Macrocyclic peptides eliciting activity with respect to hepatitis c virus

FIELD: organic chemistry, biochemistry, medicine, pharmacy.

SUBSTANCE: invention relates to macrocyclic peptides of the general formula (I): wherein W means nitrogen atom (N); R21 means hydrogen atom (H), (C1-C6)-alkoxy-, hydroxy-group or N-(C1-C6-alkyl)2; R22 means hydrogen atom (H), (C1-C6)-alkyl, CF3, (C1-C6)-alkoxy-group, (C2-C7)-alkoxyalkyl, C6-aryl or Het wherein het means five- or six-membered saturated or unsaturated heterocycle comprising two heteroatoms taken among nitrogen, oxygen or sulfur atom and wherein indicated Het is substituted with radical R24 wherein R23 means hydrogen atom (H), -NH-C(O)-R26, OR26, -NHC(O)-NH-R26, -NHC(O)-OR26 wherein R26 means hydrogen atom, (C1-C6)-alkyl; R3 means hydroxy-group or group of the formula -NH-R31 wherein R31 means -C(O)-R32, -C(O)-NHR32 or -C(O)-OR32 wherein R32 means (C1-C6)-alkyl or (C3-C6)-cycloalkyl; D means a saturated or unsaturated alkylene chain comprising of 5-10 carbon atoms and comprising optionally one-three heteroatoms taken independently of one another among oxygen (O), sulfur (S) atom, or N-R41 wherein R41 means hydrogen atom (H), -C(O)-R42 wherein R42 means (C1-C6)-alkyl, C6-aryl; R4 means hydrogen atom (H) or one-three substitutes at any carbon atom in chain D wherein substitutes are taken independently of one another from group comprising (C1-C6)-alkyl, hydroxyl; A means carboxylic acid or its alkyl esters or their derivatives. Invention relates to pharmaceutical compositions containing indicated compounds and eliciting activity with respect to hepatitis C virus and these peptides inhibit activity of NS3-protease specifically but don't elicit significant inhibitory activity with respect to other serine proteases.

EFFECT: valuable biochemical and medicinal properties of peptides.

106 cl, 9 tbl, 61 ex

 

The technical field to which the invention relates.

The present invention relates to compounds, compositions, methods of producing such compounds and to methods for treating infections caused by hepatitis C virus (HCV). The present invention relates, in particular, to new peptide analogs, pharmaceutical compositions containing such analogs and to methods of using these analogs for treatment of HCV infection.

Background of invention

Hepatitis C virus (HCV) is the major causative agent of hepatitis unrelated to hepatitis a and b, which occurs after a blood transfusion and that affects people all over the world. It is established that more than 170 million people worldwide are infected with a virus. A large percentage of carriers becomes chronically infected and many patients it leads to chronic liver disease, the so-called chronic hepatitis C. In turn, this group of patients has a high risk of the disease is such a serious disease of the liver, such as cirrhosis of the liver, liver cell cancer and in the last stage of liver disease, leading to death.

The mechanism by which the preservation of the HCV virus in the body and provides a high incidence of chronic liver disease are not yet understood. It is unknown how HCV interacts with the immune si is theme of the master and overcomes it. In addition, also have not identified the role of cellular and humoral immune responses in protection against HCV infection and disease hepatitis. There is evidence that antibodies can be used to prevent associated with blood transfusion viral hepatitis, however, the Center for disease control currently does not recommend treatment with immunoglobulins for this purpose. The lack of effective protective immune response makes it difficult to develop a vaccine or adequate preventive measures after exposure, so in the near future hopes mainly rely on anti-virus tools.

In order to identify pharmaceutical agents with proven efficacy in the treatment of HCV infection in patients with chronic hepatitis C, have conducted various clinical trials. In these tests was used interferon-alpha individually or in combination with other antiviral agents. These studies have allowed to establish that the main majority of the participants of the experiment were not detected reaction to such regimens, and of those participants who were sensitive to treatment, mostly after the end of treatment was observed relapse.

Thus, until recently, therapy with interferon (IFN) was the only one who passed affordable treatment for patients with chronic hepatitis C, has been proven clinically effective. But the duration of such treatment is small, and furthermore, treatment with interferon causes serious side-effects (i.e. retinopathy, thyroiditis, acute pancreatitis, depression), which reduces the quality of life of patients undergoing treatment. Currently, interferon in combination with ribavirin proposed for the treatment of patients who are not sensitive to IFN applied individually. This approach is currently recommended for the treatment not been previously treated patients and is the best in the treatment of HCV. However, the side effects caused by IFN, do not decrease when such joint therapy.

Thus, there is a need for the development of effective antiviral agents for treatment of HCV infection, which would be devoid of the disadvantages of existing methods of treatment based on the use of pharmaceuticals.

HCV is enclosed in a sheath of positive strand of RNA of a new virus family Flaviviridae. The genome of HCV, presents the single-stranded RNA, consists of approximately 9500 nucleotides and has a single open reading frame (ORF), which encodes a single large polyprotein, consisting of approximately 3000 amino acids. In infected cells, this prepreteenmodels in many sites of cellular and viral proteases with the formation of structural and non-structural (NS) proteins. In the case of HCV under the action of two viral proteases are formed Mature non-structural proteins (NS2, NS3, NS4A, NS4B, NS5A and NS5B). The first of them, which are not yet characterized, splits link NS2-NS3; the second is semipretioase contained in the N-terminal region of NS3 (hereinafter designated as NS3-protease), and it mediates all the subsequent cleavage occurring during transcription relative to NS3, both in CIS-orientation, at the site of cleavage of the NS3-NS4A, and in the TRANS-orientation to other sites NS4A-NS4B, NS4B-NS5A, NS5A-NS5B. The NS4A protein probably has multiple functions, acting as a cofactor for the NS3 protease and possibly contributing to the localization on the membrane NS3 and other viral replicase. The formation of the complex of the protein with NS3 NS4A, probably necessary for processing, amplification proteolytic efficiency in all sites. The NS3 protein also has nucleotidyltransferase activity and RNA-helicase activity. NS5B is an RNA-dependent RNA polymerase involved in HCV replication.

In the patent application WO 97/06804 described (-)-enantiomer nucleoside analogue of cytosine-1,3-oxathiolane (also known as 3TC)with activity against HCV. Although in earlier clinical trials for this compound detected activity against HIV and HBV, yet clincheck is the first evidence of its activity against HCV and is not revealed its mechanism of action in respect of this virus.

The overall strategy for the development of antiviral agents is inactivation of the encoded virus enzymes that are important for virus replication.

Considerable efforts made in this regard to identify compounds that inhibit the NS3-protease or RNA helicase of HCV has led to the following results:

In the patent US 5633388 described substituted by a heterocycle carboxamide and their analogues with activity against HCV. The target for these compounds is elikana activity of the NS3 protein of the virus, however there is no data about their clinical trials.

Chu and others (Tet. Lett., (1996), 7229-7232) described phenanthridine with activity against NS3 protease of HCV in vitro. No additional data on this connection will not be published.

In a research report submitted to the Ninth international conference on antiviral research (Ninth International Conference on Antiviral Research, Urabandai, Fukyshima, Japan (1996) (Antiviral Research, (1996), 30, 1, A23 (abstract 19)), it was reported thiazolidinone derivatives having inhibitory activity against HCV-protease.

Some of the research described in connection with ingibiruet action against other semiprotect, such as human leukocyte elastase. One group of such compounds is described in WO 95/33764 (Hoechst Marion Roussel, 1995). Described in this application peptides represent a morph is vinilkarbazola analogues, which structurally differ from the peptides of the present invention.

In WO 98/17679 (name VeKTex Pharmaceuticals Inc.) described inhibitors of serine proteases, in particular NS3 protease of hepatitis C.

Firm Hoffman LaRoche (WO 98/22496, US 5866684 and US 6018020) also described Hexapeptide, which proteinase inhibitors as antiviral agents for treatment of HCV infection.

etc. and Ingallinella et al. published data on the product inhibition NS4A-4B (Biochemistry (1998), 37, 8899-8905 and 8906-8914).

In WO 97/43310 in the name of Schering Corporation described consisting of 20 and 21 of the amino acid sequence of peptides with activity against NS3 protease of HCV.

In WO 98/46597 in the name of Emory University described peptides and mimetics of peptides with activity in vitro against semiprotect.

In WO 98/46630 in the name of Peptide Therapeutics Limited described depsipeptide substrate, having the ability to inhibit the NS3-protease of HCV.

Finally, in patent US 5869253 described enzymatic RNA molecules, which have the ability to inhibit the NS3-protease of HCV.

None of the above patent applications are not described cyclic peptides with activity against NS3 protease of HCV.

In WO 99/07733, WO 99/07734, WO 00/09543 and WO 00/09558 described exacerbated and Tripeptide analogues possessing the ability to inhibit the NS3-protease. However, these descriptions do not offer what are macrocyclic analogues of the present invention and presents the information may not lead to their creation.

In the application WO 99/38888 on the name of the Institute de Richerche di Biologia Moleculare (IRBM), published on August 5, 1999, describes small peptide inhibitors of the NS3-protease of HCV.

This description is not proposed and is not specified the cyclic structure of the peptides of the present invention. In addition, this description was published after the priority date of the present description.

In the application WO 99/64442 in the name of IRBM, which also published after the priority date of the present description, oligopeptides described, comprising a keto acid in position P1.

Application WO 99/50230 name VeKTex Pharmaceuticals (published 7 October 1999) also published after the priority date of this application. And in this case the publication is not even remotely refers to any cyclic peptides of the present invention.

One of the advantages of the present invention is that it claims to macrocyclic peptides, which possess inhibitory activity against NS3 protease of hepatitis C.

Another advantage of the object of the present invention is the fact that these peptides specifically inhibit the NS3-protease and do not show appreciable inhibitory activity against other semiprotect, such as elastase of human leukocytes (HLE), elastase pancreatic pigs (RRE) or bovine chymotrypsin pancreatic, or in respect of cysteinate, such as cathepsin In human liver (Cat).

And another advantage of the present invention is that it presents small peptides with low molecular weight, which have the ability to penetrate cell membranes and to inhibit NS3-protease activity in cell culture.

Another advantage of the compounds of the present invention is that they possess activity against both major genotypes detected in clinical isolates (1A and 1B), which is strong evidence that these compounds possess activity against all currently known genotypes of HCV.

Summary of the invention

The object of the present invention are the compounds of formula (I):

where

W represents CH or N,

R21denotes H, halogen, C1-C6alkyl, C3-C6cycloalkyl, C1-C6haloalkyl, C1-C6alkoxy, C3-C6cycloalkane, hydroxy, or N(R23)2where each R23independently of each other denotes H, C1-C6alkyl or C3-C6cycloalkyl; and

R22denotes H, halogen, C1-C6alkyl, C3-C6cycloalkyl, C1-C6haloalkyl, C1-C6thioalkyl, C1-C6alkoxy, C3-C6cycloalkane, C2-sub> 7alkoxyalkyl, C3-C6cycloalkyl,6- or10aryl or Het, where Het denotes a five-, six-, or semiology saturated or unsaturated heterocycle, containing one to four heteroatoms selected from nitrogen, oxygen and sulphur,

while specified cycloalkyl, aryl or Het substituted by the radical R24where

R24denotes H, halogen, C1-C6alkyl, C3-C6cycloalkyl, C1-C6alkoxy, C3-C6cycloalkane, NO2N(R25)2, NH-C(O)-R25or NH-C(O)-NH-R25,

where each R25independently of each other denotes H, C1-C6alkyl or C3-C6cycloalkyl,

or

R24denotes NH-C(O)-OR26where R26represents C1-C6alkyl or C3-C6cycloalkyl,

R3denotes hydroxy, NH2or a group of the formula-NH-R31where R31stands With6-or10aryl, heteroaryl, -C(O)-R32, -C(O)-OR32or-C(O)-other32where

R32represents C1-C6alkyl or C3-C6cycloalkyl,

D denotes consisting of 5-10 atoms, saturated or unsaturated alkylenes chain, optionally comprising one to three heteroatoms, independently from each other selected from O, S, or N-R41where

R41denotes H, C1-C6alkyl, C3- 6cycloalkyl or-C(O)-R42where R42represents C1-C6alkyl, C3-C6cycloalkyl or6- or10aryl;

R4represents H or one to three substituent on any carbon atom of the chain D, where the substituents independently of one another selected from the group including C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, hydroxy, halogen, amino, hydroxy, tigroup or C1-C6thioalkyl, and

And indicates the amide of formula-C(O)-NH-R5where

R5selected from the group including C1-C8alkyl, C3-C6cycloalkyl,6- or10aryl or7-C16aralkyl; or

And denotes a carboxylic acid, or its pharmaceutically acceptable salt or ester.

Under the scope of the present invention also subject a pharmaceutical composition comprising effective against hepatitis C virus, the amount of the compounds of formula I or its pharmaceutically acceptable salt or its complex ester in a mixture with a pharmaceutically acceptable carrier or auxiliary substance.

An important object of the invention is a method of treating infections caused by hepatitis C virus in a mammal, involving the administration to a mammal effective against hepatitis C virus quantities of the compounds of formula I, Riego therapeutically acceptable salt, or a complex ester, or the above-described composition.

Another important object of the invention is a method of inhibiting replication of hepatitis C virus by treatment of the virus inhibiting NS3-protease of hepatitis C virus by a number of the compounds of formula I, or a therapeutically acceptable salt, or a complex ester, or the above-described composition.

Another object of the invention is a method of treating infections caused by hepatitis C virus in a mammal, involving the administration to a mammal effective against hepatitis C virus amount of a composition comprising the compounds of formula I, or a therapeutically acceptable salt or ester. According to one of embodiments of the pharmaceutical compositions according to the invention include additional immunomodulator. Examples of additional immunomodulatory agents include, but are not limited to) α-, βor γ-interferons.

In an alternative embodiment, the pharmaceutical compositions of the present invention can additionally include anti-virus agent. Examples of antiviral agents include ribavirin and amantadine.

In another alternative embodiment, the pharmaceutical compositions of the present invention may optionally include other inhibitors of HCV protease.

And even on the nom embodiment, the pharmaceutical compositions according to the invention may additionally include an inhibitor of other targets in the life cycle of HCV, such as helicase, polymerase, metalloprotease or IRES.

A detailed description of the preferred embodiments

Definition

Unless otherwise stated, in the context of the present description, the concept used have the following meanings:

In those cases, when to denote the configuration of the substituent, for example, R4the compounds of formula I, using the symbol (R) or (S), the designation applies to the entire connection, not only to the Deputy.

The designation "P1, P2, and P3 in the context of the present description refers to the position of amino acid residues starting from the C-end of the peptide analogues and extending into the N end [i.e., P1 indicates the position 1 relative to the C-end, P2 denotes the position 2 relative to the C-end, etc.) (see Berger A. & Schechter I., Transactions of the Royal Society London series (1970), B257. 249-264].

In the context of the present description the term "1-aminocyclopropane acid" (ACPP) refers to the compound of the formula:

In the context of the present description, the term "vinyl-ACPP" refers to the compound of the formula:

In the context of the present description the term "Gamaliel-ACPP" refers to the compound of the formula:

The term "halo" in the context of the present description denotes halogen Deputy, selected from the group on the waiting bromine, chlorine, fluorine or iodine.

The term "C1-C6haloalkyl individually or in combination with another substituent in the context of the present description refers to an acyclic alkyl substituents with a straight or branched chain containing 1-6 carbon atoms and having one or more substituents of the hydrogen atom, which represents a halogen, selected from a range that includes bromine, chlorine, fluorine or iodine.

The term "C1-C6thioalkyl individually or in combination with another substituent in the context of the present description refers to an acyclic alkyl substituents with a straight or branched chain, containing Tilney group, such as thiopropyl.

The term "C1-C6alkyl" or "(ness.)alkyl" in the context of the present description, one or in combination with another Deputy, means an acyclic alkyl substituents with a straight or branched chain, containing from 1 to 6 carbon atoms, and includes, for example, methyl, ethyl, propyl, butyl, hexyl, 1-methylethyl, 1-methylpropyl, 2-methylpropyl, 1,1-dimethylethyl.

The term "C3-C6cycloalkyl" in the context of the present description, one or in combination with another Deputy, indicates cycloalkenyl Deputy containing from 3 to 6 carbon atoms, and includes cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

The term "unsaturated cyclo is lcil" includes, for example, substituted cyclo-hexenyl:

The term "saturated or unsaturated alkylene" in the context of the present description refers to divalent alkyl substituent obtained by removing one hydrogen atom from both ends of saturated or unsaturated aliphatic hydrocarbon straight or branched chain and includes, for example, -CH2CH2C(CH3)2CH2CH2-, -CH2CH2SN=SNSN2CH2- or-CH2With≡CLO2CH2-. Such alkyl chain optionally may include a heteroatom, such as oxygen (for example,- CH3-CH2-O-CH2-).

The concept of "1-C6alkoxy individually or in combination with another Deputy denotes the Deputy-O-C1-C6alkoxy, where alkyl has the specified higher value and contains up to six carbon atoms. The term alkoxy includes methoxy, ethoxy, propoxy, 1-methylethoxy, butoxy and 1,1-dimethylmethoxy. The last Deputy is usually referred to as tert-butoxy.

The concept of "3-C6cycloalkyl individually or in combination with another substituent in the context of the present description denotes the Deputy-O-C3-C6cycloalkyl containing from 3 to 6 carbon atoms.

The term "C1-C6alkoxyalkyl" in the Contex is e of the present description denotes Deputy C 1-C6alkyl-O-C1-C6alkyl, where alkyl has the above values and contains up to six carbon atoms. For example, methoxymethyl denotes-CH2-O-CH3.

The concept of "2-C7acyl" either individually or in combination with another Deputy represents C1-C6the alkyl group attached to a carbonyl group such as-C(O)-C1-C6alkyl.

The concept of "6- or10aryl" in the context of the present description, individually or in combination with another Deputy, means either an aromatic monocyclic system containing 6 carbon atoms or an aromatic bicyclic system containing 10 carbon atoms. For example, aryl includes phenyl or naftalina ring system.

The concept of "7-C16aralkyl" in the context of the present description, individually or in combination with another Deputy, means aryl, as defined above, associated with the alkyl group, where alkyl has the above values, and contains from 1 to 6 carbon atoms. Aralkyl includes, for example, benzyl and butylphenyl.

The term "Het" in the context of the present description, individually or in combination with another Deputy, denotes a monovalent Deputy obtained by removing hydrogen from 5-, 6 - or 7-membered saturated or unsaturated (including num is aromatic) heterocycle, containing from 1 to 4 heteroatoms selected from the series comprising nitrogen, oxygen and sulfur. Examples of suitable heterocycles include then it is carbonated, drofuran, thiophene, diazepine, isoxazol, piperidine, dioxane, morpholine, pyrimidine or

The term "Het" includes a heterocycle as defined above, is fused with one or more other cycles, which may represent a heterocycle or any other cycle. One example is the thiazole[4,5-b]pyridine.

Although it is usually referred to as the "Het", the concept of “heteroaryl” in the context of the present description strictly refers to unsaturated heterocycle, double bonds which form an aromatic system. Relevant examples of heteroaromatic systems are quinoline, indole, pyridine,

The term "pharmaceutically acceptable ester" in the context of the present description, individually or in combination with another Deputy, refers to esters of the compounds of formula I in which any of the carboxyl functions in the molecule, but preferably C-terminal function, substituted alkoxycarbonyl function:

where the fragment R of ester selected from alkyl (e.g. methyl, ethyl, n-propyl, tert-butyl, n-butyl); alkoxyalkyl (for example, methoxymethyl); al is axially (for example, acetoxymethyl); aralkyl (e.g., benzyl); aryloxyalkyl (for example, phenoxymethyl); aryl (e.g. phenyl), optionally substituted with halogen, C1-C4the alkyl or C1-C4alkoxygroup. Other suitable as prodrugs esters can be found in "Design ofprodrugs", H. Bundgaard, ed. Elsevier (1985), this publication is included in the present description by reference. Such pharmaceutically acceptable esters are usually hydrolyzed in vivo when administered to a mammal and turn into acid form of the compounds of formula I.

With regard to the above-described esters, unless otherwise stated, any present alkyl fragment preferably contains 1 to 16 carbon atoms, particularly preferably 1-6 carbon atoms. Any present in such esters, aryl fragment preferably is a phenyl group.

In particular, the esters may represent a complex C1-C16alkilany ether, unsubstituted benzyl ether or benzyl ether, substituted by at least one halogen atom, a C1-C6the alkyl, C1-C6alkoxy-, nitro-group or trifluoromethyl.

The term "pharmaceutically acceptable salt" in the context of the present description include salts derived from pharmaceutically acceptable bases. Examples of the reception of the protected grounds include choline, ethanolamine and Ethylenediamine. Under the scope of the present invention also includes salts of Na+, K+and CA++(see also "Pharmaceutical salts", Birge S.M. and others, J. Pharm. Sci., (1977), 66, 1-19, publication included in the present description by reference).

Preferred embodiments of the

R1:

Preferred embodiments of the present invention described above include the compounds of formula I in which the radical R1choose from two different diastereoisomers, where the carbon center in position 1 is R - configuration, which is represented by structures (i) and (ii):

D is a SYN orientation with respect to the amide (i), or is in a SYN orientation with respect to A (ii)

More preferably the linker D is associated with R1in a SYN orientation with respect to A, as represented by structure (ii).

R2:

Preferred embodiments of the present invention include the above compounds of formula I, where a fragment of R2means

Preferably R21denotes H, C1-C6alkyl, C1-C6alkoxy, hydroxy, chlorine or N(R23)2where R23preferably denotes N or C1-C6alkyl. More preferably R21denotes N or C1-C6ALCO is si. Most preferably, R21denotes methoxy.

Preferably R22denotes H, C1-C6thioalkyl, C1-C6alkoxy, phenyl or Het selected from the group including:

Preferably R24denotes H, C1-C6alkyl, NH-R25, NH-C(O)-R25or NH-C(O)-NH-R25or NH-C(O)-OR26

More preferably R2represents C1-C4alkoxy, phenyl or Het selected from the group including

More preferably R24denotes H, C1-C6alkyl, NH-R25, NH-C(O)-R25; or NH-C(O)-OR26.

Most preferably, R22indicates ethoxy, or Het selected from the group including:

Most preferably, R24adenotes NH-R25, NH-C(O)-R25or NH-C(O)-OR26. Most preferably, R24bdenotes N or C1-C6alkyl.

Preferably each R25independently of each other denotes H, C1-C6alkyl or C3-C6cycloalkyl. More preferably R25represents C1-C6alkyl or C3-C6cycloalkyl.

More preferably R25represents C1-C6alkyl. Preferably R26represents C1-C6and the keel.

R3:

Preferred embodiments of the present invention described above include the compounds of formula I, where a fragment of R3preferably means amide of the formula NH-C(O)-R32, urea of the formula NH-C(O)-NH-R32or a carbamate of the formula NH-C(O)-OR32. More preferably R3means of carbamate or urea. Most preferably, R3means carbamate.

Preferably R32represents C1-C6alkyl or C3-C6cycloalkyl. More preferably R32represents C1-C6alkyl or C4-C6cycloalkyl. Most preferably, R32represents tert-butyl, cyclobutyl or cyclopentyl.

D:

Preferred embodiments of the present invention include compounds of formula I, where the linker D is a saturated or unsaturated alkylenes chain consisting of 6 to 8 atoms. More preferably the linker D is a chain consisting of 7 atoms.

Preferably the chain D contains one or two heteroatoms selected from the group comprising O, S, NH, N-C1-C6alkyl or T2-C7acyl. More preferably, the chain D optionally contains one heteroatom selected from the group comprising NH or T-C2-C7acyl, most preferably N(Ac), and it is on the atom is in position 10 of the chain. Most preferably, the circuit containing the nitrogen atom is saturated.

In alternative D contains one heteroatom selected from O or S. Preferably, if the chain D contains 7 atoms, the atom of O or S is in position 9 of the chain. Preferably this circuit has a substituent R4where R4denotes N or C1-C6alkyl. More preferably R4denotes H or methyl. Most preferably, R4denotes H or 8-(S)-IU. Even more preferably the chain D is saturated. In alternative D contains one double bond between positions 11 and 12. Preferably, this double bond is in the TRANS orientation.

In an alternative embodiment, D is a saturated or unsaturated alkylenes chain containing only carbon atoms. In this case, the circuit D is preferably saturated and consists of 7 atoms. More preferably the circuit D has a substituent R4where R4denotes H, oxo, thio, hydroxy, thioalkyl, alkoxy or alkyl. More preferably R4denotes N or C1-C6alkyl. Most preferably, R4denotes H or methyl. Most preferably, R4denotes H or 10-(S)-IU.

In an alternative embodiment, D is alkylenes chain consisting solely of carbon atoms, which p is edocfile contains one double bond and includes 7 atoms. More preferably the specified double bond is located between positions 13 and 14 of the chain. Most preferably, this double bond is in a CIS-orientation. Preferably this circuit is substituted D R4where R4denotes H, oxo, hydroxy, alkoxy or alkyl. More preferably R4denotes N or C1-C6alkyl. Even more preferably R4denotes H or methyl. Most preferably, R4denotes H or 10-(S)-IU.

And:

Preferred embodiments of the present invention described above include the compounds of formula I, where a denotes a carboxylic acid.

Specific options:

Preferred embodiments of the present invention described above include the compounds of formula I, where R2denotes a Deputy representing quinoline (i.e. W denotes N);

R3denotes a group of formula-NH-C(O)-other32or-NH-C(O)-OR32where R32represents C1-C4alkyl or C4-C6cycloalkyl;

D denotes consisting of 6-8 atoms saturated or unsaturated alkylenes chain attached to R1in a SYN orientation with respect to And, optionally incorporating one or two heteroatoms, independently from each other selected from O, S or N-R41where R41represents C2-C7AC is l;

R4represents H or one to three substituent independently from each other selected from hydroxy, or C1-C6of alkyl; and

And denotes a carboxylic acid, or pharmaceutically acceptable salts or esters.

More preferred are the compounds of formula I, where R’ has the above meanings; R21denotes H or methoxy;

R22represents C1-C6alkoxy, or Het selected from the group including:

where R24adenotes H, C1-C6alkyl, NH-R25, NH-C(O)-R25, NH-C(O)-NH-R25,

where R25represents: H, C1-C6alkyl or C3-C6cycloalkyl;

or R24Adenotes NH-C(O)-OR26where R26represents C1-C6alkyl or C3-C6cycloalkyl;

and R246denotes N or C1-C6alkyl;

R3denotes a urea of the formula NH-C(O)-other32or a carbamate of the formula NH-C(O)-

OR32where R32represents C1-C6alkyl or C3-C6cycloalkyl;

D denotes containing 7 carbon atoms saturated or unsaturated alkylenes chain, optionally containing one double bond between positions 11, 12 or 13, 14.

The chain D optionally includes one heteroatom independently selected from O, S, NH, N(Me) or N(Ac); and R 4denotes N or C1-C6alkyl.

Most preferred are the compounds of formula I, where R21denotes methoxy, and R22indicates ethoxy or:

where R24Adenotes NH-(C1-C4alkyl), NH-C(O)-(C1-C4alkyl); or NH-C(O)-O-(C1-C4alkyl); and

D is a saturated chain or contains one double bond in CIS-orientation between positions 13, 14.

Finally, under the scope of the present invention is applicable to all compounds of formula I are presented in tables 1-9.

The pharmaceutical compositions of the present invention can be administered orally, parenterally or by using a device for implantation. It is preferable to oral administration or introduction by injection. The pharmaceutical compositions of the present invention may contain any conventional non-toxic pharmaceutically acceptable carriers, adjuvants or excipients. In some cases, the pH value of the composition can be adjusted with pharmaceutically acceptable acids, bases or buffers to enhance the stability of the compounds included in the composition or in the form of its introduction. In the context of the present description, the term "parenteral" includes subcutaneous, intradermal, nutrive the tion, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal route of administration by injection or infusion, as well as the introduction in the area of corruption.

The pharmaceutical compositions can take the form of a sterile preparation for injection, for example, sterile injectable aqueous or fat-soluble suspension. This suspension can be obtained using well known methods using a dispersing or wetting agents (such as, for example, tween 80) and suspendida agents.

The pharmaceutical compositions of the present invention can be administered orally in any acceptable dosage form for oral administration, including but not limited to, capsules, tablets, and aqueous suspensions and solutions. In the case of tablets for oral administration commonly used carriers include lactose and corn starch. As a rule, add the oil, such as magnesium stearate. For oral administration in capsule form acceptable diluents include lactose and anhydrous corn starch. When oral and injected water suspension, the active substance is combined with emulsifying and suspendresume agents. If necessary, can be added to certain sweeteners and/or corrigentov, and/or dyes.

Other PR is suitable fillers or carriers for the above preparative forms and compositions can be found in conventional pharmaceutical reference books for example, in "Remington''s Pharmaceutical Sciences", The Science and Practice of Pharmacy, 19th ed. Mack Publishing Company, Easton, Penn., (1995).

Doses in the range of from about 0.01 to about 100 mg/kg of body weight per day, preferably from about 0.5 to about 75 mg/kg of body weight per day of the compounds according to the invention, which is a protease inhibitor, can be used in monotherapy for the prevention and treatment mediated HCV disease. Typically, the pharmaceutical compositions according to the invention can be administered from about 1 to about 5 times a day, or in another embodiment by continuous infusion. This introduction can be used for both emergency and long-term treatment. The amount of active ingredient that may be combined with carriers to obtain a standard dosage forms, should vary depending on the host to be treated and the particular route of administration. The usual composition may contain from about 5% to about 95% active ingredient (wt.%). Preferably such compositions contain from about 20% to about 80% active ingredient.

As should be obvious to the person skilled in the art, can be lower or higher doses than those indicated above. Specific schemes dosing and treatment for any particular patient can depend on various factors, including the activity to Kratovo used connection age, body weight, General health, sex, diet, time of administration, rate of excretion, combination of drugs, the severity and characteristics of the disease, the predisposition of the patient to infection, and they are determined by the attending physician. Typically, treatment begins with small doses, significantly lower than the optimum dose of the peptide. Then the dose is increased in small portions until the optimal actions in specific conditions. Typically, most preferably the compound in this range of concentrations that provide antiviral activity, but does not have any harmful or dangerous side effects.

When the compositions of the present invention include a combination of compounds of formula I and one or more additional therapeutic or prophylactic agents, the compound and the additional agent should be present in the range of doses from about 10% to 100%, and more preferably from about 10 to 80% of the dose normally used in monotherapy.

When these compounds or their pharmaceutically acceptable salts are combined in preparative form with a pharmaceutically acceptable carrier, the resulting composition may be administered in vivo to a mammal, such as man, to inhibit the NS3 protease of HCV, or for the treatment or warned what I infection, caused by HCV virus. Such treatment may also be achieved using the compounds according to the invention in combination with agents including immunomodulators, such as α-, βor γ-interferons; other antiviral agents such as ribavirin, amantadine; other inhibitors of NS3 of HCV protease; inhibitors of other targets in the HCV life cycle, which include (but are not limited to) the helicase, polymerase, metalloprotease or internal site of the entrance to the ribosome (IRES); or a combination of both. Additional agents can be combined with the compounds according to the invention to create a single dosage form. Alternatively, these additional agents may be administered to a mammal separately as part of a multiple dosage form.

According to another variant implementation of the present invention relates to methods of inhibiting the activity of the NS3 protease of HCV in a mammal by introducing the compounds of formula I in which the substituents have the above values.

According to a preferred variant implementation of these methods can be used to reduce the activity of the NS3 protease of HCV in a mammal. If the pharmaceutical composition comprises as active substance only connection according to the invention, such methods may additionally include stage introduced the I to the mammal agent, selected from immunomodulator, anti-virus agent, an inhibitor of HCV protease, or an inhibitor of other targets in the HCV life cycle, such as helicase, polymerase, or metalloprotease. Such additional agent may be administered to the mammal prior to, concurrently or after administration of the compositions according to the invention.

According to another preferred variant implementation of these methods can be used for inhibition of virus replication in the body of a mammal. Such methods can be used for the treatment or prevention caused by HCV disease. If the pharmaceutical composition comprises as active substance only connection according to the invention, such methods can further include the stage of introduction of the agent to a mammal, selected from immunomodulator, anti-virus agent, an inhibitor of HCV protease, or an inhibitor of other targets in the HCV life cycle. Such additional agent may be administered to the mammal prior to, concurrently or after administration of the composition according to the invention.

Proposed in the present invention compositions can also be used as laboratory reagents. Applicants first proposed low molecular weight compounds that have high activity and specificity against the NS3 protease of HCV. Some of the compounds according to the present image is meniu can be a tool for systems development analysis of virus replication, create systems analysis in the animal organism and study the relationship between structure and biological activity with the goal of increasing understanding of the mechanisms associated with HCV disease.

Compounds of the present invention can also be used to eradicate or prevent viral contamination of materials, and thus they reduce the risk of infection for laboratory personnel, or medical institutions that have contact with such materials (for example, blood, tissue, surgical instruments and garments, laboratory instruments and garments, instruments and materials for blood collection).

The method

Some methods for the synthesis of acyclic intermediates for compounds of formula I described in WO 00/09543 and WO 00/09558.

Compounds of the present invention are synthesized according to the General process illustrated in schemes I, II and III (where PG denotes the corresponding protective group). [All the following diagrams D’ has the same meaning as D, but shorter 2-5 atoms].

When considering the compounds of formula I, where a denotes the N-substituted amide, vinyl-ACPP or Gamaliel ACPP (R1) are combined with the appropriate amine prior to combination with P2. Methods of this combination is well known to specialists in this field. As Izv the local experts in this field, this amide (A) is unsecured, but may have any of the above suitable substituents R5.

The reaction ring closure (macrocyclization) are carried out either by olefin metathesis (Scheme I), or when the linker contains a nitrogen atom, by reductive amination (Scheme II), or by formation of a peptide bond according to Scheme III.

These processes are described in detail below:

A. Macrocyclization by olefin metathesis

Scheme I

Scheme I:

There are several ways to implement the sequence of reactions combinations, which are well known to specialists in this field. Using as the source of the product 4-(S)-hydroxyproline, the substituent in the 4-hydroxy-group may be introduced through reaction of Mitsunobu (according to the method described by Mitsunobu,

Cunthesis 1981, January, 1-28; Rano and others, Tet. Lett. 1994, 36, 3779-3792; Krchnak, etc., Tel. Lett. 1995, 366, 6193-6196) before implementation of macrocyclization or after it. In an alternative embodiment, the Assembly can be carried out using the appropriate 4-(R)-hydroxy-substituted Proline according to the General processes described in WO 00/09543 and WO 00/09558 (below presents specific examples of these fragments).

Stages a, B, C: In General, the fragments P1, P2 and P3 can be linked by well known methods of peptide sequencing the Oia and they are generally described in WO 00/09543 and WO 00/09558.

Stage G: getting the macrocycle can be carried out by olefin metathesis using containing Ru catalyst, as described in Miller, S.J.; Blackwell, H.E.; Grubbs, R.H. J. Am. Chem. Soc. 1996, 118, 9606-9614 (a); Kingsbury, J.S.; Harrity, J.P.A.; Bonitatebus, P.J.; Hoveyda, A.H. J. Am. Chem. Soc. 1999, 121, 791-799 (b) and Huang, J.; Stevens, E.D.; Nolan, S.P.; Petersen, J.L.; J. Am. Chem. Soc. 1999, 121, 2674-2678 (in). It should also be noted that for this reaction can be used catalysts containing other transition metals such as Mo.

Stage E: optional Double bond can be recovered using standard hydrogenation methods, well known in this field. If A’ is a protected carboxylic acid, it is also appropriate to remove the protective group.

B. Macrocyclization by reductive amination (for linkers containing N)

If the linker contains a nitrogen atom, macrocyclization can be carried out by reductive amination as illustrated in Scheme II with getting inhibitors having the General structure II.

Scheme II

Stage a: Hydroporinae double bond according to the method of brown (NS Brown and B.C. Subba Rao, J. Am. Che. Soc. 1959, 81,6434-6437) with subsequent oxidation of the resulting alcohol (for example, using periodinane dess-Martin . Am. Chem. Soc. 1991, 113, 7277-7287) results sootvetstvuyuschego aldehyde.

Stage B: Hydrogenation in the presence of acid leads to the destruction of aminosidine group, followed by macrocyclization by reductive amination.

The P3 fragment used in this synthesis can be easily derived from various amino acids such as lysine, ornithine, glutamine (through reaction of Hoffmann: Weg. 1881, 14, 2725), and others; modifications of this method of synthesis is well known in this field.

Stage: Secondary amine in the linker D (obtained in stage G) optional alkylate using alkylhalogenide or acetylinic using alkyl - or arylchloranhydrides using techniques well known in the field, obtaining inhibitors having the General structure II. If A’ is a protected carboxylic acid, it is accordingly also remove the protective group.

Century Macrocyclization by the formation of lactam

On the other hand, it should be understood that these macrocyclic compounds having the General structures I and II can be synthesized by other methods.

For example, P1 and P3 can be connected to the first linker D, then subjected to the combination of P2 and the reaction of macrocyclization can lead to the formation of the lactam, which can be done in two possible what ways, as is known to experts in this field, as illustrated in scheme III.

Scheme III

Synthesis of P1

For the synthesis inhibitors having the General structures I and II require the same fragments P1:

(a) vinyl-ACPP, synthesis and secretion of which is described in WO 00/09543 and WO 00/09558, or

b) Gamaliel-ACPP (example 1, compound 1f).

Synthesis of P2

Some of the fragments P2 used in the synthesis of compounds of formula I described in WO 00/09543 and WO 00/09558.

Other fragments P2 synthesized as follows:

Synthesis of 2-"Het"-4-hydroxy-7-methoxyquinoline derived

(i) an Approach based on using as starting product of the corresponding "t"-carboxylic acid IVb

Scheme IV

The synthesis is carried out according to the modified method described by Li and others J. Med. Chem. 1994, 34, 3400-3407. Intermediate IVa, where R21means OMe (example 7, compound 7b) are obtained according to the method described by Brown and others, J. Med. Chem. 1989, 32, 807-826.

Stage A: Intermediate IVa are combined with a heterocyclic carboxylic acids IVb in basic conditions in the presence of l3to activate the carboxylate groups. To obtain inhibitors using various carboxylic acids having the General structure IVb, which either are in p is adage, or can be synthesized as shown in schemes V, VI and VII, or can be synthesized individually by using the methods described in the specific examples.

Stage B: ring Closure and subsequent dehydration is carried out in basic conditions with the quinoline having the General structure of the IVd.

(i.a). Synthesis of "Het-carboxylic acids of General formula IVb

Synthesis of 2-(substituted)amino-4-carboxylesterase derivatives (V)

Apply the modified process described in Berdikhina, etc.. Chem. Heterocycl, Compd. (translated into English) 1991, 4,427-433.

Scheme V

According to the General synthesis methodology presented in scheme V, using thioureas (Va)with different alkyl substituents (R25denotes an alkyl group), and 3-bronirovochnoy acid get different 2-alkylaminocarbonyl-4-carboxylic acid, which are formations

having a General structure of the Vc. This type of condensation reaction is widely known in this field.

In an alternative embodiment, the fragment P2, including derivatives of 2-aminosilanes thiazole, synthesized from the corresponding 2-carboxyl derivative, as shown in scheme VI, according to the method described in Unangst, R.S.; Connor, D.T. J. Heterocyc. Chem. 29, 5,1992, 1097-1100.

Scheme VI

The examples that the CSO process described in WO 00/09543 and WO 00/09558.

Synthesis of 2-carboxy-4-substituted aminothiazole derivatives of the formula VIId According to the General synthesis methodology presented in scheme VII, receive various 4-alkylaryl-2-carboxylic acid, which are compounds having the General structure VIId.

Scheme VII

Use the process described in Janusz etc., J. Med. Chem. 1998, 41, 3515-3529 with the following modifications: atitikimas (Vila) is subjected to interaction with various β-bracketname having the General structure VIIb (R24denotes an alkyl group), receiving thiazolecarboxamide acid having the General structure VIId. This type of condensation reaction is widely known in the art. Synthesis of 2-carboxy(substituted)imidazole derivatives (VIIIb). According to the General synthesis methodology presented in scheme VIII, receive various alkylimidazole-2-carboxylic acid, which are compounds having the General structure VIIIb.

Scheme VIII

Use the process described by Baird and others, J. Amer. Chem. Soc. 1996, 118, 6141-6146: alkylimidazole deprotonated using a strong base (e.g. n-BuLi) and then subjected to interaction with CO2getting carboxylic acid of formula VIIIb. This type of condensation reaction is widely known in this field.

B. Synthesis of 4-hydroxy-7-methoxy-2-(them is azolyl or pyrazolyl)quinoline

4-Hydroxy-7-R21-quinoline having in position C2 imidazolidinyl or Chi-mailny fragment, are generally according to the methodology presented in scheme IX.

Scheme IX

The synthesis of the primary intermediate product, (where R21means OMe) 4-benzyloxy-2-chloro-7-methoxyquinoline formula IXa is described in detail in example 6 (compound 6E).

Stage A: At high temperatures for replacing 2-chloro-fragment in connection IXa can be used various imidazoles, the alkyl substituted imidazoles, pyrazoles or alkyl substituted pyrazoles, resulting in a gain of compounds having the General structure IXb.

Stage B: After removal of the benzyl protective group from 4-hydroxy-quinoline fragment using standard methods of hydrogenation get quinoline derivatives having the General structure CHD.

Synthesis P3

For macrocyclization by olefin metathesis synthesize various fragments P3 containing the corresponding D-linker elongation. In General containing a terminal olefin fragments P3 synthesize for metathesis according to the following General schemes (Scheme X, XI and XII).

Synthesis of linkers from class a

This General method of synthesis used for linkers containing only atoms of carbon (no heteroatoms) (Scheme X).

Scheme X

The synthesis is carried out according to the method described by Evans and others, J. Am. Chem. Soc. 1990, 112,4011-4030.

The original carboxylic acid (Ha) are commercially available or can be obtained according to the literature methods known to experts in this field.

Stage A: Carboxylic acid Ha activate using pivaloyloxy and then subjected to the interaction with the anion chiral auxiliary 4(S)-4-(phenylmethyl)-2-oxazolidinone Evans according to the widely known method (see review: D.J.Ager and others, Aldrichimica Acta 1997, 30, 3-11 and therein references)to give compounds having the General structure b.

Stage B: Stereoselective α-sideropenia chiral midtolate, which can be obtained from compounds having the General structure b, in the presence of a base, such as KHMDS, using tritiated also widely known in this field (see review: D.J. Ager and others, Aldrichimica Acta 1997, 30, 3-11 and therein references).

Stage: After restoring α-azide catalyzed using SnCl2, protect the formed amine as its tert-BUTYLCARBAMATE, receiving the intermediate products having the General structure of Cholesterol. Such reactions are widely known in this field.

Stage G: At the end of chiral auxiliary product hydrolyzing in basic conditions, for example, in a mixture of H2O2with LiOH, receiving the amino acid linkers of the type having the General structure Heh.

Alternatively, fragments P3 having the General structure Heh, synthesized according to the method described by M.J. Burk and others, J. Am. Chem. Soc 1998, 120, 657-663, illustrated in scheme XI. These compounds differ in the number of methylene units (-CH2-) in the linker (m=1-5) and by substitution of alkyl groups in R4but they do not contain heteroatoms.

Scheme XI

Stage A: Derived monobasic acid of the formula XIb is obtained from commercially available diethyl-2-acetamidomalonate by standard hydrolysis of ester in basic terms.

Stage B: the reaction of the condensation type knoevenagel between aldehyde having the General structure of the XIc and the compound of the formula XIb in the presence of a base, such as pyridine, and acetic anhydride leads to the formation of enamide formula XId, which is an intermediate product, which is characterized by a Z-stereochemical configuration relative to the newly formed double bond, as shown in the diagram.

Stage: Regioselective and enantioselective catalytic hydrogenation of the intermediate product, representing UNAMID formula XId obtaining an intermediate product, which is the amino acid of formula Xie, carried out according to the method of Burke.

<> Stage G: Then the nitrogen acetamido derivative of formula XIe double-protect by attaching a substituent, represents a tert-BUTYLCARBAMATE, before the acetate group, and also in the form of ethyl ester, hydrolyzing the product in standard basic conditions with fragments P3 having the General structure XIf.

Synthesis of linkers from class B

This General scheme of synthesis used for obtaining linkers containing oxygen or sulfur.

Scheme XII

Stage A: appropriately protected amino acid, such as methyl ester BOC-(L)-serine, methyl ester BOC-(L)-threonine or methyl ester BOC-(L)-allochrony, alkylate using alkylated in the presence of Ag2O getting the methyl ether of the formula XIIb.

Stage B: hydrolysis of complex methyl ester in the standard basic conditions are linkers in the form of a simple ester having the General structure XIIc (X is O).

Stage: Similar, containing sulphur, derived from the same source of amino acids of formula Ha (protected accordingly, as described above) and its hydroxyl group is transformed into easily removable group (such as an intermediate product, representing toilet formula XIId), using standard methods, well known in the data is the second region.

Stage G: then cosily fragment replaces the anion thioacetate, receiving by the inversion of the chiral center on the β-carbon atom of the intermediate product, representing a complex tiefer He.

Stage D: By hydrolysis of the thioester fragment in a weak basic conditions have free thiol of the formula XIIf.

Stage E: Alkylation Colnago fragment can be easily carried out in basic conditions using Allilueva.

Stage G: In the end by hydrolysis of complex methyl ester using standard methods get sulfide analogue of formula HS (X denotes S).

Synthesis of fragment R3:

Examples of the synthesis of fragments, where R3denotes NH-R31described in WO 00/09543.

EXAMPLES

Below the invention is illustrated in the examples, not limiting its scope. Other specific methods of synthesis or separation is described, for example, in WO 00/09543 and WO 00/09558.

Temperatures are given in degrees Celsius. Interest for solutions represent % wt./about., as ratios in solutions represent a ratio of volumes, unless otherwise stated. The spectra of nuclear magnetic resonance (NMR) were recorded using a spectrometer firm Brucker at 400 MHz, chemical shifts (S) expressed in frequent./million and related to the internal deuterium solvent, unless otherwise noted. The NMR spectra of the all target compounds (inhibitors) were recorded in the solution of their salts with triperoxonane acid (TFA) in DMSO-d 6if not stated otherwise. Rapid chromatography on a column was performed on silica gel (SiO2) according to the method of Steele (W.C.Still etc., J. Org. Chem., (1978), 43, 2923). In the examples the following abbreviations are used: VP: benzyl; BOC: tert-butyloxycarbonyl {IU3SOS(O)}; BSA: bovine serum albumin; CBS: benzyloxycarbonyl; CHAPS: 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate; DBU: 1,8-diazabicyclo[5.4.0]undec-7-ene; CH2CL2=DHM: methylene chloride; DEAD: diethylazodicarboxylate; DYADS: diisopropylsalicylic; DIEA: diisopropylethylamine; DIPEA: diisopropylethylamine; DMAP: dimethylaminopyridine; DCC: 1,3-dicyclohexylcarbodiimide; DME: 1,2-dimethyloxetane; DMF: dimethylformamide; DMSO: dimethyl sulfoxide; DTT: dithiothreitol or threo-1,4-dimercapto-2,3-butanediol; DFA: diphenylphosphoryl; add: ethylenediaminetetraacetic acid; Et: ethyl; EtOH: ethanol; EtOAc: ethyl acetate; Et2O: diethyl ether; ESMS: mass spectrometry with ionization by electron beam; GATA: [hexaphosphate O-7-asobancaria-1-yl)-1,1,3,3-tetramethyluronium]; GHUR: liquid chromatography high resolution; MS: mass spectrometry; MALDI-TOF: time-of-flight mass spectrometry with matrix laser desorption sample; FAB: fast atom bombardment; LAG: alumaguard lithium; Me: methyl; Meon: methanol; MES: (2-{N-morpholino}econsultancy acid); NaHMDS: bis(trim the Telcell)sodium amide; NMM: N-methylmorpholine; NMMO: oxide N-methylmorpholine; N: N-methylpyrrolidine; WG: propyl; Succ: 3-carboxypropanoyl; pNA: 4 nitrophenylamino or para-nitroanilide; TBUF: fluoride, Tetra-n-butylamine; TBTU: tetrafluoroborate 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium; DEF: hydrochloride Tris(2-carboxyethyl)phosphine; TPA: triperoxonane acid; THF: tetrahydrofuran; TIS: triisopropylsilane; TLC: thin layer chromatography; TSE: trimethylsilylmethyl; Tris/HCl: hydrochloride Tris(hydroxymethyl)aminomethane.

FRAGMENTS P1

Example 1

Synthesis of tert-butyl(1R,2R)/(1S,2S)-1-amino-2-homoallylamines (1f):

A. To a suspension of chloride benzyltriethylammonium (5,08 g, 22.3 mmole) in 50%aqueous NaOH solution (50 ml), was added 1,2-dibromo-5-hexene (1b, 8,10 g, 33,46 mmole) and di-tert-butylmalonate (1A, 4,82 g, 22,30 mmole). The mixture was intensively stirred at RT for 16 h, then diluted with H2O and was extracted with CH2Cl2(3×50 ml). Then the organic layer was washed N2About (2×50 ml), saline/N2On (2/1,2×50 ml), dried over SO4and was evaporated. The crude residue was purified using the rapid chromatography on columns on silica gel, using as eluent a gradient from 3% to 5% tO in hexane, obtaining compound 1C with 38% (2,48 g).

1H NMR (Dl3,400 MHz): δ 1,19 (bd, J=7.9 Hz, 2H), 1,25-of 1.33 (m, 1H), of 1.46 (s, 9H), to 1.48 (s, 9H), 1,47 is 1.60 (m, 1H), 1,75-to 1.82 (m, 1H), 2,14-2,22 (m, 2H), 4,93-of 5.50 (m, 2H), 4,96 (dm, J=10,2 Hz, 1H), 5,18 (dm, J=and 17.2 Hz, 1H). ES(+) MS m/z 297 (M+N)+

B. To a suspension of tert-butoxide potassium (5.75 g, 51,25 mmole) in anhydrous diethyl ether (150 ml) at 0°, was added N2(203 μl, 11,27 mmole) and the reaction mixture was stirred at 0° within 10 minutes was Added a solution of compound 1C in diethyl ether (2,48 g in 10 ml of diethyl ether, 10.25 mmole) and the mixture was stirred at RT for 5 h the Mixture was diluted with chilled on ice N2O and was extracted with diethyl ether (3×200 ml). The aqueous layer was acidified to pH 3.5-4 using chilled on ice and 10%aqueous citric acid solution and re-extracted with tO (3×200 ml). tO-layer washed with N2About (2×100 ml), brine solution (100 ml), dried over SO4and evaporated, obtaining compound 1d with the release of 85% in terms of the number of recovered original product.

1H NMR (CDCl3, 400 MHz): δ is 1.51 (s, 9H), 1,64 by 1.68 (m, 1H), 1,68 is 1.75 (m, 1H). 1,77-of 1.88 (m, 1H), 1,96 is 2.01 (m, 1H), 2,03-2,22 (m, 3H), 5,01 (dm, J=6,4 Hz, 1H), 5,03 (dm, J=14,Hz, 1H), 5,72-of 5.83 (m, 1H).

ES(+)MS:m/z 241 (M+H)+

C. To a solution of acid 1d in anhydrous benzene (1,14 g in 25 ml of benzene, 4,74 mmole)was added et 3 n (800 μl, of 5.68 mmole), and then added diphenylphosphoryl (1.13 ml, a total of 5.21 mmole) and the mixture was heated to the temperature of reflux distilled in 3,5 o'clock Then add trimethylsilylethynyl (1,36 ml, 9,48 mmole) and continued stirring at the temperature of reflux distilled for 4 hours Then the mixture was cooled to CT, was evaporated to half its original volume, diluted with diethyl ether (30 ml) and washed with 5%aqueous solution of NaHCO3(2×30 ml), brine solution (50 ml), dried over SO4and was evaporated. The remainder in the form of oil chromatographically on silica gel, using as eluent a 10%tO in hexane, resulting in a net 1E connection with the release of 88% (1,49 g).

1H NMR (CDCl3, 400 MHz) δ of 0.03 (s, 9H), 0.91-0.99 are equivocal (m, 2H), 1,18-of 1.29 (m, 2H), 1,45 (bs, 11H), 1.56 to 1,72 (m, 2H), 2,02-to 2.18 (m, 2H), 4,12 (t, J=8,3 Hz, 2H), 4,93 (dm, J=10,2 Hz, 1H), 4,98 (dm, J=and 17.2 Hz, 1H), 5,07 (bs, 1H), 5,71-of 5.83 (m, 1H).

G. To a solution of cyclopropenes derived 1E (1.19 g, at 3.35 mmole, 30 ml of THF)was added tert-V4NF (6,7 ml in the form of a 1M solution in THF, 6.7 mmole) and the mixture is first stirred at RT for 16 h, and then heated to the temperature of reflux distilled for 15 minutes the Solvent is carefully evaporated under reduced pressure (due to high volatility of the free amine 1f it is necessary to take precautions upon evaporation of the solvent). The crude residue was re-dissolved in tO (100 ml) and washed with H2O (2×50 ml), brine solution (50 ml), dried over mg SO4and then carefully evaporated of rest ritel. The crude product 1f (representing a mixture of the two enantiomers 1f’ and 1f) was used in combination with a derivative of Proline P2 without additional purification. At this stage Express chromatography could easily be the selection RR with the desired stereochemical structure P1 (example 21, the fragment 21b).

FRAGMENTS P2

Example 2

Synthesis of BOC-4(R)-[(7-methoxy-4-chinoline)oxy]Proline (2C):

4-Hydroxy-7-methoxyquinoline (2b) was obtained according to the method described in M.W.Chun, K.K.Olmstead, Y.S.Choi, C.O.Lee, C.-K.Lee, J.H.Kirn, Lee,.J. Bioorg. Med. Chem. Lett. 1997, 7, 789. A solution of compound 2b (1.88 g, of 10.73 mmole) and DEAD (3.4 ml, 21,46 mmole) in anhydrous THF was added under stirring to a solution of protected CIS-hydroxyproline 2A (2,63 g of 10.73 mmole) and triphenylphosphine (5,63 g, 21,46 mmole) in anhydrous THF (160 ml) at 0° in an atmosphere of N2. The reaction mixture was allowed to warm up to CT and it was stirred for 14 hours Then THF evaporated and provided pure product 2C using rapid chromatography on a column, using as eluent 5%Meon in tO; yield 35% (1.5 g).

1H NMR (CDCl3, 400 MHz): δ the 1.44 (s, 9H), of 1.65 (bs, 1H), 2,34 is 2.43 (m, 1H), 2,63 was 2.76 (m, 1H), of 3.78 (s, 3H), 3.75 to 3,85 and the 3.89-3,99 (2m, 1H, 2 rotamer), 3,95 ('s. 3H), 4,51 and 4.60 (2t, J=8 Hz, 1H, 2 rotamer), of 5.15 (bs, 1H), 6,53-6,59 (m, 1H), 7,12-to 7.18 (dd, J=8,9 and 2.2 Hz, 1H), was 7.36 (d, J=2.6 Hz, 1H), 8,03 (bd, J=9,2 Hz, 1H), 8,65 (bd, J=5,1 Hz, 1H).

Example 3

Synthesis of 2-ethoxy-4-hydroxy-7-methoxyquinoline (3C)

Synthesis of methyl-pair-of ethoxyacrylate 3A was carried out according to the method described in Katzu others, J. Org. Chew., 1953, 18, 1380-1400.

General procedure for the synthesis of a derivative of quinoline 3 is a modification of the method described in Watcch and others, Indian Journal of Chemistry, 1995, Sat. In, 330-332.

A. Methyl-para-mitoxantrone 3A (3,069 g, 16,96 mmole) was dissolved in triethylorthoformate (4,7 ml, 25.4 mmole)was then added a solution of anhydrous Hcl (4 n solution in dioxane, 50 μl, 0.6 mmole). The resulting mixture was stirred at the temperature of reflux distilled for 19 hours and Then the volatile components are evaporated in vacuum, obtaining the product 3b (4,92 g, oil of amber, quantitative yield)which was used in the next stage without further processing.

B. To a solution of the substrate 3b (according to theoretical estimate 16,96 mmole) in THF (34 ml) at -78°C in an atmosphere of nitrogen was added LiHMDS (1 M solution in THF, 22 ml, 1.3 EQ.). Soon after the addition the cooling bath was removed and the mixture was stirred at ambient temperature for 1 h, after which was added another portion of LiHMDS (16 ml). The resulting mixture was stirred until complete disappearance of the original product (1 h) according to TLC (100% tO, retention time of imidate Rf=0,7, retention time of the product Rf=0,2). After this was added Hcl (4n. solution in dioxane, 10 ml) and the mixture to which is centered in a vacuum. The resulting pasty product is triturated with a mixture tO (10 ml) and aqueous solution of NaH2PO4(1M, 10 ml) and irradiated with ultrasound. The result has formed an abundant precipitate, which was collected by filtration, washed with water and dried, obtaining the desired product 3C in the form of a solid beige color (3,117 g, yield 84% for two steps, purity >99% according to analysis with IHVR).

1H NMR (400 MHz, DMSO-d) δ (part./million): 7,88 (d, J=8,9 Hz, 1H), 6,98 (br. s, 1H), 6.89 in (br. d, J=8.6 Hz, 1H), 5,94 (br. s, 1H), 4,30 (br. s, 2H), of 3.84 (s, 3H), of 1.34 (t, J=7.0 Hz, 3H).

Example 4

Synthesis of 4-hydroxy-7-methoxy-2-(3-methyl-1,2,4-oxadiazol-5-yl)quinoline(4d)

A. To a solution of 2-carbomethoxy-4-hydroxy-7-methoxyquinoline 4A (receipt described in WO 00/09543 and WO 00/09558) (1 g, the 4.29 mmole) in DMF (10 ml) under nitrogen atmosphere was added NaH (60%in mineral oil, 190 mg, 4,98 mmole). The resulting mixture was stirred at ambient temperature for 1 h, then was added dropwise a MEM chloride (455 μl, 4,98 mmole) and the resulting mixture was stirred at ambient temperature in a period of 19.5 hours the Reaction mixture was diluted with EtOAc (100 ml), washed with H2About (50 ml), Solanum solution (50 ml), dried over MgSO4concentrated in vacuum, obtaining the crude reaction product (1,37 g). It was purified using the rapid chromatography on a column, olucha product 4b (1.04 g, yield 75%) as a colourless oil.

B. To a mixture containing the newly activated molecular sieve with size 4 E (500 mg) and acetamidoxime (248 mg, at 3.35 mmole) was added THF (3 ml). The resulting mixture was stirred for 15 minutes in a nitrogen atmosphere at ambient temperature, after which the portions was added NaH (60%in mineral oil, 124 mg, 3,24 mmole). The resulting suspension was stirred at ambient temperature for 1 h, then was added ester 4b (500 mg, of 1.56 mmole) in solution in THF (5 ml). The resulting mixture was stirred at the temperature of reflux distilled for 1 h, then filtered through celite, washed with EtOAc (3 portions of 20 ml) and concentrated in vacuum. The resulting crude mixture was purified via Express chromatography (100%EtOAc)to give the product 4 (352 mg, yield 65%) as a solid white color.

Century To a simple MEM-ether 4 (170 mg, 0,493 mmole) in THF (4 ml) was added an aqueous solution of Hcl (1 ad, 1 ml). The resulting mixture was stirred at ambient temperature for 1 h, then was diluted aqueous solution Pan2RHO4(1M, 50 ml). The formed solid product was filtered, triturated with EtOAc, filtered and dried, obtaining the desired product (4d) (90 mg, 71%yield) as a solid white color. MS (ES+) 258 (M+1), (ES-) 256 (M-1).

1H NMR (400 MHz, DMSO-) δ (part./million): 8,03 (d, J=9,2 Hz, 1H), 7,38 (d, J=2.2 Hz, 1H), 7,06 (d, J=8.6 Hz, 1H), 6,85 (br. s, 1H), 3,88 (s, 3H), of 2.64 (s, 3H).

Example 5

Synthesis of 4-hydroxy-7-methoxy-2(5-methyl-1,3,4-oxadiazol-2-yl)quinoline (5e)

A. To the substrate 4b (465 mg, of 1.45 mmole) in ethanol (5 ml) was added anhydrous hydrazine (57 μl, 1.8 mmole). The resulting solution was kept at the temperature of reflux distilled for 4 h, then was concentrated in vacuum, obtaining the product 5A (704 mg, quantitative crude yield) as a solid yellow color, which was used without purification in the next stage.

B. Compound 5A (according to theoretical estimate of 1.45 mmole) in triethylorthoformate (5 ml) kept at 100-110°C in nitrogen atmosphere. Then the resulting mixture was diluted with EtOAc (100 ml), washed with aqueous saturated solution of NaHCO3(50 ml), brine solution (50 ml), dried over MgSO4concentrated in vacuo and purified using the rapid chromatography on a column (100%EtOAc). Compound 5b (359 mg, yield 61% for two steps) was obtained as yellow oil. MS (ES+) 392 (m+1), (ES-) 390 (m-1).

C. Compound 5b (333 mg, 0,852 mmole) was kept at 140°C in high vacuum for 8.5 h and purified using the rapid chromatography on a column (100%tO), receiving a mixture of compound 5b (116 mg, yield 35%, Rf=0.5) and compound 5C (138 mg, adjusted in the course of 72%, rf=0,3). To a solution of compound 5C (138 mg, 0.4 mmole) in THF (4 ml) was added an aqueous solution of Hcl (1 ad, 1 ml) and the resulting mixture was stirred to complete the reaction (30 min). THF is evaporated in vacuo and added an aqueous solution of NaH2PO4(1 M, 2 ml). The resulting suspension was irradiated by ultrasound, was filtered and the solid product was dried in high vacuum, obtaining the desired product 5d (75 mg, yield 73%) as a solid beige color. MS (ES+) 258 (m+1), (ES-) 256 (m-1).1H NMR (400 MHz, DMSO-d): δ 8,03 (d. J=9,2 Hz, 1 H), 7,39 (d, J=2.2 Hz, 1 H), 7,06 (br. d, J=8.6 Hz, 1 H), 6,85 (br. s, I H), 3,88 (s, 3H), of 2.64 (s, 3H).

Example 6

Synthesis of 4-benzyloxy-2-(chloro)-7-methoxyquinoline (6th)

A. commercially Available meta-anisidine (25 g, of 0.20 mol) in dioxane (80 ml) was cooled to 0°and added anhydrous Hcl (4n. solution in dioxane and 75 ml of 0.30 mole). Then add Et2O (500 ml) and continued stirring for 1 h then filtered received a solid beige color and was dried in vacuum, obtaining salt 6a (31,88 g, yield 98%).

B. this salt was added ethylcyanoacrylate (21,3 ml of 0.20 mol) and the mixture is placed in a flask equipped with a distillation head and a flask for collection of the product was heated to 280-300°C. To monitor the course of the reaction is collected ethanol. After accumulated 9 ml of ethanol (in theory, expect the TES number is 11.7 ml) heating was stopped, the reaction mixture was cooled to CT, the mixture was diluted with water (200 ml) - tO (200 ml), then stirred and added an aqueous solution Pan2RHO4(300 ml). After stirring for 1 h, filtering and drying the obtained product 6b (19,06 g, purity of 84.5%, yield ~50%) in a solid yellow color, which is used for the next reaction without further purification.

C. Compound 6b (11,0 g of 57.8 mmole) in DMF (100 ml) at 0° (C) was added to NaH (60%solution in mineral oil, 2,78 g, 115,6 mmole). Then the ice bath was removed and the mixture was stirred at ambient temperature for 1 h, then was added benzylbromide (7,6 ml, 63,6 mmole) and the reaction mixture was stirred for 16 h then the solution was diluted with a mixture tO (220 ml) - hexane (220 ml) and filtered the resulting solid product was washed with an aqueous saturated solution Panso3(110 ml), washed with water, hexane-tO (ratio 1:1, 100 ml) and dried in high vacuum. The result has been the product 6s (5.6 g, purity 91%, yield 35%) as a solid yellow color.

To compound 6C (2.67 g, 9,52 mmole) in acetic acid (21 ml) was added ISO-amyl nitrite (3.8 ml, 28.6 mmole), the resulting mixture was stirred at ambient temperature and observed using GHUR. After 2 h was added another portion of ISO-s is nitrite (1.3 ml, 9,52 mmole) and the mixture was left to mix for 90 h (according to GHUR product content 81%of the substrate 3%). To the resulting suspension was added water (100 ml) and then filtered. The collected solid was dried in high vacuum, receiving product 6d (2.35 g, purity 92%, yield 72%).

, To the compound 6d (1.5 g, 4,39 mmole) was added phosphorus oxychloride (13 ml, 141 mmole) and the resulting mixture was stirred at the temperature of reflux distilled for 1 h, then was diluted tO (150 ml) and the reaction was stopped by slowly added at 0°With aqueous NaOH (1 N., 150 ml), bringing the pH value to 9. Separated the two layers and the organic layer was dried over MgSO4and concentrated in vacuum, obtaining a solid brown color, which was purified using the rapid chromatography on a column (15%tO/hexane). Received the product 6E (819 mg, purity >99%, yield 62%) as a solid yellow color.

1H NMR (400 MHz, CDCl3): δ 8,07 (d, J=9,2 Hz, 1 H). 7,50-7,40 (m. 5H), 7,29 (d, J=2,5 Hz. 1 H), 7,12 (dd, J=9,2, 2.5 Hz, 1 H), 6.73 x (s, 1H), 5,26 (s, 2H), 3,92 ('s. 3H).

Example 7

Synthesis of 4-hydroxy-2-(1-imidazolyl)-7-methoxyquinoline (7b); 4-hydroxy-2-(4-methyl-1-imidazolyl)-7-methoxyquinoline(7d); 4-hydroxy-7-methoxy-2-(1-pyrazolyl)quinoline (7f); and 4-hydroxy-2-(3-methyl-1-pyrazolyl)-7-methoxyquinoline (7h).

A. Compound 6e (423 mg, of 1.41 mmole) and Idasa (400 mg, 5,88 mmole) was kept at 110°C for 20 hours and Then the mixture was diluted with EtOAc and washed with water and with brine, dried over MgSO4concentrated under reduced pressure, obtaining the compound 7a (422 mg, purity 96%, yield 90%) as a solid yellow color. Compound 7a (319 mg, 0,963 mmole) in the presence of Pd (5%/C, 64 mg) in ethanol (5 ml) and THF (5 ml) was purged with hydrogen and left in an atmosphere of hydrogen at a pressure of 1 ATA. After stirring for a period of 7.5 h at ambient temperature the reaction mixture was filtered, washed with a mixture of chloroform-methanol and concentrated, obtaining the product 7b (130 mg, purity of 97.7%, yield 56%) as a solid yellow color. MS (ES+) 242 (m+1), (ES-) 240 (m-1).

1H NMR (400 MHz, DMSO-d): δ 8,51 (s, 1H), 8,03 (d, J=8,9 Hz, 1H), to 7.93 (s, 1H), 7.23 percent (d, J=1.9 Hz, 1H), 7,15 (s, 1H), 7,12 (dd, J=9,2, 2.2 Hz, 1H), 6,92 (br. s, 1H), 3,91 (s, 3H).

B. Compound 6E (251 mg, 0,837 mmole) and 4-Mei (344 mg, 4.19 mmole) was kept at 110°C for 20 hours Then the mixture was diluted tO, washed with water and with brine, dried over MgSO4, and concentrated under reduced pressure, obtaining the crude product containing a mixture of 10:1 isomers of 4-methyl - 5-methylimidazole respectively. The main desired isomer 11S representing solid white (166 mg, purity 99%, yield 57%)was separated from the second, more polar fraction (76 mg, you are the od 23%). containing a mixture of isomers of 4 - and 5-methylimidazole, using rapid chromatography on a column (100%tO). Compound 7C (163 mg, 0,472 mmole) in the presence of Pd (5%/C, 33 mg) in ethanol (2.4 ml) and THF (5 ml) was purged with hydrogen and left in an atmosphere of hydrogen at a pressure of 1 ATA. After stirring for 18 h at ambient temperature the reaction mixture was filtered, washed with a mixture of chloroform-methanol and concentrated, obtaining compound 7d (118 mg, purity 99%, yield 98%) as a solid white color.

1H NMR (400 MHz, DMSO-d): δ 8,42 (br. s, 1H), 8,01 (d, J=9,2 Hz, 1H), to 7.64 (br. s, 1H), 7,21 (br.s, 1H),7,10(d, J=8,9 Hz, 1H), 6.89 in (br. s, 1H), 3,90 (s, 3H), of 2.20 (s, 3H).

C. Compound 6e (184 mg, 0,614 mmole) and pyrazole (209 mg, of 3.07 mmole) was kept at 110°C for 17 h Then the mixture was diluted tO and washed with an aqueous solution of NaOH (1 ad) and with brine, dried over MgSO4concentrated under reduced pressure, obtaining the crude product, which was purified using the rapid chromatography on a column (a mixture of 2:1 hexane-tO)to give compound 7E (103 mg, yield 50%) as a solid light yellow color. Compound 7E (103 mg, 0,311 mmole) in the presence of Pd (5%/C (20 mg) in ethanol (2 ml) and THF (2 ml) was purged with hydrogen and left in an atmosphere of hydrogen at a pressure of 1 ATA. After stirring for 5.5 h at ambient temperature reactionuses was filtered, washed with a mixture of chloroform-methanol and concentrated, obtaining the compound 7f(77 mg, purity 99%, yield 99%) as a solid yellow color. MS (ES+) 242 (m+1), (ES-) 240 (m-1).

1H NMR (400 MHz, DMSO-d): δ 8,72 (d, J=2.5 Hz, 1H), 8,31 (s, 1H), 8,00 (d, J=8,9 Hz, 1 H), 7,83 (br. s, 1 H), 7,43 (br. s, 1 H), 7,24 (br. s, 1 H), 7,10 (d, J=8.6 Hz, 1 H), 6,59 (br. s, 1H), 3,90 (s,3H).

, Compound 6E (217 mg, 0,724 mmole) and 4-methylpyrazole (594 mg, from 7.24 mmole) was kept at 110°C for 23 hours Then the mixture is composed of a mixture 1:1 dibenzylamino connection 7h and Antilibanus product 7g was diluted with EtOAc (2-3 ml) and filtered, obtaining pure dibenzylamine product 7h (111 mg, purity 95%, yield 54%) as a solid white color.

1H NMR (400 MHz, DMSO-d): δ 8,58 (d, J=2.6 Hz, 1H), 7,98 (d, J=9,2 Hz, 1H), 7.25 (br. s, 1H), 7,20 (s, 1H),? 7.04 baby mortality (br. d, J=9,2 Hz, 1H), 6,38 (s, 1H), with 3.89 (s, 3H), of 2.30 (s, 3H).

Example 8

Synthesis of 4-hydroxy-7-methoxy-2[4-(2-isopropylaminoethyl)]quinoline (8f)

Note: [using the same scheme of synthesis by replacing compound 8b other alkylthiophene received various 2-alkylaminocarbonyl deputies].

A. For the conversion of the meta-anisidine in connection 8A used a Protocol identical to those described in the literature: F.J.Brown etc., J. Med. Chem. 1989, 32, 807-826. However, the cleaning procedure was Modific is by, to avoid purification using chromatography. tO-phase containing the desired product was treated with a mixture containing MgSO4coal and 5 wt.% (in terms of theoretically calculated mass) silica gel. After filtration through celite, the product was ground with simple ether. Compound 8A was obtained in the form of a solid light brown color with a purity >99% (according to analysis with IHVR).

B. Suspension of isopropyltoluene (8b, 3.55 g, 30 mmol) and 3-bronirovochnoy acid (8s, 5 g, 1 EQ.) in dioxane (300 ml, 0.1 M) was heated to 80°C. After the temperature reached 80°C, the solution became transparent, and soon after this happened the deposition of the product in the form of a solid white color. After heating for 2 h, the solution was cooled to CT and the precipitate was filtered white color, getting 8d connection with a high purity (purity >98% according to NMR analysis) and 94% (7.51 g).

C. a Mixture containing carboxylic acid 8d (4,85 g, 18.2 mmole) and the aniline derivative 8A (3 g, 1 EQ.) in pyridine (150 ml, 0.12 M) was cooled to -30°With (after cooling, the clear solution turned almost in suspension). Then slowly over a period of time of 5 min was added phosphorus oxychloride (3,56 ml, 2.1 EQ.). The reaction mixture was stirred at -30°C for 1 h, the bath was removed and the reaction mixture Dawa and heated to CT. After 1.5 h the reaction mixture was poured on ice, the pH value was brought to 11 with 3h. aqueous NaOH solution, was extracted with CH2Cl2, dried over anhydrous MgSO4, filtered and concentrated in vacuum. Then the solid beige color was purified via Express chromatography (45%EtOAc in hexane)to give compound 8E in the form of a solid of light yellow color with a yield of 73% (6,07 g).

G. a Solution of tert-GFCF (2,42 g, 21.6 mmole) in anhydrous tert-VION (40 ml, 0,14M, purified by distillation from the metallic Mg) was heated to the temperature of reflux distilled. Within 5 min was added in portions compound 8E (1,8g, 5.4 mmole) and the resulting solution is dark red was stirred at the temperature of reflux distilled within 20 min (completion of the reaction was estimated using IHVR). The mixture was cooled to CT and was added Hcl (4 BC in dioxane, 1.5 EQ.). The mixture is then concentrated under vacuum in order to ensure that deleted all the amount of Hcl and dioxane, the product twice re-dissolved in CH2CL2and dried in vacuum, obtaining the resulting hydrochloride of compound 8f in a solid beige color (1,62 g, purity 93%, according to IHVR). Then the product was poured into phosphate buffer (1H. a solution of NaH2PO4, pH~4.5) and was irradiated with ultrasound. Solid beige product was filtered and dried in vacuum, n is the best compound 8f (1,38 g, yield 81%) as a solid beige color (purity 91% according to IHVR).

1H NMR (400 MHz, DMSO) δ of 8.27 (s, 1H), 8,12 (d, 1H, J=9,2 Hz), 7,97 (br.s, 1H), 7,94 (s, 1H), 7,43 (s, 1H), 7,24 (dd, 1H, J=9,2, 2.2 Hz), of 3.97 (m, 1H), 3,94 (s, 3H), 1,24 (d, 2H, J=6.4 Hz).

Example 9

Synthesis of 4-hydroxy-7-methoxy-2-[2-(4-isopropylthiazole)]quinoline (9f).

Note: using the same scheme of synthesis, in which the compound 9b was replaced by other α-bracketname received various 2-(4-alkyl)thiazolidine deputies.

A. To a solution of 3-methylbutane-2-it (8 g, 93 mmole) in Meon (100 ml) at -30°C, was added dropwise within 45 min VG2(4,79 ml, 93 mmole, 1 EQ.). Then the resulting mixture was stirred at RT for 90 minutes was Added pentane and the solution was washed with 5%aqueous solution Panso3, the organic layer was dried over anhydrous Na2SO4, filtered and concentrated in vacuum. The compound obtained 9b, representing a crude yellow oil, used without further purification. The solution containing atitikimas (9a, 1.8 g, 13.5 mmole) and derived bracelona 9b (13.5 mmole) was stirred at 70°C for 15 h then the mixture was concentrated in vacuo and then purified using the rapid chromatography on a column using as aloe is that 15%EtOAc in hexane, receiving compound 9c (740 mg, yield 28%).

B. a Solution of compound 9c (700 mg, 3.5 mmole) in a mixture of THF/Meon/N2About (ratio 3:1:1, 13 ml) was treated at RT for 5h LiOH2(148 mg, 3.5 mmole, 1 EQ.). Then the pH was brought to 6 with a 0.1 n Hcl solution and the mixture was concentrated to dryness in a vacuum, getting acid 13d, which is directly used in the next stage without additional purification.

C. a Solution containing 4-methoxy-2-aminoacetophenone (intermediate compound 8A, 570 mg, 3.45 mmole) and carboxylic acid derivative 9d (590 mg, 3.45 mmole, 1 EQ.) in pyridine (30 ml) was cooled to -20°C. Then dropwise over 5 min was added l3(0.35 ml, with 3.79 mmole, 1.1 EQ.). The resulting solution was stirred at -10°C for 2 hours the Reaction was stopped by adding N2Oh and the mixture was concentrated in vacuum. The residue was poured into a saturated aqueous solution Panso3and was extracted with EtOAc. The organic layer was dried over anhydrous MgSO4, filtered and concentrated in vacuum. The crude product was purified using the rapid chromatography on a column, using as eluent 25%tO in hexane, resulting in the received connection 9F in a solid white color (740 mg, yield 67%).

, Tert-GFCF (518 mg, 2.1 EQ.) was added to a suspension of compound 9F (700 mg, 2.2 mmole) in anhydrous tert-VION (ml). The resulting mixture was heated to 75°With in a period of 7.5 h, then the solution was cooled to CT and acidified by addition of Hcl (4 n solution of Hcl in dioxane, 2.5 ml). The mixture was concentrated in vacuum and the obtained residue was poured into 1 n solution of NaH2PO4and filtered. Then the solid product was ground with a small amount tO, was filtered and was dried in vacuum, obtaining the compound 9f in the form of a solid light beige color (270 mg, yield 41%).

1H NMR (400 MHz, DMSO-d6) δ 8,00 (br. s, Ih), 7,60 (br. s, 1H), 7,51 (br. s, 1H), 7,43 (br. s, 1 H), 7,29 (br. s, 1 H), 7,14 (br. s. 1 H), 6,95 (br. and, 1 H), 3,90 (s, 3H), 3.15 in (m, 1 H), of 1.33 (d, J 5.4 Hz, 6N).

Example 10

Synthesis of 4-hydroxy-2-(1-methyl-2-imidazolyl)-7-methoxyquinoline (10d)

A. a Solution of N-methylimidazole 10A (5 g, 61 mmol) in 100 ml THF was cooled to -78°C. dropwise within 15 min was added n-BuLi (24,4 ml of 2.5 M solution in Et2O, 1 EQ.). The resulting mixture was stirred for 90 min at -78°C, after which the portions was poured into an excess of solid CO2. The heterogeneous mixture was stirred for 2 h and then allowed it to warm up to CT. Added 1 n solution of Hcl to achieve pH 5, separated water layer and liofilizirovanny. Thus obtained residue was extracted with tO (to remove salts), dried (Na2SO4), filtered and concentrated under reduced on the no. The result was obtained 6.2 g (yield 80%) of solid product 10b white.

B. a Solution containing 4-methoxy-2-aminoacetophenone 8A (394 mg, 2,39 mmole) and a derivative of carboxylic acid 10b (301 mg, 1 EQ.) in pyridine (10 ml), cooled to -20°C. Then dropwise over 5 min was added l3(244 μl, 1.1 EQ.). The resulting solution was stirred at -10°and for 2.5 hours Then added water and the mixture was concentrated under reduced pressure. The residue was poured into a saturated solution Panso3and was extracted with EtOAc. The organic phase was dried (MgSO4), filtered and concentrated under reduced pressure. The product was purified by chromatography on silica gel (25%tO/Hex)to give 530 mg (yield 81%) as a solid 10C pale yellow color.

C. To a suspension of the substrate 10C (500 mg, 1.8 mmole) in 8 ml of tert-VION was added tert-GFCF (431 mg, 2.1 EQ.). Then the resulting mixture was heated to 75°C for 7 h, the Solution was allowed to cool overnight to room temperature and was added 2.5 ml of Hcl (4 n solution in dioxane). The mixture was concentrated under reduced pressure and the obtained residue was diluted with EtOAc. Added 1N. the NaOH solution to reach pH 7. Separated organic phase was dried (MgSO4), filtered and concentrated under reduced pressure, getting 145 mg of compound 10d (yield 31%) as a solid substance svetlorozovogo color. 1H NMR (400 MHz, DMSO-d): δ to 7.99 (d, J=8,9 Hz, 1H), 7,49 (s, 1H), 7,37 (s, 1H), 7,18 (s, 1H), 6,92 (d, J=8,9 Hz, 1H), of 6.31 (s, 1H), a 3.87 (s,3H), of 3.84 (s,3H).

Example 11

Synthesis of 4-hydroxy-2-(1-pyrrolyl)-7-methoxyquinoline (11b)

A. a Solution containing the substrate (obtained from compound 6 by hydrogenolysis of benzyl group in the presence of 5%Pd/C in a solution of ethanol-THF) (1 g, a 5.25 mmole) and 2,5-dimethoxytetrahydrofuran (0.68 ml, 1 EQ.) in glacial acetic acid was heated under reflux for 4.5 h and then was allowed to cool to CT. The mixture is then concentrated under reduced pressure. The residue was diluted with methanol and was added 1 N. NaOH (aqueous solution) to achieve pH 7. The product was purified by chromatography on silica gel (3%Meon/CH2CL2the residue was previously adverbially on silica gel). As a result received 140 mg (yield 13%) of compound 11b in the form of a solid white color.

1H NMR (400 MHz, DMSO-d): δ 7,98 (d, J=9,2 Hz, 1H), to 7.64 (s, 2H), 7,18 (d, J=2.5 Hz, 1H), 7,05 (br. d, J=7.9 Hz, 1H), 6,88 (br. s, 1H), 6,32 (s, 2H), 3,90 (s, 3H).

Example 12

Synthesis of 4-hydroxy-7-methoxy-2-(6-methyl-2-pyridyl)quinoline (12d)

A. 6-Methilpiralidone acid 12A (411 mg, 3.0 mmole) and SOCl2(0,520 ml, 7.2 mmole, 2.4 EQ.) boiled under reflux for 2 h in benzene (5 ml). The solvent and excess SOCl was removed from the reaction mixture under vacuum and the residue triturated with pentane. The formed solid product was filtered and the filtrate was concentrated, obtaining the acid chloride 12b (500 mg, 2.6 mmole).

B. To a solution of the crude acid chloride 12b in CH2CL2(5 ml) at 0°C, was added a solution of aniline 8a (344 mg, 2,08 mmole), DIPEA (1,45 ml, 8,35 mmole) and DMAP (61 mg, 0.5 mmole) in CH2CL2(10 ml). The reaction mixture was stirred at RT for 16 h the Volatile components were removed in vacuo, the residue was dissolved in EtOAc and the solution washed with 5%Panso3(2), N2Oh and salt solution. Then the organic layer was dried over MgSO4and concentrated in vacuum. The mixture was purified using the rapid chromatography on a column, using as eluent a mixture of EtOAc/ hexane (1:2)to give amide 12C (490 mg, yield 82%).

C. To a suspension of amide 12C (490 mg, 1,71 mmole) in tert-VION (10 ml), was added tert-GFCF (410 mg, of 3.43 mmole) and the mixture was stirred at 75°C for 6 h and then at RT for 16 h Then the mixture was poured into phosphate buffer (175 ml, pH 7) and stirred for 30 minutes, the Solid product was twice washed with ethyl acetate. The organic phase was washed with brine, dried over MgSO4and concentrated in vacuum. The obtained solid product was washed with EtOAc, getting a quinoline derivative 12d (263 mg, yield 58%).1H NMR: (CDCl3400 MG what): 5 of 2.68 (s, 3 H), of 3.94 (s, 3 H), 6,85-6,88 (2d, J=8,68 and 9.5 Hz, 2 H)6,94 (dd, J=8,9 and 2.2 Hz, 1 H), 7,27 (dd, J=6,7 and 1.9 Hz, 1 H), 7,73-7,79 (m, 2 H). of 8.28 (d, J=8,9 Hz, 1 H), and 10.3 (br s, 1 H).

Example 13

Synthesis of 4-hydroxy-7-methoxy-2-(5-methoxy-2-pyridyl)quinoline (13d)

A. To a solution of compound 13A (623 mg, of 3.73 mmole) in Meon, was added NaOH (2M, 4,70 ml) and the reaction mixture was stirred at RT for 2 h Then the solution was acidified with Hcl (6 N., 2.2 ml) and concentrated, obtaining compound 13b, which is used in the next stage without purification.

B. To a solution of the crude compound 13b (~of 3.73 mmole) in pyridine (25 ml), was added 8A (500 mg, 3,03 mmole) and the solution was cooled to -25°C, after which was added l3(0.35 ml, of 3.73 mmole). The reaction mixture was stirred at -10°C for 1 h and then at 0°C for 2 hours Then the mixture was poured into H2O and was extracted with EtOAc (2-3x). The combined organic layers were washed with 5%Panso3and with brine, dried over MgSO4and concentrated in vacuum. The crude product was purified using the rapid chromatography on a column, using as eluent a mixture of tO/hexane (1:2), resulting in a received 13C amide (617 mg, yield 55%).

C. To a suspension of amide 13C (617 mg, of 2.05 mmole) in anhydrous tert-VION (10 ml) was added tert-GFCF (490 mg, 4,11 mmole) and the mixture was stirred at 75°C for 6 h and then at RT during the 16 hours The reaction mixture was poured into phosphate buffer (175 ml, pH 7) and stirred for 30 minutes the Resulting solid product was filtered and triturated with EtOAc, receiving a derivative of quinoline 13d (250 mg, yield 43%).1H NMR: (DMSO, 400 MHz): δ 3,86 (s, 3 H), of 3.94 (s, 3 H), 6,72 (bs, 1 H)6,91 (dd, J=8,9 and 1.9 Hz, 1 H), 7,54 (d, J=1.9 Hz, 1 H), 7,60 (dd, J=8,9 and 2.9 Hz, 1 H), 7.97 (d, J=8,9 Hz, 1 H), 8,21 (d, J=8.6 Hz, 1 H), 8,48 (d, J=1.9 Hz, 1 H).

Example 14

Synthesis of 4-hydroxy-7-methoxy-2-(oxazol-5-yl)quinoline (14C):

A. Protected quinoline derivative 4b of example 4 (3.8 g, 11.8 mmole) was dissolved in CH2CL2(60 ml) and cooled to -78°after which very slowly for 15 min was added hydride diisobutylaluminum (DIBAL) (7.9 ml, 1 EQ., 1.5 M solution in toluene). After stirring for 80 min was added an additional portion of DIBAL (5.5 ml, 0.7 equiv., 1.5 M solution in toluene). After stirring at -78°even for 2 h, the reaction was carefully stopped with methanol (4 ml) at -78°and then was poured into an aqueous solution of Rochelle salt (1 N. K-Na tartrate). A thick paste was stirred with CH2CL2(300 ml) for 2 h before until the mixture became transparent. The phases were separated and the organic phase was dried (MgSO4), filtered and concentrated, gaining a solid white color. By cleaning with Express chromatography (SiO2, 230-40 mesh) using a mixture of 50%EtOAc/hexane received aldehyde 14a in the form of a solid white color (2.5 g, yield 73%).

B. To a suspension of2CO3(48 mg, 0,34 mmole) in Meon (7 ml) under stirring was added toluensulfonate (66 mg, 0.34 in mmole). The reaction mixture was heated to 45°and added the aldehyde 14a (0.10 g, 0.34 in mmole). The reaction mixture was heated to 80°C for 16 h and then concentrated to dryness in vacuo. After making purification using Express chromatography (SiO2, 230-400 mesh mesh) required oxazol 14b (0,089 g, yield 80%). MS: 331,0 (M+H)+.

Century MEM-protected hydroxyquinolin 14b was dissolved in THF (3 ml) and was treated with aqueous solution of Hcl (1 ad, 1 ml). The reaction mixture was stirred for 30 min at RT, then concentrated to dryness in vacuo. The residue was treated with phosphate buffer (3 ml, 1 n solution, pH 4.5) and stirred, after which the product was filtered, washed with distilled water and dried overnight in high vacuum (60°C, 16 h). Required hydroxyquinolin 14C in the form of a solid reddish-brown (0,065 g, yield 100%). MS: 242.9 (M+H)+.

1H NMR (DMSO-d6): δ 8,65 (s, 1H), 8,02 (bs, 1H), of 7.97 (d, J=8,9 Hz, 1H), 7.19 (s, 1H), 6,93 (d, J=7.9 Hz, 1H), 6.42 per (bs, 1H), a 3.87 (s, 3H). ES (+) MS: m/z 242,9 (M+N)+.

Fragments of the PEPTIDE LINKER (P3)

Example 15

Synthesis of (2S)-N-BOC-aminona-8-enoeou acid (15g)

A. To a solution available is the sale of diethyl-2-acetamidomalonate 15A (100 g, and 0.46 mol) in dioxane (500 ml) was added dropwise over 30-45 min aqueous solution of sodium hydroxide (1 M, 1 EQ., 460 ml). The resulting mixture was allowed to stand for 16.5 h, then the dioxane is evaporated in vacuo, the aqueous solution was extracted with three portions of 300 ml of ethyl acetate and acidified to pH 1 using concentrated Hcl. The solution was left to crystallize in the bath with a mixture of ice-water. After getting a small number of crystals, and the mixture was irradiated with ultrasound, which formed the abundant precipitate. After filtering and drying in vacuum were obtained compound 15b (totals 62.52 g, yield 72%) as a solid white color.

B. To stir with a magnetic stirrer emulsion containing a commercially available 7-octene-1,2-diol 15C (25 g, 0,173 mol) and H2O (100 ml)in round bottom flask with a volume of 1 l, were added within 20 min (weak exothermic reaction) aqueous solution of periodate sodium (40,7 g, 0,190 mol, 1.1 EQ., in 475 ml of N2About). The resulting mixture was stirred at room temperature for 1 h (completion of the reaction was confirmed by TLC). The mixture is then poured into a separating funnel and the aqueous layer was separated from the organic layer. An aqueous solution saturated with NaCl, decanted and again separated from the organic fraction. Two organic fractions were combined, dried over sulfate n the Tria and was filtered through a cotton plug (Pasteur pipette), receiving compound 15d (15,135 g, colorless oil, yield 78%). The aqueous solution was extracted with CH2CL2, dried over anhydrous MgSO4, and concentrated in vacuo (without heating at the boiling point of heptane 153° (C)receiving an additional amount of compound 15d (1,957 g, colorless oil, yield 10%). The total yield was 88%.

Century solid ethyl-2-acetamidomalonate 15b (EUR 7.57 g, 40 mmol) for 1 min was added 6-heptenal 15d (4,48 g, 40 mmol) in a solution of pyridine (32 ml, 10 EQ.). The resulting solution was cooled in a bath at 10°and for 4 min was added acetic anhydride (12 ml, 3.2 EQ.). The resulting orange solution was stirred for 3 h at RT and was added another portion of ethyl-2-acetamidomalonate 15b of 2.27 g). The resulting mixture was stirred at room temperature for 11 hours and Then added to ice (60 ml) and the solution was stirred for 1.5 h, after which the mixture was diluted with 250 ml of water and was extracted with two portions of simple ether. The ether solution washed with 1 N. Hcl solution, a saturated solution of Na-HCO3, dried over PA2SO4concentrated and purified using the rapid-chromatography (40%tO/hexane)to give compound 15th (4.8 g, yield 50%) as oil is light yellow in color.

, Degassed (by barbatii with argon for 30 min) to a solution of Z-ethyl-2-acetamido-2,8-is indienet 15th (scored 8.38 g, 35 mmol) in anhydrous ethanol (70 ml) was added (S,S)-Et-DUPHOS Rh(COD)OTf (51 mg, S/C=496). The mixture was placed in an atmosphere of hydrogen at a pressure of 30 lbs./square inch (after 4 cycles of vacuum-N2) and was stirred by using a Parr shaker for 2 h the Resulting mixture was evaporated to dryness, obtaining the crude compound 15f, which was used in the next stage without purification.

D. To a solution of crude (S)-ethyl 2-acetamido-8-nonenoate 15f (7,3 g of 30.3 mmole) in THF (100 ml) was added BOC2On (13,2 g, 2 EQ.) and DMAP (740 mg, 0.2 EQ.) and the reaction mixture was stirred at the temperature of reflux distilled for 2.5 hours and Then the greater part of the solvent, representing THF, evaporated, the crude mixture was diluted with CH2CL2and washed with 1 N. Hcl to remove DMAP. Then the organic layer was extracted with saturated aqueous Panso3, dried over anhydrous Na2SO4and concentrated in vacuum. After that, the crude product was diluted with THF (50 ml) and water (30 ml), was added LiOH·N2On (2,54 g, 2 EQ.) and the resulting mixture was stirred at RT for 25 h (completion of the hydrolysis was confirmed by TLC). The reaction mixture was concentrated in vacuo to remove most of the solvent, representing THF and diluted with CH2CL2. The resulting solution was washed for 1H. a solution of Hcl, dried the over anhydrous Na 2SO4and concentrated in vacuum. To remove small amounts of impurities and excess Boc2O the crude product was purified via Express chromatography (using as eluent a gradient of solvent a 100%hexane to 100%EtOAc). The result has been specified in the header connection 15g with high purity in the form of oil is light yellow in color (of 5.82 g, 71%yield).1H NMR (DMSO, 400 MHz): δ 7,01 (d, J=8 Hz, 1H), 5,79 (tdd, Jt=6,7 Hz, Jd=17,0, 10,2 Hz, 1H), 5,00 (md, Jd=17,0 Hz, 1H), 4,93 (md, Jd=10,2 Hz, 1H), 3,83 (m, 1H), 2,00 (q, J=6.9 Hz, 2H), 1,65-1,5 (m, 2H), 1,38 (s, 9H), 1,35-to 1.21 (m, 6H).

Primera

An alternative method of synthesis of (2S)-N-BOC-aminona-8-ene acid (15g):

A. To a suspension of finely chopped magnesium turnings (0.55 g, 22.5 mmole) in anhydrous THF (30 ml)containing dibromoethane (0.1 ml), under stirring was added dropwise within 15 min (the reaction is slightly exothermic) 8-bromo-1-octene (15h, 2,52 ml, 15 mmol). After 30 min, the mixture was heated to 38°C for 1 h and then was cooled to -78°C, after which it was added via cannula to the excess amount of solid CO2. The mixture was diluted with diethyl ether (100 ml) and the solution was washed with brine (2×50 ml), dried over MgSO4and was evaporated. Received the crude product in the form of oil, which was purified by chromatogr is the philosophy on silica gel, using as eluent 15%EtOAc in hexane, resulting in the received connection 15i exit 62% (1.44 g).

1H NMR (CDCl3, 400 MHz) δ of 1.31-1.42 (m, 6H), 1,60 was 1.69 (m, 2H), 2,02-of 2.09 (m, 2H), 2,35 (t, J=8,3 Hz, 2H), 4,99 (dm, J=10.0 Hz, 1H), 5,04 (dm, J=17,0 Hz, 1H), 5,75 and 5.86 (m, 1H).

B. To intensively stirred solution of carboxylic acid 15i (1,36 g, 8.7 mmole) in anhydrous THF (70 ml) at -78°C was added via syringe in anhydrous conditions protestirovanny immediately before the reaction E3N (1.6 ml and 11.3 mmole) and pivaloate (1,18 ml, 9.58 mmole). The mixture was stirred at -78°C for 15 min and then at 0°C for 45 minutes the Mixture was again cooled to -78°and then transferred via cannula to an anhydrous solution of the lithium salt of 4(S)-4-(phenylmethyl)-2-oxazolidinone in THF at -78°C; previously received the reagent constituting the lithium salt of oxazolidinone, by slowly adding n-BuLi (2.00 M solution in hexane, the 7.85 ml, 15.7 mmole) in a solution of oxazolidinone (2,78 g, 15.7 mmole) in THF (20 ml) at -78°C.

The reaction mixture was stirred at -78°C for 15 min and then at RT for 1.5 h To complete the reaction was stopped using an aqueous solution of sodium bisulfate (100 ml of 1 M solution) and THF evaporated before reaching ½ of its original volume. The residue was extracted with EtOAc (2×150 ml) and obyedinenie the e organic layers were washed with 5%solution Panso 3(3×50 ml), brine solution (2×50 ml), dried over MgSO4and was evaporated. The resulting crude product in the form of oil was chromatographically on silica gel, using as eluent 15%EtOAc in hexane, resulting in the received connection 15j exit 68% (1.88 g).

1H NMR (CDCl3, 400 MHz) δ of 1.35 to 1.47 (m, 6N), 1,67-of 1.74 (m, 2H), 2,02-of 2.09 (m, 2H), 2,65 (dd, J=13,4 and 9.9 Hz, 1H), 2,84-to 3.02 (m, 2H), and 3.31 (dd, J=13,4 and 3.2 Hz, 1H), 4.13-4,22 (m, 2H), 4,62-4,71 (m, 1H), is 4.93 (d, J=10,2 Hz, 1H), 5,00 (dd, J=17,2 and 1.6 Hz, 1H), 5,75 of 5.84 (m, 1H), 7.18 in-7,38 (m, 5H).

C. To a solution of KHMDS (0.8 M solution in THF, 22 ml of 17.5 mmole) in anhydrous THF (50 ml) at -78°under stirring was added via cannula a solution derived acid 15j (3.25 g, 10.30 mmole) in anhydrous THF (40 ml) at -78°C. the Mixture was stirred at -78°C for 45 minutes To this mixture was added a solution trailside (3,67 g, 11,85 mmole) in anhydrous THF (40 ml) at -78°C. the Mixture was stirred at -78°C for 3 min, then the reaction was stopped with acetic acid (5 ml). After that, the mixture was stirred at RT for 1 h 45 min and finally at 40°C for 15 minutes Large part of the THF evaporated. The residue was dissolved in EtOAc (100 ml) and the organic solution washed with N2O (50 ml), 5%solution of NaHCO3(3×50 ml) and with brine (50 ml), (MgSO4) and evaporated. The resulting product in the form of oil was chromatographically on silica gel, using ka is este eluent hexane/CH 2CL2(1/1), resulting in the received connection 15k (2,47 g, yield 67%).

1H NMR (CDCl3, 400 MHz) δ 1,32-of 1.45 (m, 6H), of 1.45 to 1.6 (m, 1H), 1,75-1,88 (2, 2H, rotamer), 2,01-2,11 (m, 2H), 2,82-2.87 (m, 1H), 3,33 (dd, J=13,4 and 3.2 Hz, 1H), 4,10-4,28 (m. 2H), to 4.62-4.72 in (m, 1 H), 4,90-of 5.05 (m, 3H), 5,73-5,88 (m, 1 H), 7,17-7,38 (m, 5H).

G. To a solution of anhydrous SnCl2(2,61 g, 13.8 mmole) in anhydrous Meon (80 ml) under stirring was added at 0°via cannula a solution of azide 15k (2,45 g, 6.9 mmole) in anhydrous Meon (20 ml). The mixture was stirred at RT for 4 h Evaporated Meon and the obtained foamy product was dissolved in a mixture of dioxane/H2On (100 μl/20 μl) and treated Vos2Oh (3.0 g, of 13.8 mmole) and Panso3(2,89 g, 34.5 mmole) (if necessary, the pH was brought to 8 with additional quantities Panso3) and the mixture was stirred at RT for 16 h part of the dioxane evaporated (~50%) and the residue was twice extracted with EtOAc. The organic solution was washed with brine (2×50 ml), dried and evaporated. The obtained residue was chromatographically on silica gel, using as eluent 20-25%EtOAc in hexane, receiving the connection 151 (1.75 g, yield 60%).

1H NMR (CDCl3, 400 MHz) δ 1,27-of 1.53 (m, 6H), of 1.46 (s, 9H), of 1.80 (m, 1H), 2.00 in of 2.08 (m, 1H), 2,80 (t, J=12.1 Hz, 1H), 3,34 (d, and 14.3 Hz, 1H), 4,17-to 4.23 (m, 2H), 4,60-of 4.66 (m, 1 H), is 4.93 (d, J=10,2 Hz, 1 H), of 5.05 (dd, J=17,2 and 1.9 Hz, 1 H), 5,13 (bs, I H), 5,38-5,43 (m. 1H), 5,74 of 5.84 (m, 1H), 7,22 and 7.36 (m, 5H).

D. To a solution of N-Boc-what Assenova derived 151 (1,74 g, Android 4.04 mmole) in a mixture of THF/N2O (75 ml/15 ml) was added under stirring at 90°With H2About2(30% vol./wt., of 2.05 ml, 16.2 mmole) and LiOH·H2O (0.34 g, 8.1 mmole) and the solution was stirred at 0°C for 1 h the Reaction was stopped by adding Na2SO3(2.24 g in N2About, 15 ml, 17.8 mmole). The pH value was brought to 4-5 with 10%aqueous citric acid solution and the mixture was diluted with EtOAc. The aqueous fraction was extracted once more EtOAc and the organic solution washed twice with brine, dried and evaporated. The residue was chromatographically on silica gel, using as eluent 20%hexane in EtOAc, which received the free carboxylic acid 15g (0,76 g, yield 70%). This connection in all respects was identical to the compound of example 15.

Example 16

Synthesis of methyl ester (2S)-N-BOC-amino-5-Oksanen-8-enoeou acid (16d):

This synthesis process is based on a method described in T. Tsuda and others, J. Am. Chem. So.,1980, 102,6381-6384.

A. To a solution of monoallyl ester of malonic acid (1.50 g, of 10.4 mmole) in anhydrous THF (20 ml) in an atmosphere of N; at -78°with vigorous stirring for 5 min was added dropwise n-Bu2Mg (0.9 M solution in hexane, 5.8 ml, 5.2 mmole). Then the thick suspension was stirred at RT for 1 h and evaporated to dryness (vacuum suction in the atmosphere 2). Solid magnesium salt 16b was dried in vacuum for 1 h

A derivative of glutamic acid 16A is first mixed with 1,1’-carbonyldiimidazole (1,65 g of 10.21 mmole) in anhydrous THF and the mixture was stirred at RT for 1 h in order to activate the free acid fragment. Then the activated derivative of glutamic acid was introduced through the cannula into a solution of a magnesium salt 16b and the resulting reaction mixture was stirred at RT for 16 h Then diluted tO and the organic solution was washed with 0.5 N. chilled on ice Hcl, washed with brine, dried and evaporated. The obtained residue was chromatographically on silica gel, using as eluent 35-40%tO in hexane, resulting in the received connection 16C (1.85 g, yield 53%).

1H NMR (CDCl3, 400 MHz) δ the 1.44 (s, 9H), 1.85 to 1,95 (m, 1 H), 2,12-2,22 (m, 1H), 2,58-to 2.74 (m, 2H), 3,48 (s, 2H), 3,74 (s, 3H), 4,24-4,34 (m, 1H), to 4.52 (dm, J=5.7 Hz, 2H), 5,09 (m, 1H), 5.25-inch (dm, J=10,2 Hz, 1H), 5,34 (dm, J=and 17.2 Hz, 1H), 5,91 (m, 1H).

B. To a solution of tetrakis(triphenylphosphine)Pd (0) (0,116 g, 5 mol., 0.1 mmole) in anhydrous DMF (7 ml) under stirring was added with a syringe in an atmosphere of N2complex fluids 16C (0,687 g, 2 mmole) in anhydrous DMF (3 ml). The mixture was stirred at RT for 3.5 h DMF is evaporated under reduced pressure and the residue was diluted with EtOAc (20 ml). tO the solution was washed with 0.5 N. chilled on ice Hcl (5 ml)salt Rast is a PR (10 ml), was dried and evaporated. The residue was chromatographically on silica gel, using as eluent 15-20%EtOAc in hexane, resulting in the received connection 16d (0,253 g, yield 42%).

1H NMR (CDCl3, 400 MHz) δ the 1.44 (s, N), 1,84-of 1.94 (m, 1H), 2,08-2,22 (m, 1H), 2,33 (dd, J=14,0 and 7.3 Hz, 2H), 2,45 is 2.55 (m, 4H), 3,74 (s, 3H), 4,28 (bm, 1H), 4,98 (dm, J=10,2 Hz, 1 H), 5,03 (dm, J=and 17.2 Hz, 1 H), 5,00-5,10 (m, 1H), 5,74-to 5.85 (m, 1H).

Example 17

Synthesis of (2S,5R)-N-Boc-2-amino-5-metalion-8-enoeou acid (17f)

A, B, C, G commercially Available (R)-(+)- citronellal 17A first turned in the derived amino acid 17b, using the same stage of the synthesis, which were previously described in example 15 for the conversion of aldehyde 15d in the intermediate product, which is the amino acid 15f.

D. Compound 17b (0,675 g, 5.6 mmole) was dissolved in a mixture of tert-VION/acetone/N2About (1:1:1, 18 ml) and placed in an ice bath (0°). Consistently added NMMO (0,789 g, 6,74 mmole, 1.2 EQ.) and OsO4(2.5 wt.% in tert-VION, 0.7 ml, 0,067 mmole, 0,012 EQ.) and the reaction mixture was stirred at RT for 4 h the Greater part of the acetone was removed by evaporation under vacuum and then the mixture was extracted with EtOAc. After that, the organic layer washed with N2O and brine solution, dried over anhydrous MgSO4and was evaporated to dryness. With the rapid chromatography on a column using as eluent a 1%EOH in EtOAc received diol 17c with a high purity with a yield of 77% (0,575 g).

E. To a solution of 17c (0,575 g of 1.73 mmole) in THF/N2About (1:1, 20 ml) at 0°With added NaIO4(0,48 g, 2.25 mmole, 1.3 EQ.) and the reaction mixture was stirred at RT for 3.5 h Then the greater part of the solvent, representing the THF was removed by evaporation in vacuo and the remaining mixture was extracted with EtOAc (2×100 ml). Then the combined organic layers washed with 5%aqueous citric acid solution (2×20 ml), 5%aqueous solution Panso3(20 ml) and with brine (2×50 ml), then EtOAc solution was dried over anhydrous MgSO4and was evaporated to dryness in vacuum. The intermediate product, which represents an aldehyde 17d (0,47 g of the crude product) was used in the next stage without additional purification.

G. To a solution of RH3RSN3VG (925 mg, 2.6 mmole) in anhydrous toluene (15 ml) was added KHMDS (0.5 M solution in toluene, 5.2 ml, 2.6 mmole) and the resulting yellow suspension was stirred at RT for 30 min in N2 atmosphere. After this time the first suspension was cooled to 0°C, was added via syringe a solution of aldehyde 17d (0,47 g of 1.73 mmole dissolved in 15 ml of anhydrous THF) and the mixture was allowed to warm up to CT. After stirring at RT for 1 h large part of the THF was removed by evaporation in a vacuum to the mixture was added EtOAc (100 ml) and the organic layer washed with N2O (30 ml), 5%in denim solution of NaHCO 3(30 ml) and with brine (30 ml). Then tO solution was dried over anhydrous MgSO4and was evaporated to dryness in vacuum. After purification using rapid chromatography on a column of silica gel using as eluent a mixture of hexane:tO (3:2) was isolated a pure compound e exit 63% (0.29 grams) for the last two stages.

For connection 17f was carried out by hydrolysis of a complex of ethyl ether and the simultaneous replacement of N-acetyl protective group on a group of Vos in the intermediate product a using the same procedure described for the conversion of compound 15f in connection 15g (17f, 310 mg, quantitative yield).1H NMR (Dl3, 400 MHz): δ to 0.88 (d, J=6.4 Hz, 3H), 1,18 of 1.28 (m, 2H), of 1.35 to 1.48 (m, 3H), 1,45 (s, N), 1,64-1,74 m. 1H). 1,81-1,89 (m, 1H), 1,94-2,12 (m, 2H), 4,28 (bd, J=~3,2 Hz, 1H),4,93 (dm, J=11,1 Hz, 1H), 5,00 (dm, J=16,8 Hz, 1 H), 5,74 of 5.84 (m, 1H).

Example 18

Synthesis of N-BOC-O-allyl-(L)-threonine (18d)

A. BOC-(L)-threonine 18a (500 mg, 2,28 mmole) was partially dissolved in a mixture of CH2Cl2/MeOH (8 ml/0.5 ml, respectively) at 0°C. was Slowly added a solution of diazomethane in diethyl ether until then, until he had pronounced yellow color, indicating the presence of excess diazomethane. After evaporation of the solvents was obtained methyl ester 18b in the form of a turbid oil white (0.534 g).

B. Then the intermediate product is t 18b (311 mg, of 1.33 mmole) was dissolved in anhydrous diethyl ether (8 ml), was added Ag2O (341 mg, about 1.47 mmole) and activated directly before this, 4A molecular sieves (1 g). At the end of the reaction flask was added allergodil (134 μl, about 1.47 mmole) and the mixture was stirred at the temperature of reflux distilled. After 20 h and 30 min was added to two portions of Allilueva each time for 45 μl, of 0.50 mmole) and continued stirring in total for 36 hours and Then the mixture was filtered through celite and purified using the rapid chromatography on a column of silica gel, using as eluent a mixture of tO/hexane (1:4), resulting in a received 73 mg (yield 27%) of compound 18C in the form of a clear oil.

1H NMR (CDCl3, 400 MHz): δ to 1.21 (d, J=6.0 Hz, 3H), 1,45 (s, 9H), of 3.75 (s, 3H), 3,82-a 3.87 (m, 1H), 3,99-4,07 (m, 2H), 4,29 (dd, J=9.5 and 2.5 Hz, 1H), 5,14 (dm, J=10.5 Hz, 1H), total of 5.21 (dm, J=and 17.2 Hz, 1H), 5,75 of 5.84 (m, 1H).

C. the Union, which represents an ester 18C (99 mg, 0,362 mmole) was dissolved in a mixture of THF/Meon/N2About (2:1:1,4 ml) was added LiOH·H2O (61 mg, of 1.45 mmole). The solution was stirred at RT for 2 h and then acidified with 1 n Hcl solution to reach pH ~3, after which the solvent was removed in vacuum. The resulting compound 18d as oil used without further processing for the synthesis of macrocyclic inhibitors.

Example 19

Synthesis of (2S,38)-N-BOC-2-amino-(mercaptoethyl)butane acid (19th)

A. Compound 19a (9.1 mmole) was dissolved in pyridine (5 ml) and the solution was cooled to 0°C in an ice bath, was added in small portions taillored (2.3 g, 11.8 mmole, 1.3 EQ.) and relational the mixture was stirred at RT for 24 hours After this time the reaction mixture was distributed between diethyl ether (300 ml) and N2O (100 ml). Then the ether layer was washed with 0.2 N. a solution of Hcl (6×100 ml) and with brine (100 ml), dried over anhydrous MgSO4, filtered and concentrated to dryness in vacuo. After purification of the crude product using rapid chromatography on a column using as eluent a mixture of hexane/tO (gradient from 8:2 to 7:3) was isolated derived Totila 19b with the release of 85% (3,05 g).

B. To a solution of intermediate 19b (775 mg, 2 mmole) in anhydrous DMF (2.5 ml) was added thioacetic potassium (365 mg, 3.2 mmole, 1.6 EQ.) and the reaction mixture was stirred at RT for 24 h Then a large part of the DMF is evaporated in vacuo and the remaining mixture was distributed between EtOAc and H2O. the Aqueous layer was re-extracted with EtOAc, the combined organic layers were washed with brine, dried over anhydrous MgSO4and was evaporated to dryness. By purification of the crude product using rapid chromatography on a column using as eluent a mixture of exon/tO (in the ratio 4:1) was isolated compound 19s with yields of 80% (465 mg).

C. To a solution of complex tiefer 19s (465 mg) in a mixture of N2O/tO (ratio 3:5, 8 ml) was added a 0.2 M aqueous solution of NaOH (2.4 ml) and the mixture was stirred at RT for 1.5 h and Then added allride (0,292 ml, 3.2 mmole, 2 EQ.) and continued stirring at RT for 30 min the Reaction mixture was concentrated to half of its original volume and then was extracted with EtOAc. The aqueous layer was acidified to pH~3 with water of 0.5 n Hcl solution and re-extracted with EtOAc. The combined organic layers were washed with brine, dried over anhydrous MgSO4and was evaporated to dryness in vacuum. The reaction mixture contained at least four product; all of these products were isolated using rapid chromatography on a column of silica gel, using as eluent a mixture of hexane/tO (gradient from 9:1 to 3:1). The structure of the least polar compounds (TLC Rf=0,68 in a mixture of hexane/tO, 4:1) was consistent with the desired product 19d (83 mg, yield 18%).

1H NMR (CDCl3, 400 MHz): δ 1,24 (d, J=7,0 Hz, 3H), of 1.46 (s, 9H), 3,13-3,19 (m, 2H), 3,24-3,29 (m, 1H), of 3.77 (s, 3H), 4,50 (dd, J=8.6 and 3.8 Hz, 1H), 5,12 (d, J=12,4 Hz, 1H), 5,15 (dd, J=18,4 and 1.3 Hz, 1H), 5,22 (bd, J=7,6 Hz, 1H), 5,75-to 5.85 (m, 1H).

, The complex Solution of methyl ether 19d (83 mg, 0,287 mmole) in a mixture of Meon/N2O (3:1, 4 ml) was mixed with aqueous solution of NaOH (0,2 N., 1.3 ml, of 0.26 mmole) for 24 h at RT and for 1 h at 40° C. the Reaction mixture was acidified with cold aqueous solution of Hcl (0,5 N. Hcl at 0°C, pH 4-5), Meon was removed in vacuum and the remaining aqueous mixture was extracted with EtOAc. The organic solution was dried over MgSO4and was evaporated to dryness, obtaining the 19th connection. The 19th connection used for the final synthesis inhibitors without any additional purification.

Example 20

Synthesis of (S)-N-BOC-2-amino-3-methyl-3-(1-mercapto-4-butenyl)butane acid (20C)

A. L-penicillamine 20A (448 mg, 3 mmole) was dissolved in a mixture of DMF/DMSO (ratio 5:1,6 ml), was added 4-bromantan ones (0.46 ml, 4.5 mmole, 1.5 EQ.) and CsOH•H2O (1.0 g, 6 ml, 2 EQ.) and the reaction mixture was stirred at RT. After 24 h, to the reaction mixture were added Vos2On (820 mg, of 3.75 mmole, 1.25 EQ.) and continued stirring for 12 hours then DMF was removed in vacuo, the remaining mixture was diluted with cold 0,5 N. aqueous solution of Hcl, leading to pH~4-5 and then was extracted with tO (2×50 ml). The organic layer was washed with brine (2x), dried over anhydrous MgSO4and was evaporated to dryness, obtaining the crude carboxylic acid 20b.

B. Purification of compound 20b was difficult, so the crude product is first treated with diazomethane to obtain the corresponding complex methyl ester 20C, and then purified using expressrotary on column using as eluent a mixture of hexane/tO (9:1), resulting in a received 190 mg (yield 20%) of the pure complex methyl ester 20C.

1H NMR (400 MHz, CDCl3): δ to 1.35 (s, 3H), of 1.37 (s, 3H), of 1.44 (s, 9H), 1,59-to 1.67 (m, 2H), 2,11-2,17 (m, 2H), of 2.51-2,60 (m, 2H), 3,74 (s, 3H), 4,29 (d, J=8.6 Hz, 1H), 4,98 (dm, J=10.5 Hz, 1H), 5,03 (dm, J=19 Hz, 1H), 5,35 (bd, J=7 Hz, 1H), 5,72-of 5.83 (m, 1H).

Century After this ester was dissolved in a mixture of THF/Meon/N2O (2:2:1, 5 ml), was added LiOH•H2O (50 mg, 2.0 mmole, 2 EQ.) and the reaction mixture was stirred at 40°C for 4 h to hydrolyze ester 20C to acid 20b. The reaction mixture was acidified to 0.5 N. Hcl to pH 4-5, THF and the Meon evaporated completely and the remaining aqueous solution was extracted with tO. tO layer was dried over anhydrous MgSO4and was evaporated to dryness, obtaining the connection 20b, which was used in the subsequent synthesis of macrocyclic inhibitors without further purification.

Intermediate acyclic dipeptides and tripeptides

The General method of carrying out reactions in combination mortar and concrete examples of their implementation are described in WO 00/09543 and WO 00/09558.

These methods were used to synthesize intermediate dipeptide 26C, 30a and tripeptides 23a, 24A, 31A, 32A and 33a.

Example 21

Synthesis of acyclic Tripeptide e

A. To a solution containing a derivative of Proline 21 a (received and is commercially available BOC-4(R)-hydroxyproline and 4-chlorhydrin according to the method described in WO 00/05543 and WO 00/09558) (1,32 g, 3,68 mmole) and the crude Gamaliel ACPP 1f (-3,35 mmole) in CH2CL2(10 ml)was sequentially added NMM (1,21 ml, 10.05 mmole) and GATA (1,53, was 4.02 mmole) and the suspension was stirred at RT for 18 hours after this time the solvent is evaporated and the crude reaction mixture was re-dissolved in EtOAc (30 ml). The solution was washed with 5%aqueous solution of NaHCO3(2×10 ml), brine solution (10 ml), dried over MgSO4and was evaporated. The crude product was purified by chromatography on silica gel, using as eluent 8%diethyl ether in EtOAc, resulting in the required diastereoisomer of compound 21b with a yield of 20% (absolute stereomicroscope structure was not determined).

1H NMR (CDCl3, 400 MHz): δ of 0.93 and 1.01 (t, J=8,3 Hz, 1H, rotamer in the ratio 3:7), to 1.14 and 1.35 (m, 2H), of 1.44 (s, 9H), 1,45 (s, 9H), 1,50-to 1.82 (m, 4H), 2,08-of 2.24 (m, 2H), 2,32 (bs, 0,7H), 2,63 (bs, 0.75H), with 2.93 (bs, 0.75H), and 3.16 (m, 0.25H), of 3.77 (bs, 1,5H), 3,88 (bs, 0,5H), 4,4-4,55 (m, 1H), to 4.98 (d, J=10,2 Hz, 1H), to 5.03 (dd, J=17,2 and 1.6 Hz, 1H), 5,24 (bs, 1H), 5,75-5,88 (m, 1H), 6,57 and 6.78 (2bs, 1H, 2 rotamer). 7,42-7,58 (m, 3H), 7,63-7,73 (m, 2H), 8,04 (d, J=8,3 Hz, 1H), 8,11 (d, J=8,3 Hz, 1H), total of 8.74 (d, J=5,1 Hz, 1H).

B. To a solution of dipeptide 21b (137 mg, 0,248 mmole) in anhydrous CH2CL2solution was added Hcl in dioxane (4M, 4 ml) and the mixture was stirred at RT for 1.5 h and Then the solvent is evaporated and the residue was dried in high vacuum, receiving St. the free amino acid. The mixture was dissolved in a mixture of diethyl ether /Meon (3 μl/2 ml) and was treated with a slight excess of diazomethane dissolved in diethyl ether. After 30 min the excess diazomethane was removed by addition of Hcl (4M solution in dioxane) and the mixture was evaporated to dryness, obtaining hydrochloride connection C, which was used in the next stage without any treatment.

C. To a suspension of the crude dipeptide s (0,23 g, 0.48 mmole) in CH2CL2(25 ml) was sequentially added with stirring (2S)-N-BOC-aminoet-6-envoy acid 21d (0,151 g of 0.62 mmole), NMM (210 μl, at 1.91 mmole) and GATA (0,236 g of 0.62 mmole) and the mixture was stirred at RT for 16h (if necessary after 1 h the pH value was brought to ~8 using NNM). CH2CL2evaporated, the residue was dissolved in EtOAc (50 ml) and the organic solution washed with 5% Panso3(2×20 ml), brine solution (2×20 ml), dried and evaporated. The crude compound was chromatographically on silica gel (50 ml, 2% EtOH/EtOAc)to give compound 21 (0,139 g, yield 46%).

1H NMR (CDCl3400 MHz processor, the ratio of rotamers 6:1) chemical shifts of the main rotamer δ 1,21-of 1.27 (m, 1H), 1,36 (s, N), 1,45-1,81 (4m, 7H), 2,20-2,22 (m, 4H), 2,28-is 2.37 (m, 1H), 2,90-2,99 (m, 1H), 3,66 (s, 3H), 3,94-3,98 (m, 1H), 4,29 (bd, J=9.9 Hz, 1H), 4,46-4,50 (m, 1H), to 4.81 (dd, J=8,3 and 5.4 Hz, 1H), 4.92 in-of 5.06 (m, 4H), 5,16 (d, J=8,3 Hz, 1 H), lower than the 5.37 (m, 1 H), 5,70 of 5.84 (m, 2H), PC 6.82 (d, J=5,1 Hz, 1 H), 7,47-of 7.55 (m, 2H), 7.71 (dt, J=7.0 and 1.3 Hz, 1H), 8,03 (d, J=8,6 the C, 1H), 8,17 (d, J=8.0 Hz, 1H), 8,78 (d. J=5,1 Hz, 1H).

Macrocyclic peptides

Example 22

The General procedure of macrocyclization using olefin metathesis

In all cases tripeptidyl diene was dissolved in CH2CL2at a concentration of 0.01 M and the solution was deoxyribose by ozonation argon (~1 h to 500 ml). Solution was added catalyst (5-30 mol. %dissolved in a small amount of degassed CH2CL2) and the reaction mixture was stirred at the temperature of reflux distilled until then, until the entire original product will not turn in the final product(s) according to the analysis by TLC and GHUR. Then the crude reaction mixture was concentrated almost to dryness and filtered through a short pad of silica gel, elwira first using CH2CL2to remove the greater part of the catalyst, and then with EtOAc for elution of all macrocyclic products(a) (mostly in the form of an individual stereoisomer). The crude products of each reaction were analyzed using chiral GHUR on CHIRALCEL OJ-R column (obtained from the firm Chiral Technologies Inc, ⊘ 0,46×15 cm)using an isocratic solvent mixture of 70% N2About+0.06% of the TPA - 30% SN3SP+0,06% TFA at 205 nm. Basic macrocyclic products were fully characterized using data obtained with the use of1H, COSY, OCSY and ROESY NMR analyses, to confirm their structure and stereochemical structure.

Example 23

Synthesis of macrocyclic intermediate product (23b)

A solution of diene 23a (4.0 g, 7,88 mmole) in anhydrous CH2CL2(800 ml, Aldrich company. anhydrous) was deoxyribose by ozonation AG for 2 hours Then was added Hoveyda catalyst (262 mg, 0,434 mmole, of 5.5 mol.%) in the form of solids and the reaction mixture is boiled under reflux in an atmosphere supplied from a cylinder AG. 28 hours and evaporated the solution is red-orange color to obtain amorphous solids and then purified using the rapid chromatography on a column of silica gel. As the initial solvent system used 10%EtOAc in CH2CL2. As soon as I finished the elution of the catalyst from the column, the solvent was replaced with pure EtOAc. The process of elution of the catalyst from the column was visible due to its color. Macrocyclic product 23b was isolated as a colourless foam and re-dissolved in a mixture of CH2CL2/hexane (~1:2). After evaporation of solvent received the product as white powder (3,362 g, yield 89%).

1H NMR (CDCl3, 400 MHz): δ 1,20-1,50 (m, 6H), USD 1.43 (s, 9H), 1,53 (dd, J=9.5 and a 5.4, 1H), 1,61 is 1.70 (t, 1H), 1,76-1,90 (m,2H), 2.05 is-of 2.26 (m, 4H), of 2.45 (d, J=14,3, 1H), to 3.67 (s, 3H), 3,71 (d, J=11,1, 1H), 3,90 (dd, J=11,1 and 4.3, 1 is), 4,43-a 4.53 (m, 2H), amounts to 4.76 (d, J=8,6, 1H), 4,86 (bd, J=9,8, 1H), 5,20-5,23 (m, 2H), to 5.57 (dt, J=7,0 and 9.8, 1H), 7,32 (bs, 1H).

Example 24

Synthesis of macrocyclic intermediate product (24b):

A solution of diene 24A (2.76 g, 3,82 mmole) in anhydrous CH2Cl2(600 ml, anhydrous) was deoxyribose by ozonation AG for 1.5 hours using a cannula solution was added Hoveyda catalyst (117 mg, 0,19 mmole, of 0.05 EQ.) in anhydrous and degassed CH2Cl2(8 ml) and the reaction mixture was stirred at the temperature of reflux distilled in an atmosphere of AG supplied from a cylinder. After 20 h the reaction mixture was reacted for approximately 50%, was added to the second portion of catalyst (117 mg) and continued stirring for 16 hours Then the solution was concentrated to a volume of ~100 ml, was applied on the top surface of the bed of silica gel (6×10 cm) and were primarily allocated catalyst by elution with CH2CL2. Compound 24b was washed out of the bed of silica gel using 3%Meon in EtOAc and re-purified using the rapid chromatography on a column, using as eluent a mixture of EtOAc/hexane (2:1), resulting in a received solid white color with a faint olive tinge (yield 70%) (1.85 g, purity 94% according to analysis with IHVR).

1H NMR (400 MHz, DMSO-d6) δ 8,69 (s, 1H, 8,13 (d, J=9,2 Hz, 1H). 7,50-7,44 (m. 2H), 7,17 (dd, J=9,2, 2.2 Hz, 1H),? 7.04 baby mortality (d, J=6,4 Hz, 1H), ceiling of 5.60-to 5.56 (m, 1H), 5,52 (dd, J=9,2 Hz, 1H), 5.25-inch (dd, J=9,2 Hz, 1H), 4,59 (d. J=11 Hz, 1H), of 4.44 (dd, J=9,2 Hz. 1H). 4,05-3,98 (m, 1H), 3,94 (s, 3H), 3,92 (s, 3H), 3,89-3,82 (m, 1H), 3,55 (s, 3H), 2,64 of $ 2.53 (m, 1 H), 2,46 (d, J=7,3 Hz, 1H), 2.40 a-2,31 (m, 1H), 2,21 (dd, J=8,9 Hz, 1H), 1,78-of 1.65 (m, 2H), 1.55V (dd, J=4,8 Hz, 1H), of 1.485 (dd, J=4,8 Hz, 1H), 1.41 to of 1.30 (m, 7H), of 1.16 (s, 9H). MS; es+: 795,4 (M+N)+.

Example 25

The synthesis of compounds 202 and 203 (table 2)

A. Diene derived 21e (0,130 g, 0,205 mmole) was subjected to cyclization using catalytic amounts dichloride bis(tricyclohexylphosphine)benzyladenine IV (catalyst Grubba above) (52 mg, 0,064 mmole) in CH2CL2(60 ml) at the temperature of reflux distilled for 2 h and after chromatography on silica gel (50 ml, 3%EtOH/EtOAc) received compound 25A (60.1 mg, yield 48%).

1H NMR (CDCl3, 400 MHz) δ 1,22-of 1.30 (m, 2H), of 1.35 (s, 9H), 1,44 to 2.35 (m, 13H), of 3.07-3.14, and 3,16-3,24 (2m, 1H, ratio of rotamers 1:3), of 3.69 (s, 3H), 3.96 points-Android 4.04 (m, 1H0, 4,42-4,50 (m, 1H), 4,95-5,04 (m, 1H), of 5.05-of 5.15 (m, 1H), 5,20-and 5.30 (m, 1H), 5,55-the 5.65 (m, 1H), 6.75 in-6,79 (2d, J=5.4 Hz, 1H, ratio of rotamers 1:3), was 7.36 (s, 1H). 7,46 is 7.50 (m, 1H), 8,03 (d, J=8,3 Hz, 1H), 8,13 and 8,17 (2d, J=8.0 Hz, 1H, ratio of rotamers 1:3), 8,77 (d, J=5,1 Hz, 1H).

Century Ester fragment of macrocyclic compounds 25A (0,0156 g, 0.026 mmole) hydrolyzed using LiOH•H2O (8,7 mg, 0,206 mmole) in a mixture of THF/Meon/N2O (4 ml /2 ml /2 ml ). The crude product was purified using GHUR with reversed-phase 18-column type Whatman (PaKTisil 10,ODS3) 50/2 .4 cm, using a gradient solvent of 5%aqueous solution of CH3CM to 100%CH3The joint VENTURE, resulting in a net connection 202 in the form of an amorphous solid white (11.8 mg).

1H NMR (DMSO, 400 MHz): δ of 1.12 (s, 9H), 1,20-1,24 (m, 2H), 1.32 to-1,40 (m, 3H), 1,58-of 1.62 (m, 2H), 1,68-of 1.78 (m, 3H), 1,95-2,02 (m, 1H), 2,08-to 2.18 (m, 2H), 2,42 at 2.59 (m, 2H), 3,97-4,00 (bd, J=9.8 Hz, 2H), 4,47 (t, J=8.6 Hz, 1H), 4,58 (d, J=11.8 Hz, 1H), 5,22-of 5.29 (m, 1H), 5,46 is 5.54 (m, 1H), to 5.66 (s, 1H), 7,12 (d, J=6.0 Hz, 1H), 7,49 (d, J=3.5 Hz, 1H), 7,68 (dd, J=7,3 Hz, 1H), 7,98 (dd, J=7,0 Hz, 1H), 8,08 (d, J=8,3 Hz, 1H), 8,21 (s, 1H), 8,35 (d, J=8,3 Hz, 1H), remaining 9.08 (d, J=5 Hz, 1H).

C. Macrocyclic compound 25A (20 mg, 0,033 mmole) in anhydrous CH2CL2(1 ml) was stirred in the presence of 4 M Hcl in dioxane (5 ml) for 1 h the Mixture was evaporated and carefully dried. The residue was re-dissolved in CH2CL2/DMF (3 ml /1 ml) and was treated with NMM (14,5 μl, 0,132 mmole) and acetic anhydride (7,0 μl, 0,073 mmole) and stirred at RT for 14 h the Mixture was evaporated and dried in high vacuum. Then the residue was dissolved in a mixture of THF/Meon/N2O (4 ml/2 ml/2 ml) and stirred overnight with LiOH·2H2O (11 mg, 0,264 mmole). The remainder allocated after acidification to pH 3 using 1N. chilled on ice Hcl, purified using GHUR with reversed-phase column With a 18, using a gradient solvent 0-40% aqueous solution of CH3JV (0,06% TFA), resulting in allocated net connection 23 in the form of an amorphous solid white (12 mg).

1H NMR (50 mm PA2RHO4buffer, pH=6,0, 600 MHz): δ 1,22-of 1.27 (m, 2H), 1,38 was 1.43 (m, 2H), 1,58-of 1.64 (m,2H), 1,67 to 1.76 (m, 2H), 1.77 in-of 1.84 (m, 1H), 1,92 of 1.99 (m, 1H), 2,22-of 2.08 (m, 1H), 2,12-of 2.27 (m, 1H), 2,22-of 2.27 (m, 1H), 2,60-2,67 (m, 1H, Pro-β’), 2,83-2,89 (m, 1H, Pro-β), 4,32 (dd, J=12.1 and 3.5 Hz, 1H, Pro-δ’), to 4.41 (dd, J=12.1 and 7,3 Hz, 1H), 4,56 (bd, J=8.0 Hz, 1H, Pro-δ), to 4.62 (dd, J=8,9 Hz, 1H, Pro-α), 5,40-5,46 (m, 1H), 5,55-5,61 (m, 1H), 5,73 (bs, 1H, Pro-γ), 7,41 (d, J=6.3 Hz. 1H), to 7.64 (bs, 1H, ACPP-NH), 7,80 (dd, J=7.9 Hz, 1 H), 8,03 (dd, J=8.0 Hz, 1H), 8,07 (d, J=9.5 Hz, 1H), 8,16 (d, J=7 Hz, 1H, AcNH). at 8.36 (d, J=8,3 Hz, 1H), 8,90 (d, J=6.0 Hz, 1H).

Example 26

The synthesis of compounds 508 (table 5)

A. a Solution of the BOC-protected L-glutamine 26a (4,93 g, 20 mmol) and diacetate yogashala (7,73 g, 24 mmole, 1.2 EQ.) in a mixture tO/CH3SP/N2About (2:2:1, 60 ml) was stirred under 16°C for 1 h and at 20°C for 3 h Then the reaction mixture was diluted with H2O (20 ml), the solvents tO and CH3CN was removed in vacuo and the remaining aqueous mixture was extracted with diethyl ether (3×50 ml) and tO (50 ml) to remove most of the impurities. Then the aqueous layer (containing the intermediate product in the form of amine) was concentrated to dryness, the remaining product was re-dissolved in 10%PA2CO3(30 ml), cooled to 0°C in an ice bath and slowly added (~10 min) solution benzylchloride (3,3 ml, 20.4 mmole, of 1.02 EQ.) in dioxane (40 ml). The reaction mixture was stirred at 0°With whom for 1 h and at RT for 2 hours

Then the mixture was diluted with H2O (50 ml), was extracted with cold (~5° (C) diethyl ether (3×50 ml), acidified with 4 M Hcl to pH 3-4 and extracted with EtOAc (3×50 ml). The combined organic layers were dried over anhydrous MgS4and was evaporated to dryness in vacuum. The crude product was purified using the rapid chromatography on a column, using as eluent a mixture of tO/hexane/Asón (7:2,9:0,1), resulting in the received connection 26b with a total yield of 43% (3.04 from g).

B. Intermediate product, which represents a dipeptide 26C (250 mg, 0,41 mmole), compound 26b (171 mg, 0,49 mmole, 1.2 EQ.) and HA TU (185 mg, 0,49 mmole, 1.2 EQ.) was dissolved in CH2Cl2(6 ml) was added DIPEA (0,29 ml of 1.62 mmole, 4 EQ.). The reaction mixture was stirred at RT for 14 h, then CH2CL2evaporated in vacuum and the crude product was re-dissolved in EtOAc. tO the solution washed with aqueous 5%solution Panso3and with brine, dried over anhydrous MgSO4and was evaporated to dryness. After purification of the crude product using rapid chromatography on a column using as eluent a mixture of EtOAc/hexane (4:1) received connection 26d exit 98% (338 mg).

C. a Solution of compound 26d (335 mg, 0,394 mmole) in THF (5 ml) was cooled to 0°and the solution was added BH3in dimethyl sulfide (0,12 ml of 10 M solution, 1.2 mmol who, 3 EQ.). The reaction mixture was allowed to warm up to CT and it was stirred for 1 h and Then it was again cooled to 0°With, then slowly over 15 min was added an aqueous solution of NaOH (0.8 ml of a 2.5 M solution of 1.97 mmole, 5 EQ.), and then slowly added (~15 min) aqueous solution of H2O; (0.8 ml 8.8 M solution, 6.9 mmole, 17,5 EQ.). The reaction mixture was allowed to warm up to CT and it was stirred for 1 h then the reaction mixture was acidified to pH ~4, in order to neutralize the excess NR3then added an aqueous solution Panso3bringing the pH to ~9-10 were removed in vacuum THF and the crude product was distributed between the H2O and EtOAc. The aqueous layer was re-extracted with EtOAc, the combined organic layers were washed with brine, dried over anhydrous MgSO4and was evaporated to dryness in vacuum. The crude product was purified using the rapid chromatography on a column, using as eluent a mixture of tO/hexane/NH4HE (8:2:0.5), resulting in a net connection 26th with the release of 57% (192 mg).

G. To a solution of compound 26th in CH2Cl2(8 ml) was added periodical dessa-Martin (195 mg, 97%, of 0.33 mmole, 1.5 EQ.) and the reaction mixture was stirred at RT for 1.5 h the Reaction was stopped by adding an aqueous solution PA2S2O3(3 ml of a 5%solution)was then added saturated aqueous shall actor NaHCO 3(5 ml) and the mixture was stirred at RT for 15 minutes At the end of the crude reaction mixture was extracted with tO, the organic layer washed with aqueous 5%solution of NaHCO3and with brine, dried over anhydrous MgSO4and was evaporated in vacuum, getting 188 mg of aldehyde 28f, which was used in the next stage without further purification.

D. a Solution containing the compound 26f(188 mg, 0.22 mmole), CH3CO2N (38 μl) and PD(OH)2(25 mg) in ethanol (5 ml)was stirred at RT in the atmosphere of H2at atmospheric pressure for 16 hours, after this period of time the vessel was added an additional amount of the gaseous H2Pd(OH)2(180 mg) and CH3CO2N (154 ml) and continued stirring for another 24 h and Then the mixture was filtered and the solvent evaporated to dryness, the crude macrocyclic product was purified using the rapid chromatography on a column, using as eluent a mixture of l3/Meon/Asón (10:2:1), resulting in the received connection 26g c output ~30% (48 mg).

That is, the Mixture containing the compound 26g (22 mg, 0,031 mmole), DIPEA (27 μl, 0,155 mmole, 5 EQ.) and acetic anhydride (8,7 μl, 0,093 mmole, 3 EQ.) in CH2CL2(5 ml)was stirred at RT for 16 h Then CH2CL2was removed under vacuum, was added a mixture of THF/Meon/N2About (2:2:1, 5 ml) and LiOH·2H2O(13 mg, at 0.31 mmole, 10 EQ.) carried out the hydrolysis reaction for 68 h at RT and for 2 h at 50°C. Then the reaction mixture was acidified (pH ~4) and purified using GHUR with reversed phase, obtaining the target compound 508 (~6 mg, yield ~26% for the last 2 stages).

1H NMR (DMSO, 400 MHz) for compounds 508 (mixture of rotamers, identified on the basis of COSY, TOCSY and ROESY NMR analysis): δ of 1.18 (s, 9H), 1,09-1,85 (overlapping m, 11N), of 1.95 (s, 3H), 2,30 (m, 1H), 2.63 in (m, 1H), 3,18-4,14 (overlapping m, 6H), of 3.96 (s, 3H), of 4.44 (m, 1H), 4.62, and 4,69 (2d, J=11.8 Hz, 1H, rotamer), of 5.82 (bs, 1H), 7,20 (m, 2H), 7,53 (bs, 1H). to 7.67 (bs, 4H), 8,19 (bs, 3H), 8,61 (s, 1H).

Example 27

Synthesis of saturated macrocyclic derivative (27A)

A. Unsaturated macrocyclic derivative 23b (3.50 g, 7,30 mmole) was dissolved in tO (30 ml) was added 700 mg (20 wt.%) 5%Rh on alumina. The mixture was stirred in an atmosphere of gaseous N2at atmospheric pressure and at RT for 1.5 hours after this time period analysis using GHUR confirmed that the original product is completely turned into two products, the desired product 27A and a minor product (8% of the total mass), which was later identified as compound 27b formed in the opening cyclopropanol rings. The reaction mixture was filtered and concentrated, gaining solid light green is the first color (3,47 g). The solid was subjected to co-evaporation with tO to completely remove tO (presence tO influence the course of the reaction in the next stage). It was found that the separation of compounds 27A and 27b by using chromatography is difficult, therefore, was chosen as an alternative method, based on the difference in relative velocities of hydrolysis of their respective fragments complicated methyl ester.

B. the Crude mixture of compounds 27A and 27b (3,47 g) was dissolved in a mixture of THF:Meon (1:1, 20 ml)was added aqueous LiOH solution·H2O (24 mg in 5 ml of H2O, 8% squ.) and the reaction mixture was stirred at RT for 16 h (completion of the hydrolysis by-product 27b with the formation of the corresponding acid 27 was determined by analysis with IHVR). The reaction mixture was concentrated in vacuo to remove most of THF and Meon and distributed between the H2O(100 ml) and EtOAc (300 ml). The organic layer was washed with 0.5 N. NaOH (3×100 ml), brine solution (100 ml), 10%aqueous citric acid solution (2×100 ml), brine solution (100 ml), dried over anhydrous MgSO4, filtered and concentrated to dryness. The result was obtained the desired product 27A with a high purity (purity >90% according to analysis with IHVR) in the form of foam light green color with a total yield of 93% (3.28) for the two stages.

1N is Mr: (400 MHz, CDCl3): δ 1,1-of 1.38 (m, 13H), of 1.42 (s, N)and 1.51-1.57 in (m, 1H), 1,63-1,67 (dd, J=8.0 and 5.1 Hz, 1H), 1,81-to 1.87 (m, 1H), 1,92 of 1.99 (m, 1H), 2,02-of 2.08 (m, 1H), 2,62 (d, J=14 Hz, 1H), 3,4 (d, J=8,3, 1H), 3,65 (s, 3H), 4,01 (dd, J=10,8 and 4.1 Hz, 1H), 4,42-4,48 (m, 1H), 4,51-4,55 (m, 1H), 4,87 (d, J=8.6 Hz, 1H), 5,14 (d, J=8.6 Hz, 1H), 7,97 (br s, 1H).

Example 28

Synthesis of compound No. 741 (table 7)

A derivative of quinoline 8f was added to the obtained before this macrocyclic compound 23b through reaction of Mitsunobu. A derivative of quinoline 8f(30 mg, 0,095 mmole) was dissolved in THF, and then added a macrocyclic compound 23b (of 45.6 mg, 1 EQ.) and h3(49,8 mg, 2 EQ.). The resulting mixture was cooled to 0°C. Then was added dropwise DYADS (or 37.4 μl, 2 EQ.). The solution was stirred for 1 h at 0°C, and then stirred at room temperature overnight. After that, the mixture was diluted with EtOAc (15 ml), washed with saturated solution of NaHCO3(15 ml)and then with brine. The solution was dried over MgSO4, filtered and concentrated in vacuum. Received 202 mg of the product as a yellow oil. The product was purified using the rapid chromatography on silica gel (100%EtOAc). After cleaning, the product still contained a byproduct of DYADS. The resulting product contained 55 wt.% the desired product, so the output must be 62%.

Intermediate product in the form of ester (46 mg, is 0.06 mmole) was dissolved in sm is si THF/Meon/N 2About (ratio 2:1:1, 2 ml), was added LiOH·H2O (20 mg, 0.48 mmole) and the solution was stirred at RT. After 16 h the analysis of the reaction mixture using GHUR showed that the hydrolysis was completed. Organic solvents were removed in vacuo and the remaining crude product is dissolved in DMSO, and purified using GHUR with reversed phase column type C18, receiving net inhibitor 741.

1H NMR (400 MHz, DMSO-d6) δ (part./million): 8,67 (s, 1H), 8,29-to 8.14 (m, 2H), 8,08-of 7.97 (m, 1H), to $ 7.91 for 7.78 (m, 1H), 7,74 (s, 1H), 7,31-7,20 (m, 1H), 7,10 (d, J=5.7 Hz, 1H), of 5.82-5,71 (m, 1H), 5,58-vs. 5.47 (m, 1H), 5,32-5,23 (m, 1H), 4,74 with 4.64 (m, 1H), 4,55-4,47 (m, 1H), 4,23-4,06 (m, 1H), 4.04 the-of 3.94 (m, 1H), of 3.97 (s, 3H), 3,92-of 3.85 (m, 1H), 2,70 is 2.55 (m, 2H), 2,53-of 2.36 (m, 2H), 2,20-of 2.09 (m, 1H), 1,80-of 1.62 (m, 2H), 1.56 to USD 1.43 (m, 2H), of 1.42-1.29 (m, 6H), 1.27mm (d, J=3.2 Hz, 3H), 1,25 (d, J=2,9 Hz, 3H), of 1.12 (s, 9H).

MS: 763,1(M+1), 761,1 (M-1).

Example 29

Synthesis of compound 205 (table 2)

To a solution of macrocyclic compounds 25A (21 mg, 0.035 mmole) in a mixture of tert-butanol/N2O) (1.5 ml/1.5 ml) at 0°C, was added a solution of OsO4in tert-butanol (0,36 ml, 35% wt./about., 0.035 mmole) and the mixture was stirred at RT for 1 h the Mixture was diluted with EtOAc (20 ml) and the organic solution washed with 5%NaHCO3(2×10 ml). salt solution (2×10 ml), dried and evaporated to dryness. The crude compound was dissolved in a mixture of THF/Meon/N2About (3 ml / 1.5 ml / 1.5 ml) and stirred in the presence of LiOH·H2O (13 mg, 0.28 and the mole) for 16 hours The mixture was acidified to pH 4 with 0.5 to N. chilled on ice Hcl, evaporated and purified using GHUR with reversed phase column type C18, using a gradient solvent from H2On (0,06% TFA) to 40%aqueous solution of CH3CN (0,06% TFA). Allocated diol 205 having the SYN-configuration, in the form of an amorphous solid white color with a high degree of purity.

Connection # 205:1H NMR (DMSO, 400 MHz): δ a 1.01 (s, 9H), 1.06 a-of 1.30 (m, 9H), 1,48 by 1.68 (m,3H), 1,78-of 1.88 (m, 1H), ≈2,2-2,5 (2m, 2H), 3,78-3,82 (m, 1H), 3,86-3,90 (m, 1H), 4,39 (t, J=8,9 Hz, 1H), br4.61 (d, J=11,4 Hz, 14), the ceiling of 5.60 (bs, 1H, Pro-γ), 7,03 (d, J=6.0 Hz, 1H), 7,40 (bs, 1H), 7,58 to 7.62 (m, 1H), 7,87-to $ 7.91 (m, 1H), 8,00 (d, J=8,3 Hz, 1H), 8,24 (d, J=8.6 Hz, 1H), at 8.60 (s, 1H), 8,99 (bs, 1H).

EMC (form negative ionization): m/z 625 (M-N)-.

Example 30

The synthesis of compounds 214 and 218 (table 2)

A. a Solution of complex ketoanalogue ether 16d (0,180 g, 0.6 mmole) in a mixture of Meon/N2About (5 ml/2 ml) was stirred at RT in the presence of LiOH·H2O (50 mg, 1.2 mmole) for 1 h, the Solution was acidified to pH 6 with 0.5 to N. chilled on ice Hcl and evaporated most of the Meon. Then the residue was dissolved in tO (30 ml) and the solution was washed with 0.5 N. chilled on ice Hcl (10 ml), brine solution (10 ml), dried and evaporated. Then the crude residue was re-dissolved in CH2Cl2(10 ml) and subjected to interaction with FR what gentom P1-P2 30A (of 0.337 g, 0.6 mmole) in the presence of GATA (233 mg, 0,612 mmole) and DIPEA (420 μl, 2.4 mmole) for 16 h at RT. The reaction mixture was chromatographically on silica gel, using as eluent a mixture of tO/hexane (1/1), resulting in the isolated pure compound 30b (0,370 g, yield 83%, purity >95% according to analysis with IHVR).

1H NMR (CDCl3, 400 MHz) δ of 1.41 (s, 9H), 1,45-and 1.54 (m, 1H), 1,58-of 1.62 (m, 1H), 1,73-to 1.77 (m, 1H), 1,86 is 1.91 (m, 1H), 2,16 (dd, J=17.8 and 8.6 Hz, 1H), 2.26 and-2,43 (2m, 2H), 2,46-of 2.58 (m, 2H), 2,64-of 2.81 (m, 1H), 2,85 of 2.92 and 2.95-3,03 (2m, 1H, mixture of rotamers in a ratio of 1:3), to 3.67 (s, 3H), of 3.95 (s, 3H), 4,10-4,18 (m, 1H), 4,20-4,30 (m, 1H), 4,40-4,55 (m, 1H), 4.80 to 4,88 (m, 1H), 4.92 in-5,10 (m, 2H), 5,14 (dd, J=10.2 and 1,Hz, 1H), 5,24 is 5.38 (m, 4H), 5,42 is 5.54 (m, 1H), 5,68 and 5.86 (m, 2H),? 7.04 baby mortality-7,14 (m, 2H), 7,42-to 7.64 (m, 5H), 7,92-to 8.12 (m, 3H).

B. Dien 30b (0,370 g of 0.49 mmole) was subjected to cyclization in the presence of a catalyst comprising a dichloride, bis(tricyclohexylphosphine)benzyladenine(IV) (0.125 mg, 0.15 mmole) in CH2Cl2(freed by distillation from Sao2and degassed with argon for 30 min) at a temperature of reflux distilled for 2 hours After rapid chromatography on a column of silica gel using as eluent a mixture of EtOAc/hexane (3/1) received the compound in the form of a mixture of stereoisomers (30C and 30d, the ratio of 1:1) to yield 35% (0.124 g).

1H NMR mixture of compounds 30C and 30d (CDCl3, 400 MHz) δ the 1.44 (s, 4H) and of 1.37 (s, 4H), to 1.60 (m, 2H)and 1.83 (m, 0,5H), for 2.01 (m, 1H), 2,09 (m, 1H),2,42 (m, 5H), by 2.73 (m, 2H), 3,26 (m, 0,5H), of 3.69 (s, 1,5H), 3,76 (s, 1,5H), of 3.96 (s, 3H), 4,10 (m, 1H), 4,24 (m, 0,5H), 4,10 (m, 0,5H), 4,58 (m, 1H), 4,73 (m, 1H), 4,89 (m, 0,5H), equal to 4.97 (m, 0,5H), and 5.30 (m, 0,5H), 5,44 (m, 2H), 5,64 (m, 1H), 7,1-a 7.0 (m, 3H), 7,47 (m, 4H), 8,08-7,98 (m, 3H).

In that, the Hydrolysis of methyl esters 30C and 30d (24 mg, 0,033 mmole) was carried out in a mixture of THF/Meon/N2About (1 ml/0.5 ml/0.5 ml) using LiO•H2O (11 mg, 0,246 mmole) for 16 h at RT. After this time the mixture was acidified to pH 4-5 and chromatographically using GHUR with reversed phase column type With 18 using a gradient solvent from H2O (0,06% TFA) to 50%-aqueous solution of CH3CN (0,06% TFA). From a mixture of two compounds were allocated the required connections 214 and 218 having a high degree of purity (purity 94% according to analysis with IHVR), with a yield of 15% (3 mg).

Connection 214:1H NMR (DMSO, 400 MHz) δ to 1.15 (s,9H), 1,48-and 1.54 (m, 2H), 1,65-of 1.74 (m, 1H), 1,77-of 1.85 (m, 1H), 2,12 was 2.25 (m, 4H), 2,27-of 2.34 (m, 1H), 2,61 of 2.68 (m, 1H), 2,87 (bt, J=11.5 Hz, 1H), 3,92 (dd, J=9.2 and 1.5 Hz, 1H, Pro-5), of 3.97 (s, 3H, -och3). 4,14-4,20 (m, 1H), to 4.52 (t, J=7.8 Hz, 1H, Pro-α), of 4.66 (d, J=11.8 Hz, 1H, Pro-α), the 5.45 (t, J=9.9 Hz, 1H), 5,51-to 5.58 (m, 1H), of 5.82 (bs, 1H, RHS-γ), to 7.09 (d, J=6.0 Hz, 1H, BocNH), 7,26 (bs, 1H), 7,53 (s, 1H), to 7.67 (bs, 3H), 8,16 (d, J=2 Hz, 1H), 8,18 (s, 1H), 8,83 (s, 1H, ACPP-NN).

Compound 218:1H NMR(DMSO MHz): δ 1,06-1,10 (m, 1H), l,18(s,9H), of 1.52-1.55V (m, 1H), 1,62 and 1.80 (m, 1H), 2,10-2,68 (overlap, N), 3,90 (bd, J=8,3 Hz. 1H), 3.96 points (s, 3H, och3), 4,20-4,27 (m, 1H), 4,58-4,63 (m, 1H, Pro-δ), of 4.66 (dd, J=8,3 Hz, 1H, Pro-α) 4,88 (dd, J=10,2 Hz, 1H), 5,18-of 5.26 (m, 1H), 5,73-5,79 (m, 1H, Pro-γ), 7,01 (d, J=6,4 Hz, 1H), 7.23 percent (bs, 1H), 7,50 (bs, 1H), 7,66 (bs, 3H), to 8.20 (bs, 2H), 8,53 (s, 1H).

Example 31

Synthesis of compound 209 (table 2)

A. Dien 31A (249 mg, 0,330 mmole) was dissolved in 30 ml anhydrous CH2Cl2and the solution was degirolami with argon for 15 minutes, the Catalyst comprising a dichloride, bis(tricyclohexylphosphine)benzyladenine (IV) (82 mg, 0,100 mmole) was dissolved in 3 ml of anhydrous and degassed CH2Cl2and was added to a solution of diene. The reaction mixture was stirred at the temperature of reflux distilled for 2 h in an atmosphere of N2. The solution was concentrated and purified using the rapid chromatography on a column of receiving the connection 31b in the form of a solid brown color with a 71% yield (171 mg).

1H NMR (CDCl3, 400 MHz): δ 1,22-of 1.44 (m, 10H), of 1.42 (s, 9H), 1.66-of 1.74 (m, 1H), 1,87-of 1.97 (m,2H), 2,13-of 2.28 (m,3H), 2,32-2,39 (m, 1H), is 3.08-and 3.16 (m, 1H),3,41 (s, 3H), 4,07-4,22 (m, 3H), 4,28-4,34 (m, 1H), 4,58 with 4.64 (m, 1H), 4,95-4,99 (m, 1H), 5,22-of 5.29 (m, 2H), 5,38-5,43 (m, 1H), 5,48-to 5.56 (m, 1H), 7,00 for 7.12 (m, 3H), 7,43-of 7.55 (m, 4H), 7,97-8,11 (m, 3H).

ES(+)MS: m/z 727,4 (M+N)+.

B. Compound 31b (0,117 mmole) was stirred in a solution of Hcl (1 ml 4n. solution in dioxane) for 30 min and concentrated to dryness. The solid product was dissolved in CH2CL2(2 ml) and sequentially added Et3N (82 μl, 0,585 mmole) and tert-utilizationa (35 mg, 0,351 mmole). After stirring Ave is in CT for 20 h the mixture was concentrated to dryness and the crude compound is used on the final hydrolysis step without further purification.

C. Connection s (85 mg, 0,117 mmole) was dissolved in a mixture of THF/Meon/N2About (2 ml /1 ml/ 1 ml), was added LiOH·N2On (39 mg, 0,936 mmole) and the solution was stirred for 20 h at RT. After this period of time was added acetic acid (1 ml) and the solution was concentrated to remove the Meon and THF. After purification of the crude product using GHUR with reversed phase column type 18 was isolated a pure compound 209 (25 mg, yield ~31%).

1H NMR (DMSO, 400 MHz): δ was 1.04 (s, 9H), 1,15-1,24 (m, 2H), 1,30-1,40 (m, 5H), 1,44-is 1.51 (m, 2H), 1,54 by 1.68 (m, 1 H), 1,75-of 1.88 (m, 1H), 2,18 (dd, J=17,2 and 8.5 Hz, 1H), 2,32 at 2.45 (m, 1H, Pro-β), 2,54-2,62 (m, 1H), 2,65 of 2.68 (m, 1H, Pro-β), 3,91 (dd, J=11.1 and 3.5 Hz, 1H, Pro-δ), of 3.96 (s, 3H, -och3), 4,17-to 4.23 (m, 1H), 4,47 (dd, J=8,6, 1H, Pro-α), 4,67 (bd, J=7.9 Hz, 1H, Pro-δ), and 5.30 (dd, J=9.5 Hz, 1H), 5,52 (bdd, J=19 and 8.3, 1H), of 5.68 (s, 1H), 5,78 (bs, 1H, Pro-γ), 5,94 (bs, 1H), 7,21 (bs, 1H), 7,51 (bs, 1H), 7,66 (bs, 4H), 8,19 (s, 2H), 8,40 (d, J=7 Hz, 1H), 8,61 (s, 1H, ACPP-NH).

ES(+)MS: m/z 698,3 (M+H)+.

Example 32

The synthesis of compounds 404 and 407 (table 4)

A. Dien 32A (84 mg, of 0.11 mmole) was dissolved in anhydrous CH2Cl2(11 ml) and the solution was degirolami for 15 minutes by means of a jet of argon. A catalyst comprising a dichloride, bis(tricyclohexylphosphine)benzyladenine(IV) (19 mg, is 0.023 mmole) was first dissolved in 1 ml of degassed CH2CL2and then it was transferred via cannula to the reaction flask. React the mixture was stirred for 2 h at the temperature of reflux distilled. Then the solvent was removed in vacuum and the reaction mixture was purified using the rapid chromatography on a column, using as eluent a mixture of tO/ hexane (1:1), resulting in a received macrocyclic compound 32b in the form of a yellow oil (33 mg, yield 41%).

B. Intermediate product, which represents an ester 32b (33 mg, of 0.045 mmole), was dissolved in a mixture of THF/MeOH/H2O (2:1 ratio:1.2 ml), was added LiOH·H2O (8 mg, of 0.18 mmole) and the solution was stirred at RT. After 16 h the analysis of the reaction mixture using GHUR showed that the hydrolysis is not complete. So I added an additional amount of LiOH·H2O (4 mg, and 0.09 mmole) and the solution was stirred at RT in total within 36 hours At the end the solution was acidified small aliquot quantity of acetic acid, the organic solvents were removed in vacuo and the remaining crude product was purified using GHUR with reversed phase column type With 18, while purely inhibitor 404.

1H NMR(DMSO, 400 MHz): δ 1.21 (d, J=6.0 Hz, 3H, Me), of 1.36(s, 9H, BOC), 1,1-1,4 (3m, 3H), of 1.66 (m, 1 H), of 1.80 (m, 1 H), 2,10 (m, 2H), 2.57 m (m, 2H), 3,90 (m, 4H), 4,47 (bd, J=a 12.7 Hz, 1H), 4,58 (bd, J=of 7.3, 1H), 4,66 (dd, J=8.0 Hz, 1H), to 5.57 (m, 1H), to 5.66 (m, 1H), of 5.83 (bs, 1H), 6,18 (bd, J=6,9 Hz, 1H), 7,25 (bd, J=7,3 Hz, 1H), 7,56 (bs, 1H), of 7.70 (m, 4H), by 8.22 (bd, J=2,9 Hz, 2H), 8,29 (bs, J=9,2 Hz, 1H).

Century Inhibitor 404 (15 mg, 0,021 mmole) was dissolved in ethanol (2 ml) was added 10%Pd/C (2 mg). The mixture is eremetical in an atmosphere of hydrogen at RT for 16 hours After filtration, the mixture was purified using GHUR with reversed phase column type With 18 receiving the inhibitor 407 in the form of a solid white color (10 mg, yield 66%)

1H NMR (DMSO, 400 MHz): δ 1,04 (m, 1H), 1,17 (d, J=6.0 Hz, 3H), of 1.35 (s, 9H), 1.25 and about 1.75 (m, 12 H), 2,32 at 2.45 (m, 1 H), 3,40-to 3.50 (m, 2 H), 3,74-a 3.83 (m, 1H), 3,85-3,93 (m, 1H), of 3.97 (s, 3H), 4,27 is 4.36 (dd, J=21,1 and 8.6 Hz, 1H), 4,54 (dd, J=7.95 and of 7.95 Hz, 1H), 5,64 (d, J=8,3 Hz, 1H), of 5.82 (brs, 1H), 7,27-7,33 (m, 1H), 7,53-EUR 7.57 (bs, 1 H), 7,60-7,74 (m, 4 H), 8,13-of 8.27 (m, 3 H), 8,30-8,35 (br s, 1H).

Example 33

The synthesis of compounds 824 (table 8)

A. Compound 33a (~0.55 mmole) was dissolved in CH2CL2(100 ml) and the solution was carefully degirolami, then add a portion of the Hoveyda catalyst (17 mg, 0,028 mmole, of 0.05 EQ.). Then the solution was stirred at the temperature of reflux distilled for 5 hours the Reaction mixture was concentrated and purified using the rapid chromatography on a column using a gradient solvent of CH2CL2/tO (from 3:2 to 2:3 ratio), resulting in the received connection 33b with the release of 72% (194 mg).

B. To a solution containing compound 33b (70 mg, 0,142 mmole), 2-ethoxy-4-hydroxy-7-methoxyquinoline 3C (63 mg, 0,284 mmole, 2 EQ.) and RH3R (186 mg, of 0.71 mmole, 5 EQ.) in anhydrous THF (15 ml)slowly over 20 min was added at 0°DYADS (140 μl, of 0.71 mmole, 5 EQ.). The reaction mixture was allowed to warm up to CT and it was stirred at RT for 2.5 h Then THF in perivale in vacuum and posisinya product was purified using the rapid chromatography on a column using a gradient solvent of hexane/tO (from 7:3 to 1:1). Allocated net connection 33 with the output 73% (72 mg).

C. Compound 33C (72 mg, 0.104 g mmole) was mixed with CH2CL2(5 ml) and 4 m solution of Hcl in dioxane (5 ml) and the mixture was stirred at RT for 1.5 h to remove the protective groups are BOC and obtain hydrochloride of the intermediate product 33d. The crude reaction mixture was evaporated to dryness in vacuo, and dried under vacuum to ensure complete removal of Hcl, and the product was used in the next stage without purification.

G. To a solution of Cyclopentanol (29 μl, of 0.32 mmole) in THF (10 ml) was added dropwise a solution of phosgene in toluene (1,93M, 274 μl, 0,528 mmole) and the mixture was stirred at RT for 2 h to obtain cyclopentylpropionate reagent. After this time approximately half of the solvent was removed by evaporation in vacuo, the remaining solution was light yellow color was diluted by adding CH2Cl2(5 ml) and again concentrated to half the original volume in order to ensure complete removal of all excess phosgene. Then the above solution cyclopentylpropionate reagent was diluted in THF (10 ml), cooled to 0°and was added to the solid connection 33d (0.104 g mmole) at 0°C. To the reaction mixture was added Et2N (75 μl, 0,534 mmole, 5.2 EQ.) and prod liali stirring at 0° C for 1.5 h the Solvent was removed in vacuum and the crude product was purified using the rapid chromatography on a column, using as eluent a mixture of EtOAc/hexane (1:1), resulting in the received connection e with almost quantitative yield (75 mg).

D. Hydrolysis of complex methyl ester was performed by reacting compound e (75 mg, or 0.11 mmole) with LiOH·H2O (35 mg, from 0.84 mmole, 8 EQ.) in a solvent mixture of THF/Meon/N2About (ratio 2:2:1, 7.5 ml) at 50°within 2.5 hours After completion of the hydrolysis, the mixture was acidified to pH 4.5 and the solvents evaporated to dryness in a vacuum. The crude product was purified using preparative GHUR with reversed phase column type With 18 using a gradient solvent from H2O to 58%aqueous solution of CH3CN (containing 0.06% of TFA), which received the inhibitor No. 824 in the form of an amorphous solid white (45 mg, yield 65%).1H NMR for Na+-salt of compound No. 824 (DMSO, 400 MHz): δ to 0.88 (d, J=6,7 Hz, 3H), 0.95 to-1,70 (overlapping resonance peaks, 17H), to 1.37 (t, J=7 Hz, 3H), 2,00-2,10 (m, 1H), 2,10-of 2.33 (m, 3H), 2,38 is 2.44 (m, 1H), 3,80-of 3.85 (m, 1H), 3,85 (s, 3H), was 4.02-4,08 (m, 1H), 4,42 (q, J-7 Hz, 2H), 4,35-of 4.44 (m, 1H), 4,50 (d, J=10,8 Hz, 1H), 4,63 (bs, 1H), 5,28 (dd, J=9.5 Hz, 1H), 5,38 (bs, 1H), 5,42-5,49 (m, 1H), 6,37 (s, 1H), 6.87 in (dd, J=8,9 and 2.2 Hz, 1H), 7,07 (d, J 2.2 Hz, 1H), 7,28 (d, J=7,0 Hz, 1H). of 7.90 (d, J=8,9 Hz, 1H), to 8.57 (s, 1H).

Example 34

The synthesis of compounds 812 (table 8)

A. To a solution containing macrocyclic intermediate product 23b (of 13.05 g of 27.2 mmole, 1.0 EQ.), RH3P (of 14.28 g, 54,4 mmole, 2.0 EQ.) and 2-carboxymethoxy-4-hydroxy-7-methoxyquinoline (WO 00/09543 and WO 00/09558) in (6.67 g, 28.6 mmole, of 1.05 equiv.) in THF (450 ml)was added dropwise over 15 min at 0°DYADS (10,75 ml, 54,6 mmole, 2.0 EQ.). Then the ice bath was removed and the reaction mixture was stirred at RT for 3 h After transformation of the original product in the final products, the solvent is evaporated in vacuum, the remaining mixture was diluted with EtOAc, washed with saturated solution of Panso3(2) and with brine (1x), the organic layer was dried over anhydrous MgSO4, filtered and evaporated to dryness. After rapid chromatography on a column of received net connection 34a; the column was suirable has snachala a mixture of hexane/tO (50:50), and then l3/tO (95:5) to remove by-products PH3RO and DYADS, the process of elution of impurities was observed by TLC. In conclusion, the desired product 34a was suirable from the column using a mixture l3/tO (70:30). Typically, the phase chromatography should be repeated 2-3 times to ensure that the connection 34a could be selected with a high degree of purity in a solid white color with a total yield of 68% (12.8 g, 99.5% purity according to Ana who studied with IHVR).

B. To a solution of the BOC-protected intermediate 34a (1,567 g) in CH2CL2(15 ml) was added 4 N. Hcl in dioxane (12 ml) and the reaction mixture was stirred at RT for 1 o'clock [in the case when the reaction began to form a thick gel, was added 10 ml of CH2Cl2]. After removal of the protective group of the solvents evaporated to dryness, obtaining solid yellow and pasty product. The mixture was re-dissolved in approximately 5%solution of a Meon in CH2Cl2and re-evaporated to dryness in vacuum, obtaining the connection 34b in the form of a solid yellow color, which is used in the next stage without any treatment.

C. To a solution of Cyclopentanol (614 μl, 6,76 mmole) in THF (15 ml) was added dropwise a solution of phosgene in toluene (1,93 M, 5,96 ml, 11,502 mmole) and the mixture was stirred at RT for 2 h, the receiving agent (z), representing cyclopentyl-chloroformiate. After this period of time approximately half of the solvent was removed by evaporation in vacuo, the remaining solution was light yellow color was diluted by adding SN2CL2(5 ml) and concentrated to half of its original volume in order to ensure the removal of all excess phosgene. Then obtained above cyclopentylpropionyl the reagent rasb ulali THF (15 ml) and added to unindividualized 34b. The mixture was cooled to 0°C in an ice bath, the pH value was brought to ~8,5-9 by adding Et3N (by adding dropwise and the reaction mixture was stirred at 0°C for 1 h, after this time the mixture was diluted with EtOAc, washed with water (1x), saturated solution of NaHCO3(2x), N2On (2x) and with brine (1x). The organic layer was dried over anhydrous MgSO4, was filtered and was evaporated in vacuum, obtaining a foam with a light amber color. After purification using rapid chromatography on a column (using as eluent a gradient solvent of 30%hexane to 20%hexane in EtOAc) received connection 34C in the form of a white foam with a yield of 80% (1.27 g) and a purity of >93%.

, Dimethyl ether Complex 34C (1,17 g) was dissolved in a mixture of THF/Meon/N2O (20 ml, the ratio of 2:1:1) was added aqueous NaOH solution (1.8 ml, 1 BC, 1 EQ.). The reaction mixture was stirred at RT for 1 h, after which it was evaporated to dryness, obtaining the sodium salt 34d in a solid white color (~of 1.66 mmole). Connection 34d used in the next stage without purification.

D. the Crude sodium salt 34d (of 1.66 mmole) was dissolved in THF (17 ml)was added Et3N and the mixture was cooled to 0°C in an ice bath. Was added dropwise to Isobe-telharmonium (322 μl, 2.5 mmole) and the mixture was stirred at 0°C for 75 minutes after this period is Yes time was added diazomethane (15 ml) and continued stirring at 0° C for 30 min and then for 1 h at RT. A large part of the solvent was removed by evaporated to dryness in vacuo, the remaining mixture was diluted with EtOAc, washed with saturated solution of Panso3(2), N2O (2) and with brine (1x), dried over anhydrous MgSO4that was filtered, evaporated to dryness, obtaining the connection a foam light yellow (1.2 g, ~of 1.66 mmole). The intermediate product, representing diazoketone e used in the next stage without purification.

E. Diazoketone e (1,2g, of 1.66 mmole)dissolved in THF (17 ml) was cooled to 0°C in an ice bath. Was added dropwise an aqueous solution NVG (48%, 1,24 ml) and the reaction mixture was stirred at 0°C for 1 h Then the mixture was diluted with EtOAc, washed with saturated solution of NaHCO3(2x), H2O (2x) and brine solution (1×), the organic layer was dried over anhydrous MgSO4, was filtered and was evaporated to dryness, obtaining an intermediate product that is a β-Bratton 34f, foam light yellow (~1,657 mmole).

G. To a solution of bracelona 34f (600 mg, 0,779 mmole) in isopropanol (5 ml) was added thiourea (118 mg, of 1.55 mmole) and the reaction mixture was placed in an oil bath, preheated to 75°where it was stirred for 1 h Then the isopropanol was removed under vacuum and the product was dissolved in EtOAc (100 ml). The solution was washed with saturated RA is tworoom Panso 3and with brine, the organic layer was dried over anhydrous Na2SO4, was filtered and was evaporated, obtaining the crude product 34g (522 mg) as a solid reddish-brown color. This product is used at the final stage without any further purification.

H. the Crude methyl ester 34g (122 mg, 0,163 mmole) was dissolved in a mixture of THF/Meon/N2About (2:1 ratio:1,4 ml) and was subjected to saponification using LiOH•H2O (89 mg, and 2.14 mmole). The hydrolysis reaction was carried out for 12-15 h at RT. Then the solvent was removed in vacuum and the crude product was purified using GHUR with reversed phase column type With 18 using a gradient solvent of 10%of CH3SP in N2O to 100%CH3The joint VENTURE, resulting in the received connection 812, representing an inhibitor of HCV protease, in a solid yellow color (24 mg, total yield in the conversion of the intermediate product 34f in the inhibitor 812 was 20%).

1H NMR (400 MHz, DMSO-d6) δ 8,63 (s, 1H), compared to 8.26-of 8.15 (m, 2H), 7,79 (bs, 1H), 7.72 (bs, 1H), 7,50 (bs, 2H), 7,33-7,25 (m, 2H), 5,77 (bs, 1H), 5,52 (dd, J=8,3 Hz, 1H), 5,27 (dd, J=9,2 Hz, 1H), with 4.64 (d, J=10,8 Hz, 1H), 4,50 (dd, J=8,3 Hz, 1H), 4,39-or 4.31 (m, 1H), 4,08-to 3.99 (m, 2H), of 3.94 (s, 3H), a 3.87 (d, J=9.5 Hz, 2H), 2,65 of $ 2.53 (m, 2H), 2,46-2.36 (m, 2H), 2,20-2,12 (dd, J=8.6 Hz, 1H), 1,80-of 1.64 (m, 2H), 1,63 was 1.06 (m, 14H). MC; es+:733,2 (M+H)+, es-: 731,2 (M-N)-.

Example 34a

Using Taku the same procedure that described in example 34, but exposing engagement Bratton 34f c commercially available N-methylthiophenol received connection # 811 (table 8)

1H NMR (400 MHz, DMSO-d6): δ 8,63 (s, 1H), to 8.20 (s, 1H), 8,18 (s, 1H), 8,12-to 7.93 (m, 1H), 7,88-of 7.69 (m, 2H), 7,32-7,24 (m, 2H), of 5.82-of 5.75 (m, 1H), 5,52 (ddd, J=8,1 Hz, 1H), 5,28 (dd, J=9.9 Hz, 1H), 4,67-br4.61 (m, 1H), 4,51 (dd, J=8,8 Hz, 1H), of 4.44-4,37 (m, 1H), 4,08-4,00 (m, 1H), 3.96 points (s, 3H), with 3.89 (m, 1H), 3.04 from (d, J=4,1 Hz, 3H), 2,65-is 2.37 (m, 3H), of 2.16 (m, 1H), 1.77 in.of 1.65 (m, 2H), 1,63-1,11 (m, 17H). MC; es+: 747,2 (M+H)+, es-:745,3 (M-H)-.

Example 34B

Using the same procedure described in example 34, but exposing engagement Bratton 34f c commercially available N-ethyltoluidines received connection # 810 (table 8)

1H NMR (400 MHz, DMSO-d6): δ 8,63 (s, 1H), 8,27 (bs, 1H), to 8.20 (d, J=9.0 Hz, 1H), 8,13-8,07 (m, 1H), 7,86 (bs, 1H), 7,78 (s, 1H), 7,33-7,25 (m, 2H), of 5.81 (bs, 1H), 5,54 (dd, J=8,8 Hz, 1H), 5,28 (dd, J=9.7 Hz, 1H)and 4.65 (d, J=and 12.4 Hz, 1H), 4,51 (dd, J=8,8 Hz, 1H), to 4.38 (bs, 1H), 4,03 (m. 1H), of 3.97 (s, 3H), 3,92-a 3.87 (m, 1H), 3,54-of 3.46 (m, 2H), 2,68-to 2.65 (m, 2H), 2,47-of 2.38 (m, 1H), 2,15 (dd, J=8.6 Hz, 1H), 1,78-of 1.65 (m, 2H), 1.60-to of 1.12 (m, 17H), 1,25 (t, J=7,3 Hz, 3H). MC; es+: 783,2 (M+Na)+, es-:761,2 (M+H)-.

Example 34B

Using the same procedure described in example 34, but exposing engagement Bratton 34f c commercially available N-isopropylamino received connection 822

H NMR (400 MHz,DMSO-d6) δ 8,63 (s, 1H), 8,33-8,23 (bs, 1H), 8,21 (d, J=9,2 Hz, 1H), 8,04 (d, J=8,3 Hz, 1H), 7,86 (bs, 1H), to 7.77 (s, 1H), 7,35-of 7.23 (m, 2H), of 5.81 (bs, 1H), 5,52 (dd, J=8.5 Hz, 1H), 5,27 (dd, J=9,2 Hz, 1H), 4,65 (d, J=11.8 Hz, 1H), 4,51 (dd, J=7,6 Hz, 1H), 4,37 (bs, 1H), 4,15 (bs, 1H), 4.07-3,98 (m, 2H), of 3.97 (s, 3H), 3,88 (d, J=8,9 Hz, 1H), 2,60 of $ 2.53 (m, 2H), 2,47-is 2.37 (m, 2H), 2,19-2,10 (dd, J=9,2 Hz, 1H), 1,80-1,64 (m, 2H), 1,63-of 1.29 (m, 13H), of 1.27 and 1.25 (2 x d, J=6,5 Hz, 6N), 1,23-of 1.09 (m, 2H). MC; es+:775,0 (M+H)+, es-:772,9 (M-N)-.

Example 34g

Using the same procedure described in example 34, but exposing engagement Bratton 34f c commercially available N-acetylthiocholine received connection # 809

1H NMR (400 MHz, DMSO-d6): δ to 8.62 (s, 1H), 8,30 (bs, 1H), 8,17 (d, J=8,9 Hz, 1H), 7.62mm (bs, 1H), 7,52 (bs, 1H), 7,28 (d, J=6,4 Hz, 1H), 7,21 (bs, 1H), 5,63 (bs, 1H), 5,54 (dd, J=8,1 Hz, 1H), 5,28 (dd, J=9.5 Hz, 1H), to 4.62 (d, J=12.1 Hz, 1H), 4,56-to 4.46 (m, 2H), 4,11-Android 4.04 (m, 1H), 3,95 (s, 3H), 3,93-3,88 (m, 1H), 2,62-of 2.54 (m, 1H), 2,45-of 2.36 (m, 1H), 2,22 (s, 3H), 2.21 are to 2.13 (m, 1H), 1,79 was 1.69 (m, 2H), 1,65-of 1.30 (m, 16H), 1,26-of 1.12 (m, 2H). MC; es+: 775,3 (M+H)+, es-: 773,3 (M-N)-.

Example d

To a solution of the intermediate product, representing 2-amino-4-thiazolyl 34g (0.24 g, of 0.32 mmole)in CH2CL2(5 ml) at RT was added DIPEA (0,55 ml, 3,18 mmole, 10 EQ.) and methylchloroform made (0.13 ml, 1.6 mmole, 5 EQ.). The reaction mixture was stirred for 6.5 h, after which it was concentrated in vacuum. After that, the crude isolated product was subjected to hydrolysis as description is but in example 34, with the formation of the desired carboxylic acid, receiving connection # 818

1H NMR (400 MHz, DMSO-d6): δ 8,61 (s, 1H), 8,21-8,07 (m, 2H), to 7.61-7,38 (m, 2H), 7,26 (d, J=6,4 Hz, 1H), 7,19-7,10 (m, 1H), ceiling of 5.60-vs. 5.47 (m, 2H), 5,27 (dd, J=9,2 Hz, 1H), 4.63 to-a 4.53 (m, 1H), 4,47 (d, J=7.9 Hz, 1H), 4,13-Android 4.04 (m, 1H), 3,93 (s, 3H), 3,92-a 3.87 (m, 2H), 3,79 (s, 3H), 2,42-of 2.30 (m, 2H), 2,17 (dd, J=9,2 Hz, 1H), 1,81 by 1.68 (m, 2H). 1,63-of 1.29 (m, 16H), 1,23-1,10 (m, 2H). MC; es+: 791,1 (M+H)+, es-: 789,1 (M-H)-.

Example a

Using the conditions described in example d, but applying isobutylparaben received intermediate product, representing the corresponding substituted isobutylparaben. Then the crude isolated product was subjected to hydrolysis, receiving the required No. 819

1H NMR (400 MHz, DMSO-d6): δ to 8.62 (s, 1H), of 8.47-of 8.27 (bs, 1H), 8,18 (d, J=8.6 Hz, 1H), 7,69-of 7.60 (m, 1H), 7,60-7,51 (m, 1H), 7,28 (d, J=6,7 Hz, 1H), 7,28-7,19 (m, 1H), 5,70-the ceiling of 5.60 (m, 1H), 5,52 (dd, J=8,3 Hz, 1H), 5,27 (dd, J=9.8 Hz, 1H), 4,63 (d, J=11.8 Hz, 1H), 4.53-in-of 4.44 (m, 2H), 4,10-to 3.99 (m, 1H), Android 4.04 (d, J=6,7 Hz, 2H), 3,95 (s, 3H), 3,94-a 3.87 (m, 1H), 2,65 of $ 2.53 (m, 1H), 2,46-2.34 (m, 1H), 2,16 (dd,J=8,1 Hz, 1H), 2,03 is 1.91 (m, 1H), 1,79-of 1.09 (m, 20H), of 0.95 (d, J=6,7 Hz, 6N). MC; es+: 833,2 (M+H)+, es-: 831,2 (M-N)-.

Example 35

Synthesis of compound No. 908

Using as starting product derived 27A and the same synthesis procedure described in example 34, received the following macrocycle compound No. 908 (table 9).

1H NMR (400 MHz, DMSO-d6): δ of 8.47 (s, 1H), 8,16 (d, J=10 Hz, 1H), 8,15-8,07 (m, 1H), 7,82-7,63 (m, 2H), 7,53-7.43 (m, 2H), 7,33-7,22 (m, 1H), 7,13 (d, J=7 Hz, 1H), 5,77-the 5.65 (m, 1H), 4,62-to 4.52 (m, 2H), 4,50-4,4 (m, 1H), 4,20-4,10 (m, 1H), 3,94 (s, 3H), 3,89-a 3.83 (m, 1H), 2,59 of $ 2.53 (m, 1H), 2,48-to 2.40 (m, 1H), 1,79-1,0 (m, 25H). MC; es+: 735,2 (M+H)+, es-: 733,2 (M-H)-.

Example 35A

Synthesis of compound No. 909

Using the same synthesis procedure described in example 35, but employing the appropriate N-acetylthiocholine received connection # 909 (table 9).

1H NMR (400 MHz, DMSO-d6): δ 8,53-to 8.41 (m, 2H), to 8.20 (d, J=9,2 Hz, 1H), 7,68 (bs, 1H), 7,68 (bs, 1H), 7,27 (dd, J=9,2 Hz, 1H), 7,15 (d, J=6,4 Hz,1H), 5,67 (bs, 1H), 4,65-4,50 (m, ZN), of 4.44-4,37 (m, 1H), 4,21-4,13 (m, 1H), 3.96 points (s, 3H), 3,99-3,86 (m, 1H), 2,62-2,39 (m, 2H), 2,24 (s, 3H), 1,78-to 1.67 (m, 3H), 1,67-a 1.01 (m, 22H).

MC; es+: 798,0 (M+Na)+, es-: 777,0 (M+N)+.

Example 35B

Synthesis of compound No. 910

Using the same synthesis procedure described in example 35, but employing the appropriate N-ethylthiophene received connection # 910 (table 9).

1H NMR (400 MHz, DMSO-d6): δ of 8.47 (s, 1H), 8,29 (bs, 1H), to 8.20 (d, J=9,2 Hz, 1H), 8,09 (bs, 1H), 7,87 (s, 1H), to 7.77 (s, 1H), 7,32 (dd, J=9,2 Hz, 1H), 7,14 (dd, J=6,7 Hz, 1H), 5,78 (bs, 1H), 4,58 (dd, J=8,1 Hz, 2H), 4,43 (bs, 1H), 4,18-4,12 (m, 1H), of 3.97 (s, 3H), a 3.87 (d, J=8,9 Hz, 1H), 3,55-of 3.46 (m, 2H), 2,63 of $ 2.53 (m, 1H), 2,47-to 2.41 (m, 1H), 1,78-1.00 (m. 25N), 1,25 (t, J=7,3 Hz, 3H).). MC; es+: 763,1 (M+H)+, es-: 761,1 (M-H)-/sup> .

Example 35V

Synthesis of compound No. 911

Using the same synthesis procedure described in example 35, but employing the appropriate N-isopropylamino received connection # 911 (table 9).

1H NMR (400 MHz, DMSO-d6): δ of 8.47 (s, 1H), 8,29-8,19 (m, 1H), 8,19 (d, J=9,2 Hz, 1H), 8,09 to 8.0 (m, 1H), 7,83 (bs, 1H), 7,74 (bs, 1H), 7,31 (d, J=8 Hz, 1H), 7,14 (d, J=6,4 Hz, 1H), USD 5.76 (bs, 1H), with 4.64-4.53-in (m, 2H), of 4.44 (bs, 1H), 4,22-4.09 to (m, 3H), of 3.97 (s, 3H), a 3.87 (d, J=8.6 Hz, 1H), 2,63-of 2.58 (m, 1H), 2,46-to 2.41 (m, 1H), 1,79-1,10 m. 24H), of 1.27 and 1.26 (2 x d, J=6,5 Hz, 6N). MC; es+: 777,0 (M+H)+, es-: 775,0 (M-N)-.

Example 36

Synthesis of compound No. 716

1H NMR (400 MHz, DMSO-d6): δ (part./million): to 8.62 (s, 1H), 8,13 (d, J=9,2 Hz, 1H), of 7.64-rate of 7.54 (m, 2H), 7,47 (d, J=2,6 Hz. 1H), 7,16 (dd, J=9,2, 2.2 Hz, 1H), 7,03 (d, J=6.0 Hz, 1H), 5,63 (s, 1H), 5,52 (q, J=9.9 Hz, 1H), 5,26 (t, J=8,9 Hz, 1H),4.62 (d, J=of 11.45, 1H), 4,45 (dd, J=9,2, of 8.27 Hz, 1H), was 4.02 (m, 1H) 3,93 (s, 3H), 3,7 (dd, J=7,6, 1.0 Hz, 1H), 2,66 (s, 3H), 2,55-to 2.65 (m, 1H), 2,35 at 2.45 (m, 1H), 2,17 (q, J=8.6 Hz, 1H), 1,65 is 1.75 (m, 2H), 1,5-of 1.35 (m, 7H)and 1.15(s, 9H).

MC:705(M+1),703(M-1).

Example 37

Synthesis of compound No. 717

1H NMR (400 MHz, DMSO-d6): δ (part./million): to 8.62 (s, 1H), 8,15 (d, J=8,9 Hz, 1H), 7.62mm (s, 1H), 7,49 (s, 1H), 7,19 (dd, J=9,2, 2.2 Hz, 1H), 7,02 (d, J=5.4 Hz, 1H), 5,64 (s, 1H), 5,52 (q, J=9.9 Hz, 1H), 5,26 (t, J=9,2 Hz, 1H), 4,63 (d, J=11,44, 1H), 4,45 (t, J=9,2 Hz, 1H), 3,94 (s, ZN), from 3.9 to 3.8 (m, 1H), 2,7-to 2.55 (m, 1H), 2,4-2,3 (m, 1H), 2,18 (q, J=8,9 Hz, 1H), 1,75-of 1.65 (m, 2H), 1,5-1,2 (m, 7H), to 1.14 (s, 9H).

MS:705(M+1),703(M-1).

Example 38

Si is TEZ connection No. 718

1H NMR (400 MHz, DMSO-d6): δ (part./million): of 9.55 (s, 1H), 8,63 (s, 1H), 8,43 (s, 1H), 8,13 (d, J=9,2 Hz, 1 H), 7,66 (s, 1 H), 7,46 (s, 1 H), 7,32 (d, J=2.6 Hz, 1 H), 7,10-7,07 (m, 2H), 5,64 is 5.54 (m, 1H), 5,59-of 5.48 (m, 1H), 5,33-5,23 (m, 1H), 4,73-br4.61 (m, 1H), 4,45 (dd, J=7,5, and 9.1 Hz, 1H), 4.09 to 4,00 (m, 1H), 3,92 (s, 3H), 3,93-a 3.83 (m, 1H), 2,67 is 2.55 (m, 2H), 2,53 is 2.43 (m, 1H), 2,42-2,31 (m, 1H), 2,23-2,12 (m, 1H), 1,81-of 1.66 (m, 2H), 1,52-of 1.42 (m, 2H), 1,42-1,25 (m, 6H), to 1.21 (s, 9H).

MS: 689,3 (M+1), 687,3 (M-1).

Example 39

Synthesis of compound No. 722

1H NMR (400 MHz, DMSO-d6): δ (part./million): to 9.70 (s, 1H), 8,64 (s, 1H), compared to 8.26 (s, 1H), 8,14 (d, J=9,2 Hz, 1H), 7,45 (s, 1H), 7,30 (d, J=2.5 Hz, 1H), 7,14-7,06 (m, 2H), ceiling of 5.60 is 5.54 (m, 1H), 5,58-of 5.48 (m, 1H), 5,31-5,23 (m, 1H), 4,71-to 4.62 (m, 1H), 4,49-and 4.40 (m, 1H), 4,08-to 3.99 (m, 1H), 3,92 (s, 3H), 3,92-a-3.84 (m, 1H), 2,69-of 2.54 (m, 2H), 2,53 is 2.46 (m, 1H), 2,42-2,31 (m, 1H), is 2.37 (s, 3H), 2,22 and 2.13 (m, 1H), 1,81-of 1.64 (m, 2H), 1,54-of 1.42 (m, 2H), 1,42-1.27mm(m, 6H), to 1.22 (s,9H).

MS: 703,3 (M+1), 701,3 (M-1)

Example 40

Synthesis of compound No. 733

1H NMR (400 MHz, DMSO-d6): δ (part./million): 8,75 (m, 1H), to 8.62 (s, 1H), of 8.06 (d, J=9,2 Hz, 1H), 7,88-7,87 (m, 1H), of 7.48 (s, 1H), 7,28 (d, J=2.6 Hz, 1H), 7,05-7,00 (m, 2H), 6,64-6,63 (m, 1H), 5,62-to 5.58 (m, 1H), 5,55-5,49 (m, 1H), 5,28-5,24 (m, 1H), with 4.64-br4.61 (m, 1H), 4,48-of 4.44 (m, 1H), 4,07-a 4.03 (m, 1H), 3,91 (s, 3H), 3,92-of 3.85 (m, 1H), 2,67-of 2.54 (m, 2H), 2,53 at 2.45 (m, 1H), 2,41-of 2.34 (m, 1H), 2,20-and 2.14 (m, 1H), 1,75 was 1.69 (m, 2H), 1,50 was 1.43 (m, 2H), 1.41 to -1,32 (m, 6H), of 1.17 (s, 9H).

MS: 689,3 (M+1), 687,2 (M-1).

Example 41

Synthesis of compound No. 703

1H NMR (400 MHz, DMSO-d6): δ and 8.50 (s, 1H), 8,19 (s, 1H, 8,17 (s, 1H), 8,11-of 8.00 (m, 1H), 7,88-to 7.77 (m, 1H), 7,73 (s, 1H), 7,25 (d, J=8.6 Hz, 1 H), 6,93 (d, J=6 Hz, 1H), of 5.89-of 5.68 (m, 1H), to 4.62 (d, J=11 Hz, 1H), 4.53-in (dd, J=8,3 Hz, 1H), 4,16-4,07 (m, 1H), 3.96 points (s, MN), 3,88 (bd, J=9.5 Hz, 1H), 3,53-of 3.43 (m, 2H), 2,63 is 2.51 (m, 1H), 2,46-of 2.36 (m, 1H), 1,81-of 1.62(m, 2H), 1.60-to a 1.01 (m, 15 NM), 1,24 (t, J=7,4 Hz, MN), by 1.17 (s, 9H). MS: es+:751,1 (M+N)+, es-: 749,1-(M-N)-.

Example 42

Synthesis of compound No. 734

1H NMR (400 MHz, DMSO-d6): δ (part./million): to 8.62 (s, 1H), 8,54 (s, 1H), 8,04 (d, J=9,2 Hz, 1H), of 7.70 (s, 1H), 7,43 (s, 1H), 7,24 (d, J=2.6 Hz, 1H), 7,05-6,98 (m, 2H), 5,57 is 5.54 (m, 1H), 5,55-of 5.48 (m, 1H), 5,28-5,24 (m, 1H), 4.63 to-4,59 (m, 1H), 4,47-4,43 (m, 1H), 4,13-to 3.99 (m, 1H), 3,90 (s, 3H), 3,92-a 3.83 (m, 1H), 2,67 is 2.55 (m, 2H), 2,53 is 2.46 (m, 1H), 2,43-2,31 (m, 1H), 2,22-of 2.15 (m, 1H), 2,15 (3H), 1,75 is 1.70 (m, 2H), 1,51-of 1.42 (m, 2H), 1,41 of 1.28 (m, 6H), l,17(s,9H).

MC: 703,2 (M+1), 701,3 (M-1).

Example 43

Synthesis of compound No. 738

1H NMR (400 MHz, DMSO-d6): δ (part./million): 8,64 (d, J=2.5 Hz, 1H), to 8.62 (s, 1 H), of 8.04 (d, J=9,2 Hz, 1H), 7,39 (s, 1H), 7,24 (d, J=2.5 Hz, 1H),? 7.04 baby mortality (d, J=6.0 Hz, 1H), 6,99 (dd, J=2,2, and 9.2 Hz, 1H), gold 6.43 (d, J=2.2 Hz, 1H), 5.62-to 5.57 (m, 1H), 5,56-vs. 5.47 (m, 1H), 5,31-5,22 (m, 1H), 4,65-4,56 (m, 1H), 4,45 (dd, J=7,6, and 8.9 Hz, 1H), 4,07-4,00 (m, 1H), 3,90 (s, 3H), 3,88-a-3.84 (m, 1H), 2,68-of 2.56 (m, 2H), 2,54 is 2.43 (m, 1H), 2,42-2,31 (m, 1H), 2,34 (s, MN), 2,24 with 2.14 (m, 1H), 1,80-of 1.64 (m, 2H), 1,52 was 1.43 (m, 2H), 1,43-of 1.27 (m, 6H), of 1.18(s, N).

MS: 703,2 (M+1), 701,2 (M-1)

Example 44

Synthesis of compound No. 725

1H NMR (400 MHz, DMSO-d6): δ (part./million): to 8.62 (s, 1H), 8,10 (d, J=9,2 Hz, 1H), EUR 7.57 (s, 1H), 7,49 (s, 1H), 7,35 (d, J=2.2 Hz, 1H), to 7.09-7.03 is (m, 2H), 5,65-5,61 (m, 1H), 5,55-5,49 (m, 1H), 5,28-5,24 (m, 1H), 4,62-of 4.57 (m, 1H), 4,49-of 4.45 (m, 1H), 4,08-4,01 (m. 1H), 3,93 (s, 3H), 3,92-3,86 (m, 1H), 3,20-3,14 (m, 1H), 2,65-of 2.56 (s, 1H), 2,53-2,47 (m, 1H), 2,42 to 2.35 (m, 1H), 2,22-of 2.15 (m, 1H), 1,79 by 1.68 (m, 2H), 1,50 was 1.43 (m, 2H), 1,41 of 1.28 (m, 12H), of 1.18(s,9H).

MC: 748,2 (M+1), 746,2 (M-1).

Example 45

Synthesis of compound No. 726

1H NMR (400 MHz, DMSO-d6): δ (part./million): 8,64 (s, 1H), 8,10 (d, J=9.5 Hz, 1H), 7,83-7,76 (m, 2H), 7,60 (s, 1H), 7,44-7,42 (m, 1H), 7.18 in-7,01 (m, 2H), 5,56-5,49 (m, 2H), from 5.29-5,24 (m, 1H), 4,66-4,63 (m, 1H), 4,47 was 4.42 (m, 1H), 4,28 (s, MN), 4,06-was 4.02 (m, 2H), 3,94 (s, MN), 3,93-3,86 (m, 1H), 2,66 is 2.55 (m, 2H), 2,42-2,31 (m, 2H), 2,22 with 2.14 (m, 1H), 1,79-of 1.65 (m, 2H), 1,52-of 1.27 (m, 7H), to 1.22 (s, 9H).

MS:703,2(M+1),701,3(M-1).

Example 46

Synthesis of compound No. 906

1H NMR (400 MHz, DMSO-d6): δ (part./million): 8,46 (s, 1H), of 8.06 (d, J=9,2 Hz, 1H), EUR 7.57 (s, 1H), 7,49 (s, 1H), 7,34 (m, 1H), 7,14-7,05 (m, 2H), 5,63-to 5.58 (m, 1H), 4,66-br4.61 (m, 1H), 4,54-of 4.44 (m, 2H), 4,23-4,18 (m, 1H), 3,93 (s, 3H), 3,92-3,88 (m, 1H), 3,21-3,14 (m, I H), 2,44 is 2.33 (m, 1H), 1,35 (d, J=7 Hz, 6N), 1,73-a 1.01 (m, 26H)

MC: 762,0 (M+1), 759,9 (M-1).

Example 47

Synthesis of compound No. 907

1H NMR (400 MHz, DMSO-d6): δ (part./million): 8,46 (s, 1H), 7,98 (d, J-a 8.9 Hz, 1H), to $ 7.91-7,89 (m, 2H), 7.23 percent-7,21 (m, 2H), 7,07-7,00 (m, 2H), 6,35-6,32 (m, 2H), 5,64-to 5.58 (m, 1H), 4,65-br4.61 (m, 1H), 4.53-in-4,47 (m, 2H), 4,24-4,19 (m, 1H), 3,90 (s, 3H), 3,86-a-3.84 (m, 1H), 2.40 a is 2.33 (m, 1H), 1,73-a 1.01 (m, 26H).

MC: 702,0 (M+1), 699,9 (M-1).

Example 48

Fluorogenic full analysis heterodimeric protein NS3-NS4A

the 3’non-Coding region NS2-NS5B cloned with the help Of the RT-PCR into the vector pCR® 3 (firm Invitrogen), using RNA extracted from the serum of a person infected with HCV genotype 1b (provided by Others. Bernard Willems, Hopital St-Luc, Montreal, Quebec, Canada). Then NS3-NS4A-o6nacTb (NS3-NS4AFL) was subclinically with PCR expression baseconverter vector pFastBac™HTa (firm Gibco/BRL). The sequence of vector includes the region encoding consisting of 28 residues N-terminal sequence that contains a tag of six histidine residues. For the preparation of recombinant baculovirus used the baculovirus expression system You-to-You™ (firm Gibco/BRL). Protein His-NS3-NS4AFL expressed, infecting at 27°106cell line Sf21/MA recombinant baculovirus with the factor of infection of 0.1-0.2. When expression was authentic autoproteolysis, the result was the formation of non-covalent and stable protein complex NS3-NS4A (marked as "FL"). The infected culture was collected through 48-64 h by centrifugation at 4°C. Cellular debris homogenized in a mixture containing 50 mm NaPO4, pH 7.5, 40% glycerol (wt./vol.), 2mm / β-mercaptoethanol, in the presence of a mixture of protease inhibitors. Then His-NS3-NS4AFL were extracted from the cell lysate using a 1.5% NP-40, 0,5% Triton X-100, 0.5 M NaCl and processing Dnazol. After ultracentrifugation soluble extract was diluted 4 times and binding and with a Ni-chelating column type Pharmacia Hi-Trap. His-NS3-NS4AFL was suirable in shape, having a purity >90% (which is confirmed by using LTO-page)using a gradient of imidazole from 50 to 400 mm. His-NS3-NS4AFL kept at -80°in a mixture containing 50 mm sodium phosphate, pH 7.5, 10% (wt./about.) glycerol, 0.5 M NaCl, and 0.25 M imidazole, 0.1% of NP-40. Before use it was subjected to thawing on ice and diluted.

Protease activity of His-NS3-NS4AFL was evaluated in 50 mm Tris-HCl, pH 8.0, 0,25M the sodium citrate, 0.01 percent (wt./about.) n-dodecyl-β-D-maltoside, 1 mm TCEP. Incubated 5 m substrate Anthranilic-DDIVPAbu[C(O)-O]-AMY(3-NO2)TW-OH, which was terminated internal reaction, in the presence of various concentrations of inhibitor with 1,5M His-NS3-NS4AFL for 45 min at 23°C. the Final concentration of DMSO did not exceed 5.25 percent. The reaction was stopped by adding 1 M MES, pH of 5.8. The fluorescence of the N-terminal product was determined using fluorimetry type Perkin-Elmer LS-50B, equipped with a 96-well plate reader (wavelength of excitation: 325 nm; the wavelength of emission: 423 nm).

Inhibition (%) was determined from the following equation:

100-[(number of pulsesing.-number of pulsescistrons.)/(number of pulsescounter.-number of pulsescistron.100]

Data on the dependence of inhibition on the concentration was approximatively nonlinear curve using the model of hill and the magnitude of the effective concentration at which is occurs 50%inhibition (IC 50), was calculated using the software SAS (Statistical Software System; SAS Institute, Inc. Cary, N.C.).

Example 49

Radiometric analysis of the NS3 protease of HCV

The substrate, which is used for radiometric analysis of the NS3 protease of HCV, i.e. DDIVPC-SMSYTW, is cleaved by the enzyme between residues cysteine and serine. The sequence DIVPC-SMSYTW corresponds to the natural cleavage site NS5A/NS5B, in which the cysteine residue in P2 is replaced by Proline. Peptide substrate DDIVPC-SMSYTW and label Biotin-DDIVPC-SMS[125I-Y]TW incubated with recombinantly NS3-protease in the presence of inhibitors or without them. Separating the substrate from the products was performed by adding covered Avidya agarose pellets to the mixture for analysis, followed by filtration. The amount of product SMS[125I-Y]TW detected in the filtrate (with inhibitor and without it), allows to calculate the percent conversion of the substrate and the percentage of inhibition.

A. Reagents

Tris and Tris-Hcl (high purity (UltraPure)) received from the company Life Technologies. Glycerin (UltraPure), MES and BSA were obtained from the company Sigma®. DEF received from the firm of Pierce, DMSO was obtained from the firm Aldrich® and NaOH from the company Anachemia®.

Buffer for analysis of: 50 mm Tris-HCl, pH 7.5, 30% (wt./about.) glycerol, 2% (wt./about.) CHAPS, 1 mg/ml BSA, 1 mm TCEP (TCEP was added immediately before use from a 1 M stock solution in water). Substrate: DDIVPC-SMSYTW to a final concentration of 25 μm (from a 2 mm stock solution in DMSO, stored -20°in order to avoid oxidation).

Label: restored monitorowanie substrate (Biotin-DDIVPC-SMS[125I-Y]TW (final concentration of ~1 nm).

NS3-protease type 1b HCV, the final concentration of 25 nm (from a stock solution containing 50 mm sodium phosphate, pH 7.5, 10% glycerol, 300 mm NaCl, 5 mm DTT, 0.01%of NP-40).

B. Protocol

The analysis was carried out in 96-well polypropylene plate. Each hole contained:

20 μl of substrate/label buffer for analysis;

10 µl ± inhibitor in 20% DMSO/buffer for analysis;

10 µl of the NS3-protease 1b.

On the same tablet for analysis of preparing pure control (without inhibitor and without enzyme) and control (without inhibitor).

The enzymatic reaction was initiated by adding the enzyme solution, and analyzed the mixture incubated for 60 min at 23°With careful stirring. Added 20 μl of 0,n. NaOH to stop the enzymatic reaction.

In the filtration tablet type Millipore® MADP N65 was added 20 μl covered Avidya agarose pellets (obtained from the firm of Pierce®). Mixture for analysis, which stopped the enzymatic reaction, was transferred to a filtration tablet, and incubated for 60 min at 23°With careful stirring.

The tablets were filtered using the device for vacuum filtration type Millipore® MultiScreen Vacuum Manifold Filtraion and 40 μl of the filtrate was transferred to an opaque 96-well plate, containing 60 μl of scintillation fluid per well.

The radioactivity of the effluent was calculated using counter Packard® Top-Count, using a Protocol based on the use of125I-fluid for 1 minute

Inhibition (%) was calculated from the following equation:

100-[(number of pulsesing.-number of pulsescistron.)/(number of pulsesing.-number of pulsescistron.)×100]

Data on the dependence of inhibition on the concentration was approximatively nonlinear curve using the model of hill and the magnitude of the effective concentration at which 50%inhibition (IC50), was calculated using the software SAS (Statistical Software System; SAS Institute, Inc. Cary, N.C.).

Example 50

Determination of specific activity

The specific activity of the compounds was determined against various semiprotect: elastase of human leukocytes, elastase pancreatic pig and bovine α-disease of the pancreas and one cysteinate: cathepsin In human liver. In all cases used a Protocol based on the use of 96-well plates and are specific to each enzyme chromogenic substrate. Each analysis included a pre-incubation for 1 h enzyme-inhibitor at room tempera on who ur with the subsequent addition of substrate and hydrolysis to ≈ 30% conversion, which was estimated using the tablet reader for tetrazinni microplate type UV Thermomax® or fluorescent tablet reader type Perkin-Elmer® LS50B. The concentration of the substrate was maintained at the lowest possible level in comparison with KMin order to reduce competition of the substrate. Concentration of compounds varied from 300 to 0.06 μm, depending on their activity. The final terms of each analysis were as follows:

50 mm Tris-Hcl pH 8, 0.5m PA2SO450 mm NaCl, 0.1 mm etc, 3% DMSO, 0.01% tween-20 with

[100 μm Succ-AAPF-pNA and PM α-chymotrypsin], [133 μm Succ-AAA-pNA and 8 nm elastase pigs], [133 μm Succ-AAV-pNA and 8 nm of leukocyte elastase], or

[100 mm NaHPO4pH 6, 0.1 mm etc, 3% DMSO, 1 mm TSAF, 0.01% tween-20, 4 μm Z-FR-AMC (7-amino-4-methylcoumarin) and 0.5 nm cathepsin (original enzyme activated before use in buffer containing 20 mm TSEP)].

Below as a typical example of a generalized conditions for elastase pancreatic pigs:

In flat-bottomed polystyrene 96-well plate (type Cellwells, firm Corning) was added using a hand-held intake for liquids type Biomek (firm Beckman):

40 μl of buffer for analysis (50 mm Tris-HCl, pH 8, 1 M Na2SO450 mm NaCl, 0.1 mm etc);

20 μl of an enzyme solution (50 mm Tris-HCl, pH 8, 50 mm NaCl, 0.1 mm etc, 0.02% tween-20, 40 nm elastase pancreatic pigs); and

20 μl of a solution and is hibitor (50 mm Tris-HCl, pH 8, 50 mm NaCl, 0.1 mm etc, 0.02% tween-20, 1.5 mm to 0.3 μm inhibitor, 15% vol. DMSO).

After pre-incubation for 60 min at room temperature, each well was added 20 μl of substrate solution (50 mm Tris-HCl, pH 8,0,5 M Na2SO450 mm NaCl, 0.1 mm etc, 665 μm Succ-AAA-pNA) and the reaction mixture was additionally incubated for 60 min at room temperature, after which he determined the absorbance using a tablet reader type UV Thermomax®. The rows of holes left to the control variant (without inhibitor) and for pure control (without inhibitor and without enzyme).

Spent a serial twofold dilution of the solution of the inhibitor on a separate tablet using a handheld fluid intake, using 50 mm Tris-HCl, pH 8, 50 mm NaCl, 0.1 mm etc, 0.02% tween-20, about 15. % DMSO.

All experiments on determination of the specific activity carried out in the same way.

Inhibition (%) was determined from the following equation:

[1-((UVing.UVcistrons.)/(UVcounter.UVcistrons.))]×100

Data on the dependence of inhibition on the concentration was approximatively nonlinear curve using the model of hill and the magnitude of the effective concentration at which 50%inhibition (IC50), was calculated using the software SAS (Statistical Software System; SAS Institute, Inc. Cary, N.C.).

Example 51

This analysis was performed using a cell line Huh-7, which is a line of human cells derived from hepatoma, which was co-transfectional two DNA structures:

the first (labeled NS3), expressing the part of non-structural polyprotein HCV, merged with the tTA protein after the cleavage site NS5A-NS5B in the following order: NS3-NS4A-NS4B-NS5A-(NS5B)tTA, where (NS5B) denotes the first 6 amino acids NS5B.

This polyprotein Express under the control of the CMV promoter, the second (labeled SEAP)expressing the reporter protein secreting alkaline phosphatase (SEAP) under the control of sensitive to tTA promoter.

The first design allows for the expression of polyprotein, which after cleavage of the NS3-protease released different Mature proteins. Probably the Mature viral proteins form a complex on the membrane of the endoplasmic reticulum. tTA is a fused protein, described by Gossen and Bujard (Proc. Natl. Acad. Sci. USA, 89 (1992): 5547-5551), which contains the DNA-binding domain and activator of transcription. The release of protein tTA requires dependent NS3 cleavage site cleavage NS5A-NS5B between NS5A and to themselves. This is the last splitting gives tTA able to migrate into the nucleus and Tran-aktivirovat SEAP gene. Thus, the recovery of proteolytic activity of NS3 to what should be cause to limit the scope of finding tTA cytoplasm and contribute to the reduction of SEAP activity.

For evaluation at the cellular level, other types of active connections, other than the inhibition of NS3 protease, was carried out parallel joint transfection of a construct expressing only the tTA, and the same reporter construct, in which the SEAP activity was not dependent protease NS3.

Protocol analysis: cell line Huh-7 grown in CHO-SFMII (firm Life Technologies) + 10% FCS (fetal calf serum), was co-transfectional two structures of DNA taken in the following proportions:

7 μg NS3 + 500 ng SEAP + 800 ál FuGenel (firm Boehringer Mannheim) at 4×106cells Huh-7. After incubation for 5 h at 37° C cells were washed, treated with trypsin and the sown (80,000 cells/well) in 96-well plates, containing various concentrations of test compounds. After 24-hour incubation period, the SEAP activity in the medium was determined using a set of Phospha-Light, Tropix).

Estimated percentage of inhibition of SEAP activity depending on the concentration of the compound using the software SAS, receiving the value of the IC50.

The connection table

The following table lists representative compounds according to the invention. Found that all compounds are presented in tables 1-9, possess enzymatic activity identified using the analysis described in example 48. The number marked with an asterisk () indicates the enzymatic activity, which was determined using a radiometric analysis described in example 49, to which the IC50below about 50 microns. In these enzymatic assays using the following grading: A≥1 μm; 1 μm>>0.1 µm, and With≤0.1 ám.

Some compounds were tested for their specific activity using the analysis described in example 50, and it was found that they have specificity against NS3 protease. In General, the results of analyses to determine the specificity allow to establish that the studied compounds have the following activity: HLE>300 μm; re>300 μm; α-Chym.>300 μm; Cat.>300 μm, which suggests that they have a high specificity against NS3 protease and you should expect a lack of serious side effects.

In addition, some of these compounds were tested at the cellular level in the experiment described in example 51, and they were found to have activity, with the value of the EU50was less than 10 μm, which very likely suggests that these compounds can cross the cell membrane. In particular, the compounds of tables 7, 8 and 9 were tested at the cellular level and data about their activity are summarized in the last column. The results obtained at the cellular level, use the trace is the following grades: A> 1 μm;≤1 μm. In the following tables have used the following abbreviations:

MS: mass spectrometry using electrospray; the value of m/z represents MN4except as indicated by an asterisk (*) to m/z represents MN-; AC: acetyl; WP: benzyl; BOC: tert-butoxycarbonyl Ph: phenyl: RG: propyl.

Table 1

Conn. No.The double bondStereochemical structure connection D-R1R22MSEnzymatic activity
10112,13-TRANS1R, D in SYN orientation with respect to the amidePhenyl685,8A*
102No1r, d in SYN orientation with respect to acidPhenyl687,2
103No1R D in SYN orientation with respect to the amidePhenyl687,2A*

Table 2

Conn No.R3R4The double bond Stereochemical structure connection D-R1R21R22MSFermat.
202NH-BocH11,12-TRANS1R or 1S, D in SYN orientation with respect to acidNN593,7In
203NH-acetylH11,12-TRANS1R or 1S, D in SYN orientation with respect to acidNN535,6And
205NH-Boc11-OH

12-OH CIS
No1R or 1S, D in SYN orientation with respect to acidNN627,7In
206NH-BocH13,14-CIS1R, D in SYN orientation with respect to acidNN593,7
207NH-BocH13,14-num1R, D SYN in SYN orientation with respect to acidOmeN623,7

699,8
208NH-BocN13,14-CIS1R, D SYN in SYN orientation with respect to acidOmePhenyl
209NH-C(O)-NH-tert-VIN13,14-CIS1R, D in SYN orientation with respect to acidOmePhenyl698,8
210NH-BocN13,14-CIS1S, D in SYN orientation with respect to acidOmePhenyl699,8A*
211NH2N13,14-CIS1R, D in SYN orientation with respect to acidOmePhenyl599,7
213HE (one isomer)N13,14-CIS1R, SYN orientation with respect to acidOmeN524,6In
214NH-Boc10-oxo13,14-CIS1R, D in SYN orientation with respect to acidOmePhenyl713,8
215NH-BocNNo1R, D in SYN orientation with respect to acidOmePhenylof 701.8
217NH-Boc10-HE (a mixture of diastereoisomers)13,14-CIS1R, D in SYN-or the orientation towards acid OmePhenyl715,8
218NH-Boc10-oxo13,14-CIS1R, D in SYN orientation with respect to the amideOmePhenyl713,8
219NH-AcNNo1R,D in SYN orientation with respect to acidOmePhenyl643,2
220NH-BocN13,14-CIS1R, D in SYN orientation with respect to the amideOMe706,2

1. The compound of formula (I)

where

W denotes N

R21denotes H, C1-C6alkoxy, hydroxy, or N(C1-C6alkyl)2;

R22denotes H, C1-C6alkyl, CF3With1-C6alkoxy, C2-C7alkoxyalkyl,6is aryl or Het,

where Het represents a five - or six-membered saturated or unsaturated heterocycle, containing two or three heteroatoms selected from nitrogen, oxygen and sulfur, with the specified Het substituted by the radical R24where

R24denotes H, C1-C6alkyl, C1-C6alkoxy-NH-C(O)-R26, -OR26-The other26, -NHC(O)-NH-R26, -NHC(O)-OR26and R26,

where R26denotes hydrogen, C1-C6alkyl;

R3denotes hydroxy, NH2or a group of the formula-NH-R31where

R31denotes-C(O)-R32, -C(O)-other32or-C(O)-OR32where

R32stands With1-C6alkyl or C3-C6cycloalkyl,

D denotes consisting of 5-10 atoms, saturated or unsaturated alkylenes chain, optionally comprising one to three heteroatoms, independently from each other selected from O, S, or N-R41where

R41denotes H, C1-C6alkyl, -C(O)-R42where R42stands With1-C6alkyl, C6-aryl;

R4represents H or one to three substituent on any carbon atom of the chain D, where the substituents independently of one another selected from the group include1-C6alkyl, hydroxyl or oxo;

And denotes a carboxylic acid or its complex alkalemia esters.

2. The compound of formula (I) according to claim 1, where the fragment R1choose from 2 different diastereoisomers having structure (i) and (ii)

thus D is a SYN orientation with respect to the amide (i) or D is a SYN orientation with respect to And (ii).

3. The compound of formula (I) according to claim 2, where D is attached in the SYN-orientation to a, as represented by the structural formula (ii).

4. The compound of formula (I) according to claim 1,

where

W represents N;

R21denotes H, C1-C6alkoxy, hydroxy, or N(C1-C6alkyl)2;

R22denotes H, C1-C6alkyl, C1-C6alkoxy, phenyl or Het selected from the group including

where R24denotes H or a group NH-C(O)-OR26where R26means

With1-C6alkyl.

5. The compound of formula (I) according to claim 4, where

R21denotes N or C1-C6alkoxy.

6. The compound of formula (I) according to claim 4, where R22stands With1-C4alkoxy, phenyl or Het selected from the group including

,

where R24denotes H, C1-C6alkyl, NH-R26, NH-C(O)-R26or

NH-C(O)-OR26where R26matter specified in paragraph 4.

7. The compound of formula (I) according to claim 5, where R21denotes methoxy.

8. The compound of formula (I) according to claim 6, where R22indicates ethoxy or Het selected from the group including

,

or R24adenotes NH-C(O)-OR26where R26With1-C6alkyl, and R24bdenotes N or C1-C6alkyl.

9. The compound of formula (I) according to claim 1, where R3denotes an amide of the formula NH-C(O)R32or urea of the formula NH-C(O)-NH-R32or a carbamate of the formula NH-C(O)-OR32where R32stands With1-C6alkyl or C3-C6cycloalkyl.

10. The compound of formula (I) according to claim 9, where R3denotes the urea or carbamate, where R32stands With1-C6alkyl or C4-C6cycloalkyl.

11. The compound of formula (I) according to claim 10, where R3means carbamate, and R32represents tert-butyl, cyclobutyl or cyclopentyl.

12. The compound of formula (I) according to claim 1, where D denotes containing 6-8 atoms saturated or unsaturated alkylenes chain, optionally comprising one or two heteroatoms, independently from each other selected from O, S or N-R41/sup> where R41denotes H, C1-C6alkyl or C2-C7acyl.

13. The compound of formula (I) indicated in paragraph 12, where D is optionally contains one heteroatom selected from NH and N-(C2-C7)acyl.

14. The connection indicated in paragraph 13, where the heteroatom is selected from NH and N(Ac).

15. The connection indicated in paragraph 13, where the chain D contains 7 atoms.

16. The connection indicated in paragraph 15, where the heteroatom is in position 10 chain D.

17. The connection indicated in paragraph 13, where the chain D is saturated.

18. The compound of formula (I) indicated in paragraph 12, where D denotes containing 6-8 atoms saturated or unsaturated alkylenes chain, optionally containing one heteroatom selected from O or S.

19. Connection p, where the chain D contains 7 atoms.

20. The connection according to claim 19, where the heteroatom is in position 9 of the chain D.

21. Connection claim 20, where the chain D is substituted at position 8 the radical R4where R4denotes N or C1-C6alkyl.

22. Connection item 21, where the radical R4denotes H or methyl.

23. Connection p.22, where the radical R4denotes H or 8-(S)-IU.

24. Connection item 23, where the chain D is saturated.

25. The connection according to claim 19, where the chain D contains one double bond between positions 11 and 12.

26. Connection A.25, where the double bond is in the TRANS orientation.

27. The compound of formula (I) indicated in paragraph 12, where D denotes a saturated or unsaturated Ala the Lenovo chain, containing 6-8 atoms, all of which are carbon atoms.

28. The compound of formula (I) according to item 27, where D denotes a circuit containing 7 atoms.

29. The compound of formula (I) p, where the chain D is saturated.

30. The connection clause 29, where the chain D is substituted by the radical R4where R4denotes H, hydroxyl, oxo or alkyl.

31. Connection item 30, where the radical R4denotes N or C1-C6alkyl.

32. Connection p, where the radical R4denotes H or methyl.

33. Connection p, where the radical R4denotes H or 10-(S)-IU.

34. The compound of formula (I) p, where D contains one double bond.

35. The compound of formula (I) in clause 34, where the double bond is located between positions 13 and 14 of the circuit D.

36. The compound of formula (I) p, where the double bond is in a CIS-orientation.

37. Connection p, where the chain D is substituted by the radical R4where R4denotes H, hydroxyl or1-C6alkyl.

38. The connection clause 37, where R4denotes N or C1-C6alkyl.

39. Connection § 38, where R4denotes H or methyl.

40. Connection § 39, where R4denotes H or 10-(S)-IU.

41. The compound of formula (I) according to claim 1, where a denotes a carboxylic acid.

42. The compound according to claim 1, where

W denotes N

R3denotes a group of formula-NH-C(O)-other32or-NH-C(O)-OR 32where

R32stands With1-C4alkyl or C4-C6cycloalkyl,

D denotes consisting of 6-8 atoms saturated or unsaturated alkylenes chain attached to R1in a SYN orientation with respect to And, optionally incorporating one or two heteroatoms, independently from each other selected from O, S or N-R41where R41denotes N or C2-C7acyl,

R4represents H or one to three substituent selected independently from each other hydroxy or1-C6of alkyl, and

And denotes a carboxylic acid,

or its ester.

43. The compound of formula (I) according to § 42, where

R21denotes H or methoxy,

R22stands With1-C6alkoxy or Het selected from the group including

R24adenotes NH-C(O)-OR26where

R26stands With1-C6alkyl; and

R24bdenotes N or C1-C6alkyl;

R3denotes a urea of the formula N-C(O)-other32or a carbamate of the formula N-C(O)-OR32,

where R32stands With1-C6alkyl or C3-C6cycloalkyl;

D denotes contains 7 atoms alkylenes chain, optionally containing one double bond between what ojeniyi 11 and 12 or 13 and 14,

the chain D optionally includes one heteroatom independently selected from O, S, NH, N(Me) or N(Ac), and

R4denotes N or C1-C6alkyl.

44. The compound of formula (I) according to item 43, where R21denotes methoxy, and R22indicates ethoxy or

where R24adenotes NH(C1-C4)alkyl, NH-CO-(C1-C4alkyl), NH-C(O)-O-(C1-C4alkyl), NH-C(O)-NH-(C1-C4alkyl), and

D denotes a7chain, all atoms of which are carbon and which is saturated or contains one double CIS-relationship between the clauses 13 and 14.

45. The compound according to claim 1 of the formula

includes one stereoisomer R1where the double bond, the stereochemical structure of the connection D-R1and R22have the following meanings

Conn.No.Double bond:Stereochemical structure connection D-R1:R22:
10112,13-TRANS1R, D in SYN orientation with respect to the amidephenyl
102No1R, D in SYN orientation with respect to acidphenyl
103No1R D in SYN orientation with respect to the amide phenyl

46. The compound according to claim 1 of the formula

includes one stereoisomer R1where R3, R4the position of the double bond, the stereochemical structure of the connection D-R1, R21and R22have the following meanings:

Conn No.R3:R4:The double bondStereogum. the structure of the connection D-R1:R21:R22:
1234567
202NH-BOCN11,12-TRANS1R or 1S, D in SYN orientation with respect to acidNN
203NH-acetylN11,12-TRANS1R or 1S, D in SYN orientation with respect to acidNN
1234567
205NH-BOC11 HE 12 HE CISno1R or 1S, D in SYN orientation with respect to acidNN

NH-BOC
206N13,14-CIS1R, D in SYN orientation with respect to acidNN
207NH-BOCN13,14-CIS1R, D in SYN orientation with respect to acidOMeN
208NH-BOCN13,14-CIS1R, D in SYN orientation with respect to acidOMephenyl
209NH-C(O)-NH-tert-BuN13,14-CIS1R, D in SYN orientation with respect to acidOMephenyl
1234567
210NH-BOCN13,14-CIS1S, D in SYN orientation with respect to acidOMephenyl

211NH2N13,14-CIS1R, D in SYN orientation with respect to

acid
OMephenyl
213HE (one isomer)N13,14-CIS1R, D in SYN orientation with respect to acidOMeN
214 NH-BOC10-oxo13,14-CIS1R, D in SYN orientation with respect to acidOMephenyl
215NH-BOCNno1R, D in SYN orientation with respect to acidOMephenyl
217NH-BOC10-HE (a mixture of diastereoisomers)13,14-CIS1R, D in SYN orientation with respect to acidOMephenyl
1234567
218NH-BOC10-oxo13,14-CIS1R, D in SYN orientation with respect to the amideOMephenyl

219NH-ACNno1R, D in SYN orientation with respect to acidOMephenyl
220NH-BOCN13,14-CIS1R, D in SYN orientation with respect to the amideOMe

47. The compound according to claim 1 of the formula

includes one stereoisomer R1where R3D, CTE is eochemical structure connection D - R 1, R21and R22have the following meanings:

Conn. No.R3:-D-:Stereochemical structure connection D-R1R21:R22;
123456

301NH-BOC1R or 1S, D in SYN orientation with respect to acidNN
302NH-BOC1R, D in SYN orientation with respect to the amideOMephenyl
303NH-BOC1R, D in SYN orientation with respect to the amideOMephenyl
304NH-BOC1R, D in SYN orientation with respect to acidOMephenyl
305BUT1R, D in SYN orientation with respect to acidOMephenyl
123 456
306NH-BOC1R, D in SYN orientation with respect to the amideOMephenyl

307NH-BOC1R, D Shin-

orientation in relation to

acid
OMe
308NH-AC1R, D Shin-

orientation in relation to

acid
OMet.

48. The compound according to claim 1 of the formula

where the D-R1is a SYN orientation with respect to the acid, and R4, X9and the double bond 11, 12 have the following meanings:

Conn. No.R4:X9:The double bond

11, 12:
1234
401NCH2TRANS
402NCH2CIS
403NAbout TRANS

404AboutTRANS
405AboutTRANS
406NAboutno
407Aboutno
408Aboutno
409AboutCIS
410STRANS
411SCIS
4128-(Me)29-SCIS

49. The compound according to claim 1 of the formula

where the D-R1is a SYN orientation with respect to the acid, and X10, X11and X12have the following meanings:

Conn. No.X10:X11:X12:

1234
501CH2AboutCH2
502CH2CH2CH2
503CH2CH2NH
504CH2CH2N(Me)
505CH2CH2N(CO)Me
506CH2CH2N(CO)Ph
507NHCH2CH2
508N(CO)MeCH2CH2

50. The compound according to claim 1 of the formula

where the D-R1is a SYN orientation with respect to the acid, and R21and R22have the following meanings:

Conn.No.R21:R22:
601N(Me)2
602HE(CF3)

603OMe

51. The compound according to claim 1 of the formula

where the D-R1is a SYN orientation with respect to the acid, R4X9X10X11and the double bond between positions 13 and 14 and R22have the following meanings:

Conn.No.R4:X9X10or X11:The double bond 13, 14R22:
12345
701N11-OCISphenyl
702NCH2CIS
703NCH2no
704NCH2CIS

705NCH2CIS
12345
707NCH2qi is
708NCH2CIS
709NCH2no
710NCH2no
711NCH2no
712NCH2CIS-OEt
713NCH2no
714NCH2no-OEt
715NCH2CIS

716NCH2CIS
717NCH2CIS
12345
718NCH2CIS
719NCH2CIS
720NCH2no
721NCH2no
722NCH2CIS
723NCH2no

724NCH2no
725NCH2CIS
12345
726NCH2CIS
727NCH2CIS-CH2-OMe
728N CH2CISIU
729NCH2CIS
730NCH2no
731NCH2CIS
732NCH2CIS
733NCH2CIS

734NCH2CIS
735NCH2CIS
12345
736NCH2CIS
737NCH2CIS
738NCH2 CIS
73910-(R)MeCH2noPh
74010-(S)MeCH2noPh
741NCH2CIS

52. The compound according to claim 1 of the formula

where the D-R1is a SYN orientation with respect to the acid, the double bond between positions 13 and 14 is in a CIS-orientation, and R32, R4and R22have the following meanings:

Conn.No.R32:R4:R22:
1234
801N
803n-PrNOEt
804N
805N
806 NOEt
807NOEt

808HOEt
809H
810H
811H
1234
812H
813HOEt
814H
815H
816H

817H
818H
819H
820H
1234
821H
822H
823H
82410-(R)-IUOEt

53. The compound according to claim 1 of the formula

where the D-R1is a SYN orientation with respect to the acid, and R32, R4and R22have the following meanings:

Conn.No. 32:R4:R22:
1234
901NOEt
902N
903N
904N
905N

906N
907N
908N
909N
910 N
911N
912N
913N
914N
915N

91610(R)IUOEt

54. Pharmaceutical composition having activity against hepatitis C virus, including effective against hepatitis C virus, the amount of the compounds of formula (I) and po or its therapeutically acceptable salt, or a complex ester in a mixture with a pharmaceutically acceptable carrier or auxiliary substance.

55. The pharmaceutical composition according to item 54, additionally comprising an additional immunomodulator.

56. The pharmaceutical composition according to § 55, where additional immunodeficiency is modulator selected from the group includes α-, β- and δ-interferons.

57. The pharmaceutical composition according to item 54, additionally comprising an anti-virus agent.

58. The pharmaceutical composition according to § 57, where the antiviral agent is chosen from the group comprising ribavirin and amantadine.

59. The pharmaceutical composition according to item 54, further comprising the protease of hepatitis C virus (V-protease).

60. The pharmaceutical composition according p, where the inhibitor is chosen from the group including the helicase, polymerase and metalloprotease.

61. The pharmaceutical composition according to § 55, where the immunomodulator is a α-interferon.

62. The pharmaceutical composition according p, where the antiviral agent is ribavirin.

63. Connection p.43, where R3denotes NH-C(O)-OR32where R32stands With1-C4alkyl or C4-C6cycloalkyl; D represents a chain consisting of 7 carbon atoms, which is saturated or contains a double CIS-relationship between positions 13 and 14; and R22means

where R24Adenotes NH-(C1-C4alkyl); NH-C(O)- (C1-C4alkyl); NH-C(O)-O-(C1-C4alkyl); or NH-C(O)-NH-(C1-C4alkyl).

64. Connection 208 on p.46.

65. Connection 209 on p.46.

66. Connection 214 on p.46.

67. Connection 217 on p.46.

68. Connection 408 on p.

69. Connection 508 according to § 49.

70. Connection 601 according to item 50.

71. Connection 603 in item 50.

72. Connection 702 according to § 51.

73. Connection 703 according to § 51.

74. Connection 709 according to § 51.

75. Connection 714 according to § 51.

76. Connection 715 according to § 51.

77. Connection 719 in § 51.

78. Connection 725 in § 51.

79. Connection 736 in § 51.

80. Connection 738 in § 51.

81. Connection 801 in paragraph 52.

82. Connection 809 in paragraph 52.

83. Connection 810 in paragraph 52.

84. Connection 811 in paragraph 52.

85. Connection 812 in paragraph 52.

86. Connection 814 in paragraph 52.

87. Connection 818 in paragraph 52.

88. Connection 819 in paragraph 52.

89. Connection 821 in paragraph 52.

90. Connection 822 in paragraph 52.

91. Connection 823 in paragraph 52.

92. Connection 904 in item 53.

93. Connection 909 in item 53.

94. Connection 914 in item 53.

95. Connection 916 on item 53.

96. Pharmaceutical composition having activity against hepatitis C virus containing effective against hepatitis C virus, the amount of the compounds of formula (I) according to any one of p-95 or its therapeutically acceptable salt, or a complex ester in combination with a pharmaceutically acceptable carrier or auxiliary agent.

97. The pharmaceutical composition according p, optionally including one immunomodulator.

98. The pharmaceutical composition according p where additional immunomodulator selected from the group including α-, β- and δin aharony.

99. The pharmaceutical composition according p where additional immunomodulator is a α-interferon.

100. The pharmaceutical composition according p, including additional antiviral agent.

101. The pharmaceutical composition according to item 100, where the antiviral agent is ribavirin.

102. The pharmaceutical composition according p, including additional antiviral agent.

103. The pharmaceutical composition according to 102, where the antiviral agent is ribavirin.

104. The pharmaceutical composition according p, including additional antiviral agent.

105. The pharmaceutical composition according p, where the antiviral agent is ribavirin.

106. The pharmaceutical composition according p, further comprising another inhibitor V-protease.



 

Same patents:

FIELD: organic chemistry, medicine.

SUBSTANCE: invention represents ligands MC-4 and/or MC-3 of the formula (I): , wherein X means hydrogen atom, -OR1, -NR1R1' and -CHR1R1' wherein R1 and R1' are taken among the group: hydrogen atom, (C1-C6)-alkyl and acyl; (1) each R2 is taken independently among the group: hydrogen atom, (C1-C6)-alkyl; or (2) (a) R2 bound with carbon atom that is bound with X and Z1 and substitute R5 can be optionally bound to form carbocyclic or heterocyclic ring that is condensed with phenyl ring J; or (b) R2 bound with carbon atom that is bound with ring Ar can be bound with R7 to form ring condensed with ring Ar; each among Z1, Z2 and Z3 is taken independently from the following groups: -N(R3e)C(R3)(R3a)-, -C(R3)(R3a)N(R3e)-, -C(O)N(R3d)-, -N(R3d)C(O)-, -C(R3)(R3a)C(R3b)(R3c)-, -SO2N(R3d)- and -N(R3d)SO2- wherein each among R3, R3a, R3b and R3c, R3d, R3e when presents is taken independently among hydrogen atom and (C1-C6)-alkyl; p is a whole number from 0 to 5 wherein when p above 0 then R4 and R4' are taken among hydrogen atom, (C1-C6)-alkyl and aryl; R5 represents 5 substitutes in phenyl ring J wherein each R5 is taken among hydrogen atom, hydroxy-, halogen atom, thiol, -OR12, -N(R12)(R12'), (C1-C6)-alkyl, nitro-, aryl wherein R12 and R12' are taken among hydrogen atom and (C1-C6)-alkyl; or two substitutes R5 can be bound optionally to form carbocyclic or heterocyclic ring that is condensed with phenyl ring J; q = 0, 1, 2, 3, 4 or 5 wherein when q above 0 then R6 and R6' are taken among hydrogen atom and (C1-C6)-alkyl; Ar is taken among the group consisting of phenyl, thiophene, furan, oxazole, thiazole, pyrrole and pyridine; R7 are substitutes at ring Ar wherein each R7 is taken among hydrogen, halogen atom, -NR13R13', (C1-C6)-alkyl and nitro- wherein R13 and R13' are taken among hydrogen atom and (C1-C6)-alkyl; r is a whole number from 0 to 7 wherein when r is above 0 then R8 and R8' are taken among hydrogen atom and (C1-C6)-alkyl; B is taken among -N(R14)C(=NR15)NR16R17, -NR20R21, heteroaryl ring and heterocycloalkyl ring wherein R14-R17, R20 and R21 are taken independently among hydrogen atom and (C1-C6)-alkyl; s = 0, 1, 2, 3, 4 or 5 wherein when s is above 0 then R and R9' are taken among hydrogen atom and (C1-C6)-alkyl; R10 is taken among the group consisting of optionally substituted bicyclic aryl ring and optionally substituted bicyclic heteroaryl ring; D is taken among hydrogen atom, amino- and -C(O)R11 wherein R11 is taken among the following group: hydroxy-, alkoxy-, amino-, alkylamino-, -N(R19)CH2C(O)NH2 wherein R19 represents (C1-C6)-alkyl, -NHCH2CH2OH and -N(CH3)CH2CH2OH, or its isomers, salts, hydrates or biohydrolysable ester, amide or imide.

EFFECT: valuable medicinal properties of compounds.

18 cl, 107 ex

The invention relates to compounds of the prodrugs of inhibitors dipeptidylpeptidase IV (DP IV) the General formula a-b-C, and And denotes the amino acid refers to a chemical bond between a and C or the amino acid and stable inhibitor of DP IV with the missing C-terminal phosphonate residue, which represents AMINOETHYLPIPERAZINE, aminoacetanilide or N-dipeptidyl, O-arylhydroxylamine

The invention relates to compounds of formula (1), where X and Y Is N or O; R1substituted alkyl, substituted arylalkyl or cycloalkyl; R2and R3Is h or alkyl; And a Is-C(O)-, -OC(O)-, -S(O)2-; R4- alkyl, cycloalkyl or (C5-C12)aryl; compounds of the formula (2), where X and Y are O, S or N; R1- alkyl, optionally substituted arylalkyl; R2and R3Is h or alkyl;- C(O)-; R6- Deputy, including the condensed heterocyclic rings; and compounds of the formula (3), where X and Y are O, S or N; R1- alkyl, alkylsilane, (C5-C12)arylalkyl, (C5-C12)aryl; R2and R3Is h or alkyl; R2' and R3' - N; R11, R12and E together form a mono - or bicyclic ring which may contain heteroatoms

-interleukin" target="_blank">

The invention relates to compounds of General formulaand-

< / BR>
< / BR>
where n = 0, 1, or 2, m and m' = 1 or 2; R11is

< / BR>
or

< / BR>
R2'= R2= H, R3is-CH2Ar or 5-15 membered non-aromatic monocyclic group which may contain from 0 to 2 endocycles nitrogen atoms; R4is a branched (C1-5) alkyl group; R5choose from a group comprising-C(O)R7, -C(O)OR9, -C(O)C(O)R7; R7selected from the group: phenyl, naphthyl, isoquinoline, and phenyl may be substituted with halogen, (C1-6) alkoxy, 1,2-methylenedioxy or - N(H)C(O)(C1-6)-alkyl, R9independently selected from straight line (C1-5) alkyl group, optionally substituted by phenyl; R12and R13independently selected from the group comprising-R7-C(O)-R7and-C(O)-N(H)-R7or R12and R13together form a 4-8-membered saturated cyclic group, f is -interleukin (ICE), method of inhibiting ICE activity, methods of treating or preventing IL-mediated diseases

The invention relates to the compounds of formula I or formula II, where R1denotes N(R10)(R11); R2means thio-lower alkyl; each of R3and R5independently represents CH2or C(O); R4denotes a substituted or unsubstituted dionissia alkyl, where the Deputy is CH2NHC(O)R13and he added to the specified tighrope; R6denotes the residue synthetic heteroaromatic-amino acids; R7denotes a residue of natural or synthetic-amino acids; R8IT denotes or lower alkoxy, or together with R7forms homoserine; R9denotes H; each of R10and R11, independently, is H; R12denotes a substituted or unsubstituted fragment selected from aryl, allyssia of alkyl, where the substituents are one or more lower Akilov or halogen; R13denotes lower alkyl; R18denotes H; provided that if R4denotes unsubstituted dionissia alkyl, available tigraphy of R2and R4can form a disulfide bond; or pharmaceutically acceptable salts

The invention relates to new compounds of General formula 1: R1- SO2- B - X - Z - C(O) - Y, where R1represents a (1-12C)alkyl, which optionally may be substituted CF3, (7-15C)aralkyl or Campari; represents a bond, an amino acid of formula-NH-CH[(CH2)pC(O)OH]-C(O)-, where R = 1, 2, or 3, D-3-Tiq, or L - or D-amino acid containing a hydrophobic or neutral side chain; X represents an amino acid with a hydrophobic side chain, glutamine, cyclic amino, -NR2-CH2-C(O) -, or a group:

< / BR>
where n = 2, 3 or 4, W represents CH; R3represents H, (1-6C)alkyl; Z represents a lysine or 4-aminocyclohexanol; Y represents-NH-(1-6C)alkylene-C6H5, -OR4where R4represents H, (2-6C)alkyl, or NR5R6and R5and R6independently represent H, (1-6C)alkoxy or (1-6C)alkyl, optionally substituted with halogen, or R5and R6together represent a (3-6C)alkylene, or R5and R6together with the nitrogen atom to which they are attached, represent< / BR>
where V carts is naphthyl-SO2-Asp-Pro-Lys[COCO]-OH,having anticoagulant activity; and the pharmaceutical composition having inhibitory by combinationally

The invention relates to a series peptidergic heterocyclic compounds, intermediates used in their receiving and containing pharmaceutical compositions

The invention relates to new derivatives of Proline, and more specifically to individual forms new derivative of 1-substituted N-[2-methyl-1-(TRIFLUOROACETYL)- propyl]pyrrolidin-2-carboxamide, which are inhibitors of elastase of human leukocytes (ALC), also known as elastase human neutrophils (ANC), which are important, for example, as a means of research work in pharmacological, diagnostic and related studies and in the treatment of diseases of mammals, which also involved ALC

The invention relates to medicine, specifically to pharmacology

The invention relates to medicine, namely to the treatment of HIV-infected patients

The invention relates to the field of medicine and pharmaceutical industries and can be used in the creation, production and application of therapeutic and preventive drugs

The invention relates to medicine, dermatology, specifically to methods for treating genital genital condylomatosis
The invention relates to medicine, in particular as antiviral composition for the treatment of HIV-infected patients with high viral load

The invention relates to medicine, namely to pharmaceutical compositions based on oil phase, which comprises a pharmaceutically active agent connection pianola, which is the inhibitor of retroviral protease, a mixture of diglyceride and monoglyceride in a ratio of from about 9:1 to about 6:4 (diglyceride:monoglyceride), where diglyceride and monoglyceride are esters of mono - or dimensioned fatty acids and glycerol having a chain length of 16-22 carbon atoms, one or more pharmaceutically acceptable solvents and one or more pharmaceutically acceptable surfactants

The invention relates to veterinary

The invention relates to medicine, in particular to the pharmacology and treatment of infectious diseases
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