residue according to the present invention enhances the covalent modification of cysteine with the help of the head of the group. In some embodiments of the cysteine residue according to the present invention is Cys159 of HCV protease subtype 1b or its variant, the numbering of the residues carried out in accordance with Uniprot (code Q91RS4). Residues of cysteine proteases of other genotypes and subtypes of HCV, which may be subjected to covalent modification by means of irreversible inhibitors according to the present invention, include balances below in Table 3A, where the bold and underlined "C_" refers to the residue of cysteine, which is conservative in the position equivalent to Cys159 of HCV protease subtype 1b.

Table 3A
Genotype/ subtype GHSThe plot is a typical sequence"ID Patient The sequence number
1AGHAVGLFRAAVC_TRGVAKAV_.H77.NC_004102SEQ ID NO:1
1AGHAVGIFRAAVC_TRGVAKAVCH.BID-V271.EU482858SEQ ID NO:2
1AGHAVGIFRAAVC_TRGVAKAVDE.BID-V25.EU482831SEQ ID NO:3
1AGHAVGLFRAAVC_TRGVAKAVUS.H77-H21.AF011753SEQ ID NO:4
1bGHAVGIFRAAVC_TRGVAKAVAU.HCV-A.AJ000009SEQ ID NO:5
1bGHVVGIFRAAVC_TRGVAKAVCH.BID-V272.EU482859SEQ ID NO:6
1bGHAVGIFRAAVC_TRGVAKAVJP.HCV-BK.M58335SEQ ID NO:7
1CGHAVGIFRAAVC_TRGVAKAVID.HC-G9.D14853SEQ ID NO:8
1CGHVAGIFRAAVC_TRGVAKAVIN.AY051292.AY051292SEQ ID NO:9

Genotype/ subtype GHSThe plot is a typical sequenceaID PatientThe sequence number
2AGAVGIFRAAV C_SRGVAKSIJP.AY746460.AY746460SEQ ID NO:10
2AGHAVGIFRAAVC_SRGVAKSIJP.JCH-6.AB047645SEQ ID NO:11
2AGHAVGIFRAAVC_SRGVAKSI_.G2AK1.AF169003SEQ ID NO:12
2bGHAVGLFRAAVC_ARGVAKSIJP.HC-J8.D10988SEQ ID NO:13
2bGHAVGLFRAAVC_ARGVAKSIJP.MD2b1-2.AY232731SEQ ID NO:14
2CGHAVGIFRAAVC _SRGVAKSI_.BEBE1.D50409SEQ ID NO:15
2iAHAVGIFRAAVC_SRGVAKSIVN.D54.DQ155561SEQ ID NO:16
2kGHAVGIFRAAIC_TRGAAKSIMD.VAT96.AB031663SEQ ID NO:17
3AGHVAGIFRAAVC_TRGVAKALCH.452.DQ437509SEQ ID NO:18
3AGHVAGIFRAAVC_TRGVAKALDE.HCVCENS1.X76918SEQ ID NO:19
3AGHVAGIFRAAVC_ TRGVAKALID.ps23.EU315121SEQ ID NO:20
3bGHVMGIFIAVVC_TRGVAKALIN.RG416.DQ284965SEQ ID NO:21
3bGHVVGIFRAAVC_TRGVAKALJP.HCV-Tr.D49374SEQ ID NO:22
3kGHVAGIFRAAVC_TRGVAKALID.JK049.D63821SEQ ID NO:23
4AGHAAGIFRAAVC_TRGVAKAVEG.Eg9.DQ988077SEQ ID NO:24
4AGHAAGLFRAAVC_TRGVAKAV_.01-09 .DQ41782 SEQ ID NO:25
4AGHAAGLFRAAVC_TRGVAKAV_.F753.DQ418787SEQ ID NO:26
4dGHAAGIFRAAVC_TRGVAKAV_.03-18 .DQ418786SEQ ID NO:27
4dGHAAGIFRAAVC_TRGVAKTV_.24.DQ516083SEQ ID NO:28
4fGHAVGIFRAAVC_TRGVAKAVFR.IFBT84.EF589160SEQ ID NO:29
4fGHAVGIFRAAVC_TRGVAKAVFRJFBT88.EF589161SEQ ID NO:30
5aGHVVGVFRAAVC_TRGVAKALGB.EUH1480.Y13184SEQ ID NO:31
5aGHVVGVFRAAVC_TRGVAKALZA.SA13.AF064490SEQ ID NO:32
6aGHVVGLFRAAVC_TRGVAKSLHK.6a74.DQ480524SEQ ID NO:33
6aGHVVGLFRAAVC_TRGVAKSLHK.6a77.DQ480512SEQ ID NO:34
6aGHVVGLFRAAVC_TRGVAKSLHK.EUHK2.Y12083SEQ ID NO:35
6bGHVVGLFRAAVmath display="block"> C_TRGVAKAL_.Th580.NC009827SEQ ID NO:36
6cGHVVGLFRAAVC_TRGVAKALTH.Th846.EF424629SEQ ID NO:37
6dDHVVGLFRAAVC_TRGVAKALVN.VN235.D84263SEQ ID NO:38
6eGHVVGLFRAAVC_TRGVAKAICN.GX004.DQ314805SEQ ID NO:39
6fGHAVGIFRAAVC_TRGVAKAITH.C-0044.DQ835760SEQ ID NO:40

Genotype/ subtype GHScastac typical sequence aID PatientThe sequence number
6fGHAVGIFRAAVC_TRGVAKAITH.C-0046.DQ835764SEQ ID NO:41
6gGHVVGLFRAAVC_TRGVAKALHK.HK6554.DQ314806SEQ ID NO:42
6gGHVVGLFRAAVC_TRGVAKALID.JK046.D63822SEQ ID NO:43
6hGHVAGIFRAAVC_TRGVAKSLVN.VN004.D84265SEQ ID NO:44
6iGHVAGIFRAAVC_TRGVAKSL TH.C-0159.DQ835762SEQ ID NO:45
6jGHVAGIFRAAVC_TRGVAKSLTH.C-0667.DQ835761SEQ ID NO:46
6JGHVAGIFRAAVC_TRGVAKSLTH.Th553.DQ835769SEQ ID NO:47
6kGHVAGIFRAAVC_TRGVAKSLCN.KM41.DQ278893SEQ ID NO:48
6kGHVAGIFRAAVC_TRGVAKSLCN.KM45.DQ278891SEQ ID NO:49
6kGHVAGIFRAAVC_TRGVAKSLVN.VN405.D84264SEQ ID NO:0
6lGHVAGIFRAAVC_TRGVAKSLUS.537796.EF424628SEQ ID NO:51
6mGHAVGVFRAAVC_TRGVAKSLTH.C-0185.DQ835765SEQ ID NO:52
6mGHAVGVFRAAVC_TRGVAKSLTH.C-0208.DQ835763SEQ ID NO:53
6nGHVVGIFRAAVC_TRGVAKSLCN.KM42.DQ278894SEQ ID NO:54
6nGHVVGIFRAAVC_TRGVAKSLTH.D86/93.DQ835768SEQ ID NO:55
6o GHAVGLFRAAVC_TRGVAKAICA.QC227.EF424627SEQ ID NO:56
6pGHVVGLFRAAVC_TRGVAKAICA.QC216.EF424626SEQ ID NO:57
6qGHAVGLFRAAVC_TRGVAKAICA.QC99.EF424625SEQ ID NO:58
6tGHVVGLFRAAVC_TRGVAKAIVN.TV241.EF632069SEQ ID NO:59
6tGHVVGLFRAAVC_TRGVAKAIVN.TV249.EF632070SEQ ID NO:60
6tGHVVGLFRAAV C_TRGVAKAIVN.VT21.EF632071SEQ ID NO:61
7aSHCVGIFRAAVC_TRGVAKAVCA.QC69.EF108306SEQ ID NO:62
aAn ordinary person skilled in the art will understand that any virus prone to mutations and prone to polymorphism and any of the consensus sequences of genotypes described in this text, is typical for a given genotype or subtype. Such a typical consensus sequence is available at http://hcv.lanl.gov/content/sequence/NEWALIGN/align.html.

[00190] the drug Resistance is becoming a serious problem for directed therapy. For example, have reported the development of resistance to protease inhibitors HCV who were in the development stage. These compounds include BILN 2061 and VX-950, developed by Boehringer Ingelheim and Vertex Pharmaceuticals, respectively. The chemical structure of BILN 2061 and VX-950 is shown below.

[00191] it is Known that a recent article published by Vertex Pharmaceuticals, entitled "In Vitro Resstance Studies of Hepatitis With Virus Serine Protease" facing directly to the problem of stability of the mutants seen with VX-950 and BILN 2061. Cm. Lin et al., The Journal of Biological Chemistry, Vol.279, No. 17, Issue of April 23, p.17508-17514, 2004. This article concludes that "future therapy designed for the treatment of hepatitis C, including the use of low molecular weight inhibitors of HCV enzymes, may require a combination of several drugs, as in the case with the methods of HCV treatment is applied at the present time". (See page 17513, last paragraph.)

[00192] the Resistance to specific antiviral drugs is a major factor limiting the effectiveness of many therapies directed against retroviruses or RNA-containing viruses. Contributing to the error, the nature of these viruses allows the emergence of mutations that increase resistance to drugs currently available, and drugs undergoing clinical trials. The issue of sustainability is a major obstacle encountered in the development of new inhibitors that are specific to HCV for the treatment of HCV patients.

[00193] a Recent in vitro investigation of the stability using two inhibitors of the HCV protease NS3-4A, VX-950 and BILN 2061 found that mutations selected under the action of both the first and second inhibitor led to a significant decrease in the sensitivity to the inhibitor. However, first cnie resistant mutants to BILN 2061 had full sensitivity to VX-950, and viruses carrying primary mutation resistance to VX-950, remained sensitive to BILN 2061 (Lin et al., Jour. Biol. Chem. 279(17): 17508-14, 2004).

[00194] it has been Unexpectedly found that the compounds according to the present invention inhibit the protease, at least five HCV mutants, including AT, A156S, D168V, and D168A, and R155K. This fact contrasts with the properties of other HCV protease inhibitors (e.g., VX-950 and BILN 2061), each of which inhibits the protease only two mutants. In fact, none of the drugs, known from the prior art, showed effective inhibition of the protease of all known mutants of HCV. An example is the data for BILN 2061 and VX-950, shown below in Tables 4A and 4b, published by Lin et al., as well as in other literature on the GHS, and data for compounds 1-3, obtained in accordance with the methods described below in the Examples. Without being bound to any particular theory, it is believed that the compounds according to the present invention can serve as effective inhibitors of HCV protease drug-resistant forms. While table 4b shows the activity of the substance 1-3 against three reference HCV variants (AT, A156S, D168V, and D168A), the following examples describe the other compounds according to the present invention, which has activity against these options, and the e against the fifth version of the GHS (R155K).

Table 4A
Comparative values of Ki(nm)a
BILN 2061VX-950
WT19100
AT>12009900
A156S1122900
D168V>120043
D168A>1200150
andData for wild-type were obtained during cellular analyses, and data for the mutants were obtained using biochemical methods of analysis. Cm. Lin et al., and the protocols given in this description.

Table 4b
Comparative Values IC50(nm)a
LN 2061VX-950 Compound I-1
WT44020.66
AT----3
A156S746502
D168V50901632
D168A18601938
andData for wild-type were obtained in the course of conducting cell assays, and the data for the mutants were obtained using biochemical methods of analysis. Cm. Lin et al., and the protocols given in this description.

[00195] Without being bound to any particular theory, believe that the compound of formula I is more effective for inhibiting HCV protease, or its mutant, in comparison with the corresponding compound of formula I, in which R3formula I is the parent group, and represents, for example, unbranched alkyl (such as alkyl not containing substituents), branched alkyl, cyclea the keel or alkenyl. For example, the compound of the formula I can be more effective for inhibiting HCV protease, or its mutant, in comparison with the corresponding compound of formula I, in which R3formula I is the parent group, but represents methyl, ethyl, propyl, butyl (e.g. t-butyl), unbranched or branched alkenyl, not containing substituents (for example, C1-8alkenyl), cyclohexyl or cyclopentyl.

[00196] the Compound of formula I, as described above, may have a greater activity, based on the values of IC50against the HCV protease or its mutant forms, for example AT, A156S, D168V, D168A, or other specimens, such as described in the text of this application than the corresponding compound of formula I, in which R3formula I is the parent group, but represents methyl, ethyl, propyl, butyl (e.g. t-butyl), unbranched or branched alkenyl, not containing substituents (for example, alkenyl C1-8), cyclohexyl or cyclopentyl. Found on the comparative activity of the compounds of formula I relative to the corresponding compounds of the formula I, in which R3formula I is the parent group can be determined by using the standard time-dependent methods for the analysis of, for example, the methods described below in Approx the arts. In some embodiments of the compound of formula I is significantly more active than the corresponding compound of formula I, in which R3formula I is the parent group, but represents methyl, ethyl, propyl, butyl (e.g. t-butyl), unbranched or branched alkenyl, not containing substituents (for example, C1-8alkenyl), cyclohexyl or cyclopentyl. In some embodiments of the compound of formula I is much more active, when the activity of the compounds is measured after approximately 1 minute, about 2 minutes, about 5 minutes about 10 minutes about 20 minutes about 30 minutes about 1 hour about 2 hours about 3 hours, about 4 hours, about 8 hours, about 12 hours, about 16 hours, about 24 hours or about 48 hours, relative to the corresponding compounds of the formula I, in which R3formula I is the parent group, but represents methyl, ethyl, propyl, butyl (e.g. t-butyl), unbranched or branched alkenyl, not containing substituents (for example, C1-8alkenyl), cyclohexyl or cyclopentyl. In some embodiments of the compound of formula I may possess in approximately RA, approximately 2-fold, about 5 fold, about 10 fold, about 20 fold, about 25-fold, about 50 fold, about 100 fold or even approximately 1000 times greater activity than the corresponding compound of formula I, in which R3formula I is the parent group, but represents methyl, ethyl, propyl, butyl (e.g. t-butyl), unbranched or branched alkenyl, not containing substituents (for example, C1-8alkenyl), cyclohexyl or cyclopentyl.

[00197] the Term "clinical resistance to drugs used in the present description, refers to the loss of sensitivity of the target of the drug to the treatment specified drug due to mutations of the target drug.

[00198] the Term "sustainability"used in the present description, refers to changes in the nucleic acid sequence of the wild type, which encodes the target protein, when these changes reduce or eliminate the inhibitory effect of the inhibitor on the specified target protein.

[00199] Examples of proteases, subject to inhibition by the compounds and compositions according to the present invention for inhibiting the activity are useful ways described in the text of this application, include NS3, NS3·4A or Mutan the s.

[00200] the potency of the compound used according to the present invention as an inhibitor of NS3 protease, NS3·4A or their mutants can be investigated in vitro, in vivo or by using the cell line. Methods of in vitro assays include assays, which determine the inhibition of the activity of serine proteases, and/or further functional investigation, or ATPase activity of activated NS3, NS3·4A or their mutants. Other methods of in vitro assays include measuring the ability of the inhibitor to contact NS3 or NS3·4A. The binding of the inhibitor can be measured by placing a radioactive label in the composition of the inhibitor prior to binding, separation of the complex inhibitor/NS3 or inhibitor/NS3·4A and determine the amount of bound radioactive label. Also the binding of the inhibitor can be determined using competitive analysis, during which new inhibitors are incubated with NS3 or NS3·4A associated with known ligands with a radioactive label. Detailed description of the conditions for research compound used according to the present invention as an inhibitor of NS3 or NS3'4A or their mutants below in the Examples.

[00201] Serine proteases constitute a large family of proteolytic enzymes that cleave peptide bonds in proteins. The family of serine proteases includes digestive EN zymes is s disease, trypsin, and elastase and proteases that play a role in blood coagulation. Serine proteases are "catalytic triad"including series, aspartic acid and histidine, which joint action activate series with the formation of covalent bonds with the enzyme's substrate, conducting, therefore, the hydrolysis of peptide bonds. In addition to the above, the serine protease involved in many functions, including the immune system and inflammatory processes.

[00202] the Terms "treating", "treat" or "cure"used in the present description, refers to treatment of disease, alleviation of symptoms, delay the onset, or preventing the development of diseases or disorders or one or more symptoms as described in the text of this application. In some embodiments of the treatment can be prescribed after development of one or more symptoms. In other embodiments of the treatment may be administered in the absence of symptoms. For example, the recipient can be treated before symptoms appear (for example, in the light of history and/or genetic or other predisposing factors). Treatment can be continued after elimination of symptoms, for example, in order to prevent or delay relapse.

[00203] the Compounds and compositions in accordance with the methods with which according to the present invention can be introduced in any dosage and using any mode of administration, which is effective to treat or ameliorate the severity of cancer, autoimmune diseases, neurodegenerative or neurological disease, schizophrenia, diseases of the bones, liver or heart disease. The exact dosage required to treat a particular patient will vary depending on species, age and General health of the patient, severity of infection, the specific compound, its mode of administration, and other factors. Compounds according to the present invention is preferably made in the form of a unit dosage, characterized by ease of introduction and versatility dosage. The expression "unit dosage"as used in the present description, refers to an individual, in physical terms, the unit of a compound corresponding to the treatment of the patient. However, it should be understood that the total daily volume of injected compounds and compositions according to the present invention should be determined during a visit to a therapist, in the framework of common sense, from a medical point of view. Specific to each particular patient or organism dose level depends on many factors, including the disease being treated and its severity; the specific activity of the applied compound; the age, body weight, General state of health the conditions, gender and diet, which adheres to the patient; the time of administration, mode of administration, the rate of excretion of the specific applicable connection; the duration of the treatment; drugs used in combination with the compound according to the present invention or the use of which coincides with the specific applicable connection, and other factors well known in the medical fields. The term "patient"used in the present description, refers to an animal, preferably a mammal and most preferably to humans.

[00204] the Introduction of pharmaceutically acceptable compositions according to the present invention can be realized in humans and other animals oral, rectal, parenteral, intracisternal, endovaginal, by intraperitoneal, local (for example, in the form of powders, ointments or drops), buccal, for example in the form of an oral or nasal spray, and in other ways, depending on the severity of the infection being treated. In some embodiments of compounds according to the present invention it is possible to introduce oral or parenterally in doses, comprising from about 0.01 mg/kg to about 50 mg/kg and preferably from about 1 mg/kg to about 25 mg/kg of body weight of the patient per day, one or more times a day to achieve the Oia desired therapeutic effect.

[00205] In some embodiments of the composition according to the present invention is administered to the needy in this patient once a day, every day. Without being bound to any particular theory, I believe that increased duration of action is irreversible inhibitor of HCV protease NS3 is a special advantage when administered to a patient who needs it, once a day, daily, for the treatment of diseases associated with HCV protease. In some embodiments of the composition according to the present invention is administered to a patient in need of it, at least once a day, every day. In other embodiments of the composition according to the present invention is administered to a patient in need of it, two, three or four times a day, every day.

[00206] the compounds of formula I, for example, typically provide increased duration of action when administered to a patient, compared with the corresponding compound of formula I, in which instead of the head group contains a group such as unbranched alkyl (e.g., not containing substituents alkyl), branched alkyl, cycloalkyl or alkenyl. For example, the compound of the formula I can provide increased duration of action when administered to a patient, in comparison with the corresponding compound of formula I, in part to the th group, R 3formula I instead of the head group is a group as methyl, ethyl, propyl, butyl (e.g. t-butyl), not containing substituents unbranched or branched alkenyl (for example, C1-8alkenyl), cyclohexyl or cyclopentyl.

[00207] Liquid dosage forms intended for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly used in this field, such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl ester of carbonic acid, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyleneglycol, dimethylformamide, oils (in particular, cottonseed, groundnut, corn oil, wheat germ, olive, castor and sesame oils), glycerol, tetrafluoropropyl alcohol, polyethylene glycols and esters sorbitan and fatty acids and mixtures thereof. Besides inert diluents, compositions intended for oral administration may also contain adjuvants, such as moistening agents, emulsifying and suspendresume agents, sweeteners, flavors the e and aromatic agents.

[00208] the Dosage form intended for administration by injection such as a sterile suspension in water or oil-based, intended for injection, can be produced according to methods known in this field, when appropriate dispersant, or moisturizer, or suspendida agents. Sterile dosage form intended for injection, may also be a solution for injection, suspension or emulsion in a nontoxic parenterally acceptable diluent or solvent, for example a solution in 1,3-butanediol. Among the acceptable vehicles and solvents can be listed the following examples: water, ringer's solution, U.S.. and isotonic sodium chloride solution. Also as a solvent or suspendida environment successfully apply of fatty oils. For this purpose you can use any fatty oil soft consistency, including synthetic mono - or diglycerides. Also in the manufacture of dosage forms for injection, use fatty acids such as butyric acid.

[00209] the Sterilization dosage forms intended for injection can be performed, for example, by means of filtration through inhibiting bacteria filter or by the conclusion of sterilizing agents in the form of sterile solid compositions, which prob is tenderly dissolving or dispersing before use in sterile water or other sterile environments suitable for injection.

[00210] In order to prolong the validity of the compounds according to the present invention, it is often desired is slow suction connection with the introduction of subcutaneous or intramuscular injection. This can be achieved by applying the compounds in the form of a liquid crystalline suspensions or in amorphous form, having a low solubility in water. Further, the degree of absorption of connection depends on speed of dissolution of the compound, which, in turn, can be depending on the size of the crystals and their forms. Also, as a variant of the delayed suction connection, when introduced parenterally can be achieved by dissolution or suspension of the compounds in the oil medium. Dosage forms of the compounds of slow absorption, intended for injection is obtained by making microencapsulating forms compounds in biodegradable polymers such as polylactide-polyglycolide. Depending on the ratio of the compounds and polymers and the specific nature of applied polymer can control the release rate of the connection. Examples of other biodegradable polymers include poly-orthoepy and poly-anhydrides. Dosage forms of the compounds of slow absorption, intended for injection, made by placing the operating connection in liposomes or microemulsions, compatible with body tissues.

[00211] Compositions intended for rectal or vaginalis introduction, preferably are suppositories which can be produced by mixing the compounds according to the present invention with corresponding not cause irritation fillers or carriers such as cocoa butter, polyethylene glycol or wax for suppositories, which is in a solid state at room temperature, and turning into a liquid state at body temperature and, therefore, soluble in the rectum or vaginal space, with the release of the current connection.

[00212] the Solid dosage forms intended for oral administration include capsules, tablets, pills, powders and granules. In the composition of these solid dosage forms the active compound is mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate, and/or a) fillers, such as starch, lactose, sucrose, mannitol and silicic acid, b) binders compounds, such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and gum, with hygroscopic compounds, such as glycerol, d) with agents, prednaznacenija improve raspadaemosti, such as agar-agar, calcium carbonate, potato or starch from cassava, alginic acid, certain silicates and sodium carbonate, e) with agents, retarding dissolution such as paraffin, f) with accelerators suction, for example, Quaternary ammonium compounds, g) wetting agents such as, for example, cetyl alcohol and glycerylmonostearate, h) absorbents, for example kaolin and bentonite clay, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures these compounds. In the case of capsules, tablets and pills dosage form may also contain buffering agents.

[00213] Such solid compositions can also be used as a filling in soft and hard gelatin capsules using such excipients as lactose or milk sugar, or high molecular weight glycol and similar compounds. Solid dosage forms such as tablets, coated tablets, capsules, pills and granules can be produced with coatings and shells, such as intersolubility coatings and other coatings well known in the field farmacevticheskoi products. These compositions may include a contrast agent, and they can make so that the current connection (connection) you who were woodlice exclusively or preferably in a particular part of the digestive tract, so the release could have a slower character. Examples of compositions according to the present invention, suitable for use include polymeric substances and waxes. Such compositions in solid form may also be used as filters in soft and hard gelatin capsules using such excipients as lactose or milk sugar, or high molecular weight glycols and similar compounds.

[00214] the Active compounds can also be in microencapsulating form, with one or more filler as described above. Solid dosage forms such as tablets, coated tablets, capsules, pills and granules can be prepared with coatings and shells, such as intersolubility coatings, coatings with controlled-release, and other coatings well known in the field of pharmaceutical preparations. In these solid dosage forms, the active compound can be mixed with at least one inert diluent such as sucrose, lactose or starch. These dosage forms can also contain, in accordance with normal practice, additional connections, in addition to the inert solvent, for example, lubricants and other auxiliary compounds used in the manufacture of tablets, for example, steer the magnesium and microcrystalline cellulose. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents. These dosage forms may include a contrast agent, and they can make so that the current connection (connection) was released exclusively or preferably in a particular part of the digestive tract, so that the release could have a slower character. Examples of compositions according to the present invention, suitable for use include polymeric substances and waxes.

[00215] the Dosage form of the compounds according to the present invention, intended for local or percutaneous introduction, include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalers or patches. The active compound is mixed with a pharmaceutically acceptable carrier and any preservatives or buffers, if required. Dosage forms for ophthalmic use, ear drops and eye drops are also covered by the scope of the present invention. Also according to the present invention, the application of transcutaneous patches have the added advantage consisting in a controlled release of compounds in the body. These dosage forms can be produced by dissolving or dispersing the compound in soo is a relevant environment. Also to increase revenues compounds through the skin, you can apply amplifiers absorption. Its level can be controlled by use of a membrane that is designed for this purpose, or by dispersing the compound in a polymer matrix or gel.

[00216] According to one implementation variant, the invention relates to a method for inhibiting serine protease in a biological sample, comprising the step of bringing into contact a biological sample and compounds according to the present invention, or compositions containing the specified connection.

[00217] According to another implementation variant, the invention relates to a method for inhibiting the activity of HCV protease or its mutant, in a biological sample, comprising the step of bringing into contact of the specified biological sample and compounds according to the present invention, or compositions containing the specified connection. In some embodiments of the invention relates to a method of irreversibly inhibiting the activity of HCV protease or its mutant, in a biological sample, comprising the step of bringing into contact of the specified biological sample and compounds according to the present invention, or compositions containing the specified connection.

[00218] the term "biological sample", as used in n the standing description includes, without limitation, cell cultures or extracts; biopsy material obtained from a mammal or extracts; and samples of blood, saliva, urine, feces, semen, tears, or other body fluids or extracts.

[00219] the Inhibition of the activity of HCV protease or its mutant form, in a biological sample is useful for many purposes, known to specialists in this field. Examples of these goals include, but are not limited to blood transfusion, transplantation of organs, storage of biological specimens and biological analyses.

[00220] In another embodiment implementing the present invention relates to a method of inhibiting the activity of HCV protease or its mutant in the body of a patient, comprising the step of introducing a specified patient connection according to the present invention, or compositions containing the specified connection.

[00221] According to another implementation variant, the invention relates to a method for inhibiting the activity of HCV protease or its mutant in the body of a patient, comprising the step of introducing a specified patient connection according to the present invention or compositions containing the specified connection. According to separate variants of implementation of the present invention relates to a method of irreversible inhibition of the activity of the HCV protease or its mutant in a patient's body, includes an introduction to the specified patient connection according to the present invention or compositions containing the specified connection. According to other variants of implementation of the present invention, a method for treating diseases mediated by HCV protease, or a mutant, in a patient in need of such treatment, comprising the step of introducing a specified patient connection according to the present invention or pharmaceutically effective compositions containing the specified connection. These diseases are described in detail in the text of this application.

[00222] depending on the specific condition or disease, which is subjected to treatment, in combination with the compounds or compositions according to the present invention it is possible to introduce additional therapeutic agents commonly used for the treatment of this condition. In the present description additional therapeutic agents commonly used to treat a certain disease or condition is defined as "agents suitable for treatment of a disease or condition."

[00223] In certain embodiments of a compound or composition according to the present invention is administered in combination with another inhibitor of HCV protease or its variants. In certain embodiments of a compound or composition according to this is the overarching invention is administered in combination with another antiviral agent. Such antiviral agents include, but are not limited to the following examples: immunomodulatory agents, such as α-, β - and γ-interferons, paglierani derived interferon-α and thymosin; other antiviral agents such as ribavirin, amantadine, telbivudine; other protease inhibitors of hepatitis C (inhibitors NS2-NS3 and inhibitors of the NS3-NS4A, for example, BILN 2061 and VX-950); inhibitors of other targets in the life cycle of HCV, including inhibitors of helicase and polymerase; inhibitors of internal planting ribosomes; inhibitors of viral activity of a wide range, such as the IMPDH inhibitors (e.g., mycofenolate acid and its derivatives); or combinations of any of these compounds.

[00224] In some embodiments of you can introduction a combination of 2 or more antiviral agents. Implementations can implement the introduction of a combination of 3 or more antiviral agents. In some embodiments of antiviral agents selected from ribavirin and interferon. In other embodiments of the antiviral agent is α-interferon.

[00225] Other examples of agents that can be combined inhibitors according to the present invention, include, without limitation: therapeutic agents used in the treatment of Alzheimer's disease, the same is as Aricept® and Excelon®; therapeutic agents used to treat HIV such as ritonavir; therapeutic agents used in the treatment of Parkinson's disease such as L-DOPA/carbidopa, entacapone, ropinirol, pramipexol, parlodel, pergolid, trihexyphenidyl and amantadine; therapeutic agents used in the treatment of multiple sclerosis (MS)such as beta interferon (e.g., Avonex® and Rebif®), Copaxone®, and mitoxantrone; therapeutic agents used to treat asthma such as albuterol and Singulair®; agents used in the treatment of schizophrenia, such as zyprexa, risperdal, quetiapine, and haloperidol; anti-inflammatory agents such as corticosteroids, TNF blockers (TNF), IL-1 RA, azathioprine, cyclophosphamide and sulfasalazin; immunomodulatory and immunosuppressive agents such as cyclosporine, tacrolimus, rapamycin, mycophenolate mofetil, interferons, corticosteroids, cyclophophamide, azathioprine and sulfation; neurotrophic factors such as acetylcholinesterase inhibitors, MAO inhibitors, interferons, anti-seizure drugs, blockers of ion channels, riluzole, and agents used to treat Parkinson's disease; agents used to treat cardiovascular disorders, such as beta-blockers, inhibitors EQN (ACE), diuretics, nitrates, calcium channel blockers, and statins; agents used DL the treatment of liver diseases, such as corticosteroids, cholestyramine, interferons, and anti-viral agents; agents used to treat blood disorders such as corticosteroids, agents used to treat leukemia, and growth factors; agents that prolong or improve the pharmacokinetics, such as cytochrome P450 inhibitors (for example, inhibitors of metabolic cleavage) and inhibitors of CYP3A4 (e.g., ketoconazole and ritonavir), and agents used to treat diseases that cause immune deficiency, such as gamma-globin.

[00226] In some embodiments of compounds or compositions according to the present invention is administered in combination with a monoclonal antibody or therapeutic agent, including miRNAs.

[00227] These additional agents can be entered separately from the composition containing the compound according to the present invention, as part of dosage. Also alternatively, these agents can be made part of a single dosage form, in the form of a mixture with the compound according to the present invention in a single composition. When carrying out the introduction as part of dosage two active compounds can be applied simultaneously, sequentially or with a break, usually equal five hours between receptions of connections.

[00228 the Term "combination", "in combination" and similar terms used in the present description, refers to simultaneous or sequential introduction of therapeutic agents according to the present invention. For example, the connection according to the present invention can be administered simultaneously with another therapeutic agent, or sequentially in a separate standard dosage forms or in a single standard dosage form. Accordingly, according to the present invention proposed a single standard dosage form containing a compound according to formula I, the additional therapeutic agent and a pharmaceutically acceptable carrier, adjuvant or carrier.

[00229] the Amount of both compounds, the compounds according to the present invention and additional therapeutic agent (compositions containing an additional therapeutic agent as described above), which may be connected to the material-carriers for a single dosage form will vary depending on the particular mode of administration. Preferably the compositions according to the present invention should be made in the dosage comprising 0.01-100 mg/kg body weight/day of the compounds according to the present invention.

[00230] In compositions containing an additional therapeutic agent, the decree is significant additional therapeutic agent and the compounds according to the present invention can act together. Thus, the quantitative content of the additional therapeutic agent in these compositions will be less than that required in a monotherapy applying only one therapeutic agent. In the composition of these songs, you can enter the dosage of the additional therapeutic agent, in the amount of 0.01-100 mg/kg body weight/day.

[00231] the Amount of an additional therapeutic agent in the compositions according to the present invention will be more than a number, usually injected in the composition that contains therapeutic agent as the only active compound. Preferably the quantitative content of the additional therapeutic agent in the compositions described in the text of this application, will vary from approximately 50% to 100% of the amount, usually contained in a composition containing a specified therapeutic agent as the only active compound.

Examples of compounds according to the invention

[00232] as described in the Examples below, in some illustrative embodiments of the compounds according to the present invention receive in accordance with the following General ways. It should be understood that, although the overall method describes the specific compounds according to the ACLs present invention, following the General methods as well as other methods known to experts in this field, you can apply with getting all connections, subclasses and species of each of these compounds as described in the text of this application.

[00233] the numbering of the compounds used below in the Examples corresponds to the numbering of the compounds used above in Table 3.

Example 1

[00234] The compound was obtained in accordance with steps and intermediate compounds, which are described below.

Stage 1a: the Intermediate compound 1a

[00235] To a solution of toluensulfonate acid ethyl ester (1R,2S)-1-amino-2-vinylcyclopropyl carboxylic acid (0.33 g, 1.0 mmol) and (2S,4R)-1-(tert-butoxycarbonyl)-4-(4-horizontalis-2-carbonyloxy)pyrrolidine-2-carboxylic acid (0.4 g, 1.0 mmol) in 10 ml of acetonitrile was added HATU (0.44 g, 1.2 mmol) and then DIEA (0.46 ml, 2.5 mmol) under stirring. The mixture was subjected to stirring at r.t. (room temperature) for two hours. After depletion of the parent compounds, the reaction mixture is evaporated. The precipitate was dissolved in 30 ml ethyl acetate and washed twice with water and brine, then dried over Na2SO4. After solvent removal was performed chromatography neojidannogo the product with silica gel (hexane:EtOAc=1:1). Was obtained 0.35 g of the indicated compound: MS m/z: 532.0 (M+N+).

Stage 1b: the Intermediate compound 1b

[00236] To a solution of the product obtained in step 1A (0.35 g 0.66 mmol)in 5 ml THF/Meon (1:1) was added 1N aqueous LiOH solution (2 ml, 2.0 mmol). After stirring for 10 hours at r.t. the reaction mixture was neutralized using 1.0 NHCl. Organic solvents are evaporated under vacuum and the remaining aqueous phase sekilala to pH~3 with 1.0 NHCl and were extracted using EtOAc. The organic layer was washed with saline and dried over anhydrous magnesium sulfate. After removal of the solvent was obtained 0.3 g of the specified connection: MSm/z: 526.2 (M+H+).

Step 1C: Intermediate compound 1C

[00237] To a solution of the product obtained at stage 1b (0.30 g, 0.6 mmol)in 10 ml DHM was added CDI (dialkylated) (0.16 g, 1.0 mmol) and the formed solution was stirred at 40°C for 1 hour. To the reaction mixture was added cyclopropanesulfonyl (0.18 g, 1.5 mmol) and DBU (0.16 g, 1.0 mmol). The mixture was subjected to stirring at 40°C for another 10 hours. Then solvent was removed and the residue was dissolved with the aid of EtOAc and washed aqueous NaOAc buffer (pH~5, 2×10 ml), a solution of NaHCO3and a salt solution. After drying over Na2SO4and removal of the solvent the residue is has adversely chromatography with silica gel using hexane/EtOAc(1:1~1:2). The total number of the obtained compound was 0.30 g: Rf0.1 (EtOAc:hexane=1:1), MS m/z: 605.0 (M-1).

Step 1d: Intermediate compound 1d

[00238] the Product obtained in stage 1C (0.25 g, 0.41 mmol), was dissolved in 4 NHCl in dioxane. The mixture was subjected to stirring at r.t. within 1 hour. After removal of the solvents in the mixture is poured into 10 ml DHM, then evaporated it to dryness. Procedure add DHM followed by evaporation was repeated four times to obtain a solid residue, which was directly used in the next step: MSm/z: 507.0 (M+H+).

Step 1E: Intermediate compound 1E

[00239] To a solution of the product obtained in step 1d (0.16 g, 0.28 mmol)and N-BOC-3-(Fmoc)amino-L-alanine (0.15 g, 0.35 mmol) in 5.0 ml of DMF was added HATU (125 mg, 0.33 mmol) and DIEA (130 mg, 1.0 mmol) at r.t. under stirring. After one hour, TLC analysis (thin layer chromatography) showed completion of the reaction. To the mixture was added EtOAc in a volume of 20 ml, after which the mixture was washed with buffer (pH~4, AcONa/AcOH), NaHCO3and brine and dried over MgSO4. After removal of solvent the product in the form of crude oil was subjected to chromatography with silica gel (eluent: EtOAc/hexane). The result was obtained 0.14 g of the specified connection.

Step 1f: Intermediate compound 1f

[00240] a Solution of 0.10 g of the product obtained at stage 1, in 1 ml of DMF with 12% piperidine was stirred for 1.5 hours at r.t., then evaporated to dryness under high vacuum. The residue was treated with a mixture of hexane/ether (4:1) to yield the specified connection, equal to 70 mg

Step 1g:

The compound (I-1)

(00241] (3R,5S)-1-((S)-3-acrylamide-2-(tert-butoxycarbonylamino)propanol)-5-((1R,2S)-1-(cyclopropanecarbonyl)-2-vinylcyclopropyl)pyrrolidine-3-yl 4-horizontalis-2-carboxylate: To a solution of 55 mg (0.08 mmol) of the product obtained in step 1f, 3 ml DHM containing 3 EQ. of triethylamine, drip method was added acryloyl chloride (10 μl, 0.12 mmol) at 0°C under stirring. The reaction mixture was subjected to stirring at r.t. within 1.5 hours, then diluted in 10 ml DHM. Formed solution was twice washed with saline and dried over magnesium sulfate. After removal of the solvent was obtained the crude product, which was subjected to chromatography with silica gel, and preliminary elution using a mixture of hexane/EtOAc (1:3 ~ 1:5) and then using DHM-methanol(50:1 ~ 25:1)). The total number of the obtained compound indicated in the title, was 27 mg: Rf0.4 (EA:Meon=10:1); MS m/z: 746.9 (M+H+).

[00242] using the same method an Intermediate compound 1f in which Uchali the following connections:

[00243] (3R,5S)-1-((S)-2-(tert-butoxycarbonylamino)-3-(vinylsulfonate)propanol)-5-((1R,2S)-1-(cyclopropanecarbonyl)-2-vinylcyclopropyl)pyrrolidine-3-yl 4-horizontalis-2-carboxylate Rf: 0.50 (EtOAc/MeOH 10:1); MS m/z: 805.3 (M+H+).

[00244] (3R,5S)-1-((2S)-2-(tert-butoxycarbonylamino)-3-(2-chloro-2-phenylacetamido)propanol)-5-((1R,2S)-1-(cyclopropanecarbonyl)-2-vinylcyclopropyl)pyrrolidine-3-yl 4-horizontalis-2-carboxylate: Rf: 0.50 (DHM/Meon 95:5); MS m/z: 845.2 (M+H+).

[00245] (3R,5S)-1-((S)-2-(tert-butoxycarbonylamino)-3-((E)-4-(dimethylamino)but-2-enamide)propanol)-5-((1R,2S)-1-(cyclopropanecarbonyl)-2-vinylcyclopropyl)pyrrolidine-3-yl 4-horizontalis-2-carboxylate: Rf: 0.35 (DCM/MeOH 9:1); MS m/z: 804.3 (M+H+).

[00246] in a Similar fashion, using Intermediate compounds 1d and (S)-4-(Fmoc-amino)-2-(tert-butoxycarbonylamino)butane acid was obtained following connection:

[00247] (3R,5S)-1-((S)-4-acrylamide-2-(tert-butoxycarbonylamino)butanoyl)-5-((1R,2S)-1-(cyclopropanecarbonyl)-2-vinylcyclopropyl)pyrrolidine-3-yl 4-horizontalis-2-carboxylate: Rf: 0.40 (EtOAc/MeOH 10:1); MS m/z: 761.3 (M+H+).

Example 2

[00248] The compound was obtained with the availa able scientific C with steps and intermediates, below.

Stage 2A: Intermediate compound 2A

[00249] To a solution of the product obtained in step 1d of Example 1 (0.12 g, 0.22 mmol) and N-Boc-glycine (0.054 g, 0.31 mmol) in 4.0 ml of acetonitrile was added HATU (133 mg, 0.35 mmol) and DIEA (0.12 ml, 0.66 mmol) at r.t. under stirring. The reaction mixture was subjected to stirring for 2 hours. Analysis methods LC-MS and TLC showed completion of the reaction pair. To the mixture was added 10 ml of EtOAc and was produced by washing the mixture with buffer (pH~4, AcONa/AcOH), NaHCO3and brine and drying over Na2SO4. After solvent removal was performed chromatography of the crude product by silica gel (eluent: EtOAc/hexane). The result was obtained 0.10 g of compound indicated in the heading: Rf0.2 (EtOAc); MSm/z: 664.0 (M+H+).

Stage 2b: Intermediate compound 2b

[00250] the Product obtained in stage 2A (0.10 g 0.15 mmol), was dissolved in 2 ml of 4 NHCl in dioxane and the reaction mixture was stirred for 1 hour at RT. After removal of the solvents to the mixture was added DHM in volume of 3 ml, and then evaporated the mixture to dryness. Procedure add DHM followed by evaporation was repeated three times with the formation of the compound indicated in the title Intermediate compound 2b in the form of its HCl salt (0.10 g). MS m/z: 564.0 (M+H+).

Step 2C:

[00251] (3R,5S)-1-(2-acrylamidoethyl)-5-((1R,2S)-1-(cyclopropanecarbonyl)-2-vinylcyclopropyl)pyrrolidine-3-yl 4-horizontalis-2-carboxylate: The compound was prepared by coupling Intermediate compounds 2b and acrylic acid using HATU in accordance with the description of reactions mates are for Intermediate 2A. Received 50 mg of the compound indicated in the heading: Rf0.1 (EtOAc); MSm/z: 617.9 (M+H+).

[00252] Similarly, in accordance with the method described in Example 2, were obtained the following compounds:

[00253] (3R,5S)-1-(2-((E)-but-2-unaminously)-5-((1R,2S)-1-(cyclopropanecarbonyl)-2-vinylcyclopropyl)pyrrolidine-3-yl-4-horizontalis-2-carboxylate: MS m/z: 632.0 (M+N+).

I-8

[00254] (3R,5S)-1-((R)-2-acrylamidoethyl)-5-((1R,2S)-1-(cyclopropanecarbonyl)-2-vinylcyclopropyl)pyrrolidine-3-yl-4-horizontalis-2-carboxylate: MS m/z: 632.1 (M+H+).

[00255] (3R,5S)-1-((R)-2-acrylamide-3-methylbutanoyl)-5-((1R,2S)-1-(cyclopropanecarbonyl)-2-vinylcyclopropyl)pyrrolidine-3-yl-4-horizontalis-2-carboxylate: MS m/z: 660.2 (M+H+).

[00256] (3R,5S)-1-(2-(2-acetoxybenzoic)acetyl)-5-((1R,2S)-1-(cyclopropanecarbonyl)-2-VIN who cyclopropanecarbonyl)pyrrolidin-3-yl-4-horizontalis-2-carboxylate: MS m/z: 724.0 (M+H +).

[00257] (5S)-1-((R)-2-(2-chloropyrimidine-4-ylamino)propanoic)-5-((1R,2S)-1-(cyclopropanecarbonyl)-2 - vinylcyclopropyl)pyrrolidine-3-yl-4-horizontalis-2-carboxylate: Rf: 0.35 (DHM/Meon 95:5), MS m/z: 690.3 (M+H+).

[00258] Similarly, in accordance with the method described in Example 2, were obtained the following compounds:

[00259] (5S)-1-(2-acetamido-2-(1-acrylamide-3-yl)acetyl)-5-((1R,2S)-1-(cyclopropanecarbonyl)-2-vinylcyclopropyl)pyrrolidine-3-yl-4-horizontalis-2-carboxylate: MS m/z: 715.2 (M+N+).

[00260] (5S)-1-(2-(1-acrylamide-3-yl)-2-(cyclopentanecarbonyl)acetyl)-5-((1R,2S)-1-(cyclopropanecarbonyl)-2-vinylcyclopropyl)pyrrolidine-3-yl-4-horizontalis-2-carboxylate: MS m/z: 785.2 (M+H+).

Example 3

[00261] The compound was obtained according to the steps and intermediates described below.

Stage 3A: progress connection 3A

[00262] To a solution of (S)-3-amino-2-(tert-butoxycarbonylamino)propanoic acid (2.04 g, 10 mmol), TEA (4.5 ml, 30 mmol) in 50 ml of CH2Cl2added nitrobenzenesulfonamide (2.9 g, 13.0 mmol) at RT. The mixture was subjected to stirring for 10 hours RT. The solvent was removed under vacuum, after which was added 100 ml of EtOAc. The organic layer was washed with 1 NHCl (to pH 3), water and brine. Then the organic layer was dried over Na2SO4was filtered and solvent was removed with the formation of the crude product of the Intermediate 3A (4.0 g).

Step 3b: Intermediate compound 3b

[00263] the Crude Intermediate compound 3A (2.0 g), K2CO3(1.5, 4 EQ.) was dissolved in 10 ml of DMF. MeI (0.8 ml, 4 EQ.) was added to the reaction mixture at RT. Formed and the mixture was subjected to stirring for 20 hours. The main part of the DMF was removed under vacuum and added to 100 ml of EtOAc, then washed a mixture of water and a salt solution. The organic layer was dried over Na2SO4. After removal of solvent the crude product was passed through a short column with silica gel (eluent: EtOAc/hexane) to give 1.62 g of the Intermediate compound 3b. MS m/z: 439.9 (M+Na+).

Stage 3C: Intermediate compound 3C

[00264] To a solution of Intermediate compound 3b (1.6 g, 3.8 mmol) in 10 ml THF/MeOH (1:1) was added an aqueous solution of 1 NLiOH (5.8 ml, 5.8 mmol). After stirring at r.t. within 10 hours, the reaction mixture was neutralized using 1.0 NHCl. The organic solvent evaporated under what Aquum and the remaining aqueous phase was acidentially to pH~3 with 1.0 NHCl and were extracted using EtOAc. The organic layer was washed with saline and dried over anhydrous sodium sulfate. After removal of the solvent was obtained 1.5 g of the Intermediate compound 3C. MSm/z: 402.0 (M-1 light picture display).

Stage 3d: Intermediate connection 3d

[00265] To a solution of Intermediate 1d (0.12 g, 0.20 mmol) and Intermediate 3 (0.12 g, 0.3 mmol) to 5.0 ml of anhydrous acetonitrile was added HATU (0.11 g, 0.3 mmol) and DIEA (0.14 ml, 0.9 mmol) at r.t. under stirring. Analysis methods TLC and LC-MS showed completion of the reaction mates after hours. To the reaction mixture was added EtOAc in a volume of 20 ml, and produced washing with buffer (pH~4, AcONa/AcOH), NaHCO3and saline. The organic layer was dried over Na2SO4. After solvent removal, was performed chromatography of the crude product by silica gel (eluent: EtOAc/hexane). The total number of the obtained Intermediate compound 3d was 0.10 g: Rf0.1 (EtOAc); MSm/z: 891.8 (M+H+).

Step 3E: Intermediate compound 3E

[00266] To a solution of Intermediate compound 3d (0.10 g, 0.11 mmol) in 3 ml DMF was added fenistil (30 mg, 0.26 mmol) and K2CO3(40 mg, 0.3 mmol). Formed and the mixture was subjected to stirring for 20 hours at RT. Added 30 ml of EtOAc and produced washing reactionnaire using saline and water. The organic layer was dried over Na2SO4. After solvent removal was performed chromatography of the crude product by silica gel (eluent: EtOAc/hexane) to give 0.1 g of the crude product is Intermediate compound 3E. MS m/z: 706.9 (M+N+).

Step 3f: Compound I-11

[00267] (3R,5S)-1-((S)-2-(tert-butoxycarbonylamino)-3-(N-methylacrylamide)propanol)-5-((1R,2S)-1-(cyclopropanecarbonyl)-2-vinylcyclopropyl)pyrrolidine-3-yl 4-horizontalis-2-carboxylate(I-11)acryloyl chloride (9 μl, 0.11 mmol) was added drop method at 0°C. to a solution of 0.1 g (0.1 mmol) of the product obtained in step 3E in 3 ml DHM containing 0.04 ml (0.3 mmol) of triethylamine, with stirring. The reaction mixture was subjected to stirring at r.t. within 1.5 hours, then diluted in 10 ml DHM. Formed solution was washed twice with saline and dried over magnesium sulfate. After removal of the solvent was obtained the crude product, which was subjected to purification by chromatography with silica gel with the previous elution using a mixture of hexane/EtOAc (1:3 ~ 1:5) and then using EtOAc. The total number of the obtained compound was 20 mg:

[00268] Rf0.15 (EtOAc); MS m/z: 760.9 (M+H+).1HNMR (CD3D, 400 MHz) δ 7.32 (m, 1H), 7.13-6.98 (m, 2H), 6.75 (m, 1H), 6.23 (dd, 1H, J=2.3, 16.5 Hz), 5.73 (m, 2H), 5.45-5.29 (m, 2H), 5.12 (dd, 1H, J=1.4, 10.0 Hz), 4.72 (s, 4H), 4.45 (m, 1H), 4.25-4.09 (m, 1H), 3.91 (m, 1H), 3.75-3.50 (m, 1H), 3.15 (s, 3H), 2.96 (m, 1H), 2.42 (m, 1H), 2.25 (m, 2H), 1.87 (m, 1H), 1.45-0.85 (m, 14H).

[00269] in a Similar fashion, using Intermediate compounds 3E, 2-chloroethanesulfonate and triethylamine received the following connection:

[00270] (3R,5S)-1-((S)-2-(tert-butoxycarbonylamino)-3-(N-methylphenylsulfonyl)propanol)-5-((1R,2S)-1-(cyclopropanecarbonyl)-2-vinylcyclopropyl)pyrrolidine-3-yl 4-horizontalis-2-carboxylate: Rf: 0.55 (DCM/MeOH 95:5); MS m/z: 797.3 (M+H+).

[00271] in a Similar way, using (S)-4-(Fmoc-amino)-2-(tert-butoxycarbonylamino)butane acid phase 3A instead of (S)-3-amino-2-(tert-butoxycarbonylamino)propanoic acid was obtained the following compound:

[00272] (3R,5S)-1-((S)-2-(tert-butoxycarbonylamino)-4-(N-methylacrylamide)butanoyl)-5-((1R,2S)-1-(cyclopropanecarbonyl)-2-vinylcyclopropyl)pyrrolidine-3-yl 4-horizontalis-2-carboxylate: Rf: 0.45 (DCM/MeOH 95:5); MS m/z: 775.3 (M+H+).

[00273] Similarly, in accordance with the description of the methods described in Example 3, it is possible to obtain compound I-14 using ethyl iodide in step 3b instead of methyl iodide:

(3R,5S)-1-((S)-2-(tert-butoxycarbonylamino)-3-(N-ethylacetamide)propanol)-5-((1R,2S)-1-(cyclopropylacetylene is)-2-vinylcyclopropyl)pyrrolidine-3-yl 4-horizontalis-2-carboxylate

[00274] the Compound I-15 was obtained in accordance with the description of the methods described in Example 3, using allyl bromide in step 3b instead of methyl iodide.

[00275] (3R,5S)-1-((S)-3-(N-allylamino)-2-(tert-butoxycarbonylamino)propanol)-5-((1R,2S)-1-(cyclopropanecarbonyl)-2-vinylcyclopropyl)pyrrolidine-3-yl 4-horizontalis-2-carboxylate: Rf: 0.58 (DHM/Meon 95:5); MS m/z: 787.3 (M+H+).

[00276] the Compounds below can be obtained by using as starting compound ethyl ester (1R,2S)-1-amino-2-ethylcyclopropane acid in step 1A, by following the appropriate methods described in Example 3:

(3R,5S)-1-((S)-3-(N-allylamino)-2-(tert-butoxycarbonylamino)propanol)-5-((1R,2R)-1-(cyclopropanecarbonyl)-2-ethylcyclopropane)pyrrolidin-3-yl 4-horizontalis-2-carboxylate

(3R,5S)-1-((S)-2-(tert-butoxycarbonylamino)-3-(N-methylacrylamide)propanol)-5-((1R,2R)-1-(cyclopropanecarbonyl)-2-ethylcyclopropane)pyrrolidin-3-yl 4-horizontalis-2-carboxylate

(3R,5S)-1-((S)-3-acrylamide-2-(tert-butoxycarbonylamino)propanol)-5-((1R,2R)-1-(cyclopropanecarbonyl)-2-ethylcyclopropane)pyrrolidin-3-yl 4-persoally-2-carboxyl the t

[00277] in a Similar way received the following connection:

[00278] (5S)-1-(2-(tert-butoxycarbonylamino)-3-((E)-4-(dimethylamino)-N-methylbut-2-enamide)propanol)-5-((1R,2S)-1-(cyclopropanecarbonyl)-2-vinylcyclopropyl)pyrrolidine-3-yl 4-horizontalis-2-carboxylate:

[00279] Rf: 0.45 (DHM/Meon 95:5); MS m/z: 818.5 (M+H+).

Example 4

[00280] the Connection specified in the header received in accordance with the steps and intermediates described below.

Step 4A: Interim connection 4A

[00281] the Intermediate compound 1e ethane 1e worked 4 NHCl in accordance with the method described for stage 1d, to obtain the Intermediate compound 4A in the form of its HCl salt.

MS m/z: 815.2 (M+N+).

Step 4b: Intermediate compound 4b

[00282] Cyclopentylpropionate (1.5 EQ.) added drip method at 0°C. to a solution of Intermediate 4A (1 EQ.) from step 4A, DHM containing 3 EQ. of triethylamine, with stirring. The reaction mixture was subjected to stirring at r.t. for 1.5 and then diluted in 10 ml DHM. Formed solution was twice washed with saline and dried over magnesium sulfate. After removal of solvent received neojidanni the product, which was subjected to purification by chromatography with silica gel, with the previous elution using a mixture of hexane/EtOAc (1:3 ~ 1:5) and then with EtOAc to obtain the specified connection (60-90%): MS m/z: 925.2 (M-1).

Step 4C: Compound I-19

[00283] (3R,5S)-1-((S)-3-acrylamide-2-(cyclopentanecarbonyl)propanol)-5-((1R,2S)-1-(cyclopropanecarbonyl)-2-vinylcyclopropyl)pyrrolidine-3-yl 4-horizontalis-2-carboxylate: the compound indicated in the title, were obtained on the basis of the Intermediate 4b in accordance with the description of the methods described for stage 1f and step 1g. MS m/z: 759.0 (M+H+).

[00284] Similarly, by using as the source of the corresponding intermediate compounds were obtained the following compounds:

[00285] (3R,5S)-1-((S)-2-(cyclopentanecarbonyl)-3-(N-methylacrylamide)propanol)-5-((1R,2S)-1-(cyclopropanecarbonyl)-2-vinylcyclopropyl)pyrrolidine-3-yl 4-horizontalis-2-carboxylate: Rf: 0.4 (EtOAc/Meon 20:1); MS m/z: 773.2 (M+H+).

[00286] (3R,5S)-1-((S)-2-(cyclopentanecarbonyl)-4-(N-methylacrylamide)butanoyl)-5-((1R,2S)-1-(cyclopropanecarbonyl)-2-vinylcyclopropyl)pyrrolidine-3-yl 4-horizontalis-2-carboxylate: Rf: 0.4 (EtOAc/Meon 20:1); MS m/z: 787.3 (M+H+).

[00287] (5S)-5-((1R,2S)-1-(cyclopropanecarbonyl)-2-vinylcyclopropyl)-1-(2-(4-(methoxycarbonyl)bicyclo(2.2.2]octane-1 - carboxamido)-3-(N-methylacrylamide)propanol)pyrrolidin-3-yl 4-horizontalis-2-carboxylate:

[00288] Rf: 0.45 (EtOAc/MeOH 10:1); MS m/z: 855.3 (M+H+).

Example 5

[00289] The compound was obtained according to the steps and intermediates described below.

Step 5A: Intermediate compound 5A

[00290] To a solution of Boc-L-threonine (0.44 g, 2.0 mmol) in 10.0 ml DHM added crotyl chloride (0.32 g, 3.0 mmol) at RT and then added a catalytically effective amount of DMAP and TEA (1.0 ml, 6 mmol). The reaction mixture was subjected to stirring for 10 hours at RT. The reaction was stopped by adding an aqueous solution of NaHCO3(10 ml). After 2 hours produced a slow addition of aqueous solution of 1 NHCl to pH ~3. Layer DHM collected and produced by extraction of the aqueous phase using DHM (2×10 ml). The organic layer was dried over Na2SO4was filtered and solvent was removed to obtain the crude product.

Step 5b: I-22

[00291] (3R,5S)-1-((2S,3R)-3-((E)-but-2-enolase)-2-(tert-butoxycarbonylamino)butanoyl)-5-((1R,2S)-1-(cyclopropanecarbonyl)-2-vinylcyclopropyl)pyrrolidine-3-yl-4-f is horizontalen-2-carboxylate: the Compound specified in the header, was obtained by coupling of Intermediate compounds 1d of Example 1 and Intermediate compounds 5A using HATU, in accordance with the description of reactions mates are for Intermediate compound 1E in Example 1. The total number of substances obtained from 109 mg of Intermediate 1d, was 90 mg: Rf0.5 (EtOAc); MS m/z: 774.3 (M+H+).

[00292] the following compounds were obtained in a similar way, when used as a source of Intermediate compounds 1d, by pairing with the corresponding intermediate compounds:

[00293] (3R,5S)-1-((S)-2-(tert-butoxycarbonylamino)-3-(2-(tert-butoxycarbonylamino)acryloyloxy)propanol)-5-((1R,2S)-1-(cyclopropanecarbonyl)-2-vinylcyclopropyl)pyrrolidine-3-yl 4-horizontalis-2-carboxylate: MS m/z: 862.2 (M-1).

[00294] (3R,5S)-1-((2S,3R)-3-(acryloyloxy)-2-(tert-butoxycarbonylamino)butanoyl)-5-((1R,2S)-1-(cyclopropanecarbonyl)-2-vinylcyclopropyl)pyrrolidine-3-yl 4-horizontalis-2-carboxylate: Rf: 0.4 (EtOAc); MS m/z: 760.1 (M-1).

Example 6

Compound I-25

[00295] (I-25): This compound was obtained according to the steps and intermediates described below.

Step 6A: Intermediate compound 6A

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[00296] To a solution of N-Boc-pyroglutamic acid (0.23 g, 1.0 mmol) in 10.0 ml of anhydrous THF was slowly added (2-methylprop-1-enyl)magnesium bromide (0.5 M in TF, 5 ml, 2.5 mmol) at -78°C. the Reaction mixture was subjected to stirring for 1 hour at -78°C. then was added an aqueous solution of 1 NHCl (2.5 ml) and the mixture was slowly heated to RT. The pH value was brought to ~3-4 with 1 N NHCl. Then made the removal of THF under vacuum and the remaining aqueous phase was extracted using DHM (3×15 ml), the organic layer was dried over Na2SO4produced filtration and solvent was removed to obtain the crude product.

Step 6b: I-25

[00297] (3R,5S)-1-((S)-2-(tert-butoxycarbonylamino)-7-methyl-5-exoect-6-enoyl)-5-((1R,2S)-1-(cyclopropanecarbonyl)-2-vinylcyclopropyl)pyrrolidine-3-yl 4-horizontalis-2-carboxylate: the Compound indicated in the title, was obtained by coupling of Intermediate compounds 1d of Example 1 and Intermediate compounds 6A using HATU, in accordance with the description of the reactions of conjugation, see for Intermediate compound 1E in Example 1. The total number of connections specified in the header received from 108 mg of Intermediate 1d, was 80 mg: Rf0.3 (EtOAc); MSm/z: 774.1 (M+H+).1HNMR (CD3OD, 400 MHz) δ 7.31 (dd,1H, J=13.3, 7.4 Hz), 7.13-6.98 (m, 2H), 6.18 (s, 1H), 5.74 (m, 1H), 5.38 (s, 1H), 5.32 (d, 1H, J=17.0 Hz), 5.12 (d, 1H, J=10.1 Hz), 4.72 (s, 4H), 4.48 (dd, 1H, J=17.0, 9.16 Hz), 4.29 (m, 2H), 3.89 (m, 1H), 2.93 (m, 1H), 2.60-2.35 (m, 2H), 2.22 (m. 2H), 2.10 (s, 3H), 2.02-1.75 (br, 1H), 1.88 (s, 3H), 1.46-0.80 (m, 14H).

13C NMR (CD3OD, 100 MHz):

δ 201.8, 175.3, 174.5, 170.6, 157.7, 156.9, 155.6, 141.1, 134.2, 131.2, 124.8, 119.9, 119.7, 118.6, 115.0, 1 14.8, 80.3, 76.1, 61.0, 55.0, 54.9, 54.8, 53.5, 53.3, 52.7, 50.3, 50.1, 42.6, 40.3, 35.7, 35.4, 32.1, 28.7, 28.5, 27.7, 26.9, 24.0, 20.9, 6.74, 6.47.

[00298] the following compounds were obtained when using as the source of Intermediate compounds 1d, by pairing it with the corresponding intermediate compounds, in accordance with the description for Stage 6A:

[00299] (3R,5S)-1-((S,E)-2-(tert-butoxycarbonylamino)-5-exoect-6-enoyl)-5-((1R,2S)-1-(cyclopropanecarbonyl)-2-vinylcyclopropyl)pyrrolidine-3-yl-4-horizontalis-2-carboxylate: the Total number of connections specified in the header obtained from 150 mg of Intermediate 1d, was 80 mg: Rf0.3 (EtOAc); MS m/z: 760.3 (M+H+).

[00300] (3R,5S)-1-((S)-2-(tert-butoxycarbonylamino)-6-methyl-5-exoept-6-enoyl)-5-((1R,2S)-1-(cyclopropanecarbonyl)-2-vinylcyclopropyl)pyrrolidine-3-yl 4-horizontalis-2-carboxylate: the Total number of connections specified in the header obtained from 150 mg of Intermediate 1d, was 80 mg: Rf0.4 (EtOAc); MSm/: 782.2 (M+Na +).1HNMR (CD3OD, 400 MHz) δ 7.31 (dd, 1H, J=13.3, 7.4 Hz), 7.09 (dd, 1H, J=33, 7.4 Hz), 7.0 (m, 1H), 6.12 (s, 1H), 5.82 (s, 1H), 5.74 (m, 1H), 5.39 (s, 1H), 5.31 (dd, 1H, J=1.4, 17.0 Hz), 5.12 (dd, 1H, J=10.1, 1.4 Hz), 4.73 (m, 4H), 4.48 (m, 1H), 4.32 (m, 2H), 3.90 (m, 1H), 2.91 (m, 1H), 2.42 (m, 1H), 2.22 (m, 2H), 2.01 (m, 1H), 1.90-1.85 (m, 2H), 1.84 (s, 3H), 1.40-1.02 (m, 14H).

[00301] (3R,5S)-1-((S,E)-2-(tert-butoxycarbonylamino)-6-methyl-5-exoect-6-enoyl)-5-((1R,2R)-1-(cyclopropanecarbonyl)-2-ethylcyclopropane)pyrrolidin-3-yl 4-horizontalis-2-carboxylate: the Total number of connections specified in the header obtained from 150 mg of Intermediate 1d, was 80 mg: Rf0.5 (EtOAc); MS m/z: 796.2 (M+Na+).

[00302] the following compounds were obtained when using as the source of Intermediate compounds 1d, by pairing it with the corresponding intermediate compounds, in accordance with the description for the

(3R,5S)-1-((S)-2-(tert-butoxycarbonylamino)-5-exoect-6-enoyl)-5-((1R,2S)-1-(cyclopropanecarbonyl)-2-vinylcyclopropyl)pyrrolidine-3-yl 4-horizontalis-2-carboxylate

(3R,5S)-1-((S)-2-(tert-butoxycarbonylamino)-8,8-dimethyl-5-Oksanen-6-enoyl)-5-((1R,2S)-1-(cyclopropanecarbonyl)-2-vinylcyclopropyl)pyrrolidine-3-yl 4-horizontalis-2-carboxylate

(3R,5S)-1-((S)-2-(tert-butoxy bonellii)-3,3,7-trimethyl-5-exoect-6-enoyl)-5-((1R,2S)-1-(cyclopropanecarbonyl)-2-vinylcyclopropyl)pyrrolidine-3-yl 4-horizontalis-2-carboxylate

(3R,5S)-1-((S)-2-(tert-butoxycarbonylamino)-4,4,7-trimethyl-5-exoect-6-enoyl)-5-((1R,2S)-1-(cyclopropanecarbonyl)-2-vinylcyclopropyl)pyrrolidine-3-yl 4-horizontalis-2-carboxylate

(3R,5S)-1-((S)-2-(tert-butoxycarbonylamino)-6-methyl-4-exoept-5-enoyl)-5-((1R,2S)-1-(cyclopropanecarbonyl)-2-vinylcyclopropyl)pyrrolidine-3-yl 4-horizontalis-2-carboxylate

[00303] MS m/z: 760.1 (M+H+).

[00304] the following compound was obtained using catalyzed by palladium hydrogenation of Intermediate 1d, after which the reaction was performed mates described for step 6b:

[00305] (3R,5S)-1-((S)-2-(tert-butoxycarbonylamino)-7-methyl-5-exoect-6-enoyl)-5-((1R,2R)-1-(cyclopropanecarbonyl)-2-ethylcyclopropane)pyrrolidin-3-yl 4-horizontalis-2-carboxylate: the Total number of connections specified in the header obtained from 100 mg of Intermediate 1d, was 33 mg: Rf0.5 (EtOAc); MS m/z: 776.2 (M+H+).

[00306] in a Similar way received the following connections:

(3R,5S)-1-((S)-2-(tert-butoxycarbonylamino)-6-methyl-5-exoept-6-enoyl)-5-((1R,2R)-1-(cyclopropanecarbonyl)-2-ethylcyclopropane)pyrrolidin-3-yl 4-horizontalis-2-carboxylate

(3R,5S)-1-((S)-2-(tert-butoxycarbonylamino)-6-methyl-4-exoept-5-enoyl)-5-((1R,2R)-1-(cyclopropanecarbonyl)-2-ethylcyclopropane)pyrrolidin-3-yl 4-horizontalis-2-carboxylate

I-61

(3R,5S)-1-((S)-2-(tert-butoxycarbonylamino)-5-cyclobutyl-5-oxopentanoic)-5-((1R,2R)-1-(cyclopropanecarbonyl)-2-ethylcyclopropane)pyrrolidin-3-yl 4-horizontalis-2-carboxylate

(3R,5S)-1-((S)-2-(tert-butoxycarbonylamino)-6-cyclobutyl-5-oxohexanoyl)-5-((1R,2R)-1-(cyclopropanecarbonyl)-2-ethylcyclopropane)pyrrolidin-3-yl 4-horizontalis-2-carboxylate

(3R,5S)-1-((S,E)-2-(tert-butoxycarbonylamino)-8,8-dimethyl-5-Oksanen-6-enoyl)-5-((1R,2R)-1-(cyclopropanecarbonyl)-2-ethylcyclopropane)pyrrolidin-3-yl 4-horizontalis-2-carboxylate

(3R,5S)-1-((S)-2-(tert-butoxycarbonylamino)-2-((S)-2-oxocyclopent-3-enyl)acetyl)-5-((1R,2R)-1-(cyclopropanecarbonyl)-2-ethylcyclopropane)pyrrolidin-3-yl 4-horizontalis-2-carboxylate

[00307] the following compound was obtained using catalyzed by palladium hydrogenation (24-48 hours) Intermediate compounds 1d, after which the reaction was performed mates described for step 6b:

p> (3R,5S)-1-((S)-2-(tert-butoxycarbonylamino)-7-methyl-5-exoect-6-enoyl)-5-(1-(cyclopropanesulfonyl)-1-oxohexyl-2-ylcarbonyl)pyrrolidin-3-yl 4-horizontalis-2-carboxylate: the Total number of connections specified in the header obtained from 100 mg of Intermediate 1d, was 58 mg: Ra0.5 (EtOAc); MS m/z: 800.2 (M+Na+).

Example 7

[00308] (1aR,3aS,5R,9S,16aS,Z)-11-(o-nitrophenyloctyl)-9-(tert-butoxycarbonylamino)-1A-(cyclopropanecarbonyl)-3,8-dioxo-1,1A,2,3,3A,4,5,6,8,9,10,11,12,13,14,16A-hexamethylcyclopenta[n]pyrrolo[2,1-C][1,4,8]diazacyclooctadecane-5-yl-4-horizontalis-2-carboxylate (I-45): Specified compound was obtained in accordance with steps and intermediates described below:

Stage 7a: Intermediate compound 7a

[00309] the Intermediate compound 7a was obtained by following the methods described for the synthesis of Intermediate compound 3c with the use of 5-bronet-1-ene as the alkylating agent.

Step 7b: Intermediate compound 7b

[00310] the Intermediate compound 7b was obtained by coupling of Intermediate compounds 1d of Example 1 and Intermediate compounds 7a using HATU, in accordance with the description of reactions mates are for Intermediate compound 1E in With the ore 1. MS: 946.2 (M+1).

Step 7C: compound I-45

[00311] To a solution of 540 mg of the Intermediate 7b in 150 ml of anhydrous dichloromethane was added 100 mg of 1,3-Bis(2,4,6-trimetilfenil)-4,5-dihydroimidazole-2-ilidene[2-(i-propoxy)-5-(N,N-dimethylaminomethyl)phenyl]methylindoline (II) (catalyst Jean 1B, RC-303, ZannanPharma Ltd.) under nitrogen. Formed the mixture was heated at 48°C during the night. The analysis method LC-MS showed complete conversion to the product of the reaction. The reaction solution was subjected to flash chromatography with silica gel with eluent a (heptane/EtOAcv/v 1:1 to pure EtOAc) with the formation of 140 mg of the product according to the present invention. MS: 916.3 (ES).

[00312] in a Similar fashion, by using as the source connection (S)-5-allylamino-2-tert-butoxycarbonylamino-pentanol acid phase 7a, received the following connection:

[00313] MS: 916.3 (ES-)

[00314] in a Similar fashion, by using as a starting compound ((S)-4-[But-3-enyl-(2-nitrobenzenesulfonyl)-amino]-2-tert-butoxycarbonylamino-butyric acid in step 7a, received the following connection:

[00315] MS: 916.3 (ES-)

Example 8

[00316] (1aR,3aS,5R,9S,16aS,Z)-11-acryloyl-9-(tert-butoxycarbonylamino)-1A-(cyclopropanecarbonyl)-3,8-dioxo-1,1A,2,3,3A,4,5,6,8,9,10,11,12,13,14,16A-hexade is hydrocyclone[n]pyrrolo[2,1-C][1,4,8]diazacyclooctadecane-5-yl-4-horizontalis-2-carboxylate(I-35): the Indicated compound was obtained in accordance with steps and intermediates, below:

[00317] Step 8A: Intermediate compound 8A:

[00318] the Intermediate compound 8A was obtained by treatment of compound I-45 from Example 7 thiophenols in accordance with the methods described for step 3e. MS: 733.3 (M+1).

[00319] the Stage 8b: Compound I-35:

[00320] The compound was obtained by treatment of an Intermediate compound 8A acryloyl chloride in accordance with the method described for step 3f. Rf0.2 (5% Meon in DCM); MS m/z: 787.3 (M+H+).

[00321] in a Similar way when exposed to the Intermediate connection 8A chloroacetyl chloride (1.2 equiv.) received the following connection:

(1aR,3aS,5R,9S,16aS,Z)-11-chloroacetyl-9-(tert-butoxycarbonylamino)-1A-(cyclopropanecarbonyl)-3,8-dioxo-1,1A,2,3,3A,4,5,6,8,9,10,11,12,13,14,16A-hexamethylcyclopenta[n]pyrrolo[2,1-C][1,4,8]diazacyclooctadecane-5-yl-4-horizontalis-2-carboxylate (I-45)

[00322] MS: 831.2 (M+Na+).

[00323] in a Similar way when exposed to the Intermediate connection 8A chloroacetyl chloride (3.0 EQ.), received the following connection:

[00324] MS: 907.2 (M+Na+).

[00325] in a Similar fashion, by using as starting compounds I-53 and I-54 phase 8A, received the following connection:

(1aR,3aS,5R,9S,16aS,Z)-12-acryloyl-9-(tert-butoxycarbonylamino)-1A-(cyclopropanecarbonyl)-3,8-dioxo-1,1A,2,3,3A,4,5,6,8,9,10,11,12,13,14,16A-hexamethylcyclopenta[n]pyrrolo[2,1-C][1,4,9]diazacyclooctadecane-5-yl 4-horizontalis-2-carboxylate

[00326] MS: 785.3 (ES).

(1aR,3aS,5R,9S,16aS,Z)-13-acryloyl-9-(tert-butoxycarbonylamino)-1A-(cyclopropanecarbonyl)-3,8-dioxo-1,1A,2,3,3A,4,5,6,8,9,10,11,12,13,14,16A-hexamethylcyclopenta[e]pyrrolo[1,2-a][1,4,10]diazacyclooctadecane-5-yl 4-horizontalis-2-carboxylate

[00327] MS: 787.3 (ES+), 785.3 (ES).

Example 9 (I-40)

[00328] The compound was obtained according to the steps and intermediates described in the following scheme:

2,2-dimethylhex-5-enyl carbonochloridic

[00329] Methyl-2,2,12,12-tetramethyl-4,9-dioxo-3,10-dioxa-5,8-desiccated-15-ene-7-carboxylate (Intermediate compound 9-1). To a solution of 381 mg of methyl 2-amino-3-(tert-butoxycarbonylamino)propanoate hydrochloride and 1 ml of Et3N in 10 ml of anhydrous THF, was added 1.5 mmol of 2,2-dimethylhex-5-eilerson of chloridate under stirring. Formed and the mixture was subjected to stirring at r.t. during the night, and then concentrated. The residue was subjected to chromatography in a column of silica gel with a mixture of heptane/EtOAc (v/v 7/1) as eluent, to obtain 50 mg of colorless oil (90%). 1HNMR (400 MHz, CDCl3d 5.82 (m, 1H), 5.64 (br s, 1H), 5.02 (dd, 1H, J=13.2, 1.8 Hz), 4.95 (d, 1H, J=8.2 Hz), 4.83 (br s, 1H), 4.39 (br d, 1H, J=6.0 Hz), 3.83 (br s, 2H), 3.80 (s, 3H), 2.55 (m, 2H), 2.02 (m, 2H), 1.43 (s, 9H), 1.35 (m, 2H), 0.91 (s, 6H).

[00330] 2,2,12,12-tetramethyl-4,9-dioxo-3,10-dioxa-5,8-desiccated-15-ene-7-carboxylic acid (Intermediate compound 9-2). To a mixture of 500 mg of the intermediate 8-1 in a mixed solvent Meon-THF (5 ml-5 ml), was added 420 mg of LiOH-H2O, then a 5 water under stirring. The reaction mixture was subjected to stirring at r.t. within 30 minutes, after which sekilala using 12 ml of 1.0 NHCl and were extracted using 60 ml of dichloromethane. The organic layer was washed using saline and dried over MgSO4. After filtration and concentration as a product according to the present invention was obtained 460 mg of a viscous oil (95%). LC-MS: m/z=357.2 (ES).

[00331] (2S,4R)-1-tert-butyl-2-methyl-4-(4-promisingly-2-carbonyloxy)pyrrolidine-1,2, in primary forms (Intermediate compound 9-3). To a solution containing 0.5 g of (2S,4R)-1-tert-butyl-2-methyl-4-hydroxypyrrolidine-1,2-in primary forms (2 mmol) in 5 ml of N,N-dimethylacetamide, was added 370 mg of carbonyl diimidazole (1.1 EQ.). The reaction mixture was heated at 60°C for 1 hour, after which was added 400 mg of 4-promisingly (2 mmol). The reaction was continued at 60°C over night. After Oh what adenia was produced by extraction of the reaction mixture with 50 ml of EtOAc and washing with water and brine, then was dried over Na2SO4. After filtration and concentration the residue was purified using flash chromatography with silica gel, using a mixture of heptane/EtOAc (v/v 5/2), with formation of a solid 75 mg connections, white (79%). LC-MS: m/z=369.2 (ES+, M+1 - Boc).

[00332] (2S,4R)-1-tert-butyl-2-methyl-4-(4-finalizando-2-carbonyloxy)pyrrolidine-1,2, in primary forms (Intermediate compound 9-4). Under Ar, a solution of 745 mg of the intermediate 8-3 (1.58 mmol) in 30 ml of degassed toluene was added 170 mg of palladium tetranitroaniline, and then 700 μl of vinyl tributyltin (2.4 mmol). The reaction mixture was heated at 100°C during the night. After cooling, the solvent was removed under reduced pressure and the residue was purified using flash chromatography with silica gel using a mixture of heptane/EtOAc (v/v 9/1 - 3/1) as eluent, with the formation of 533 mg solid connections, white (81%). LC-MS: m/z=317.2 (ES+, M+1-BOC).

[00333] (2S,4R)-methyl-1-((S)-3-(tert-butoxycarbonylamino)-2-((2,2-dimethylhex-5-enyloxy)carbylamine)propanyl)-4-(4-vinyl-2,3-dihydro-1H-inden-2-carbonyloxy)pyrrolidine-2-carboxylate (Intermediate compound 9-5). From the composition of the Intermediate 9-4 remove the protective group BOC using 4 NHCl in dioxane according to the present description given in Example 1. 288 mg specified the CSOs de Vos intermediate compound (0.82 mmol) was added to the mixture, contains 358 mg of the Intermediate 9-2 (1.0 mmol), 400 mg of HATU (1.05 mmol), 300 μl of DIPEA in 5 ml TF, with stirring. After stirring over night the reaction mixture was dissolved in 60 ml of EtOAc and washed with saturated sodium bicarbonate, dissolved HCl and dried over Na2SO4. After concentration and purification by the method of flash chromatography silica gel with a mixture of heptane/EtOAc (1/1) received 485 mg solid connections, white (90%). LC-MS: m/z=557.3 (ES+, M+1-BOC).

[00334] Macrocyclic Intermediate connection 9-6. Under the N2a diluted solution of 485 mg of the intermediate 9-5 and 150 mg of catalyst Jean 1B stirred in 100 ml of degassed dichloroethane at 50°C during the night. After the reaction mixture was passed through a short column and suirable using a mixture of heptane/EtOAc (v/v 1/3). Then a concentrated fraction was subjected to pre-HPLC purification with the formation of 312 mg of the solid compound of a brownish color (70%). LC-MS: m/z=529.2 (ES+, M+1-BOC).

[00335] Macrocyclic Intermediate connection 9-8. 25 mg of the intermediate 9-6 was subjected to hydrolysis in 1 ml Meon and 1 ml of THF, using 1 ml of 1.0 NLiOH, for 1 hour. After this was added 1.2 ml of 1.0 NHCl and made the extraction of the reaction mixture using 30 ml of dichloromethane. The organic layer was washed sole is the first solution, and dried over MgSO 4. After filtration and concentration the residue was re-dissolved in 1.5 ml of anhydrous acetonitrile, followed by adding 25 mg of HATU, 200 μl of DIPEA and then 13 mg of the intermediate 9-7. After stirring at r.t. within 20 minutes the reaction mixture was concentrated and directly after that was purified using prep-HPLC to obtain 20 mg of the solid compound in yellow (61%). LC-MS: m/z=727.2 (ES+, M+1-BOC, 825.2 (ES).

[00336] I-40: To a solution of 20 mg of the intermediate 9-8, stirred in 1 l of dichloromethane, was added 1.5 ml of 4.0 MHCl in dioxane. After 30 minutes the reaction mixture was concentrated. To the precipitate was added 1.5 ml of acetonitrile, 200 μl DIPEA, 20 mg of acrylic acid and 50 mg of HATU. After stirring at r.t. within 30 minutes the reaction mixture was concentrated and was purified by the method prep-HPLC to obtain 14.0 mg solid connections, white (68%). LC-MS: m/z=779.3 (ES), 781.2 (ES+)

[00337] in a Similar fashion, using Intermediate compounds 3c instead of Intermediate compound 3A as a source, you may get the following connection:

[00338] in a Similar fashion, by using saturated form Intermediate compound 9-6, you may get the following connection:

[00339] LC-MS: m/z=781.3 (ES), 783.2 (ES+).

[00340] in a Similar way, after hydrogenases and as an Intermediate connection 9-7, and the Intermediate 9-8 before pairing, you may get the following connection:

Example 10

[00341] the northward-macrocycle I-45. The specified connection was obtained in accordance with steps and intermediates described in the following scheme:

[00342] (S)-2-benzyl-1-(2,2-dimethylhex-5-enyl)-5-oxopyrrolidin-1,2, in primary forms (Intermediate compound 10-1): At 0°C to a solution of 440 mg of benzyl ether 5-oxo-pyrrolidin-2-carboxylic acid (2 mmol), 300 μl of triethylamine, 300 mg of N,N-dimethylaminopyridine (2.2 mmol) in 7 ml of dichloromethane was added 2 mmol of 4-pentenyl-1 mlhloropiramina under stirring. Then the reaction mixture was warmed up to r.t. and was subjected to stirring for 24 hours. The precipitate formed after concentration was dissolved in 40 ml of EtOAc, washed 6 ml of 1.0 N aqueous HCl salt solution, and dried over anhydrous sodium sulfate. After that evaporated organic solvent under reduced pressure and the residue was purified using flash chromatography with silica gel using a mixture of heptane/EtOAc (/4/1-2/1), to obtain 430 mg unpainted oil as an intermediate compound 10-1 (58%).

[00343] 2-((2,2-dimethylhex-5-enyloxy)carbylamine)-7-methyl-5-exoect-6-ANOVA Ki the lot (Intermediate compound 10-2). Using 13 mg of Pd(OAc)2, 24 ál Et3H, 278 μl of Et3SiH made dibenzylamine, in 2 ml of dichloromethane at r.t. within 30 minutes. After filtration, the concentrated residue was subjected to Grignard reagent (2.5 EQ.) at -78°C for 2 hours. After stopping the reaction with dilute acid, the product was extracted using dichloromethane and dried over MgSO4. The concentrated product is an Intermediate compound 10-2 according to the present invention, which is directly used in the next step.

[00344] In application of the General methods described in Example 9, was produced by the removal of the Boc protective group of the composition of the Intermediate 9-4 using 4 N HCl, then was carried out by pairing it with an intermediate 10-2 connection with the formation of Intermediate compounds 10-3. The intermediate connection 10-3 subjected to the reaction of olefinic exchange when using the catalyst of verification or catalyst Jean with the formation of macrocyclic Intermediate 10-4. After carrying out basic hydrolysis using LiOH was performed by pairing the acidic form intermediate compounds 10-4 with an Intermediate connection 9-7 to obtain compound I-55. MS: m/e=806.3 (ES-).

[00345] in a Similar way it is possible to obtain compound I-56 using saturated form Intermediate is about connections 9-7:

Example 11

Expression and purification of single-stranded peptide of HCV protease (wt)

[00346] the single-Stranded proteolytic domain (NS4A21-32-GSGS-NS33-631) cloned into cells pET-14b (Novagen, Madison, WI) and transformed into DH10B cells (Invitrogen). Formed in the plasmid was transferred into Escherichia coli BL21 (Novagen) for protein expression as described previously (1, 2). Briefly, cultures were grown at 37°C in LB medium containing 100 μg/ml ampicillin, until the value of optical density at 600 nm (OD 600)equal to 1.0 by adding isopropyl-β-D-thiogalactopyranoside (IPTG) to 1 mm. After an additional incubation period at 18°C for 20 hours to harvest the bacterial cells by centrifugation at 6000×g for 10 minutes and resuspendable in lytic buffer containing 50 mm Na3PO4, pH 8.0, 300 mm NaCl, 5 mm 2-mercaptoethanol, 10% glycerol, 0.5% Igepal CA630, and a mixture of protease inhibitors consisting of 1 mm phenylmethylsulfonyl fluoride, 0.5 mg/ml leupeptin, Pertamina and 2 mm benzamidine. Cells were literally method of freezing-thawing and subsequent destruction by ultrasound. Cell debris was separated by centrifugation at 12000×g for 30 minutes. The supernatant was subjected to further purification by passing through a 0.45-μm filter (Corning), and then through a HiTrap chelating column, NiSO 4(Amersham Pharmacia Biotech). Bound protein was suirable using a solution of imidazole in 100-to 500 mm linear gradient. Selected fractions were subjected to chromatography in Ni2+column and were analyzed by 10% sodium-dodecyl-sulfate (SDS)-polyacrylamide gel. Purified protein was separated by electrophoresis in 12% SDS-PAGE (polyacrylamide) gel, and then transferred to nitrocellulose membrane. After this protein was examined by the method of Western blotting with monoclonal antibodies to NS3. Proteins were visualized using a set of chemiluminescence (Roche) antibodies goat antibodies to mouse conjugated to horseradish peroxidase (Pierce) as secondary antibodies. Taking protein aliquots and store them at -80°C.

Example 12

Cloning and expression of drug-resistant mutants of HCV protease A156S, AT, D168A, and D168V variants C159S

[00347] using PCR method was obtained mutant DNA fragments NS4A/NS3 and cloned them into the expression vector pet. After transformation into competent cells BL21 expression was induced using IPTG for 2 hours. Proteins merge labeled with histidine, was purified by the method of affinity chromatography and then by the method of exclusion chromatography.

Example 13

[00348] Analytical buffer: 2% CHAPS, 50 mm Tris pH 7.5, 50% glycerol, 2 mm M-2235 (Bachem) substrate. To 50 μl was added to 49 μl of analytical what about the buffer, 1 μl (1U) serine protease HCV (Bioenza). Incubated for 20 minutes at room temperature. Record the results on the tablet was produced as in 350/460 nm (excitation/emission), the fluorescent apparatus for reading microplates and has been read with 1-minute intervals with obtaining kinetic curve.

[00349] the Enzyme endured the effects of 1% DMSO and 2% methanol. In other experiments of compounds compounds were dissolved in 10 times in pure DMSO with 20% methanol (10% DMSO and 20% methanol). The solution of these compounds was added to the reaction mixture (not exceeding 10% of the final volume of the reaction). The final concentration of organic solvents were 1% DMSO and 2% methanol.

Example 14

Additional research protocols

Method:

[00350] the Compounds investigated to assess the antiviral activity and cytotoxicity of compounds in vitro using HCV RNA replicons. The study used a cell line ET (luc-ubi-neo/ET), which is a cell line human hepatoma Huh7 containing the replicon HCV RNA with a stable luciferase reporter (Luc) and three cell mutations, bearing the adaptive nature of culture. Levels of HCV RNA directly measured by applying virus-specific TaqMan RT-PCR:

Direct primer: ACGCAGAAAGCGTCTAGCCAT (SEQ ID NO:63)

Reverse primer: TCTCACCGGTTCCGCAGA (SEQ ID NO:64)

sample: [6-FAM]-CCTGGAGGCTGCACGACACTCAT-[TAMRA] (SEQ ID NO:65)

[00351] the cell Line EM were grown in modified nutrient medium, Dulbecco (DMEM) with 10% fetal bovine serum (FBS), 1% penicillin-streptomycin (pen-strep), 1% glutamine, 250 μg/ml G418 in 5% incubator with CO2at 37°C. All reagents for culturing received Mediatech (Manassas, VA). The cells were trypsinization (1% trypsin:EDTA) and was applied in an amount of 5·103cells per well on 96-well analytical plates (Costar) for measuring the number of cells (cytotoxicity) or antiviral activity. Made preparations to 3-fold concentrations, 6 holes each, and perform the analysis in DMEM, 5% FBS (FBI), 1% pen-strep, 1% glutamine. As a positive control in each assay used interferon Alfa-2b human (PBL Biolabs, New Brunswick, NJ). After 72 hours after making preparations cells were still subconfluent. Antiviral activity was measured by analyzing the activity of luciferase synthesized with replicon using Steady-Glo Luciferase Assay System (Promega, Madison, WI) according to manufacturer's instructions. The number of cells in each well was determined by applying a reagent CytoTox-1 (Promega). The connection profile was obtained by calculating the corresponding values of the EU50(effective concentration at which viral replication is inhibited by approximately 50%), E is 90(effective concentration at which viral replication is inhibited by approximately 90%), IC50(the concentration at which cell survival is reduced to 50%) and SI50(index selection: EU50/IC50). The values of the IC50for some compounds are given below in Table 5.

Method: the study of HCV protease, the method RPAF

[00352] To identify inhibitors of the HCV protease NS3/4A was used quantitative method based on resonance energy transfer fluorescence (CHEF, FRET). The analysis used a synthetic peptide for RPAF obtained on the basis of customers cutting NS5A/5B HCV, HCV protease to evaluate the activity of compounds against protease carried out by measuring the splitting activity of the complex. Synthetic peptide, containing in its composition area connection NS5A-5B (NH2-EDVVCCSMSYK-COOH), contributed label Dabcyl (dimethylaminoazobenzene acid) and Edans N - and C-ends, respectively (Invitrogen, Carlsbad, CA). For the measurement of IC50the analyzed compounds was carried out by measuring fluorescence. The two form a fluorophore pair, fluorescence quenching, and form CAF inside the structure of the intact peptide. Fluorescence is restored after cleavage of the peptide detecting CHEF, a complex of proteinase HCV NS3/4A (100 ng/ml), and continuous metering is in the spectrum of excitation/emission = 340/490 nm.

Example 15

Analysis of the HCV Protease Method RPAF carried out on the Enzyme Wild-Type and Mutant NS3/4A 1b (IC50)

[00353] to obtain the values For IC50such as values for 239 connection shown above in Table 4. The Protocol is a modified analysis based CHEF (v_02)derived from In Vitro Resistance Studies of HCV Serine Protease Inhibitors, 2004, JBC, vol. 279, No. 17, p.17508-17514. Specific effectiveness of the compounds was measured against mutants A156S, AT, D168A and D168V proteolytic enzyme HCV NS3/4A 1b in the following ways:

[00354] 10X solution of proteolytic enzyme NS3/4A, Bioenza (Mountain View, CA) and 1.13X 5-FAM/QXL™520 of the substrate peptide CHEF Anaspec (San Jose, CA) were placed in 50 mm HEPES, pH 7.8, 100 mm NaCl, 5 mm DTT and 20% glycerol. 5 µl of each enzyme was subjected to pre-incubation in 384-well black raw microtiter tablets Corning (#3573) (Corning, NY) for 30 minutes at 25°C with 0.5 μl of 50% DMSO and compounds according to the present invention, dissolved in parallel in 50% DMSO. The reaction of the protease was started with the addition of 45 μl of substrate for CHEF and within 120 minutes measured λex487/λem514 using monochromators Quad4in the tablet reader Synergy4, BioTek (Winooski, VT). According to the results of each analysis dynamic curves obtained for each well was assessed by linear characteristics of the reaction and the statistical value is reported compliance (R 2absolute sum of squares). Initial speed (from 0 minutes to 30+ minutes) for each reaction was determined by the slope of the plot of relative fluorescence units of time (in minutes), and then build a graph of the concentration of inhibitor to estimate the IC50on the basis of log[Inhibitor] vs Response model Variable Bias in GraphPad Prism, GraphPad Software (San Diego, CA).

[00355] Table 5 shows the activity of some compounds according to the present invention in the analysis method BEF. The numbers of the compounds correspond to the numbers of the compounds in Table 3. Of the compounds which are defined as "A", showed IC50≤10 nm; compounds whose activity is defined as "In", showed IC50>10 nm and ≤100 nm; compounds whose activity is defined as "C", showed IC50>100 nm and ≤1000 nm; compounds whose activity is defined as "D", showed IC50>1000 nm and <10000 nm; and compounds whose activity is defined as "E", showed IC50≥10000 nm.

Table 5
Data Enzymatic Activity For common Compounds
The analyzed CompoundsThe enzyme/Analysis(I-1)Wild typeAnd
HCV A156SAnd
HCV ATAnd
HCV D168AAnd
HCV D168VAnd
(I-6)Wild typeAnd
HCV A156SAnd
HCV ATAnd
HCV D168AIn
HCV D168VIn
(I-7)Wild typein
D168AD
(I-11)Wild typeAnd
D168AIn

Example 16

The analysis method RPAF HCV Protease Wild-Type and Mutant Enzyme NS3/4A 1b (IC50_APP).

[00356] To obtain the values of "seemingly" IC50(IC50_APP), see what's below in Table 6, followed the following Protocol, without being bound to any particular theory, it is believed that IC50_APPin comparison with the values of the IC50can provide more applicable indicators of inhibition in a time-dependent, and, therefore, better describe the binding capacity. The Protocol is a modified method of analysis based on CEF (v_03), designed to assess the effectiveness of the compounds of the order and profiles of sustainability in relation to the HCV protease NS3/4A 1b wild-type and mutants C159S, A156S, AT, D168A. D168V, R155K in the following way: 10X solution of proteolytic enzyme NS3/4A, Bioenza (Mountain View, CA) and H 5-FAM/QXL™520 FRET substrate peptides, Anaspec (San Jose, CA) were placed in 50 mm Tris-HCl, pH 7.5, 5 mm DTT, 2% CHAPS, and 20% glycerol. 5 µl of each enzyme were made in 384-well black microtiter tablets Corning (#3575) (Corning, NY) after applying 0.5 μl of 50% DMSO and compounds dissolved in parallel, in 50% DMSO. The reaction of the protease were started immediately after addition of the enzyme with the addition of 45 μl of substrate CHEF, and measurement was carried out for 60-90 minutes at λex485/λem520 on the tablet reader Synergy4, BioTek (Winooski, VT). According to the results of each analysis dynamic curves obtained for each well was assessed by linear characteristics of the reaction and the statistical values of conformity (R2, 95% trust the considerable intervals, the absolute sum of squares). Initial speed (from 0 minutes to 15+ minutes) for each reaction was determined by the slope of the plot of relative fluorescence units of time (in minutes), and then build the graph depending on the concentration as a percentage of the values for the controls in the absence of inhibitor and enzyme to assess perceived value IC50on log[Inhibitor] vs Response Model Variable Bias in GraphPad Prism, GraphPad Software (San Diego, CA).

[00357] table 6 shows the activity of some compounds according to the present invention in the analysis method BEF. The numbers of the compounds correspond to the numbers of substances in Table 3. Of the compounds which are defined as "A", showed IC50≤10 nm; compounds whose activity is defined as "In", showed IC50>10 nm and ≤100 nm; compounds whose activity is defined as "C", showed IC50>100 nm and ≤1000 nm; compounds whose activity is defined as "D"showed IC50>1000 nm and <10000 nm; compounds whose activity is defined as "E"showed IC50≥10000 nm.

Table 6
Data Enzymatic Activity of Typical Compounds
Analyzed the Connection The enzyme/AnalysisInhibition
(I-1)Wild typeAnd
HCV A156SAnd
HCV ATIn
HCV D168A
HCV D168VIn
Analysis of the replicon2In
Analysis of the replicon2In1
(I-2)Wild typeAnd
HCV AAnd
HCV ATAnd
HCV D168AAnd
HCV D168VAnd
Analysis of the replicon2In
Analysis of the replicon2With1
(I-5)Wild typeAnd
HCV AAnd
HCV ATIn
HCV D168A
HCV D168VIn
Analysis of the replicon2In
Analysis of the replicon2With1
(I-6)Wild typeAnd
HCV AAnd
HCV ATAnd
HCV D168A
HCV D168V
Analysis of the replicon

Analyzed the ConnectionThe enzyme/AnalysisInhibition
Analysis of the replicon2D1
(I-7)Wild type
HCV A
HCV AT
HCV D168AD
HCV D168VD
(I-8)Wild typeAnd
HCV AAnd
HCV ATAnd
HCV D168AAnd
HCV D168VIn
(I-9)Wild typeAnd
HCV A156SAnd
HCV ATAnd
HCV D168AIn
HCV D168V
(I-10)Wild type
HCV A
HCV ATD
D
HCV D168VD
(I-11)Wild typeAnd
HCV AAnd
HCV ATAnd
HCV D168AIn
HCV D168VIn
HCV R155KIn
Analysis of the replicon2And
Analysis of the repliconIn1
(I-13)Wild typeAnd
HCV AAnd

Analyzed the ConnectionThe enzyme/AnalysisInhibition
HCV ATAnd
HCV D168A
HCV D168V In
Analysis of the repliconIn
Analysis of the replicon2With1
(I-19)Wild typeAnd
HCV AAnd
HCV ATAnd
HCV D168AIn
HCV D168VIn
(I-20)Wild typeAnd
HCV AAnd
HCV ATAnd
HCV D168AIn
HCV D168VIn
Analysis of the replicon2In
Analysis of the repliconC1
(I-22)Wild typeAnd
HCV AAnd
HCV ATIn
HCV D168A
HCV D168V
HCV R155KIn
Analysis of the replicon2In
Analysis of the replicon2In1
(I-23)Wild typeAnd
HCV AAnd
HCV ATAnd
HCV D168AAnd
HCV D168VIn
Analysis of the replicon2In

And
Analyzed the ConnectionThe enzyme/AnalysisInhibition
Analysis of the replicon2With1
(I-24)Wild type And
HCV AAnd
HCV ATAnd
HCV D168AAnd
HCV D168VAnd
Analysis of the replicon2In
Analysis of the replicon2With1
(I-25)Wild typeAnd
HCV AAnd
HCV ATIn
HCV D168A
HCV D168V
HCV R155KIn
Analysis of the replicon2And
Analysis of the replicon2In1
(I-26)Wild typeAnd
HCV AAnd
HCV AT
HCV D168AAnd
HCV D168VAnd
(I-27)Wild typeAnd
HCV AAnd
HCV ATAnd
HCV D168AAnd
HCV D168VAnd
Analysis of the replicon2In
Analysis of the replicon2With1
(I-32)Wild typeAnd
HCV A156SAnd
HCV D168AIn

Analyzed the ConnectionThe enzyme/AnalysisInhibition
HCV R155KIn
HCV S
HCV D168VIn
HCV ATAnd
(I-33)Wild typeAnd
HCV A156SIn
HCV D168AD
HCV R155K
(I-35)Wild typeAnd
HCV AAnd
HCV D168AIn
HCV R155KIn
Analysis of the repliconWith3
Analysis of the repliconC4
(I-36)Wild typeAnd
HCV AAnd
HCV D168AAnd
HCV R155KAnd
Analysis of the repliconAnalysis of the repliconWith4
(I-37)Wild typeAnd
HCV AAnd
HCV D168AIn
HCV R155KIn
Analysis of the repliconWith3
Analysis of the repliconD4
(I-39)Wild typeAnd
HCV A156SAnd
HCV D168A
HCV R155KIn
(I-40)Wild typeAnd

Analyzed the ConnectionThe enzyme/AnalysisInhibition
HCV A156SAnd
HCV D168AIn
HCV R155KAnd
(I-42)Wild typeAnd
HCV A156SAnd
HCV D168AIn
HCV R155KIn
(I-44)Wild typeAnd
HCV A156SIn
HCV D168AD
HCV R155KD
(I-45)Wild typeAnd
HCV A156SAnd
HCV D168AAnd
HCV R155KAnd
Analysis of the repliconIn3
Analysis of the repliconWith4
(I-46)Wild type And
HCV A156SAnd
HCV D168AIn
HCV R155KAnd
Analysis of the repliconD3
Analysis of the repliconD4
(I-47)Wild typeAnd
HCV A156SAnd
HCV D168AIn
HCV R155KIn
Analysis of the repliconIn3
Analysis of the repliconC4
(I-48)Wild typeB
HCV A156SB

td align="center">
Analyzed the ConnectionThe enzyme/AnalysisInhibition
HCV D168A
HCV R155K
(I-49)Wild type
HCV A156SC
HCV D168AD
HCV R155KD
Analysis of the repliconD3
Analysis of the repliconD4
(I-50)Wild typeIn
HCV A156S
HCV D168AD
HCV R155KD
Analysis of the repliconD3
Analysis of the repliconD4
(I-51)Wild typeAnd
HCV A156SAnd
HCV D168AIn
HCV R155KIn
Analysis of the repliconWith3
Analysis of the repliconD4
(I-52)Wild typeIn
HCV D168AD
Analysis of the repliconD3
Analysis of the repliconD4
(I-53)Wild typeAnd
HCV A156SAnd
HCV D168AAnd
HCV R155KAnd
Analysis of the repliconIn3
Analysis of the repliconWith4
(I-54)Wild typeAnd
HCV A156SAnd

Analyses the Connection has been created The enzyme/AnalysisInhibition
HCV D168AIn
HCV R155KIn
Analysis of the repliconWith3
Analysis of the repliconE4
(I-55)Wild typeAnd
HCV A156SAnd
HCV D168AIn
HCV R155KIn
1Denotes the value of the IC90(nm).
2The data obtained in the analysis described in Example 14.
3Denotes the value of the EU50(nm). The data obtained in the analysis described in Example 26.
4Denotes the value of the EU90(nm). The data obtained in the analysis described in Example 26.

Example 17

[00358] Performed mass spectrometric analysis of VG is wild-type or variant C159S in the presence of the analyzed compounds according to the invention. 100 pmol HCV wild type (Bioenza CA) were incubated with the analyzed substance in the course of 1 hour and 3 hours at 10-fold excess (1-3) in relation to protein. Selected aliquots of the sample volume of 1 μl (total volume of 4.24 ml), diluted with 10 µl of 0.1% of TN, and then using the tip of a pipette ZipTip C4 was applied directly onto the MALDI target using sinadinovic acid as a matrix for desorption (10 mg/ml in 0.1%TN:acetonitrile 50:50). Analyses were performed on a mass spectrometer Shimadzu Biotech Axima TOF2(Shimadzu Instruments) with time-of-flight laser ionization and desorption from the matrix (MALDI-TOF). Using the same methods above analyzed 100 mmol mutant HCV protease C159S for 3 hours at 10-fold excess of (I-3) with respect to the protein.

Example 18

Modification of Cys159 of HCV protease wild-type using the method tripticase cleavage

[00359] HCV incubated with the analyzed compound according to the present invention I-3 for 3 hours before tripticase cleavage. As the alkylating agent after incubation of the compounds used iodoacetamide. For tripticase hydrolysates were diluted aliquots of the samples with a volume of 2 μl (0.06 ág/ál) 10 ál of 0.1% of TN, and then using the tip of a pipette ZipTip C4 was applied directly onto the MALDI target using alpha-cyano-4-hydroxy-phenylacrylate acid is as a matrix (5 mg/ml in 0.1%TN:acetonitrile 50:50).

[00360] For analysis trypticase hydrolysates device installed in the mode of reflection with the use of rapid output 1800. The calibration was performed using standard Laser Biolabs Pep Mix (1046.54, 1296.69, 1672.92, 2093.09, 2465.20). For analysis CID/PSD was chosen peptide using cursors to install a temporary adjustment of the ion gates and fragmentation was observed when the power of the laser radiation above about 20%, as the gas used for collision in the CID, was Not used. Calibration for fragments was performed using calibration fragmentation P14R for Reflectron with a curved field.

Example 19

[00361] As shown in Figures 1 and 2, mass spectrometric analysis of HCV wild type in the presence of the analyzed compounds according to the present invention I-1 and I-25 were carried out in accordance with the following Protocol: NS3/4ABFC wild-type (wt) were incubated for one hour with a 10-fold excess of the analyzed compounds in relation to protein. Selected aliquots of the sample volume of 2 μl, was diluted in 10 ál of 0.1% of TN, and then using the tip of a pipette ZipTip C4 was applied directly onto the MALDI target using sinadinovic acid as a matrix for desorption (10 mg/ml in 0.1%TFA:acetonitrile 50:50). To measure the mass of the intact protein, the device was installed in the linear regime using rapid pickup is as 24500, and the standard for the calibration of the instrument was a protein.

[00362] As shown in Figure 1, in comparison with the protein in the absence of the substance according to the invention, the protein pre-incubated with compound I-1 reacted significantly with the formation of new particles at MW 25218 Yes that is approximately 751 Yes heavier and comparable to the mass of compound I-1 component 747 Yes.

[00363] As shown in Figure 2, after 1 hour of reaction, observed a transition to a new peak at MN+ 25,240 Yes, that is 773 Yes harder, and comparable to the mass of compound I-25.

[00364] Compounds I-2, I-5, I-6, I-8, I-9, I-10, I-13, I-19, I-20, I-22, I-23, I-24, I-26 and I-27 was investigated in a similar way in accordance with the methods described in Example 17, and in the course of the studies observed a significant modification of the NS3/4ABFC wild-type.

Example 20

[00365] As shown in Figures 4, 5, 6 and 7, carried out mass-spectrometric analysis of mutants of HCV in the presence of compound I-11. Mutants of HCV (A156S), (R155K), (D168A), (AT) and (D168V) were incubated for 3 hours at 10× excess of the analyzed compounds according to the present invention in relation to protein. Aliquots of the samples with a volume of 2 μl was diluted in 10 ál of 0.1% of TN, and then using the tip of a pipette ZipTip C4 was applied directly onto the MALDI target using sinadinovic acid as a matrix for desorption (10 mgml 0.1%TN:acetonitrile 50:50). To measure the mass of the intact protein, the device was installed in the linear regime with the use of rapid output 24500, and the standard for the calibration of the instrument was a protein.

[00366] As shown in Figure 4, for the mutant HCV (D168V) observed a complete metamorphosis after three hours of reaction. The difference in weight between the new forms and unreacted mutant is comparable to the mass of compound I-11.

[00367] As shown in Figure 5, for the mutant HCV(A) observed a complete metamorphosis after three hours of reaction. The difference in weight between the new forms and unreacted mutant is comparable to the mass of compound I-11.

[00368] As shown in Figure 6, for the mutant HCV (R155K) observed a complete metamorphosis after three hours of reaction. The difference in weight between the new forms and unreacted mutant is comparable to the mass of compound I-11.

[00369] As shown in Figure 7, for the mutant HCV (AT) observed a complete metamorphosis after three hours of reaction. The difference in weight between the new forms and unreacted mutant is comparable to the mass of compound I-11.

[00370] the Compounds I-19 and I-24 investigated similarly. In accordance with the methods described in Example 18, and in the course of the studies observed a significant covalent modification of NS3/4A HCV D168A.

Example 21

Culture to etoc

[00371] Huh-luc/neo-ET, Huh7-Lunet received in ReBLikon Gmbh (Heidelberg, Germany). Cells were grown in a modified method of Dulbecco environment Needle (DMEM; Invitrogen), with the addition of 2 mm L-glutamine, nonessential amino acids, 100 μl of penicillin/ml, 100 μg streptomycin/ml and 10% fetal bovine serum. G418 (Geneticin; Invitrogen) was added to a final concentration of 400 μg/ml Huh7-Lunet were grown in the absence of G418.

Example 22

Mutant constructs

[00372] the Design bearing clinically significant mutations were created by site-directed mutagenesis plasmid pFK-I389-luc-ubi-neo-NS3-3'ET (ReBLikon Gmbh (Heidelberg, Germany), using a Kit for Site-Directed Mutagenesis QuickChange II (Stratagene, La Jolla, CA) according to manufacturer's instructions and using the primers described below in table 7.

Table 7
Sequences of primers used to generate cell lines carrying the replicon
NS3-A156S-FGCTGTGGGCATCTTTCGGTCTGCCGTGTGCACCCGAGGGSEQ ID NO:66
NS3-A156S-RCCCTCGGGTGCACACGGCAGACCGAAAGATGCCCACAGCSEQ ID NO:67
NS3-A156T-FSEQ ID NO:68
NS3-A156T-RCCCTCGGGTGCACACGGCAGTCCGAAAGATGCCCACAGCSEQ ID NO:69
NS3-D168A-FGGGGTTGCGAAGGCGGTGGCCTTTGTACCCGTCGSEQ ID NO:70
AGTCT
NS3-D168A-RAGACTCGACGGGTACAAAGGCCACCGCCTTCGCAACCCCSEQ ID NO:71
NS3-D168V-FGGGGTTGCGAAGGCGGTGGTCTTTGTACCCGTCGAGTCTSEQ ID NO:72
NS3-D168V-RAGACTCGACGGGTACAAAGACCACCGCCTTCGCAACCCCSEQ ID NO:73
NS3-C159S-FATCTTTCGGGCTGCCGTGAGCACCCGAGGGGTTGCGAAGSEQ ID NO:74
NS3-C159S-RCTTCGCAACCCCTCGGGTGCTCACGGCAGCCCGAAAGATSEQ ID NO:75
NS3-R155K-FCACGCTGTGGGCATCTTTAAGGCTGCCGTGTGCACCCGASEQ ID NO:76
NS3-R155K-RTCGGGTGCACACGGCAGCCTTAAGATGCCCACAGCGTG SEQ ID NO:77

Example 23

Transcription in vitro

[00373] In vitro transcripts ±circuits HCV was created in accordance with the Protocol described in Lohmann V, et al., J. Virol., 77:3007-3019, 2003. For transcription ±circuits HCV RNA, plasmid DNA (pFK I341 PI-Luc/NS3-3'/ET, obtained in ReBLikon Gmbh (Heidelberg, Germany), was sequentially subjected to AseI and Sca1. After splitting restrictase were extracted DNA using phenol and chloroform, besieged by ethanol and dissolved in water in the absence of RNase. The reaction mixture for in vitro transcription consisted of 80 mm HEPES (pH 7.5), 12 mm MgCl2, 2 mm spermidine, 40 mm dithiothreitol, 3.125 mm concentrations of each nucleoside triphosphate, 1 mm Rnasin. For the reaction took 5 µg restricciones plasmid DNA and 80 ál of RNA polymerase T7 (Promega). After 2 hours incubation polymerase at 37°C was added an additional 40 μl of T7 and the reaction mixture incubated for a further 2 hours. Termination of transcription was performed by adding 1 μl of Gnkazy in the absence of RNase (Promega) per μg of plasmid DNA, after which incubated the mixture for 30 minutes at 37°C. After extraction with phenol and chloroform, RNA was besieged by using isopropanol and dissolved in water in the absence of RNase. The concentration was determined by measuring optical density at 260 nm (OD260) and the integrity of RNA was checked put the m electrophoresis in denaturing agarose gel.

Example 24

Transfection of full-length HCV genome and selection of stable cell lines

[00374] 7×104Huh7-Lunet cells were cultured overnight on 12-hole tablet, the next day was transfusional 1 µg RNA per well with a set of cool simple point pointers rubber Tx (Madison, WI). The transfection was carried out in accordance with the manufacturer's instructions, and 24 hours after transfection cells were subjected to analysis of luciferase or selection using G418 (400 μg/ml) to obtain stable cell lines.

Example 25

Inhibition Autoproteolysis Protease

[00375] Cells Huh-7-Luc-Neo-ET were placed in a Medium to examine the Structure of the Replicon (RPMI with 5% of the FBI, IX non-essential amino acids and penicillin/streptomycin) with a density of cells 1×105of cells per well on 12-well plates. After eight hours, the medium was removed and replaced by 1 ml of medium containing the analyzed compound according to the present invention (5-hole connection) and 0.02% DMSO, after which the cells were returned to the incubator overnight. After sixteen hours 1 hole for each connection and 1 untreated wells were washed FBI, then the cells were literally and scraped in 30 μl of Buffer for Cell Extraction (Biosource, Camarillo, CA) supplemented with Complete Protease Inhibitor (Roche, Indianapolis, IN). Remaining wells washed twice by the FBI and then filled Environment for the Analysis of the structures of The Replicon and placed back in the incubator. Cells were washed once every hour by removing old medium and replace it with a new environment and literally and collected after 4, 12, 24 and 48 hours after the first collection of cells.

[00376] the cell Lysates were separated using SDS-Page (4-20%) and transferred to the membrane Immobilon-P PVDF (Millipore Corporation, MA) and were blotting using monoclonal antibodies to NS3 (Bioenza, CA). The blots were scanned using an infrared scanner Odyssey Licor and the band is FL, and the cleavage products were calculated separately using Licor software supplied with the scanner. The cleavage product was calculated as a percentage of the total content of NS3 in each sample relative to the control DMSO so that the control DMSO reflects 100% of the activity.

Results and discussion

[00377] In the result of inhibiting the activity of protease, autoproteolysis does not occur, and the only detectable protein particles are choloenzymes. After 16 hours of continuous exposure of cells carrying the replicon, the compound is an inhibitor of NS3 products autoproteolysis not found in processed samples, but they were easily detected in untreated cells carrying the replicon used as a control. Extended validity was demonstrated by exposure of cells carrying the replicon, the effects of protease inhibitors for 16 hours, after which the connection according to the SNO present invention was removed and cells carrying the replicon, were subjected to repeated cleaning for a few hours. Covalent irreversible inhibitors of NS3 showed a sustained inhibition of internal autoproteolytic activity of NS3 in for up to 48 hours, while applying a reversible connection autoproteolytic protease activity recovered rapidly (Figure 8).

[00378] Figure 8 shows that the inner products autoproteolysis NS3 inhibited when exposed to cells carrying the replicon, Compound I-47 for 16 hours.

[00379] Figure 9 shows an irreversible covalent inhibitor (compound I-11) NS3 protease, showed prolonged inhibition activity of the NS3 protease in wild-type cells carrying the replicon, which measured the largest autoproteolysis, after removal of the specified connection. Connection incubated with cells carrying the replicon, for 16 hours, after which the connection was deleted (time point 0). Even during the period up to 48 hours after removal of irreversible covalent inhibitor of NS3 degree of inhibition autoproteolytic activity of NS3 has reached at least 50%, while using the drug with reversible effect, VX-950, almost complete return of activity was observed after 4 hours after removal of the specified drug.

[00380] In the Figure 10 shows ebrahimy covalent inhibitor (compound I-25) NS3 protease, showing extended inhibiting the activity of the NS3 protease in wild-type cells carrying the replicon, which measured the largest autoproteolysis, after removal of the specified connection. The specified connection incubated with cells carrying the replicon, for 16 hours, after which the connection was deleted (time point 0). Even during the period up to 48 hours after removal of irreversible covalent inhibitor of NS3, the degree of inhibition autoproteolytic activity of NS3 has reached at least 40%, while using the drug with reversible effect, VX-950, almost complete return of activity was observed after 4 hours after removal of the drug.

Example 26

Analysis using Luciferase system

[00381] the Compounds according to the present invention was investigated to assess the antiviral activity and cytotoxicity of compounds during the analysis activity of luciferase synthesized with replicon. In the analysis of the used cell line ET (luc-ubi-neo/ET), which is a cell line Huh7 human hepatoma containing RNA replicon HCV sustainable reporter luciferase (Luc) and mutations bearing adaptive character for cells. Line of ET cells were cultured in an incubator with 5% CO2at 37°C in modified nutrient medium, Dulbecco (DMEM) supplemented with 2 mm L-glutamine, replace the s amino acids, 100 μl of penicillin/ml, 100 μg streptomycin/ml and 10% fetal bovine serum. G418 (Geneticin; Invitrogen) was added to a final concentration of 400 μg/ml

[00382] All reagents for cell culture were obtained from Invitrogen (Carlsbad). The cells were trypsinization (1% trypsin:EDTA) and placed in concentration of 5×103of cells per well on 96-well analytical plates (Costar) with the aim of measuring the number of cells (cytotoxicity) or research antiviral activity. The analyzed compounds were added in 3-fold concentration of 6 holes each and the analysis was carried out in DMEM, 5% FBI, 1% pen-strap, 1% glutamine, 1% non-essential amino acids. As a positive control in each run was used interferon Alfa-2b human (PBL Biolabs, New Brunswick, NJ). Upon completion of 72 hours of processing cells, after making the analyzed compounds, cells were all subconfluent. Antiviral activity was measured by analyzing the activity of luciferase synthesized with replicon, when applied Systems Analysis Luciferase Steady-Glo (Promega, Madison, WI) according to manufacturer's instructions. The number of cells in each well was determined using Analysis of Titer Blue Cells (Promega). The connection profile was obtained by calculating the corresponding values of the EU50(effective concentration at which the degree of inhibition of virus replication reaches priblizitelen is 50%), EU90(effective concentration at which the degree of inhibition of virus replication reaches approximately 90%), IC50(the concentration that reduces cell viability by approximately 50%) and SI50(selective index: EC50/IC50).

[00383] Although described several variants of realization of the present invention, it is obvious that the basic examples can be modified to create other variants of implementation, providing for the use of the compounds and methods according to the present invention. Thus, it should be understood that the scope of the invention defined by the attached formula, and not the individual variants of the implementation presented in the examples.

1. The compound of formula I:

or pharmaceutically acceptable salt of the compounds where:
R1and R1'independently represent hydrogen or C1-6aliphatic group, or R1and R1'together form a 3-7-membered carbocyclic ring, optionally substituted C1-6the alkyl or C2-6alkenyl;
R2Arepresents-NHSO2R2;
R2represents a C3-7cycloalkyl;
each R independently represents hydrogen or C1-6aliphatic group, optionally substituted by oxopropoxy or halogen;
R3 is a head group-L-Y, L-Y is selected from the following:
(a) L is a bivalent linear or branched C2-8hydrocarbon chain, where L contains at least one double bond and one or two additional methylene unit in L is independently replaced by a functional group selected from-NRC(O)-, -N(R)SO2-, -OC(O)- or-C(O)-, where at least one double bond is in the alpha-beta position relative to the carbonyl or-SO2-functional group; and Y represents hydrogen or C1-6aliphatic group; or
(b) L is a bivalent linear or branched C2-8hydrocarbon chain, where L contains at least one double bond, and at least one methylene link in L is replaced by a functional group selected from-C(O)-, -NRC(O)-, -N(R)SO2- or-OC(O)-, where at least one double bond is in the alpha-beta position relative to the carbonyl or-SO2-functional group, and one or two additional link in L optionally and independently replaced by-O-, -N(R)- or-C(O)-; and Y represents hydrogen or C1-6aliphatic group; or
(C) L is a bivalent linear or branched C2-8hydrocarbon chain, where L contains at least one double bond, and at the ore one methylene link in L is replaced by a functional group, selected from-C(O)-, where at least one double bond is in the alpha-beta position relative to the carbonyl functional group, and one or two additional methylene unit in L optionally and independently replaced by-O-, -N(R)- or-C(O)-; and Y represents hydrogen or C1-6aliphatic group; or
(h) L is a bivalent linear or branched C2-8hydrocarbon chain, where L contains at least one alkylidene double bond, and at least one methylene link in L is replaced by a functional group selected from-OC(O)- or-C(O)-, where at least one double bond is in the alpha-beta position relative to the carbonyl functional group, and one or two additional methylene unit in L optionally and independently replaced by-O-, -N(R)- or-C(O)-; and Y represents hydrogen or C1-6aliphatic group; or
(l) L is a covalent bond, and Y is selected from:
(ii) C2-6alkenyl, replaced by exography and possibly substituted with halogen; or
(iii)2-6the quinil, replaced by oxopropoxy; or
(iv) a saturated 3-4 membered heterocyclic ring containing 1 heteroatom selected from nitrogen, where the specified ring is substituted by 1-2 groups Re; or
(IX) a partially unsaturated 3-6-membered carbocycles the ring, where the specified ring is substituted by 1-4 groups Re; or
(m) L represents-C(O)-, and Y is selected from:
(i) C1-6of alkyl, substituted with halogen; or
(ii) C2-6alkenyl; or
(n) L represents-N(R)C(O)-, and Y is selected from:
(i) C1-6of alkyl, substituted with halogen; or
(ii) C2-6alkenyl; or
(xiii) a 6-membered aromatic ring, where the specified ring is substituted by 1-4 groups Re; or
(o) L is a bivalent C1-8saturated or unsaturated linear or branched hydrocarbon chain; and Y is selected from:
(ii)2-6alkenyl, replaced by oxopropoxy; or
(iii)2-6the quinil, replaced by oxopropoxy; or
(R) L represents-CH2-, -NH-, -C(O)- or-CH2NH-; and Y is selected from:
(i)- (CH2CL; or
(ii) C2-6alkenyl, replaced by exography and possibly substituted with halogen; or
(iii) C2-6the quinil, replaced by oxopropoxy; or
(iv) a saturated 3-4 membered heterocyclic ring containing 1 heteroatom selected from nitrogen, where the specified ring is substituted by 1-2 groups Re; or
(vii) a saturated 3-6 membered carbocyclic ring, where the specified ring is substituted by 1-4 groups Re; or
(xiii) a 6-membered aromatic ring containing 0-2 nitrogen, where the specified ring is substituted by 1-4 groups Re;
where each Reindependently SEL is an out-Q-Z, oxo, NO2or halogen:
Q represents a divalent unsaturated linear or branched C1-6hydrocarbon chain, with one or two methylene chain in Q optionally and independently replaced by-O - or-C(O)-, and
Z represents hydrogen or C1-6aliphatic group;
or R3and R1together with in between the atoms form a possibly substituted saturated or unsaturated 12-18 membered ring containing from 2 to 6 heteroatoms, which are independently selected from nitrogen, oxygen or sulfur, and the formed ring contains-L-Y; or
R3and the ring formed by R1and R1'together with in between the atoms, form a possibly substituted, saturated or unsaturated 12-18 membered ring containing from 2 to 6 heteroatoms, which are independently selected from nitrogen, oxygen or sulfur, with the specified ring contains head group-L-Y;
R4represents H, -NHC(O)R5, -NHC(O)OR6or a group of the side chain of a natural or unnatural amino acids, and the side chain is a methyl or isopropyl;
each R5independently represents a C1-6aliphatic group or break bicikl, substituted -(CH2)0-4C(O)R°, R° represents a C1-6aliphatic group;
R 6
represents a C1-6aliphatic group; and
Rzrepresents; or R4and Rztogether with in between the atoms form a possibly substituted saturated or unsaturated 16-22 membered ring containing 2 to 6 heteroatoms, which are independently selected from nitrogen, oxygen, or sulfur;
where the specified 16-22 membered ring possibly substituted (Rm)pwhere:
p is an integer from 1 to 6 inclusive; and
in each case, Rmindependently represents a C1-6aliphatic group; and
in each case, Ryindependently selected from halogen or C1-4aliphatic group; and
m is an integer from 0 to 4 inclusive;
s is an integer from 0 to 4 inclusive;
t is an integer from 0 to 4 inclusive;
where the sum of s and t is not equal to zero.

2. The compound according to claim 1, wherein Y is selected from the following groups:




where each Reindependently represents a halogen.

3. The compound of formula I:

or pharmaceutically acceptable salt of the compounds where:
R1and R1'independently represent Soboh is hydrogen or C 1-6aliphatic group, or R1and R1'together form a 3-7-membered carbocyclic ring, optionally substituted C1-6the alkyl or C2-6alkenyl;
R2arepresents-NHSO2R2;
R2represents a C3-7cycloalkyl;
each R independently represents hydrogen or C1-6aliphatic group, optionally substituted by oxopropoxy or halogen;
R3represents-L-Y, where:
L represents a divalent saturated or unsaturated linear or branched C1-8hydrocarbon chain, one, two, or three methylene link in L optionally and independently replaced by-C(O)-;
Y is a 3-10-membered monocyclic saturated or partially unsaturated ring; and
R4represents H, -NHC(O)R5, -NHC(O)OR6or a group of the side chain of a natural or unnatural amino acids, and the side chain is a methyl or isopropyl;
each R5independently represents a C1-6aliphatic group or break bicikl, substituted -(CH2)0-4C(O)R°, R° represents a C1-6aliphatic group;
R6represents a C1-6aliphatic group; and
Rzrepresents; or R4/sup> and Rztogether with in between the atoms form a possibly substituted saturated or unsaturated 16-22 membered ring containing 2 to 6 heteroatoms, which are independently selected from nitrogen, oxygen, or sulfur;
where the specified 16-22 membered ring possibly substituted (Rm)pwhere:
p is an integer from 1 to 6 inclusive; and
in each case, Rmindependently represents a C1-6aliphatic group; and
in each case, Ryindependently selected from halogen or1-4aliphatic group; and
m is an integer from 0 to 4 inclusive;
s is an integer from 0 to 4 inclusive;
t is an integer from 0 to 4 inclusive;
where the sum of s and t is not equal to zero.

4. The compound according to claim 3, wherein Y is selected from the following groups:




where each Reindependently represents a halogen.

5. The compound according to claim 1, characterized in that:
L represents a divalent linear or branched C2-8hydrocarbon chain, and L contains at least one double bond, and at least one methylene link in L is replaced by a functional group selected from-NRC(O)-, -N(R)SO2-, OC(O)- or-C(O), where at least one double bond is in the alpha-beta position relative to the carbonyl or-SO2-functional group, and one or two additional methylene unit in L optionally and independently replaced by-O-, -N(R)- or-C(O)-;
Y represents hydrogen or C1-6aliphatic group.

6. The compound according to claim 5, characterized in that:
L represents a divalent linear or branched C2-8hydrocarbon chain, and L contains at least one double bond, and at least one methylene chain in the structure of L is replaced by a functional group selected from-C(O)-, -NRC(O)-, -N(R)SO2- or-OC(O)-, where at least one double bond is in the alpha-beta position relative to the carbonyl or-SO2-functional group, and one or two additional methylene unit in L optionally and independently replaced by-O-, -N(R)- or-C(O)-; and
Y represents hydrogen or C1-6aliphatic group.

7. The connection according to claim 6, wherein L represents a divalent linear or branched C2-8hydrocarbon chain, and L contains at least one double bond, and at least one methylene chain in the structure of L is replaced by a functional group selected from-C(O)-, where at least one double bond is in alphabeta position in relation to the specified functional group, and one or two additional methylene unit in L optionally and independently replaced by-O-, -N(R)- or-C(O)-.

8. The connection according to claim 6, wherein L represents a divalent linear or branched C2-8hydrocarbon chain, and L contains at least one double bond, and at least one methylene chain in the structure of L is replaced by a functional group selected from-OC(O)-, where at least one double bond is in the alpha-beta position with respect to a specified functional group.

9. The compound according to claim 5, characterized in that L is a-NRC(O)CH=CH-, -NRC(O)CH=CHCH2N(CH3)-, -NRC(O)CH=CHCH2O-, -NRSO2CH=CH-, -NRSO2CH=CHCH2-, -NRC(O)C(=CH2)CH2- or-CH2NRC(O)CH=CH-.

10. The connection according to claim 9, wherein L represents-NHC(O)CH=CH-, -NHC(O)CH=CHCH2N(CH3)-, -NHC(O)CH=CHCH2O-, -NHSO2CH=CH-, -NHSO2CH=CHCH2-, -NHC(O)C(=CH2)CH2- or-CH2NHC(O)CH=CH-.

11. The compound according to claim 5, wherein L represents a divalent linear or branched C2-8hydrocarbon chain, and L contains at least one alkylidene double bond, and at least one methylene chain in the structure of L is replaced by a functional group selected from-C(O)-, -NRC(O)-, -N(R)SO2- or-OC(O)-, where at least one dual is th link is in the alpha-beta position relative to the carbonyl or-SO 2-functional group, and one or two additional methylene unit in L can be independently replaced by-O-, -N(R)- or-C(O)-.

12. The compound of formula I:

or pharmaceutically acceptable salt of the compounds where:
R1and R1'independently represent hydrogen or C1-6aliphatic group, or R1and R1'together form a 3-7-membered carbocyclic ring, optionally substituted C1-6the alkyl or C2-6alkenyl;
R2arepresents-NHSO2R2;
R2represents a C3-7cycloalkyl;
each R independently represents hydrogen or C1-6aliphatic group, optionally substituted by oxopropoxy or halogen;
R3represents-L-Y, where L is a covalent bond, -CH2-, -NH-, -C(O)-, -CH2NH -, or a bivalent, saturated or unsaturated, linear or branched C1-8hydrocarbon chain, and Y is selected from C2-6the quinil, replaced by oxopropoxy;
R4represents H, -NHC(O)R5, -NHC(O)OR6or the side group of a natural or unnatural amino acids, and the specified side chain is a methyl or isopropyl;
each R5independently represents a C1-6aliphatic group or a connected m is a stick bicikl, substituted -(CH2)0-4C(O)R°, R° represents a C1-6aliphatic group;
R6represents a C1-6aliphatic group;
Rzrepresents; or R4and Rztogether with in between the atoms form a possibly substituted, saturated or unsaturated 16-22 membered ring containing 2 to 6 heteroatoms, which are independently selected from nitrogen, oxygen, or sulfur;
where the specified 16-22 membered ring possibly substituted (Rm)pwhere:
p is an integer from 1 to 6 inclusive; and
in each case, Rmindependently represents a C1-6aliphatic group; and each Ryindependently selected from halogen or1-4aliphatic group; and
m is an integer from 0 to 4 inclusive;
s is an integer from 0 to 4 inclusive;
t is an integer from 0 to 4 inclusive;
where the sum of s and t is not equal to zero

13. The compound of formula I:

or pharmaceutically acceptable salt of the compounds where:
R1and R1'independently represent hydrogen or C1-6aliphatic group, or R1and R1'together form a 3-7-membered carbocyclic ring, optionally substituted C1-6al the sludge or 2-6alkenyl;
R2arepresents-NHSO2R2;
R2represents a C3-7cycloalkyl;
each R independently represents hydrogen or C1-6aliphatic group, optionally substituted by oxopropoxy or halogen;
R3represents-L-Y, where L represents-CH2-, -NH-, -C(O)- or-CH2NH-; and Y is selected from
(i)- (CH2Cl; or
(ii)2-6alkenyl, replaced by exography and possibly substituted with halogen; or
(iii)2-6the quinil, replaced by oxopropoxy; or
(iv) a saturated 3-4 membered heterocyclic ring containing 1 heteroatom selected from nitrogen, with the specified ring is substituted by 1-2 groups Re; or
(vii) a saturated 3-6 membered carbocyclic ring, where the specified ring is substituted by 1-4 groups Re; or
(XIII) a 6-membered aromatic ring containing 0-2 nitrogen atom, where the specified ring is substituted by 1-4 groups Re;
R4represents H, -NHC(O)R5, -NHC(O)OR6or the side group of a natural or unnatural amino acids, and the specified side chain is a methyl or isopropyl;
each R5independently represents a C1-6aliphatic group, or connected by a bridge Bicycle, substituted -(CH2)0-4C(O)R°, R° represents a C1-6aliphati the mini-group;
R6represents a C1-6aliphatic group;
Rzrepresents; or R4and Rztogether with in between the atoms form a possibly substituted saturated or unsaturated 16-22 membered ring containing 2 to 6 heteroatoms, which are independently selected from nitrogen, oxygen, or sulfur;
where the specified 16-22 membered ring possibly substituted (Rm)pwhere:
p is an integer from 1 to 6 inclusive; and
in each case, Rmindependently represents a C1-6aliphatic group; and each Ryindependently selected from halogen or1-4aliphatic group; and
m is an integer from 0 to 4 inclusive;
s is an integer from 0 to 4 inclusive;
t is an integer from 0 to 4 inclusive;
where the sum of s and t is not equal to zero.

14. The connection 13, wherein Y is selected from the following groups:




where each Reindependently represents a halogen.

15. The compound according to claim 1, where R3selected from the following groups:











16. The compound according to claim 1, where the specified compound has the formula II-a or formula II-b:

or pharmaceutically acceptable salt of the compounds.

17. A compound selected from the group consisting of:





















or their pharmaceutically acceptable salt.

18. Pharmaceutical composition for inhibiting hepatitis C virus containing an effective amount of a compound according to claims 1-17, and pharmacy is is automatic acceptable adjuvant, the carrier or solvent.

19. The composition according to p in combination with an additional therapeutic agent.

20. The composition according to claim 19, where the specified additional therapeutic agent is an antiviral agent.

21. A method of inhibiting the activity of the protease of the hepatitis C virus or a mutant form of the specified enzyme in a biological sample or in a patient that includes the step of bringing into contact mentioned biological sample with a compound according to claims 1 to 17 or a composition according p.

22. The method according to item 21, wherein the activity of the protease of the hepatitis C virus or a mutant form of the specified enzyme is irreversibly inhibited by covalent modification conservative cysteine residue at the position equivalent to residue Cys159 protease of hepatitis C virus subtype 1b.

23. The method according to item 21, wherein the activity of the protease of the hepatitis C virus or a mutant form of the specified enzyme is irreversibly inhibited by covalent modification of the remainder of Cys159.

24. The method according to item 21, wherein the activity of the protease of the hepatitis C virus or a mutant form of the specified enzyme is irreversibly inhibited by covalent modification of the balance Cys16.

25. A method of treating diseases mediated by a protease of hepatitis C virus in a patient, comprising the step of introducing a given patient, the effective amount of the compound according to claims 1 to 17 or the composition of p.

26. The method according A.25, characterized in that the introduction is carried out once a day.

27. The method according to p. 25, wherein the disease is a hepatitis C.

28. The method according A.25, characterized in that the said compound irreversibly inhibits the protease of hepatitis C virus by covalent modification conservative cysteine residue at the position equivalent to the position of rest of Cys159 protease of hepatitis C virus subtype 1b.

29. The method according to p, characterized in that the genotype or subtype protease of hepatitis C virus selected from the group consisting of 1a, 1b, 1c, 2a, 2b, 2c, 2i, 2k, 3A, 3b, 3k, 4A, 4d, 4f, 5A, 6A, 6b, 6C, 6d, 6e, 6f, 6g, 6h, 6i, 6j, 6k, 6l, 6m, 6n, 6o, 6p, 6q, 6t and 7a.

30. The method according to clause 29, wherein the genotype or subtype protease of the hepatitis C virus is a 1A, 1b, 2a or 3A.

31. The method according A.25, including the stage of irreversible inhibition of the protease of hepatitis C virus by covalent modification of the balance Cys16 protease of hepatitis C.

32. The conjugate formed by reaction of the compound according to claim 1 with a protease of hepatitis C.

33. Conjugate on p, characterized in that the conjugate is formed by covalent binding between the substituent R3and sulfhydryl group of a cysteine protease of the hepatitis C virus, where cysteine is a Cys16 protease of hepatitis C.

34. To jugat on p, characterized in that the conjugate is formed by covalent binding between the substituent R3and sulfhydryl group of a cysteine protease of the hepatitis C virus, where cysteine is conservative in the position equivalent to Cys159 protease of hepatitis C.

35. The compound according to claim 1, where Rzrepresents.

36. The compound according to claim 1, where R4and Rztogether with in between the atoms form may contain substituents saturated or unsaturated 16-22 membered ring containing from 2 to 6 heteroatoms, which are independently selected from nitrogen, oxygen or sulfur.

37. Connection p, characterized in that the structure of the compounds correspond to the formula I-e or I-f:

or pharmaceutically acceptable salt of the compounds, where
p is an integer from 0 to 6 inclusive; and
each of Rmindependently represents: halogen, -OR°; -CN; -N(R°)2; or may contain substituents group selected from the following: C1-6aliphatic group and C3-7cycloalkyl; and
R° is a possibly containing substituents group selected from C1-6aliphatic groups.

38. The compound according to claim 1, wherein R1and R1'together they form the prob is tenderly containing substituents 3-7-membered carbocyclic ring.

39. Connection § 38, wherein R1and R1'together form:
,
where R° is a possibly containing substituents group selected from C1-6aliphatic groups.

40. Connection § 39, wherein R° is a vinyl.

41. Connection § 39, wherein R° represents ethyl.



 

Same patents:

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SUBSTANCE: invention relates to compounds, which can be used as inhibitors of protease of hepatitis C virus, to pharmaceutical compositions, which contain said compounds, and to methods of their application for treatment of diseases mediated by protease of hepatitis C virus.

EFFECT: obtaining compounds, which can be used as inhibitors of protease of hepatitis C virus.

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15 cl, 33 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to compounds, which can be used as inhibitors of protease of hepatitis C virus, to pharmaceutical compositions, which contain said compounds, and to methods of their application for treatment of diseases mediated by protease of hepatitis C virus.

EFFECT: obtaining compounds, which can be used as inhibitors of protease of hepatitis C virus.

37 cl, 22 dwg, 7 tbl, 34 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention described the macrocyclic compounds of formula wherein the radical values are presented in the patent claim. The above compounds are serine protease inhibitors wherein serine protease is hepatitis C virus (HCV) NS3 protease. This invention also discloses pharmaceutical compositions having antiviral activity against HCV, containing the claimed compounds, and one or more pharmaceutically acceptable carriers, as well as a method for preparing these compositions. The present invention describes a method of inhibiting the hepatitis C virus replication in a host, and a method of inhibiting activity of hepatitis C virus serine protease.

EFFECT: there are also presented a method of treating or preventing HCV infection, and a method of treating, preventing or ameliorating one or more symptoms of a liver disease or disorder associated with HCV infection in an individual.

69 cl, 39 ex, 8 tbl, 4 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to compounds of formula (I) a pharmaceutically acceptable salt thereof wherein each dash line (represented as ---) represents a double bond; X represents N or CH; R1a and R1b independently represent hydrogen or C1-6-jalkyl; L represents -O-; R2 represents hydrogen; R3 represents hydrogen or C1-6-alkyl; R4 represents quinolinyl substituted by one, two or three substitutes specified in C1-6-alkyl, C1-6-alkyloxy, thiazolyl or pyrazolyl, wherein said thiazolyl or pyrazolyl are substituted on any carbon atom by C1-6-alkyl; n is equal to 3, 4, 5 or 6; p is equal to 1 or 2. The invention refers to a pharmaceutical composition possessing the properties of KS3/4a-protease HCV inhibitors, containing a carrier, and an virally effective amount of the compound of formula (I) as an active ingredient. The method for preparing the compound of formula (I), wherein the above method involves forming an amide bond of an intermediate product (2a) and sulphonylamide (2b), as presented by the diagram, wherein G represents a group Also, the invention refers to alternative methods for preparing the compound of formula (I).

EFFECT: there are presented macrocyclic compounds possessing inhibitory activity on hepatitis C virus (HCV) replication.

13 cl, 1 tbl, 17 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: application describes prodrugs being 2-amino-6-({[2-(4-chlorophenyl)-1,3-thiazol-4-yl]methyl}thio)-4-[4-(2-hydroxyethoxy)-phenyl]pyridine-3,5-dicarbonitryl derivatives, and a method for preparing them.

EFFECT: invention can find application in treating and/or preventing cardiovascular diseases.

8 cl, 4 tbl, 18 ex

FIELD: chemistry.

SUBSTANCE: disclosed is a method of producing pure crystalline D-isoglutamyl-D-trytophan which involves a step of removing protection from essentially pure N-tert-butoxycarbonyl-D-isoglutamyl-D-tryptophan or diester thereof to yield essentially pure D-isoglutamyl-D- tryptophan. An amorphous ammonium alt of D-isoglutamyl-D- tryptophan (1:1) is also disclosed. Also disclosed is a method of producing a pure monoammonium salt of D-isoglutamyl-D-tryptophan from essentially pure N-tert-butoxycarbonyl-D- isoglutamyl-D-tryptophan. Disclosed is a compound H-D-Glu-(γ-D-Trp-OR2)-α-OR1 and pharmaceutically acceptable acid addition salts thereof. Disclosed is a solid pharmaceutical composition and use thereof as an immunodepressant or anti-psoriasis agent.

EFFECT: improved method.

51 cl, 14 ex, 8 dwg, 1 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to organic chemistry, namely new 3,8-diaminotetrahydroquinoline derivatives of formula (1a) or to their pharmaceutically acceptable salts wherein X represents CH2, C=O or CH-OR; m is 1 or 2; Ar represents a phenyl group or a 5-merous or 6-merous aromatic heterocyclic group having one element specified in S and N, (wherein the phenyl group may be substituted by 1-2 halogen atoms); each R1 and R2 represents a hydrogen atom; R3 represents a C1-C6 alkyl group or indolyl-C1-4 alkyl group (the indolyl group is optionally substituted by a C1-C6 alkyl group or a halogen atom), n is 0; R4 and R5 which may be identical or different, each represents a hydrogen atom or a C1-C6 linear or branched alkyl group; each R6 and R7 represents a hydrogen atom; and R represents a hydrogen atom. Also, the present invention refers to a drug preparation and a pharmaceutical composition of the basis of the compound of formula (1a), to the compound of formula (F1), to a method for preparing an intermediate compound (e).

EFFECT: there are prepared new 3,8-diaminotetrahydroquinoline derivatives which possess high GHS-R antagonist activity.

10 cl, 1 tbl, 124 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to crystalline modifications: 1 (polymorphous form F), 2 (polymorphous form I) and 3 (polymorphous form X) of monosodium salt of D-isoglytamyl-D-tryptophan (1:1) characterised by powder X-ray pattern peaks presented in the application materials, as well as to pharmaceutical compositions containing them. The invention describes their use for treating various diseases and body conditions of at least one autoimmune diseases specified in a group consisting of psoriasis, atopic dermatitis and rheumatoid arthritis.

EFFECT: present invention describes the methods for producing the declared crystalline modifications of monosodium salt of D-isoglytamyl-D-tryptophan (1:1).

42 cl, 4 ex, 9 dwg

FIELD: chemistry.

SUBSTANCE: disclosed is a method of producing lipodipeptides based on L-glutamic acid or L-glutamine and L-ornithine, L-lysine or L-arginine. L-glutamic acid or L-glutamine derivatives esterified with fatty alcohol residues are obtained by fusing an amino acid with a corresponding alcohol in the presence of a strongly acidic ion-exchange resin in H+ form. Amino groups of L-ornithine, L-lysine or L-arginine are protected and then activated with carboxyl groups. Further, a reaction takes place between esterified derivatives of L-glutamic acid or L-glutamine and the protected derivatives of L-ornithine, L-lysine or L-arginine to form lipodipeptides. The protective groups are then removed.

EFFECT: invention simplifies the process at the esterification step, reduces reaction temperature and reaction time to 2 hours.

3 ex

FIELD: chemistry.

SUBSTANCE: invention relates to novel polyfunctional fullerene C60 amino acid derivatives of formula (1) , wherein R is H, mono- or dinitroxyC1-6alkyl, maleinimide; N-Z denotes a α, β, γ, ω-amino acid fragment of general formula where m=2-5 and M is a nitroxyC1-6alkyl group, a C1-6alkyl group or an alkali metal salt, having biological activity, as well as methods for production thereof and a method for covalent bonding of fullerene derivatives with SH-containing proteins. The invention also relates to the use of nitroxyalkyl-N-(fullerenyl)amino acids as nitrogen monoxide donors and to use of nitroxyalkyl-N-(fullerenyl)amino acids as quick-acting vasodilatators for antihypertensive therapy. The invention also relates to a method of inhibiting a metastasis process and a method of enhancing antileukemic activity of cyclophosphamide. Disclosed nitroxyalkyl-N-fullerenyl amino acid derivatives have an effect on coronary, contractile and pumping ability of the isolated heart of Vistar rats and are quick-acting vasodilatators which reduce arterial pressure and heart rate and cause relaxation of coronary vessel with less depressive effect on myocardial function compared to nitroglycerine.

EFFECT: disclosed compounds considerably intensify antileukemic activity of cyclophosphamide, increase chemosensitising activity when combined with cyclophosphamide.

9 cl, 8 ex, 3 tbl, 3 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to a 1-(1-adamantyl)ethylamine derivative (remantadin) of general formula: wherein R is a functional group of the amino acid residue (I-IV) or the lipoic acid residue (V). (I), (II), (III), (IV), (V) which possess selective antiviral activity in relation to the influenza A strains, including the viral strains resistant to action of remantadin. The compounds wherein R means the groups (l),(ll),(lll) and (V) are novel.

EFFECT: prepared compounds may be promising as substances as a part of the antiviral drugs and applicable for creating new antiviral drugs.

2 cl, 2 dwg, 3 ex

FIELD: medicine.

SUBSTANCE: 5-10 Pletnev drops are administered orally twice a day for 60-90 days according to the following scheme: on the first day in the morning, Pletnev drops No. 5 and in the evening Pletnev drops No. 7, on the second day in the morning, Pletnev drops No. 30 and in the evening Pletnev drops No. 4, or No. 44, or No. 68; Pletnev drops No. 5 represent the wild strawberry leaves infusion; Pletnev drops No. 7 is the greater plantain leaves infusion; Pletnev drops No. 30 is the bilberry shoot infusion; Pletnev drops No. 4 is the wild strawberry leaves and foalfoot leaves infusion; Pletnev drops No. 44 is the wild strawberry leaves and wormwood herb infusion; Pletnev drops No.68 is the common yarrow herb and wild strawberry leaves infusion in 95% ethanol with taking substance 15-25 mg per 1 ml of the infusion.

EFFECT: invention provides high-effective immune therapy of viral hepatitis C with no side effects.

13 tbl, 3 ex

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