Enzyme-inhibiting compounds

FIELD: chemistry.

SUBSTANCE: invention relates to peptide-based compounds containing three-member rings containing a heteroatom, which efficiently and selectively inhibit specific activity of N-terminal nucleophilic (Ntn) hydrolase, bonded with a proteasome. The peptide-based compounds contain epoxide and are functionalised at the N-end.

EFFECT: peptide-based compounds exhibit anti-inflammatory properties and cell proliferation inhibition, oral administration of said peptide-based proteasome inhibitors is possible owing to bioavailability thereof.

23 cl, 14 ex

 

The prior art inventions

Degradation of proteins in eukaryotes is mainly mediated pathway of ubiquitin, in which proteins are targeted for degradation attached to the 76 amino acid polypeptide of ubiquitin. After targeting ubiquitinated proteins then serve as substrates for the 26S proteasome, a multicatalytic protease, which breaks proteins into short peptides by the action of its three major proteolytic activities. While having the main function involved in intracellular protein metabolism-mediated proteasome degradation also plays a key role in many processes, such as the inclusion of the main complex tissue compatibility (MHC) class I, apoptosis, cell division and NF-κB activation.

The 20S proteasome is a 700 kDa complex of multicatalytic protease cylindrical shape, composed of 28 subunits arranged in four cycles, which plays an important role in regulating cell growth, enabling the main complex tissue compatibility class I, apoptosis, processing of antigens, activation of NF-κB and transduction of Pro-inflammatory signals. In yeast and other eukaryotes 7 different α-subunits form the external cycles and 7 different β-subunits contain internal loops. the α-Subunit serve as a binder is x points for regulatory complexes 19S (PA700) and 11S (PA28), as well as as a physical barrier for the inner proteolytic chamber, formed by two cycles of β-subunits. Thus,in vivobelieve that the proteasome exists as 26S particle ("26S proteasome"). Experimentsin vivoshowed that inhibition shaped 20S proteasome can be easily correlated with the inhibition of the 26S-proteasome. Splitting aminobenzoic of proposedvalue β-subunits during the formation of particles affects aminobenzene threonine residues, which serve as the catalytic nucleophiles. Thus, the subunit responsible for the catalytic activity of the proteasome, have aminobenzamide nucleophilic residues, and these subunits belong to the group of N-terminal nucleophilic (Ntn) hydrolases (in which the N-terminal residue is, for example, Cys, Ser, Thr, and other nucleophilic members). This group includes, for example, acylase penicillin G (PGA), acylase penicillin V (PVA), amidotransferase (GAT) glutamine (PRPP) and bacterial glucosylceramidase. In addition to the ubiquitously expressed β-subunits, higher vertebrates also possess three β-subunits induced by γ-interferon (LMP7, LMP2 and MECL1), which replace their usual inclusion, X, Y, and Z, respectively, thus altering the catalytic activity is proteasome. By using different peptide substrates identified three main proteolytic activity of the 20S proteasome eukaryotes: the chymotrypsin-like activity (CT-L), which splits large hydrophobic residues; trypsin-like activity (T-L), which breaks down the basic residues; and peptidylglutamyl peptide gidrolizutaya activity (PGPH), which cleaves acid residues. The proteasome is credited with two additional less characteristic activity: BrAAP activity, which cleaves the branched chain amino acids; and SNAAP activity, which cleaves small neutral amino acids. The main proteolytic activity of the proteasome, apparently, achieved through a variety of catalytic points, because inhibitors, point mutations in the β-subunit exchange and β-subunits, including γ-interferon, in various degrees changes the activity.

In recent years, the proteasome becomes an attractive target for therapeutic intervention in cancer, immune and autoimmune diseases, inflammation, ischemic conditions, neurodegenerative disorders and other diseases. To date, FDA approved proteasome inhibitor is bortezomib (VELCADETM), but currently, clinical trials are several other is of proteasome inhibitors. While all of these therapeutic proteasome inhibitors currently injected IV (intravenous). Clinical use of proteasome inhibitors in the treatment of hematological malignancy, such as myeloma and lymphoma, partially limited by the need for frequent introduction IV and can be improved oral (PO) administration. However, due to the nature of these peptide molecules systemic effect following RO-introduction of said inhibitors is limited by several factors, including gastric pH, gastric and intestinal peptidases, aspirators, biliary excretion and intestinal and hepatic metabolic activity.

The methods used to overcome the ability of the peptides to enzymatic cleavage and improve absorption into the bloodstream from the digestive tract include analogues, which have a structure less like peptides, and are reduced in size. Such methods assume successful if the analogue peptide reaches a satisfactory level in the blood after oral administration, or, in the case of inhibitors of the proteasome, if proteasome activity in blood satisfactorily reduced.

The above technology is applicable to obtain analogues of the peptide-epoxyketone, inhibitors of the proteasome, providing them with oral bioavailability.

The essence of izobreteny the

The invention relates to the class of molecules known as α',β'-epoxy peptides and α',β'-aziridine peptides. Assume that the original molecules are effectively irreversibly and selectively contacted with the N-terminal nucleophilic (Ntn) hydrolases and can specifically inhibit the individual activity of enzymes with multiple catalytic activity.

In that case, if you think just to get rid of denaturirovannykh and necklacing proteins, the proteasome now recognize as a component of the proteolytic mechanism that regulates the level of various intracellular proteins by degradation of the signal-dependent manner. However, of great interest are identifying reagents that can specifically affect the activity of the proteasome and other Ntn-hydrolases and thus be applicable as samples to study the role of these enzymes in biological processes. Here described, synthesized and investigated compounds, which are aimed at Ntn-hydrolases. Proposed and declared peptide-epoxides and peptide-aziridine, which can effectively selectively and irreversibly inhibit the individual activity of the proteasome.

Unlike some other inhibitors on the basis of the peptides described herein peptide-epoxides and peptide-aziridine not involve a significant ing is berbania neproteinogennyh proteases, such as trypsin, chymotrypsin, cathepsin, papain and calpain, in concentrations up to 50 μm. At higher concentrations, inhibition can be observed, but rather as a competitive and nepobedimyy process, if the inhibitor only competes with the substrate. New peptide-epoxides and peptide-aziridine also involve the inhibition of activation of NF-κ and stabilization of p53 levels in cell culture. Moreover, it is assumed that these compounds have anti-inflammatory activity. Thus, these compounds may represent a unique molecular probes, which are universal to study the features of Ntn-enzyme in normal biological and pathological processes.

In one aspect the invention provides inhibitors, consisting of three-membered cycle containing a heteroatom. These inhibitors can inhibit the catalytic activity of the enzymes of the N-terminal nucleophilic hydrolases (for example, proteasome 20S or 26S proteasome when the inhibitor is present at a concentration below about 50 microns. Regarding the 20S proteasome these inhibitors hydrolases inhibit the chymotrypsin-like activity of the 20S proteasome, if inhibitor is present at a concentration below about 5 microns, and did not inhibit trypsin-like activity or PGPH activity of the 20S proteasome, if prists who meet at a concentration of below about 5 microns. Nucleoside hydrolases can represent, for example, peptide-α',β'-epoxyketone or peptido-α',β'-aziridine, and the peptide can be tetrapeptides. The peptide may include branched or unbranched side chains, such as hydrogen, C1-6-alkyl, C1-6-hydroxyalkyl,1-6-alkoxyalkyl, aryl, C1-6-aralkyl,1-6-alkylamide,1-6-alkylamino,1-6-carboxylic acid, With1-6-ether carboxylic acid, With1-6-alkylthiol or simple1-6-alkylthiomethyl, for example isobutyl, 1-naphthyl, phenylethyl and 2-phenylethyl. Α α'-carbon of α',β'-epoxyketone or α',β'-aziridination can be a chiral carbon atom, such as carbon (R) or β-configuration, as defined here.

In another aspect the invention provides pharmaceutical compositions including a pharmaceutically acceptable carrier and a pharmaceutically effective amount of an inhibitor of hydrolases, which weaken the symptoms of neurodegenerative diseases such as Alzheimer's disease), muscular dystrophy, cancer, chronic infectious diseases, fever, muscle atrophy, denervation, nerve disorders, nerve damage, starvation and immune-related conditions, and other.

In another aspect, the invention provides compounds and pharmaceutical compositions that bid the vent oral.

In another aspect the invention provides an anti-inflammatory composition.

In another aspect, the invention provides techniques for: inhibiting or reducing HIV infection in a subject, the impact on the level of gene expression of the virus in the subject; changes in the number of antigenic peptides produced by the proteasome in the body; determine whether regulated cellular, evolutionary or physiological process or product in the body proteolytic activity specific Ntn-hydrolases; treatment of Alzheimer's disease in a subject; reducing the rate of muscle protein degradation; reduce the rate of degradation of intracellular protein in a cell; reducing the rate of degradation of p53 protein in the cell; inhibiting the growth of malignant tumors, related to p53, the subject; inhibiting inclusion antigen in cells; suppression of the immune system of the subject; inhibiting the degradation of IκB-α in the body; reducing the content of NF-κB in a cell, muscle, organ or subject; the effect on cyclin-dependent cycles of eukaryotic cells; treatment of proliferative diseases in a subject; effects on proteasome-dependent regulation of oncoproteins in the cell; treatment of growth of malignant tumors in a subject; treating the subject of apoptosis related p53; and screening of proteins, produc the generated N-terminal nucleophilic hydrolases. Each of these methods provides for the introduction or contacting an effective amount of the composition containing the proposed inhibitors hydrolases, subject, cell, tissue, organ or organism.

Other features and advantages of the invention will be understood from the subsequent detailed description and from the claims.

Detailed description of the invention

The invention includes compounds used as inhibitors of enzymes. These compounds generally applicable to the inhibition of enzymes containing a nucleophilic group at the N-Terminus. For example, the activity of enzymes or subunits of enzymes containing in the side chain of the N-terminal nucleophilic amino group, such as (group) threonine, serine, or cysteine, can be successfully ingibirovany described here inhibitors of enzymes. The activity of enzymes or subunits of enzymes containing diaminotoluene nucleophilic group at the N-Terminus, such as, for example, a protective group or a carbohydrate group, can also be successfully ingibirovany described here inhibitors of enzymes.

Outside the context of a specific theory of action believe that these N-terminal nucleophiles Ntn form adducts with epoxy functional group described here inhibitors of enzymes. For example, suppose that in the β5 subunit/re2 proteasome 20S N-terminal threonine irreversibly forms of morpholino or piperazine derivatives - adduct as a result of interaction with the peptide-resin or peptide-aziridines, such as described below. The formation of this adduct is a splitting to break the cycle of epoxide or aziridine.

Options for implementation, including these groups associated with the α'-carbon stereochemistry of the α'-carbon (carbon, forming part epoxy or aziridine cycle) can be (R) or (S). The invention is based in part on the structural and functional information, which proves the following preferred stereochemical relationships. It is noted that the preferred connection can have multiple stereocentres marked top-bottom (or β-α, where β, as used here, indicates (location) above the plane (molecules)), or the ratio of (R)-(S) (i.e. it is not required that each stereocenter compounds corresponded to the preferred state). In some preferred embodiments, the implementation of the stereochemistry of the α'-carbon is (R), then there is an atom X is β, or (immediately) above the plane of the molecule.

From the point of view of stereochemistry to determine the absolute stereochemistry follow the rules of Cahn-Ingold-Prelog (Cahn-Ingold-Prelog). These rules are described, for example, inOrganic Chemistry,Fox and Whitesell; Jones and Bartlett Publishers, Boston, MA (1994); section 5-6, pages 177-178, and section 5-6 are shown as links. The peptides can have a repetitive structure of the main C the PI with the side chains extending from the links of the main chain. Typically, each link in the main chain contains an associated side chain, although in some cases the side chain represents a hydrogen atom. In other embodiments, the implementation not every link in the main chain contains an associated side chain. Peptides that are applicable to peptide-epoxides or peptide-aziridine have two or more link in the main chain. In some embodiments, implementation, applicable for inhibition of chymotrypsin-like (CT-L) activity of the proteasome, are from two to four links of the main chain, and in some preferred embodiments, the implementation for the inhibition of CT-L there are three main link chain.

Side chains extending from the links of the main chain may represent a side chain of natural aliphatic or aromatic amino acids, such as hydrogen (glycine), methyl (alanine), isopropyl (valine),second-butyl (isoleucine), isobutyl (leucine), phenylmethyl (phenylalanine), and a side chain comprising the amino acid Proline. Side chains can also be a branched or unbranched aliphatic or aromatic groups such as ethyl,npropyl,n-butyl,tert-butyl and aryl-substituted derivatives such as 1-phenylethyl, 2-phenylethyl, (1-naphthyl)methyl, (2-naphthyl)methyl, 1-(1-naphthyl)ethyl, 1-(2-naphthyl)ethyl 2-(1-naphthyl)ethyl, 2-(2-naphthyl)ethyl and the like compounds. Aryl groups can be optionally substituted branched or unbranched1-6-alkyl groups or substituted alkyl groups, acetyl and the like, or an additional aryl groups or substituted aryl groups such as benzoyl and the like. As substituents on the side chain can also be used heteroaryl and heterocyclyl group. Heteroaryl groups are nitrogen-, oxygen - and sulfur-containing aryl group, such as thienyl, benzothiazyl, naftotiekis, thianthrene, furyl, pyranyl, isobenzofuranyl, bromanil, pyrrolyl, imidazolyl, pyrazolyl, pyridyl, pyrazinyl, indolyl, purinol, chinosol and the like. Heterocyclyl groups are tetrahydrofuran, piperidine, piperazine, pyrrolidine, morpholine, lactones, lactams, and the like.

In some embodiments, implementation of the peptide-epoxides or peptide-aziridine can be introduced polar or charged residues. For example, can be entered amino acids of natural origin, such as hydroxycobalamine (Thr, Tyr, Ser) or sulfur-containing (Met, Cys), as well as non-essential amino acids, such as taurine, carnitine, citrulline, cystine, ornithine, norleucine and others. Unnatural substituents on the side chain with a charged or polar members could the t to represent such as, for example, With1-6is an alkyl chain or6-12-aryl group with one or more hydroxy-, short-chained alkoxy, sulfide, thio-, carboxyl, ester, phosphono-, amido or amino groups, or designated alternates, substituted by one or more halogen atoms. In some preferred embodiments, the implementation in the side chain of the peptide link is at least one aryl group.

In some embodiments, the implementation of the links of the main chain are amide units [-NH-CHR-C(=O)-], where R represents a side chain. This design does not exclude amino acids of natural origin, Proline, or other unnatural cyclic amino, which may be defined by experts in the field of technology.

In other embodiments, implementation of the links of the main chain are N-alkylated amide units (for example, N-methylated, and the like), analogs of olefins (in which one or more amide bonds substituted olefinic bonds), analogs of tetrazole (which tetrazolyl cycle uses a CIS-configuration of the main circuit), or a combination of these connections main circuit. In other embodiments, the implementation of the α-carbon of amino acid modified α-alkyl Deputy, for example aminoadamantane acid. In some the additional options exercise of the side chains locally modified for example, by ΔEor ΔZdegidro-modification, in which between the α - and β-atoms of the side chain double bond is present, or, for example, by ΔEor ΔZcyclopropane modification, in which between the α - and β-atoms of the side chain is present cyclopropyl group. In other additional embodiments, the implementation with the use of amino acid groups can be applied to D-amino acids. Additional options for implementation may include the cyclization of the side chains to the main chain, the formation of a disulfide bond, the formation of lactam, asosiasi and other modifications, discussed in “Peptides and Mimics, Design of Conformationally constraind by Hruby and Boteju, “Molecular Biology and Biotechnology: A Comprehensive Desk Reference” ed. Robert A. Meyers, VCH Publishers (1995), pp. 658-664, shown here as a reference.

One aspect of the invention relates to compounds of structure of formula (I) or their pharmaceutically acceptable salts:

in which

L is selected from C=O, C=S and SO2preferably the C=O;

X is selected from O, S, NH and N-C1-6-alkyl;

Z is absent, With1-6-alkyl or C1-6-alkoxygroup preferably absent;

each of R1, R2and R3independently from each other selected from hydrogen, C1-6-alkyl, C1-6-alkenyl,1-6-quinil,1-6-hydroxyalkyl, C1-6 -alkoxyalkyl, aryl, C1-6-aralkyl, heteroaryl, heterocyclyl,1-6-geterotsiklicheskie,1-6-heteroalkyl, carbocyclic and C1-6-carbonylmethyl;

R4selected from hydrogen, C1-6-aralkyl and C1-6of alkyl;

R5is heteroaryl; and

R6and R7independently from each other selected from hydrogen, C1-6-alkyl and C1-6-aralkyl.

In some embodiments, the implementation of R1, R2and R3independently from each other selected from hydrogen, C1-6-alkyl, C1-6-hydroxyalkyl,1-6-alkoxyalkyl,1-6-aralkyl,1-6-geterotsiklicheskie,1-6-heteroalkyl and C1-6-carbonylmethyl. In some embodiments, the exercise of any of R1, R2and R3independently from each other represents C1-6-alkyl selected from methyl, ethyl, propyl, isopropyl, butyl, sec-butyl and isobutyl. In some embodiments, the exercise of any of R1, R2and R3independently from each other represents C1-6-hydroxyalkyl. In some such embodiments, the exercise of any of R1, R2and R3independently from each other selected from hydroxymethyl and hydroxyethyl, preferably hydroxymethyl. In some embodiments, the exercise of any of R1, R2and R3nez the performance of each other represents C 1-6-alkoxyalkyl. In some such embodiments, the exercise of any of R1, R2and R3independently from each other selected from methoxymethyl and methoxyethyl, preferably methoxymethyl. In some embodiments, the exercise of any of R1, R2and R3independently from each other represent a1-6-heteroalkyl. In some such embodiments, the exercise of any of R1, R2and R3independently from each other selected from imidazolidinyl, parasaissetia, triazolylmethyl and pyridylmethyl, preferably imidazol-4-ylmethyl, thiazole-4-ylmethyl, 2-pyridylmethyl, 3-pyridylmethyl or 4-pyridylmethyl. In some embodiments, the exercise of any of R1, R2and R3independently from each other represents C1-6-aralkyl. In some such embodiments, the exercise of any of R1, R2and R3independently from each other selected from phenylmethyl (benzyl) and phenylethyl, preferably phenylmethyl. In some embodiments, the exercise of any of R1, R2and R3independently from each other represents C1-6-carbocyclic. In some such embodiments, the exercise of any R1is cyclohexylmethyl. In some embodiments, the implementation of all R1, R2and R3different. In some embodiments, the implementation of the NY two of R 1, R2and R3the same. In some embodiments, the implementation of all R1, R2and R3the same.

In some embodiments, the implementation of at least one of R1and R2selected from C1-6-hydroxyalkyl and C1-6-alkoxyalkyl. In some such embodiments, the implementation of at least one of R1and R2is alkoxyalkyl. In some such embodiments, the implementation of at least one of R1and R2selected from methoxymethyl and methoxyethyl.

In some embodiments, the implementation of R3selected from C1-6-alkyl and C1-6-aralkyl, preferably1-6-alkyl. In some such embodiments, the implementation of R3selected from methyl, ethyl, isopropyl, sec-butyl and isobutyl. In some such embodiments, the implementation of R3represents isobutyl. In some alternative embodiments, the implementation of R3selected from phenylmethyl and phenylethyl, preferably phenylmethyl.

In some embodiments, the implementation of R4, R6and R7independently from each other selected from hydrogen and methyl, and preferably represent hydrogen.

In some embodiments, the implementation of R5represents 5 - or 6-membered heteroaryl. In some such embodiments, the implementation of R5selected from isoxazol, izote the ash, furan, thiophene, oxazole, thiazole, pyrazole or imidazole, preferably from isoxazol, furan or thiazole.

In some embodiments, the implementation of R5is a bicyclic heteroaryl. In some such embodiments, the implementation of the bicyclic heteroaryl selected from benzisoxazole, benzoxazole, benzothiazole and benzisothiazole.

In some embodiments, the implementation of L represents C=O, Z is absent and R5represents isoxazol-3-yl or isoxazol-5-yl. In some preferred embodiments, such implementation, if isoxazol-3-yl substituted, it is substituted at least at the 5-position. In some preferred embodiments, the implementation, if isoxazol-5-yl substituted, it is substituted at least at the 3-position.

In some embodiments, the implementation of L represents C=O, Z is absent and R5represents unsubstituted isoxazol-3-yl.

In some embodiments, the implementation of L represents C=O, Z is absent and R5represents a substituted isoxazol-3-yl. In some such embodiments, the implementation of R5represents isoxazol-3-yl, substituted by the Deputy, is selected from C1-6-alkyl, C1-6-alkoxygroup,1-6-alkoxyalkyl,1-6-hydroxyalkyl, balance carboxylic acid, aminocarboxylate,1-6-alkylamino is carboxylate, (C1-6-alkyl)2aminocarboxylate,1-6alkylcarboxylic,1-6-heteroalkyl,1-6-aralkyl,1-6-geterotsiklicheskie and C1-6-carbonylmethyl. In some preferred embodiments, such implementation R5represents isoxazol-3-yl, substituted by the Deputy, is selected from methyl, ethyl, isopropyl and cyclopropylmethyl.

In some embodiments, the implementation of L represents C=O, Z is absent and R5represents isoxazol-3-yl, substituted 4 - to 6-membered nitrogen-containing1-6-heterocyclization. In some such embodiments, the implementation of R5represents isoxazol-3-yl, substituted by azetidinol, preferably azetidin-1-Iletisim. In some alternative embodiments, such implementation L represents C=O, Z is absent and R5represents isoxazol-3-yl, substituted radicalin which W represents O, NR, or CH2and R represents H or C1-6-alkyl. In some such embodiments, the implementation of W represents O.

In some embodiments, the implementation of L represents C=O, Z is absent and R5represents isoxazol-3-yl, substituted 5-membered nitrogen-containing1-6-heteroalkyl, such as parasailer, imidazoline is l, triazole-5-ylmethyl, preferably 1,2,4-triazole-5-ylmethyl.

In some embodiments, the implementation of L represents C=O, Z is absent and R5represents isoxazol-3-yl, substituted C1-6-alkoxygroup or1-6alkoxyalkyl, preferably a methoxy group, ethoxypropane, methoxymethyl or methoxyethyl.

In some embodiments, the implementation of L represents C=O, Z is absent and R5represents isoxazol-3-yl, substituted C1-6-hydroxyalkyl, preferably hydroxymethyl or hydroxyethyl.

In some embodiments, the implementation of L represents C=O, Z is absent and R5represents isoxazol-3-yl, substituted by a residue of carboxylic acid, aminocarboxylate,1-6-alkylaminocarbonyl, (C1-6-alkyl)2aminocarboxylate or1-6-alkylcarboxylic. In some such embodiments, the implementation of R5replaced by methylcarbazole or ethylcarboxylate, preferably by methylcarbazole.

In some embodiments, the implementation of L represents C=O, Z is absent and R5represents unsubstituted isoxazol-5-yl.

In some embodiments, the implementation of L represents C=O, Z is absent and R5represents a substituted isoxazol-5-yl. In some such embodiments, R 5represents isoxazol-5-yl, substituted by the Deputy, is selected from C1-6-alkyl, C1-6-alkoxygroup,1-6-alkoxyalkyl,1-6-hydroxyalkyl, balance carboxylic acid, aminocarboxylate,1-6he alkylaminocarbonyl, (C1-6-alkyl)2aminocarboxylate,1-6alkylcarboxylic,1-6-heteroalkyl,1-6-aralkyl,1-6-geterotsiklicheskie and C1-6-carbonylmethyl. In some preferred embodiments, such implementation R5represents isoxazol-3-yl, substituted by the Deputy, is selected from methyl, ethyl, isopropyl and cyclopropylmethyl.

In some embodiments, the implementation of L represents C=O, Z is absent and R5represents isoxazol-3-yl, substituted 4 - to 6-membered nitrogen-containing1-6-heterocyclization. In some such embodiments, the implementation of R5represents isoxazol-5-yl, substituted by azetidinol, preferably azetidin-1-Iletisim. In some alternative embodiments, such implementation L represents C=O, Z is absent and R5represents isoxazol-3-yl, substituted by a radical of the formulain which W represents O, NR, or CH2and R represents H or C1-6-alkyl. In some such embodiments, the OS is enforced W represents O.

In some embodiments, the implementation of L represents C=O, Z is absent and R5represents isoxazol-5-yl, substituted 5-membered nitrogen-containing1-6-heteroalkyl, such as parasailer, imidazoline, triazole-5-ylmethyl, preferably 1,2,4-triazole-5-ylmethyl.

In some embodiments, the implementation of L represents C=O, Z is absent and R5represents isoxazol-5-yl, substituted C1-6-alkoxygroup or1-6-alkoxyalkyl, preferably a methoxy group, ethoxypropane, methoxymethyl or methoxyethyl.

In some embodiments, the implementation of L represents C=O, Z is absent and R5represents isoxazol-5-yl, substituted C1-6-hydroxyalkyl, preferably hydroxymethyl or hydroxyethyl.

In some embodiments, the implementation of L represents C=O, Z is absent and R5represents isoxazol-3-yl, substituted by a residue of carboxylic acid, aminocarboxylate,1-6-alkylaminocarbonyl, (C1-6-alkyl)2aminocarboxylate or1-6-alkylcarboxylic. In some such embodiments, the implementation of R5replaced by methylcarbazole or ethylcarboxylate, preferably by methylcarbazole.

In some embodiments, the implementation of the compound of formula I selected from the

One aspect of the invention relates to a medical device comprising the composition described herein, which contains the inhibitor having the structure of formula I. In one of the embodiments the composition is administered in a medical device. In some embodiments, the medical device is a polymeric matrix containing a gel, or a ceramic matrix and the inhibitor. The specified polymer may be natural or synthetic. In some embodiments, the implementation of the specified gel acts as a depot medication, adhesive, weld, barrier, or seal.

Another aspect of the invention relates to a medical device containing a substrate having a surface on which is located the inhibitor having the structure of formula I. In one of the embodiments the inhibitor is placed directly on the medical device. In another embodiment, the coating is placed in a way, to coating consisting of a polymer matrix or ceramic matrix with an inhibitor having a structure of formula I, were dispersed or dissolved.

In one embodiment, the medical device is a coronary, cerebrovascular, or peripheral biliary stent. More specifically, the stent of the present invention is a expanding stent. In the case of the coating matrix containing the inhibitor having the structure of formula I, the matrix of elastic to fit to the compressed or expanded state of the specified expanding stent. In another embodiment of this invention, the stent has at least a portion which can be inserted or implanted into a patient's body, and the plot has a surface which is adapted to impact on the patient's body and in which at least part of the surface covered by the inhibitor having the structure of formula I, or floor consists of a matrix containing dispersed or dissolved inhibitor having the structure of formula I. an Example of a suitable stent is proposed in U.S. patent No. 4733665 provided here as a reference.

In another embodiment, a medical device of the present invention is a surgical instrument, such as a vascular implant, nutribase is a great device, surgical sealant or vascular substrate. More specifically, a medical device of the present invention is a catheter, the implantable access port of the vessel, a Central venous catheter, arterial catheter, a vascular implant, motricity balloon pump, seam, ventricular auxiliary pump, drug eluting barrier, adhesive, vascular Cape, extras/perissodactyl substrate, filter blood or filter adapted for placement in a blood vessel coated with an inhibitor having a structure of formula I, either directly or matrix containing the inhibitor having the structure of formula I.

In some embodiments, the implementation of the intraluminal medical device is coated with an inhibitor having a structure of formula I, or a coating consisting of a biologically tolerant matrix and inhibitor having the structure of formula I dispersed in the polymer, said device having an internal surface and an external surface that has a coating on at least part of the inner surface, outer surface, or both.

In some embodiments, the medical device can be used to prevent restenosis after angioplasty. The medical device may also be applicable for the treatment of various illness is evani and States implementing localized injection of the inhibitor, having the structure of formula I. These diseases and conditions include restenosis, inflammation, rheumatoid arthritis, tissue damage caused by inflammation, hyperproliferative disease, severe or arthritic psoriasis, muscular dystrophy, chronic infectious diseases, abnormal immune response, status, affecting sensitive blood cells, damage related ischemic conditions, and viral infection and proliferation. Examples of diseases and conditions that are treated with the use of medical devices of the present invention, a covered drug, are atherosclerosis, acute coronary syndrome, Alzheimer's disease, cancer, fever, muscle weakness (atrophy), denervation, occlusion of the vessel, sudden attack, HIV infection, nerve damage, kidney failure, associated with acidosis, and liver failure. See, e.g., Goldberg, U.S. patent No. 5340736.

The term "Cx-y-alkyl" refers to substituted or unsubstituted hydrocarbon groups, including alkyl groups with normal chain alkyl groups, branched chain, containing from x to y carbon atoms in the chain, including halogenoalkane groups such as trifluoromethyl and 2,2,2-triptorelin etc.0-alkyl means hydrogen, where the group is in the end position the Institute, if the connection is internal. The terms "C2-yalkenyl" and "C2-y-quinil" refers to substituted or unsubstituted unsaturated aliphatic groups analogous in length and possible substitution to the above alcelam, but containing at least one double or triple bond, respectively.

The term "alkoxygroup" refers to an alkyl group containing attached thereto an oxygen atom. Representative alkoxygroup include a methoxy group, ethoxypropan, propoxylate, tert-butoxypropan and the like. "Plain air" consists of two hydrocarbon covalently linked an oxygen atom. Accordingly, the Deputy of alkyl, which is a simple alkilany ether is or corresponds to alkoxygroup.

The term "C1-6-alkoxyalkyl" is a C1-6is an alkyl group substituted by alkoxygroup, forming a simple ether.

The term "C1-6-aralkyl", as used here, refers to C1-6is an alkyl group, substituted aryl group.

The terms "amine" and "amino" are known in the art and refer to both unsubstituted and substituted amines and their salts, such as members, which can be represented by the General formula:

in which each of R9, R10and R10'independently from each other represents hydrogen, alkyl, alkenyl, -(CH2)m0R8or R9and R10together with the N atom to which they are attached, form a heterocycle containing 4 to 8 atoms in the cycle; R8represents aryl, cycloalkyl, cycloalkenyl, heterocyclyl or polycyclic and m is zero or an integer from 1 to 8. In preferred embodiments, the implementation of only one of R9and R10can be a carbonyl, e.g., R9, R10and the nitrogen together do not form an imide. In an even more preferred embodiment, each of R9and R10(and optionally R10) independently of one another represent hydrogen, alkyl, alkenyl or -(CH2)m-R8. In some embodiments, implementation of the amino group is a primary, representing a protonated form with PKand≥7,00.

The term "amide" and "amido" is known in the art as aminosilanes carbonyl and includes a group that can be represented by the General formula:

in which R9and R10is as defined above. Preferred embodiments of the amide does not include imides which may be unstable.

The term "aryl", as used here, includes 5-, 6 - and 7-membered substituted or unsubstituted monocyclic aromatic group in which each atom of the CEC is and represents carbon. The term "aryl" also includes polycyclic ring systems containing two or more cycle in which two or more atoms) of carbon are common to two adjacent cycles in which at least one cycle is aromatic; for example, other cycles can be cycloalkyl, cycloalkenyl, cycloalkenyl, arily, heteroaryl and/or heterocyclyl. Aryl groups include benzene, naphthalene, phenanthrene, phenol, aniline and the like.

The terms "carbocycle" and "carbocyclic", as used here, refers to non-aromatic substituted or unsubstituted cycles, in which each atom of the cycle is the carbon. The terms "carbocycle" and "carbocyclic" also include polycyclic ring systems containing two or more cycle in which two or more carbons are common to two adjacent cycles in which at least one of the cycles is carbocycle, for example, other cycles can be cycloalkyl, cycloalkenyl, cycloalkenyl, arinami, heteroaryl and/or heterocyclyl.

The term "carbonyl" is known in the art and includes such groups, which can be represented by the General formula:

in which X represents a bond or (atom) oxygen or sulfur and R11is dorod, alkyl, alkenyl, -(CH2)m-R8or a pharmaceutically acceptable salt; R11'represents hydrogen, alkyl, alkenyl, -(CH2)m-R8where R8is as defined above. If X represents oxygen, and R11and R11'are not (atoms) of hydrogen, the formula represents an "ester". If X represents oxygen, and R11means hydrogen, the formula represents a "carboxylic acid".

As used here, "enzyme" can be any molecule that partially or fully reminiscent of protein, which provides the catalytic reaction path. Such enzymes can be natural enzymes fused enzymes, proenzymes, apparments, denaturirovannyj enzymes, farnesiani enzymes, ubiquitination enzymes, enzymes, acylated fatty acids, geranylgeranylpyrophosphate enzymes, enzymes associated GPI, enzymes associated with lipid, prenisolone enzymes, natural or artificial mutant enzymes, enzymes with modifications in the side or main chain enzymes with leading sequences and enzymes, complex with nebulophone substances such as proteoglycans, proteoliposome. Enzymes can be obtained by any means, including prirodno the expression, promoted expression, cloning, various syntheses of peptides in solution and in the solid phase and similar methods known to experts in this field of technology.

The term "C1-6-heteroalkyl", as used here, refers to C1-6is an alkyl group, substituted heteroaryl group.

The term "heteroaryl" includes substituted or unsubstituted 5 - to 7-membered cyclic structure, more preferably 5 - to 6-membered ring cyclic structures which include one to four heteroatoms. The term "heteroaryl" also includes polycyclic ring systems containing two or more cycle in which two or more atoms) of carbon are common to two adjacent cycles in which at least one cycle is heteroaromatic, for example, other cycles can be cycloalkyl, cycloalkenyl, cycloalkenyl, arily, heteroaryl and/or heterocyclyl. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, isoxazol, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazin, pyridazine and pyrimidine, and the like.

The term "heteroatom"as used here, means any element other than carbon and hydrogen. Preferred heteroatoms are nitrogen, oxygen, phosphorus and sulphur.

The terms "heteros who was cilil" or "heterocyclic group" refer to substituted or unsubstituted non-aromatic 3 - to 10-membered cyclic structures, more preferably 3 - to 7-membered cycles, cyclic structures which include one to four heteroatoms. The terms "heterocyclyl" or "heterocyclic group" also include polycyclic system containing two or more cycle in which two or more atoms) of carbon are common to two adjacent cycles in which at least one of the cycles is heterocyclic, for example, other cycles can be cycloalkyl, cycloalkenyl, cycloalkenyl, arinami, heteroaryl and/or heterocyclyl. Heterocyclic groups include, for example, tetrahydrofuran, piperidine, piperazine, pyrrolidine, morpholine, lactones, lactams, and the like.

The term "C1-6-heteroseksualci", as used here, refers to C1-6is an alkyl group, substituted heterocyclyl group.

The term "C1-6-hydroxyalkyl" is a C1-6is an alkyl group substituted by a hydroxy-group.

As used here, the term "inhibitor" is meant to describe a compound that blocks or reduces the activity of the enzyme (for example, inhibition of proteolytic cleavage of the standard fluorogenic peptide substrates, such as suc-LLVY-AMC, Box-LRR-AMC and Z-LLE-AMC, inhibition of the different catalytic activities of the proteasome 20S). The inhibitor can dei is painted with a competitive, unbeatable or uncompetitive inhibition. The inhibitor can bind reversibly or irreversibly and therefore the term includes compounds that are suicide substrates of the enzyme. The inhibitor may modify one or more points on or near the active point of the enzyme, or it may cause a conformational change in another place of the enzyme.

As used here, the term "orally bioavailable" is intended to describe the connection, put the mouse (number) 40 mg/kg or less, 20 mg/kg or less, or even 10 mg/kg or less that one hour after oral administration inhibits the activity of the proteasome CT-L in blood at least about 50%, at least about 75% or even at least about 90%.

As used here, the term "peptide" includes not only standard amido-linked (connection) with the standard α-substituents, but all applicable peptidomimetics with other modified linkages, the side chains of non-natural origin and modifications of the side chain, as detailed below.

The term "polycyclic" or "polycyclic" refers to from two to more cycles (for example, cycloalkyl, cycloalkenyl, cycloalkenyl, anilam, heteroaryl and/or heterocyclyl), in which two or more carbon atoms are on the common for two adjacent cycles, for example, the cycles are "condensed cycles". Every cycle polycycle may be substituted or unsubstituted.

The term "warning" is known in the art and as used by the state, such as a local recurrence (e.g., pain), a disease such as cancer, complex syndromes, such as heart failure or any other medical condition, is well understood in the art and involves the injection of a composition which reduces the frequency or deferreth the onset of symptoms of a medical condition in a subject relative to a subject who has not received the composition. Thus, prevention of cancer includes, for example, reducing the number of detectable cancerous tumors in the group of patients who received prophylactic treatment, relative to untreated control group and/or deferreth the appearance of detectable cancerous tumors in the treated group compared with the untreated control group, for example, when statistically and/or clinically significant amount. Prevention of infection includes, for example, reducing the number of diagnoses of infections in the treated group compared with the untreated control group and/or delay the onset of symptoms of infection in the treated group compared with the untreated control group. the warning pain includes, for example, reduction in force or, alternatively, delay pain attacks experienced by the subjects in the treated group compared with the untreated control group.

The term "prodrug" includes compounds that under physiological conditions become therapeutically active agents. A common method of making prodrugs is an introduction to selected groups that are hydrolyzed under physiological conditions to release the desired molecules. In other embodiments, implementation of the prodrug is converted by the action of an enzyme of an animal host.

The term "prophylactic or therapeutic treatment known in the art and includes an introduction to the owner of one or more appropriate compositions. If it lead to clinical manifestation of the unwanted condition (e.g., disease or other unwanted state of an animal host), then the treatment is prophylactic (i.e., it protects the host against developing the unwanted condition), whereas if it is administered after manifestation of the unwanted condition, the treatment is therapeutic (i.e., it expects to reduce, alleviate or stabilize the coming undesirable condition or its side effects).

The term "proteasome", as used here, refers to the inclusion of the immune and onstituting proteasome.

The term "substituted" refers to parts (molecules)containing substituents replacing a hydrogen on one or more carbon atoms of the main chain. It should be understood that "substitution" or "substituted" includes an implied condition that the said substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the result of the substitution is a stable compound, e.g., one that does not undergo spontaneous transformation, such as rearrangement, cyclization, elimination, etc. As used here, the term "substituted" is considering to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heteroclites, aromatic and nonaromatic substituents of organic compounds. For the corresponding organic compound can be one or more and same or different substituents. For the purposes of this invention, the heteroatoms such as nitrogen may have hydrogen substituents and/or any described herein permissible substituents of organic compounds which satisfy the valences of the heteroatoms. Substituents can be, for example, halogen, hydroxyl, a carbonyl (such as ka is boxel, alkoxycarbonyl, formyl or acyl), thiocarbonyl (such as a complex tiefer, thioacetal or thioformate), alkoxyl, phosphoryl, phosphate, phosphonate, phosphinate, amino group, aminogroup, amidinopropane, aminogroup, cyano, a nitro-group, asiagraph, sulfhydryl, allylthiourea, sulfate, sulfonate, sulfamoyl, sulfamidihappo, sulfonyl, heterocyclyl, aralkyl or aromatic or heteroaromatic group. Specialist in the art should understand that group, substituted on the hydrocarbon chain, themselves, if appropriate, can be substituted.

"Therapeutically effective amount" of the compounds in relation to this method of treatment refers to the amount of compound(s) in the product, if its introduction as part of a desired dosage regimen (to a mammal, preferably human) facilitated the symptom, facilitated condition, or slows the onset of disease conditions according to clinically acceptable standards for the disorder or condition that must be cured, or (achieved) cosmetic purposes, for example, when a reasonable relation to the benefits/risk inherent in any medical treatment.

The term "simple thioether" refers to an alkyl group as defined above containing attached thereto sulfur. In preferred embodiments, Khujand is to establish "a simple tiefer" represents an-S-alkyl. Representative simple thioester group include metalcorp, ethylthiourea and the like.

As used here, the term "treatment" or "therapy" includes a return to the former condition, the reduction or relief of symptoms, clinical presentation and underlying pathology of the condition thus to improve or stabilize the condition of the subject.

Selectivity for proteasome 20S

Offered here is a proteolytic enzyme applicable partly because they inhibit the activity of the 20S proteasome. Additionally, unlike other inhibitors of the 20S proteasome, the proposed connection of vysokoselektivnye against the 20S proteasome in comparison with other enzymes proteases. That is, these compounds exhibit selectivity for the 20S proteasome in excess of other proteases, such as cathepsins, calpain, papain, chymotrypsin, trypsin, tripeptidylpeptidase II. Selectivity of inhibitors of enzymes for the 20S proteasome is such that at a concentration of below about 50 microns inhibitors of the enzyme have shown inhibition of catalytic activity of the 20S proteasome, although not show inhibition of catalytic activity of other proteases, such as cathepsins, calpain, papain, chymotrypsin, trypsin, tripeptidylpeptidase II. In preferred embodiments implement ing bitory enzymes exhibit inhibition of catalytic activity of the 20S proteasome in a concentration of below about 10 microns, although not detect inhibition of catalytic activity of other proteases at the indicated concentrations. In even more preferred embodiments, the implementation enzyme inhibitors show inhibition of catalytic activity of the 20S proteasome concentration below about 1 μm, although not show inhibition of catalytic activity of other proteases at the indicated concentrations. Determination of the kinetics of the enzyme is proposed in patent application U.S. serial number 09/569748, example 2, and Stein and others,Biochem.(1996), 35, 3899-3908.

The selectivity of the chymotrypsin-like activity

Particular embodiments of compounds described herein, inhibiting the enzyme, optionally applicable due to their potential effectiveness and selectivity in the inhibition of the chymotrypsin-like activity of the 20S proteasome compared to the trypsin-like and PGPH activities. The chymotrypsin-like activity of the 20S proteasome is characterized by the cleavage of the peptides in the immediate vicinity of large hydrophobic residues. In particular, the chymotrypsin-like activity of Ntn-hydrolases can be determined by splitting the standard substrate. Examples of such substrates are well-known experts in this field. For example, can be used derived lateralisation. Determination of the kinetics of the enzyme is proposed is in the patent description U.S. serial number 09/569748, example 2, and Stein and others,Biochem.(1996), 35, 3899-3908.

The use of enzyme inhibitors

Biological consequences of inhibiting the proteasome numerous. Suggest that inhibition of the proteasome prevents and cures many diseases, including, but not limited to, proliferative diseases, neurotoxic/degenerative diseases, Alzheimer's disease, ischemic condition, inflammation, immune-related diseases, HIV, cancer, rejection of an organ transplant, septic shock, inhibition of the incorporation of the antigen, the decrease in expression of the viral gene, parasitic infections, conditions associated with acidosis, macular degeneration, pulmonary disease, muscular dystrophy, fibrotic diseases, bone diseases and disorders of hair growth. Therefore, the composition of the proteasome inhibitor, such as orally available class of molecules of peptide-epoxy-ketones, as described herein, provides a means to treat patients with these conditions.

The composition of the proteasome inhibitor can be used to treat conditions that are directly mediated proteolytic function of the proteasome, such as muscular dystrophy, or mediated indirectly via proteins which are processed by the proteasome, such as NF-κB. Particle of the proteasome with the rapid elimination of the post-translational processing of proteins (for example, enzymes that cause cell regulation (for example, cell cycle, gene transcription and metabolic pathways), intercellular communication and immune responses (e.g., presence of antigen). Specific examples discussed below include β-amyloid protein and regulatory proteins, such as cycline, and the transcription factor NF-κB.

At the cellular level after treatment of cells with different proteasome inhibitors have been described accumulation polyubiquitinated proteins, morphological changes of cells and apoptosis. Proteasome split many proteins in maturing reticulocytes and growing fibroblasts. In cells deprived of insulin or serous fluid, the rate of proteolysis is approximately doubled. Inhibition of the proteasome reduces proteolysis, thus reducing the loss of muscle protein, as well as the load of nitrogen to the kidneys or liver. One aspect of the invention relates to the treatment of cachexia and muscle dystrophy. Compounds of the invention can be used to treat conditions such as cancer, chronic infectious diseases, fever, paralysis of muscles (atrophy) and denervation, nerve damage, obesity, renal failure, associated with acidosis, and liver failure. See, e.g., Goldberg, U.S. patent No. 5340736. Therefore, some embodiments of breath is retene cover composition: decrease the rate of muscle protein degradation in a cell; reduce the speed of the degradation of intracellular protein; reducing the rate of degradation of p53 protein in the cell; inhibiting the growth of cancer related P52. Each of these methods involves contacting cells (in vivoorin vitrofor example, the muscles of the subject with an effective amount of a pharmaceutical composition containing proposed here inhibitor of the proteasome.

Intracellular proteolysis generates small peptides to provide in T-lymphocytes to induce immune response, mediated sit class I Immune system screens autologous cells that are infected by a virus or have undergone oncogenic transformation. In some embodiments implementing the invention relates to a method of inhibiting the incorporation of antigen into the cell, providing the impact on the cell described here are the links. The proteasome inhibitors of the invention can be used to treat conditions related to the immune system, such as allergies, asthma, rejection of organ/tissue (graft versus host) and autoimmune diseases, including, but not limited to, lupus, rheumatoid arthritis, psoriasis, multiple sclerosis and inflammatory bowel disease (such as ulcerative colitis and Crohn's disease). Thus, in some embodiments, implementation of the invention relative is seeking ways to suppress the immune system of a subject, providing an introduction to the subject effective amounts of compounds described herein, the inhibitor of the proteasome.

In some embodiments implementing the invention relates to a method of changing the set of antigenic peptides produced by the proteasome or other Ntn with multicatalytic activity. For example, if the PGPH activity of the 20S proteasome selectively ingibirovany, another set of antigenic peptides will be produced by the proteasome and presented in the molecules sit on the surface of cells, which will be produced and presented either in the absence of any inhibition of the enzyme, or, for example, selective inhibition of the chymotrypsin-like activity of the proteasome.

Another aspect of the invention refers to the use of the proposed compositions of the proteasome inhibitor for the treatment of neurodegenerative diseases and conditions, including, but not limited to, stroke, ischemic damage to the nervous system, nerve injury (e.g., percussive cerebral disorders, spinal cord injury, and traumatic damage to the nervous system), multiple sclerosis and other immune-mediated neuropathies (e.g., Guillain-Barre syndrome and its variants, acute motor Narita neuropathy, acute inflammatory demitrious polyneuropathy, and Fisher syndrome), comprehensive HIV / AIDS-dementia, axoneme, diabetic neuropathy, Parkinson's disease, Huntington's disease, multiple sclerosis, bacterial, parasitic, fungal and viral meningitis, encephalitis, vascular dementia, mixed infarction dementia, dementia of Lewy body dementia of the frontal lobe, such as the disease Peak, subcortical dementia (such as Huntington's or progressive supranuclear palsy), the syndrome of hereditary cortical atrophy (such as primary aphasia), metabolic-toxic dementia (such as chronic hypothyroidism or B12 deficiency and dementia caused by infections (such as syphilis or chronic meningitis).

Alzheimer's disease is characterized by extracellular deposits of β-amyloid protein (β-AR) in senile plaques and cerebral blood vessels. Β-AR represents a peptide fragment of 39 to 42 amino acids, derived from the amyloid precursor protein (APP). Known at least three isoforms of APP (695, 751 and 770 amino acids). The alternate connection mRNA generates isoforms; a normal process affects the part of the sequence of β-AR, preventing thus the generation of β-AR. Consider that abnormal activity of the proteasome in protein provides an abundance of β-AR in the brain with Alzheimer's disease. The enzyme, giving RDAs, rats contains about ten different subunits (22-CD). The 25 kDa subunit contains an N-terminal sequence of H-Gln-Asn-Pro-Met-X-Thr-Gly-Thr-Ser, which is identical to the β-subunit of excessive pain (makabali, macropain) person (Kojima, S. And others,Fed. Eur. Biochem. Soc.,(1992) 304:57-60). The enzyme that produces the RDA, breaks down communication Gln15-Lys16in the presence of calcium ion, the enzyme also cleaves the bond Met1-Asp1and communications Asp1-Ala2releasing the extracellular domain of β-AR.

Therefore, one aspect of the invention relates to a method of treatment of Alzheimer's disease, providing an introduction to the subject an effective amount of the proposed composition of the proteasome inhibitor. Said treatment includes a reduction in the rate of production of β-AR, reducing the rate of formation of plaques of β-AR, the reduction in the rate of generation of β-AR and the reduction of clinical symptoms of Alzheimer's disease.

Fibrosis represents an excessive and sustained formation of scar tissue in the hyperproliferative growth of fibroblasts and is associated with activation of TGF-β-signaling pathway. Fibrosis causes excessive deposition of extracellular matrix, and can occur in virtually any tissue or across several different tissues. Typically, the level of intracellular signaling protein (Smad), which activates the transcription of target genes after TGF-β stimulation, is regulated and the efficiency of the proteasome (Xu and others, 2000). However, in cancer and other hyperproliferative diseases observed accelerated degradation of TGF-β signaling components. Thus, in some embodiments implementing the invention relates to a method of treatment of hyperproliferative diseases, such as diabetic retinopathy, macular degeneration, diabetic nephropathy, glomerulosclerosis, IgA nephropathy, cirrhosis, atresia of bile ducts, congestive heart failure, scleroderma, fibrosis, induced by irradiation, and pulmonary fibrosis (idiopathic pneumosclerosis, collagen vascular disease, sarcoidosis, interstitial lung disease and exogenous pulmonary disease). Treatment of burn victims often prevents fibrosis, therefore, in some embodiments implementing the invention relates to local or systemic injection of inhibitors for the treatment of burns. Wound healing after surgery is often associated with disfiguring scars, which can be prevented by inhibition of fibrosis. Thus, in some embodiments implementing the invention relates to a method of preventing or reducing scarring.

Some inhibitors of the proteasome block as the degradation and processing ubiquitination NF-κBin vivoandin vitro.The proteasome inhibitors also block the degradation of IκBA and activation of NF-κB (Palombella and others, Cell(1994) 78:773-785; Traenckner, and others,EMBO J.(1994) 13:5433-5441). One aspect of the invention relates to a method for inhibiting the degradation of IκB-α, involving contacting a cell with connection described here. In some embodiments implementing the invention relates to a method for reducing the cellular content of NF-κB in a cell, muscle, organ or subject, involving contacting a cell, muscle, organ or entity described in this connection, an inhibitor of the proteasome.

NF-κB is a member of the Rel family proteins. Family Rel transcriptomic protein activators can be divided into two groups. The first group requires proteolytic processing and includes P50 (NF-κB1, 105 kDa) and P52 (NF-κ2, 100 kDa). The second group does not require proteolytic processing and includes P65 (RelA, Rel (c-Rel and RelB). Members of the Rel family can be formed both Homo - and heterodimer; NG-κB, for example, represents heterodimer road P59-R65. After phosphorylation and ubiquitination of IκB and R two proteins are degraded and treated with the formation of active NF-κB, which moves from the cytoplasm to the nucleus. Ubiquitination R also exposed purified proteasomes (Palombella and others,Cell(1994) 78:773-785). Active NF-κB forms a stereospecific complex gene-amp with other transcriptional activators and the example HMGI(Y)inducyruya selective expression of a specific gene.

NF-κB regulates the genes included in the immune and inflammatory response, and mitotic phenomena. For example, NF-κB is required for the expression of κ-light chain gene of the immunoglobulin gene of the α-chain of the receptor for IL-2, complex gene main biocompatibility of class I and a number of genes encoding cytokines, such as IL-2, IL-6, factors that stimulate granulocyte colony, and IFN-β (Palombella and others,Cell(1994) 78:773-785). In some embodiments implementing the invention relates to methods of influence on the levels of expression of IL-2, MHC-I, IL-6, TNFα, IFN-β or any of the other previously mentioned proteins, each method provides an introduction to the subject an effective amount of compositions described herein inhibitor of the proteasome. Complexes, including P50, are a quick mediators of acute inflammatory and immune responses (Thanos, D, and Maniatis, T.,Cell(1995) 80:529-532).

Excessive formation of lipopolysaccharide (LPS) - induced cytokines, such as TNFα, consider Central to the processes associated with septic shock. Moreover, it is usually assumed that the first stage in the activation of cells LPS involves the binding of LPS with specific membrane receptors. Subunit of the α - and β complex of the proteasome 20S identified as proteins that bind LPS, demonstrating that the signal transduction, inducir the bathroom LPS, may be an important therapeutic target in the treatment or prevention of sepsis (Qureshi, N. and others,J. Immun.(2003) 171:1515-1525). Therefore, in some embodiments, the implementation of the composition of the proteasome inhibitor can be used to inhibit TNFα c for the prevention and/or treatment of septic shock.

NF-κB is also involved in expresii cell adhesion genes that encodes E-selectin, P-selectin, ICAM, and VCAM-1 (Collins, T.,Lab.Invest.(1993) 68:499-508). In some embodiments implementing the invention relates to a method of inhibiting cell adhesion (e.g., cell adhesion mediated by E-selectin, P-selectin, ICAM or VCAM-1), involving contacting a cell with (or introduction to a subject) an effective amount of a pharmaceutical composition containing proposed here inhibitor of the proteasome.

NF-κB also specifically binds to the amplifier/promoter HIV. Compared to mac239 Nef regulatory protein of HIV Nef pbj14 differs by two amino acids in the region, which controls the binding of the protein kinase. Consider that the protein kinase enhances phosphorylation of IκB, causing degradation of IκB the ubiquitin-proteasome path. After degradation, NF-κB is released into the nucleus, inducing the transcription of HIV (Cohen, J.,Science(1995) 267:960). In some embodiments implementing the invention relates to a method of inhibiting or is Eisenia HIV infection in a subject or reducing the expression level of the gene of the virus, each method provides an introduction to the subject an effective amount of compositions described herein inhibitor of the proteasome.

Viral infections contribute to the pathology of many diseases. State of the heart, such as chronic myocarditis and dilated cardiomyopathy are associated with Coxsackieviruses (coxsackievirus) B3. In comparative genomic analyses microregions hearts of infected mice, which was chronic myocarditis, were uniformly adjusted subunit-specific proteasome (Szalay and others, Am. J. Patol. 168;1542-52, 2006). Some viruses use the ubiquitin-proteasome system at the stage of introduction of the virus, when the virus is released from the endosome into the cytosol. Virus mouse hepatitis (MHV) belongs to the family Coronaviridae, which also includes some coronaviruses acute respiratory syndrome (SARS). Yu and Lai (J. Virol. 79:644-648, 2005) showed that treatment of cells infected with MHV, a proteasome inhibitor, leads to reduction of virus replication, correlating with a reduced titer of the virus compared to untreated cells. Hepatitis b virus human (HBV), a member of the virus family Hepadnaviridae, similarly requires for reproduction proteins encoded by the virus shell. Inhibition route of degradation in the proteasome causes a reduction in the amount of secreted protein shell (Simsek the other, J. Virol. 79:12914-12920, 2005). In addition to HBV for secretion, morphogenesis and pathogenesis way ubiquitin-proteasome degradation can also use other hepatitis viruses (a, C, D and E). Accordingly, in some embodiments implementing the invention relates to a method of treating viral infections, such as SARS or hepatitis a, b, C, D and E, providing the contacting of the cells with (or introduction to a subject) an effective amount of the proposed connection.

Ischemia and reperfusion injury leads to hypoxia, a condition in which there is a deficiency of oxygen reaching the body tissues. The specified condition causes increased degradation of Iκ-Bα, thereby leading to activation of NF-κB (Koong and others, 1994). It has been shown that many damage caused by hypoxia can be reduced by introduction of an inhibitor of the proteasome (Gao and others, 2000; HLW and others, 2001; Pye and others, 2003). Therefore, some embodiments of the invention relates to a method of treating an ischemic condition or reperfusion damage, introducing the subject in case of need of such treatment an effective amount of the proposed connection, an inhibitor of the proteasome. Examples of such conditions or injuries include, but are not limited to, acute coronary syndrome (susceptible platelets), occlusive lesion and the criteria (heart, cerebral, peripheral artery and occlusion of the vessel), atherosclerosis (coronary sclerosis, coronary artery disease), heart attacks, heart failure, pancreatitis, myocardial hypertrophy, stenosis and restenosis.

Other transcription factors eukaryotes that require proteolytic processing, General transcription factor TFIIA, additional protein of herpes simplex VP16 (factor host cell), the factor 2 regulatory protein of the virus, inducing IFN, and the regulatory protein 1, binding element, membrane-bound Sterol.

In some embodiments implementing the invention relates to methods of excitation of cyclin-dependent cycles of the cells of eukaryotes, providing for action on the cells (in vitroorin vivo) the proposed composition of the proteasome inhibitor. Cycline represent proteins involved in cell cycle regulation. Proteasome involved in the degradation of tsiklonov. Examples of tsiklonov are mitotic cycline, G1-cycline and cyclin C. Degradation tsiklonov enables cells to go from one stage of the cell cycle (e.g., mitosis) and enter into another (e.g., division). Believe that all cycline associated with P34cdc2protein kinase or related kinases. Signal sighting proteolysis is aimed at amino acid 42-RAALGNISEN-50 (the side the destruction). Obviously, cyclin converted into a form that is unprotected from ubiquitinate, or that during mitosis is activated cyclin-specific ligase (Ciechanover, A., Cell (1994) 79:13-21). Inhibition of the proteasome inhibits the degradation cyclina and, consequently, inhibits cell proliferation, for example, in malignant tumors associated with cyclin (Kumatori, and others, Proc. Natl. Acad. Sci. USA (1990) 87:7071-7075). One aspect of the invention relates to a method of treatment of the subject has a proliferative disease (such as cancer, psoriasis or restenosis), providing an introduction to the subject an effective amount of the proposed composition of the proteasome inhibitor. The invention also relates to a method of treatment of a subject associated with cyclin inflammation, providing an introduction to the subject an effective amount of the proposed composition of the inhibitor of the proteasome.

Additional embodiments of the invention relate to methods of excitation-dependent proteasome regulation of oncoproteins and methods for treating or inhibiting the growth of malignant tumors; each method involves effects on the cell (in vivofor example, the subject has, orin vitrothe suggested composition of the proteasome inhibitor. Proteins of HPV-16 and HPV-18, derived E6, stimulate ATP - and ubiquitin-dependent conjugation and money is adalu p53 in untreated lysates reticulocytes. It is shown that recessive oncogenic p53 invalid when the temperature accumulates in cell lines with mutated thermolabile E1. Elevated levels of p53 leads to apoptosis. Examples of proto-oncoproteins, degraded by the ubiquitin system, include c-Mos, c-Fos and c-Jun. In some embodiments implementing the invention relates to methods of treatment of apoptosis-related p-53, providing an introduction to the subject an effective amount of the proposed composition of the inhibitor of the proteasome.

In some embodiments, implementation of the proposed composition can be used to treat parasitic infections, such as infections caused by protozoan parasites. Believe that the proteasome these parasites are involved mainly in the activity of differentiation and replication of cells (Paugam and others, Trends Parasitol. 2003, 19(2):55-59). Moreover, it is shown that the genus dysenteric amoeba loses the ability inzistiranja being treated with proteasome inhibitors (Gonzales and others, Arch. Med. Res. 1997, 28, Spec no. : 139-140). In some of these embodiments, the implementation of the protocols the introduction of the compositions of the inhibitor calpaine applicable for the treatment of human parasitic infections caused by protozoan parasites selected from the genus Plasmodium (including P. falciparum, P. vivax, P. malariae and P. ovale, which cause malaria), of the genus Trypanosoma (including T. ruzi, which calls the AET Chagas disease, and T. brucei, which causes African trypanosomiasis), of the genus Leishmania (including L. amazonesis, L. donovani, L. infantum, L. mexicana, and so on), Pneumocystis carinii pneumonia (simplest, as it is known, causes pneumonia (patients) AIDS and others with depressed immune system), Toxoplasma gondii, Entamoeba histolytica, Entamoeba invanens and Giardia lamblia. In some embodiments, implementation of the proposed composition of the proteasome inhibitor is applicable for the treatment of parasitic infections in animals and cattle caused by the protozoan parasites selected from Plasmodium hermani, Cryptosporidium sps., Echinococcus granulosus, Eimeria tenella, Sarcocystis neurona and Neurpspora crassa. Other compounds applicable as proteasome inhibitors in the treatment of parasitic diseases, described in the patent WO 98/10779 provided here as a reference.

In some embodiments, the implementation of the composition of the proteasome inhibitor inhibit proteasome activity in parasites without regeneration in the erythrocytes and leukocytes. In some such embodiments, the implementation of the long half-period of the existence of hemocytes can provide prolonged protection against recurrent actions of parasites. In some embodiments, the implementation of the composition of the proteasome inhibitor can provide prolonged chemoprophylaxis against future infections.

It is also shown that inhibitors that bind the proteasome 20S, stimulate images is the bone in cultures of bone bodies. Moreover, if these inhibitors are systematically injected mice, some proteasome inhibitors increase bone volume and rate of formation of bone by more than 70% (Garrett, I. R., and others,J. Clin. Invest.(2003) 111:1771-1782), proving that the ubiquitin-proteasome mechanism regulates the differentiation of osteoblasts and bone formation. Therefore, the proposed composition of the proteasome inhibitor can be applied in the treatment and/or prevention of diseases associated with bone loss such as osteoporosis.

Inhibition of the proteasome is already established as a therapeutic strategy for the treatment of cancer, in particular multiple myeloma. However, on the basis of both models,in vitroandin vivoyou can predict that it may serve as a strategy against the other (types of) cancer, in particular zlokacestvennosti associated with gem, and solid tumors. Therefore, some embodiments of the invention relate to a method of cancer treatment involving the administration to a subject, in need of such treatment, an effective amount of the proposed connection, an inhibitor of the proteasome.

Introduction

The composition obtained as described herein may be introduced in various forms, depending on the disorder to be treated, and the age, condition and body weight of the patient, is well known in medicine. For example, if the compound is administered orally, they can be made in the form of tablets, capsules, granules, powders or syrups; or for parenteral administration they may be made in the form of solution for injection (intravenous, intramuscular or subcutaneous), preparation for drip infusion or suppositories. For insertion through the eye mucous membrane they can be made in the form of eye drops or eye ointments. The above mentioned formulation can be obtained by conventional means, and if desired, the active ingredient can be mixed with any conventional additive or excipient such as a binder, dezintegriraat agent, a sliding agent, a corrective agent, solubilizer, suspenders auxiliary agent, an emulsifier, a covering agent, a cyclodextrin and/or a buffer mixture. Although the dosage may vary depending on symptoms, age and body weight of the patient, nature and severity of the disorder to be treated or prevented, the route of administration and dosage forms of the drug, usually for an adult daily dose of from 0.01 to 2000 mg, and it can be entered as a single dose or in separate doses. The amount of active ingredient that may be combined with a carrier substance for obtaining RA is new dosage forms, usually equal to the number of connections, which provides a therapeutic effect.

The precise time of administration and/or amount of composition that will yield the most effective results in terms of efficacy of treatment in this patient, dependent upon the activity, pharmacokinetics and bioavailability of a particular compound, physiological condition of the patient (including age, gender, type and stage of the disease, General physical condition, responsiveness to a given dosage and type of drug therapy), route of administration, etc. But these instructions can be used as a key for fine adjustment of treatment, such as determining the optimal time and/or quantity of the introduction, which requires no more than routine experimentation, consisting in monitoring the subject and the regulation of the dose and/or time.

The phrase "pharmaceutically acceptable" is used here to refer to such ligands, compounds, compositions and/or dosage forms which are, within reasonable medical assessment, are suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic reactions or other problems or complications, commensurate with a reasonable ratio of benefit/risk.

The phrase "pharmaceutically acceptable carrier", as used here, means supplied with the Ki acceptable substance, composition or filler, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating substance. Each carrier must be "acceptable" in the sense of compatibility with other ingredients of the finished formulation and not harm the patient. Some examples of substances which can serve as pharmaceutically acceptable carriers include; (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as maize starch, potato starch, and substituted or unsubstituted β-cyclodextrin; (3) cellulose, and its derivatives, such as carboxymethylcellulose sodium, ethylcellulose and cellulose acetate; (4) powdered tragakant; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and waxes for suppositories; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters, such as etiloleat and tillaart; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) saline; (18) ringer's solution (Ringer's); (19) ethyl alcohol; (20) phosphate buffer solution; and (21) other is s non-toxic compatible substances, used in pharmaceutical finished formulation. In some embodiments, the implementation of the pharmaceutical compositions of the present invention aerogene, i.e. do not cause a noticeable temperature increase with the introduction of the patient.

The term "pharmaceutically acceptable salt" refers to a relatively non-toxic additive salts of the inhibitor(s) with inorganic and organic acid. These salts can be obtainedin situduring the final isolation and purification of the inhibitor(s) or individual interaction purified(s) of the inhibitor(s) in the form of its free base with a suitable organic or inorganic acid and allocation of the resulting salt. Representative salts include the hydrobromide, hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, naftilan, mesilate, glucoheptonate, lactobionate, salt laurylsulphate and salts of amino acids and the like. (See, for example, Berge and others, (1977) “Pharmaceutical Salts”,J. Pharm. Sci,66: 1-19.)

In other cases, the inhibitors that are applicable in the methods of this invention may contain one or more acidic functional groups and, thus, capable of forming pharmaceutically acceptable salts with pharmaceutically reception is the subject bases. The term "pharmaceutically acceptable salts" in these instances refers to the relatively non-toxic additive salts of the inhibitor(s) with an inorganic or organic base. These salts can also be obtainedin situduring the final isolation and purification of the inhibitor(s) or individual interaction purified(s) of the inhibitor(s) in the form of its free acid with a suitable base such as the hydroxide, carbonate or bicarbonate pharmaceutically acceptable metal cation, with ammonia, or with a pharmaceutically acceptable organic primary, secondary or tertiary amine. Representative alkali or alkaline earth salts include salts of lithium, sodium, potassium, calcium, magnesium and aluminum and the like. Representative organic amines, applicable for the formation of additive salts with bases include ethylamine, diethylamine, Ethylenediamine, ethanolamine, diethanolamine, piperazine and the like (see, for example, Berge et al.,supra).

The compositions may also be available wetting agents, emulsifying agents and slip agents such as sodium lauryl sulfate and magnesium stearate, as well as dyes, antiadhesive, covering tools, sweeteners, flavors and fragrances, preservatives and antioxidants.

Examples of pharmaceutically acceptable antioxid now include: (1) water soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; (2) oil-soluble antioxidants, such as ascorbyl palmitate, bottled hydroxyanisol (BHA), bottled hydroxytoluene (BHT), lecithin, propylgallate, alpha-tocopherol and the like; and (3) agents that form chelates with metals, such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid and the like.

The finished formulation suitable for oral administration may be in the form of capsules, starch wafers, pills, tablets, pellets (using a flavored basis, usually sucrose and gum or tragakant), powders, granules, or in the form of a solution or suspension in aqueous or non-aqueous liquid, or as an emulsion oil-in-water or water-in-oil, or as an elixir or syrup, or as pastilles (using an inert matrix, such as gelatin and glycerol or sucrose and gum), and/or liquid for rinsing the mouth and the like, each of which contains a predetermined amount of inhibitor(s) as an active ingredient. The composition may be introduced in the form of a bolus, electuary or paste.

In solid dosage forms for oral administration (capsules, tablets, pills, coated tablets, p is Rosco, the granules and the like), the active ingredient is mixed with one or more pharmaceutically acceptable carriers, such as sodium citrate or dicalcium phosphate and/or one of the following: (1) fillers or dry diluents, such as starch, cyclodextrin, lactose, sucrose, glucose, mannitol, and/or silicic acid; (2) binders, such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and/or gum; (3) humectants, such as glycerol; (4) dezintegriruetsja agents, such as agar-agar, calcium carbonate, potato starch or starch from cassava, alginic acid, certain silicates and sodium carbonate; (5) agents, retarding dissolution such as paraffin; (6) absorption accelerators, such as Quaternary ammonium compounds; (7) humectants, such as, for example, acetylated alcohol (acetyl alcohol and glycerol monostearate; (8) absorbents, such as kaolin and bentonite clay; (9) moving substances such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof; and (10) coloring agents. In the case of capsules, tablets and pills, the pharmaceutical compositions may also contain buffering agents. Solid compositions of a similar type may also be produced as fillers in soft and hard gelatin capsules using such napolnitel the th, as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.

Tablets may be made by compression or extrusion, possibly with one or more auxiliary ingredients. Molded tablets can be obtained using a binder (such as gelatin or hydroxypropylmethylcellulose), moving substances, inert diluent, preservative, dezintegriruetsja substances (for example, starch glycolate, sodium or transverse cross-linked sodium carboxymethyl cellulose), surface-active or dispersing agent. Molded tablets can be obtained by compressing in a suitable machine a mixture of powdered(s) of the inhibitor(s), hydrated(ing) an inert liquid diluent.

Tablets and other solid dosage forms such as tablets, capsules, pills and granules, may be scored or obtained with coatings and shells, such as intersolubility coatings and other coatings well known in the art for the manufacture of finished pharmaceutical preparative forms. They can also be made in such a way as to provide slow or controlled release of the active ingredient, using, for example, hydroxypropylmethylcellulose in different proportions to provide the desired rate of visualaid the tion, other polymer matrices, liposomes and/or microspheres. They can be sterilized, for example, by filtration through a filter that retains bacteria, or the introduction of sterilizing agents in sterile solid compositions which can be dissolved in sterile water or some other sterile environment for injection immediately before use. These compositions may also contain fogging agents and can provide a composition that releases only the active(s) ingredient(s), or preferably in some part of the gastrointestinal tract, possibly with a delay. Examples of coating compositions which can be used include polymeric substances and waxes. The active ingredient, if appropriate, may also be microencapsulating form with one or more of the above fillers.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredient, the liquid dosage forms may contain inert diluents commonly used in the art, such as, for example, water or other solvents, soljubilizatory and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethylcarbonate, ethyl acetate, benilov the th alcohol, the benzyl benzoate, propylene glycol, 1,3-butyleneglycol, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor and sesame oils), glycerol, tetrahydrofuranyl alcohol, polyethylene glycols and esters of sorbitol with fatty acids and mixtures thereof.

Besides inert diluents, the oral compositions can also contain adjuvants, such as moisturizers, emulsifiers and suspendresume agents, sweeteners, flavorings, colorants, fragrances and preservatives.

Suspensions, in addition to active(th) ingredient(s)may contain suspendresume agents, for example ethoxylated isostearyl alcohols, polyoxyethylenated and esters of sorbitol, microcrystalline cellulose, Metagalaxy aluminum, bentonite, agar-agar and tragakant and mixtures thereof.

Ready preparative forms for rectal or vaginal injection can be presented in the form of suppositories, which can be obtained by blending one or more inhibitor(s) with one or more suitable non-irritating fillers or carriers, representing, for example, cocoa butter, polyethylene glycol, wax suppositories or a salicylate, and which is solid at room temperature, but liquid at body temperature and therefore melt in the rectal or vaginal cavity and release and the administrative agent.

Ready preparative forms, which are suitable for vaginal administration, where appropriate, also include uterine rings, tampons, gels, pastes, foams or aerosol preparations containing these media, as is known in the technique.

Dosage forms for local and percutaneous injection of inhibitor(s) include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and forms for inhalation. The active ingredient may be mixed under sterile conditions with a pharmaceutically acceptable carrier and any preservatives, buffers, or gases-displacers that may be required.

Ointments, pastes, creams and gels may contain, in addition to the inhibitor(s) excipients, such as animal and vegetable fats, oils, waxes, paraffins, starch, tragakant, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide or mixtures thereof.

Powders and aerosol preparations may contain in addition to the inhibitor(s) fillers such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and powders polyamides or mixtures of these substances. Aerosol preparations can additionally contain conventional gas propellant, such as chlorofluorocarbons, and volatile unsubstituted hydrocarbons, such is as butane and propane.

Alternatively, the inhibitor(s) may(may) be introduced in the form of an aerosol. Do this by preparing an aqueous aerosol, liposomal preparation or solid particles containing the composition. Can be used non-aqueous suspension (for example, fluorocarbon gas propellant). The preferred ultrasonic inhalers because they minimize the effect of the agent on the shear (shear), which can lead to degradation of the connection.

Typically, an aqueous aerosol is made by preparing an aqueous solution or suspension of the agent together with conventional pharmaceutically acceptable carriers and stabilizers. The carriers and stabilizers vary in accordance with the requirements of a particular composition, but typically include twin (Tweens), pluronic (Pluronics), esters of sorbitol, lecithin, cremophor (Cremophors), pharmaceutically acceptable co-solvents, such as glycol, innocuous proteins like serum albumin, oleic acid, amino acids such as glycine, buffers, salts, sugars or sugar alcohols. Aerosols are usually obtained from isotonic solutions.

Transdermal patches have the added advantage of providing controlled delivery of the inhibitor(s) in the body. These dosage forms can be manufactured by dissolving or dispersing agent under Odesa environment. To improve the penetration of the inhibitor(s) through the skin can also be used amplifiers absorption. Speed specified permeation can be controlled either by the introduction of a membrane that controls the rate or dispersion of inhibitor(s) in a polymer matrix or gel.

The pharmaceutical compositions of this invention suitable for parenteral administration comprise one or more inhibitor(s) in combination with one or more pharmaceutically acceptable sterile aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, or sterile powders which may be converted into sterile injectable solutions or dispersions prior to use, which may contain antioxidants, buffering agents, bacteriostatic factors, dissolved substances, which contribute to the creation of isotonicity with the blood of the intended recipient, or suspendresume agents or thickeners.

Examples of suitable aqueous and nonaqueous carriers which may be used in the pharmaceutical compositions of the invention include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol and the like), and suitable mixtures of vegetable oils, such as olive oil, and injectable organic esters, such as tiolet. The proper fluidity can be maintained, for example, the use of coating substances, such as lecithin, the required particle size in the case of dispersions and by the use of surface-active substances.

These compositions may also contain adjuvants such as preservatives, humectants, emulsifiers and dispersing agents. The prevention of the action of microorganisms can be ensured by the inclusion of various antibacterial and antifungal agents such as parabens, chlorobutanol, fenolcarbonove acid and the like. It is also desirable to enter in the composition of agents that regulate the tone, such as sugars, sodium chloride and the like. Advanced by the inclusion of agents which delay absorption such as aluminum monostearate and gelatin, injectable pharmaceutical form can be given (property) prolonged absorption.

In some cases, to renew the medication, it is desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. For example, slow the absorption of parenteral entered drug form is accomplished by dissolving or suspendirovanie medicines in the oil filler.

Injectable depot forms are made by creating microencapsulating matrices inhibitor(s) in biodegradable is alimera, such as polylactide-polyglycolide. The rate of release can be adjusted depending on the ratio of drug and polymer and the nature of competitive used polymer particles. Examples of other biodegradable polymers include poly(orthoevra) and poly(anhydrides). Injectable depot compositions can also be obtained by the introduction of the drug in liposomes or microemulsions that are compatible with the tissue of the body.

Drugs agents can be given orally, parenterally, topically or rectally. Of course, they can be given in forms appropriate for each route of administration. For example, they are administered in the form of tablets or capsules, by injection, inhalation, eye lotions, ointments, suppositories, injection; topical by lotion or ointment; and rectal by suppositories. Oral administration is preferred.

The phrases "parenteral administration" and "introduced parenterally"as used here, refers to the ways that differ from enteral and local administration, usually by injection, and include, but are not limited to, intravenous, intramuscular, intraarterial, vnutriobolochechnoe, intracapsular, intraorbital, intracardiac, vnutrepenialnye, intraperitoneal, transtracheal, subcutaneous, subepidermal, intra-articular, subcapsular,subarachnoid, intraspinally and intragrain injections and infusions.

The phrase "system introduction, introduced systemically", "peripheral introduction" and "put perifericheskie", as used here, means the introduction of a ligand, drug, or other substance other than directly into the Central nervous system, so it is included in the system of the patient and, thus, is subject to metabolism and other like processes, for example, subcutaneous administration.

Specified(s) inhibitor(s) can be injected into humans or animals for therapy by any suitable administration, including oral, nasal, as, for example, using an aerosol of the drug, rectal, intrawaginalno parenteral vnutripolostno and topically in the form of powders, ointments or drops, including hominids and the hyoid.

Regardless of the method of introduction of the inhibitor(s) (e) may(may) be used(s) in a suitable hydrated form, and/or (C) pharmaceutical compositions of the present invention, the formed(s) in a pharmaceutically acceptable dosage forms by conventional means, well known to the experts in this field of technology.

The actual dose levels of active ingredients in the pharmaceutical compositions of this invention may be varied so as to obtain the number of active ing is edient, which is effective to achieve the desired therapeutic response in a particular patient, compositions and mode of introduction without toxicity to the patient.

The proposed concentration of the compound in pharmaceutically acceptable mixture will vary depending on several factors, including the dose of a compound, which must be entered, pharmacokinetic characteristics used(s) connection(s) and method of administration. Typically, the compositions of this invention can be represented in the form of an aqueous solution for parenteral administration containing about 0.1-10% wt./about. offered here is a connection along with other substances. Typical doses are in the range from about 0.01 to about 50 mg/kg of body weight per day, divided nor 1-4 doses. Each separate dose may contain the same or different compounds of the invention. Dosage should be effective amount depending on several factors, including overall health of the patient, ready preparative form and route of administration selected(s) connection(s).

Another aspect of the invention provides a complex therapy, if administered one or more therapeutic agents with inhibitors of the proteasome. Such combination therapy can be achieved by simultaneous, sequential or razdelnopolye individual components of therapy.

In some embodiments, the implementation of the connection of the invention is administered together with one or more other inhibitor(s) of the proteasome.

In some embodiments of the invention, the compound of the invention is administered in conjunction with chemotherapy sredstva. Suitable chemotherapeutic sredstva may include natural products such as Vinca alkaloids (i.e. vinblastine, vincristine and vinorelbine), paclitaxel, epileptogenesis (i.e. etoposide, teniposide), antibiotics (dactinomycin (actinomycin D)daunorubicin, doxorubicin and idarubitsin), anthracyclines, mitoxantrone, bleomycin, plicamycin (mithramycin) and mitomycin, enzymes (L-asparaginase which systemically metabolizes asparagine and deprimere cells that lack the ability to synthesize their own asparagine), antithrombotic agents; antiproliferative/antimitoticescoy alkylating agents such as nitrogen mustards (mechlorethamine, cyclophosphamide and analogs, melphalan, chlorambucil), ethylenimines and methylmelamine (hexamethylmelamine, thiotepa), alkyl sulphonates (busulfan), nitrosoanatabine (carmustin (BCNU) and analogs, streptozocin), trazeni-dacarbazine (DTIC); antiproliferative/antimitoticescoe antimetabolites, such as folic acid analogs (methotrexate), pyrimidine analogues (fluorouracil, floxuridine and tarabin), the purine analogues and related inhibitors (mercaptopurine, tioguanin, pentostatin and 2-chloromethoxypropyl; aromatase inhibitors (anastrozole, exemestane and letrozole); and coordination complexes of platinum (cisplatin, carboplatin), procarbazine, hydroxyurea, mitotane, aminoglutetimid; inhibitors discontiuation (HDAC); hormones (i.e. estrogen) and agonists of hormones such as hormone agonists, releasing luteinizing hormone (LHRH) (goserelin, leuprolide and triptorelin). Other chemotherapeutic agents can be mechlorethamine, camptothecin, ifosfamide, tamoxifen, raloxifene, gemcitabine, navelbine or any options analogs and derivatives.

In some embodiments, the implementation of the connection of the invention is administered in conjunction with the cytokine. Cytokines include, but are not limited to, interferon-γ, -α and-β, interleukins 1 through 8, 10 and 12, the factor stimulating colonies of granulocytes monocytes (GM-CSF), TNF-α and-β and TGF-β.

In some embodiments, the implementation of the connection of the invention is administered in conjunction with the steroid. Suitable steroids may include, but are not limited to, 21-acetoxyphenyl, alclometasone, algestone, amcinonide, beclomethasone, betamethasone, budesonide, chloroprednisone, clobetasol, clocortolone, cloprednol, corticosterone, cortisone, cortisol, deflazacort, desonide, desoximetasone, dexamet the zones, diflorasone, diflucortolone, difluprednate, enoxolone, flashcart, fluchloralin, flumetazon, flunisolide, acetonide fluoqinolona, fluocinonide, butylfluorene, fluocortolone, fluometuron, acetate flaperon, acetate of fluprednidene, fluprednisolone, flurandrenolide, fluticasone propionate, formocortal, halcinonide, propionate halobetasol, halobetasol, hydrocortisone, etabonate loteprednol, mazipredon, Madison, meprednisone, methylprednisolone, mometasone furoate, paramethasone, prednicarbate, prednisolone, 25-diethylaminoacetate prednisolone, prednisolone-nutrifaster, prednisone, prednesol, prednisone, rimexolone, tixocortol, triamcinolone, triamcinolone acetonide, benetone triamcinolone, hexacetonide triamcinolone and their salts and/or derivatives.

In some embodiments, the implementation of the connection of the invention is administered in conjunction with immunotherapy agent. Appropriate immunotherapy agents may include, but not limit, the MDR modulators (verapamil, valspodar, biricodar, tariquidar, laniquidar), rapamycin, mycophenolate-mofetil, cyclophoshamide, cyclosporine, thalidomide, and monoclonal antibodies. Monoclonal antibodies can be either free or conjugated, such as rituximab, tositumomab, alemtuzumab, daclizumab, epratuzumab, ibritumomab-tiuxetan, gemtuzumab-ozogamicin, b is carisoma, cetuximab, erlotinib and trastuzumab.

EXAMPLES

Example 1

Scheme 1: synthesis of compounds 010

Connection (003):

To a cooled to 0°C. a solution of methyl ester of N-BOC-serine (001) (2.5 g, of 11.4 mmol), hydrochloride benzyl ester of L-alanine (002) (3.3 grams, or 11.4 mmol), NAWT (2.5 g, 18.2 mmol) and HBTU (6,9 g, 18,24 mmol) in tetrahydrofuran (400 ml) for 10 minutes was added a solution of N,N-diisopropylethylamine (8.0 ml of 45.6 mmol) in tetrahydrofuran (50 ml). The next 5 hours and the mixture was stirred at room temperature. A large part of the solvent was removed under reduced pressure and the obtained residue was diluted with ethyl acetate (300 ml). Then the solution was washed with saturated aqueous (solution) sodium bicarbonate (2×50 ml) and with brine (100 ml). The organic layers were dried over sodium sulfate and filtered through celite-545. The solvents were removed under reduced pressure and the residue was purified flash chromatography (hexane and ethyl acetate), the desired compound (003) (4.4 g) was isolated and okharakterizovali LC/MS (LCRS (MH) m/z: 457,23).

Connection (004):

To a cooled to 0°C. a solution of (003) (5,14 g, 11,25 mmol) in tetrahydrofuran (100 ml) was added 10%Pd/C (500 mg). The resulting mixture was stirred under the hydrogen pressure of 1 atmosphere for 4 hours. The mixture was filtered through celite-545 and cake on the filter is washed with tetrahydrofuran, the m The organic filtrate was concentrated under reduced pressure and kept under high vacuum for 2 hours, obtaining (004), as was confirmed by LC/MS (LCRS (MH) m/z: 367,18), which was used without further purification.

Connection (006):

To a solution of (005) (synthesis of (005) refer to patent application U.S. serial No. 11/131688) (3,9 g, 13 mmol) in triperoxonane acid (50 ml) was added 10%Pd/C (600 mg). The resulting mixture was stirred under the hydrogen pressure of 1 atmosphere for 6 hours. The mixture was filtered through celite-545 and cake on the filter was washed with dichloromethane (200 ml). The filtrate was concentrated under reduced pressure and kept at high vacuum during the night, receiving (006), as was confirmed by LC/MS (LCRS (MH) m/z: 172,13), which was used without further purification.

Connection (007):

To a cooled to 0°C. a solution of (004) and (006), HOBT (2.5 g, 18 mmol) and HBTU (6,9 g, 18 mmol) in tetrahydrofuran (400 ml) for 10 minutes was added a solution of N,N-diethylethanolamine (8 ml, 46 mmol) in tetrahydrofuran (50 ml). The next 5 hours and the mixture was stirred at room temperature. A large part of the solvent was removed under reduced pressure and the residue was diluted with ethyl acetate (400 ml). The resulting solution was washed with saturated aqueous (solution) sodium bicarbonate (2×100 ml) and with brine (100 ml). The organic layers were dried over sulfate intothree and filtered through celite-545. The solvents were removed under reduced pressure and the residue was purified flash chromatography (hexane and ethyl acetate), obtaining (007) (3.5 g), as was shown by LC/MS (LCRS(MH) m/z: 520,29).

Connection (008):

To a cooled to 0°C solution (007) (320 mg, 0,616 mmol) in dichloromethane (10 ml) was added triperoxonane acid (10 ml) and the resulting solution was stirred at the same temperature over the next hour. The organic layers were concentrated under reduced pressure and then kept under high vacuum for 2 hours, obtaining (008), which was confirmed by LC/MS (LCRS(MH) m/z: 420,24), which was used without further purification.

Connection (010):

To a cooled to 0°C. a solution of (008), 5-methylisoxazol-3-carboxylic acid (009) (94 mg, of 0.74 mmol), NOT (135 mg, 1.0 mmol) and HBTU (350 mg, 1.0 mmol) in tetrahydrofuran (100 ml) for 5 minutes was added a solution of N,N-diisopropylethylamine (0.5 ml, 2.5 mmol) in tetrahydrofuran (2 ml). The next 5 hours and the mixture was stirred at room temperature and then was diluted with ethyl acetate (200 ml). It was further washed with saturated aqueous (solution) sodium bicarbonate (2×10 ml) and with brine (10 ml) and the organic layers were dried over sodium sulfate and filtered through celite-545. The solvents were removed under reduced pressure and the residue was purified HPLC (aqueous ammonium acetate and acetonitrile), receiving (010) (195 mg), as shown by LC/MS (LCRS (MH) m/z 529,26); inhibition of >90% of the proteasome CT-L at 40 mg/kg RO.

Example 2

Scheme 2, the synthesis of example 023

Connection (013):

To a cooled to 0°C. a solution of N-Boc-L-leucine (011) (2.6 g, 11 mmol), hydrochloride benzyl ester of L-phenylalanine (012) (2.9 g, 10 mmol), NAWT (1.7 g, 11 mmol) and HBTU (3.9 g, 11 mmol) in tetrahydrofuran (200 ml) for 5 minutes was added a solution of N,N-diisopropylethylamine (4,9 ml, 30 mmol) in tetrahydrofuran (10 ml). The next 5 hours and the mixture was stirred at room temperature, after which it became homogeneous. Then it was diluted with ethyl acetate (300 ml) and washed with saturated aqueous (solution) sodium bicarbonate (2×50 ml) and with brine (100 ml). The organic layers were dried over sodium sulfate and filtered through celite-545. The solvents were removed under reduced pressure and the residue was purified flash chromatography (hexane and ethyl acetate), obtaining (013) (4.4 g), as shown by LC/MS (LCRS (MH) m/z: 469,26).

Connection (014):

To a cooled to 0°C. a solution of (013) (4,32 g, 9,24 mmol) in tetrahydrofuran (100 ml) was added 10%Pd/C (500 mg). The resulting mixture was stirred under the hydrogen pressure of 1 atmosphere for 4 hours. The mixture was filtered through celite-545 and cake on the filter was washed with tetrahydrofuran. The organic filtrate was concentrated under reduced pressure and kept at high vacuum, receiving 014) (3.5 g), as was confirmed by LC/MS (LCRS (MH) m/z: 378,22), which was used without further purification.

Connection (015):

To a cooled to 0°C. a solution of (014) (3.5 g, 9,24 mmol) and (006) (2.4 g, 11 mmol), NAWT (1.7 g, 11 mmol) and HBTU (3.9 g, 11 mmol) in tetrahydrofuran (200 ml) for 5 minutes was added a solution of N,N-diisopropylethylamine (4,9 ml, 30 mmol) in tetrahydrofuran (10 ml). The next 5 hours and the mixture was stirred at room temperature, as a result it became homogeneous. Then it was diluted with ethyl acetate (400 ml) and washed with saturated aqueous (solution) sodium bicarbonate (2×100 ml) and with brine (100 ml). The organic layers were dried over sodium sulfate and filtered through celite-545. The solvents were removed under reduced pressure and the residue was purified flash chromatography (hexane and ethyl acetate), and the desired compound (015) (5.0 g) was isolated and okharakterizovali LC/MS (LCRS (MH) m/z: 532,33).

Connection (016):

To a cooled to 0°C. a solution of (015) (5.0 g, 9,40 mmol) in dichloromethane (50 ml) for 5 minutes was added triperoxonane acid (20 ml) and the resulting solution was stirred at the same temperature over the next hour. The organic layers were concentrated under reduced pressure and then kept under high vacuum, receiving (016), which was confirmed by LC/MS (LCRS (MH) m/z: 432,33), which was used without further purification.

Connection (018):

Crestore methyl ester 5-methyl-3-isoxazolecarboxylic acid (017) (14.1 g, 100 mmol) in carbon tetrachloride (500 ml) at room temperature was added N-bromosuccinimide (23 g, 130 mmol) and benzoyl peroxide (2.5 g, 10 mmol). The resulting mixture was stirred overnight in an argon atmosphere at 80°C. the Reaction mixture was cooled and diluted with 500 ml of dichloromethane and washed with saturated aqueous (solution) sodium bicarbonate (3×100 ml). The aqueous phase was extracted with 200 ml dichloromethane and the combined organic layers were washed with brine and dried over MgSO4. The solvent was removed and the residue was purified flash chromatography (hexane and ethyl acetate), obtaining (018) (7.9 g), which okharakterizovali LC/MS (LCRS (MH) m/z: 219,05).

Connection (019):

To a cooled to 0°C solution (018) (12 g, 55 mmol) in tetrahydrofuran (20 ml) was added aqueous lithium hydroxide (35 ml, 4n.). The resulting mixture during the night was stirred at room temperature. Then acidified with hydrochloric acid (2 BC) until pH = 1 and extracted with tetrahydrofuran (3×200 ml). The combined organic layers were washed with brine (10 ml), dried over sodium sulfate and filtered. The solvent was removed and the residue was subjected to lyophilization, receiving (019)(8,2 g), which was confirmed by LC/MS (LCRS (MH) m/z: 205,95), which was used without further purification.

Connection (020):

Solution (019) (6.0 g, 30 mmol), benzyl alcohol (3.5 ml) and para-colorswitch the slots (1.1 g, 6 mmol) in toluene (100 ml) was stirred overnight at 100°C. Then left to cool, was diluted with 300 ml of ethyl acetate and washed with saturated aqueous (solution) of sodium bicarbonate. The aqueous phase was then extracted with 200 ml of ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate and filtered. The solvent was removed and the residue was purified flash chromatography (hexane and ethyl acetate), obtaining (020) (5.8 g), which okharakterizovali LC/MS (LCRS (MH) m/z: 295,98).

Connection (021):

Solution (020) (2.0 g, 6.8 mmol) and research (3.0 ml) in tetrahydrofuran (50 ml) was stirred at room temperature for two hours. Then the solvent was removed and the residue was purified flash chromatography (hexane and ethyl acetate), obtaining (021) (820 mg), which okharakterizovali LC/MS (LCRS (MH) m/z: 303,13).

Connection (022):

To a solution of (021) (400 mg, of 1.32 mmol) in tetrahydrofuran (40 ml) was added 10%Pd/C (100 mg) and the mixture was stirred at room temperature under hydrogen pressure at one atmosphere for 2 hours. Then it was filtered through celite and concentrated, obtaining (022), which was confirmed by LC/MS (LCRS (MH) m/z: 213,08) and used without further purification.

Connection (023)

To a cooled to 0°C solution (016) (139 mg, 0.3 mmol) and (022) (70 mg, 0.4 mmol), NOT (70 mg, 0.5 mmol) and HBTU (170 mg, 0.5 mmol) in tetrahydrofuran 50 ml) was added a solution of N,N-diisopropylethylamine (0.5 ml, 2.5 mmol) in tetrahydrofuran (5 ml). The next 5 hours and the mixture was stirred at room temperature, as a result it became homogeneous. Then it was diluted with ethyl acetate (200 ml) and washed with saturated aqueous (solution) sodium bicarbonate (2×10 ml) and with brine (10 ml). The organic layers were dried over sodium sulfate and filtered through celite-545. The solvents were removed under reduced pressure and the residue was purified HPLC (aqueous ammonium acetate and acetonitrile)to give compound (023) (125 mg), which okharakterizovali LC/MS (LCRS (MH) m/z: 626,35); inhibition >80% proteasome CT-L at 40 mg/kg RO.

Example 3

Scheme 3: synthesis of example 029

Connection (025):

To a cooled to 0°C. a solution of Fmoc-Val-OH (024) (348 mg, 1.6 mmol) in dichloromethane (4 ml) was added MSNT (474 mg, 1.6 mmol) and N-Mei (of 0.13 ml, 1.6 mmol). After the mixture became homogeneous, was added to the resin NMRV (400 mg, 0.32 mmol). The resulting reaction mixture was shaken for two hours at room temperature. The resin was filtered off, washed with N,N-dimethylformamide (3×10 ml) and dichloromethane (3×10 ml) and left to air dry, receiving (025).

Connection (026):

The resin (025) were placed in a 20%solution of piperidine in N,N-dimethylformamide (20 ml) and the resulting mixture was shaken for 1 hour at room temperature. Resin AHP crystal growth is microvilli and washed with N,N-dimethylformamide (3×10 ml) and dichloromethane (3×10 ml).

To a cooled to 0°C. a solution of Fmoc-Ser(OMe)-OH (546 mg, 1.6 mmol) in N,N-dimethylformamide (4 ml) was added NOWT (245 mg, 1.6 mmol), HBTU (606 mg, 1.6 mmol) and N,N-diisopropylethylamine (0.6 ml, 3.2 mmol). After the mixture became homogeneous, was added to the resin. The resulting mixture was left to shake things up during the night. Then the resin was filtered off, washed with N,N-dimethylformamide (3×10 ml) and dichloromethane (3×10 ml) and left to air dry, receiving (026).

Connection (027):

The resin (026) were placed in a 20%solution of piperidine in N,N-dimethylformamide (20 ml) and the resulting mixture was shaken for 1 hour at room temperature. The resin was filtered and washed with N,N-dimethylformamide (3×10 ml) and dichloromethane (3×10 ml).

To a cooled to 0°C solution of 5-methylisoxazol-3-carboxylic acid (009) (162 mg, 1.6 mmol) in N,N-dimethylformamide (4 ml) was added NOWT (245 mg, 1.6 mmol), HBTU (606 mg, 1.6 mmol) and N,N-diisopropylethylamine (0.6 ml, 3.2 mmol). After the mixture became homogeneous resin was added and the resulting reaction mixture was left to shake things up during the night. Then the resin was filtered off, washed with N,N-dimethylformamide (3×10 ml) and dichloromethane (3×10 ml) and left to air dry, receiving (027).

Connection (028):

To the resin (027) was added a 50%solution triperoxonane acid in dichloromethane (10 ml) and the resulting mixture of astable and shake for 30 minutes. The resin then was filtered and washed with dichloromethane (3×10 ml). Volatile (medium) was removed under reduced pressure, obtaining (028)who okharakterizovali LC/MS (LCRS (MH) m/z: 328,14) and used without further purification.

Connection (029):

To a cooled to 0°C solution (028) and (006) (117 mg, 0.4 mmol), NOT (70 mg, 0.5 mmol) and HBTU (170 mg, 0.5 mmol) in tetrahydrofuran (50 ml) was added a solution of N,N-diisopropylethylamine (0.5 ml, 2.5 mmol) in tetrahydrofuran (5 ml). The next 5 hours and the mixture was stirred at room temperature, causing it became homogeneous. Then it was diluted with ethyl acetate (200 ml) and washed with saturated aqueous (solution) sodium bicarbonate (2×10 ml) and with brine (10 ml). The organic layers were dried over sodium sulfate and filtered through celite-545. The solvents were removed under reduced pressure and the residue was purified HPLC (aqueous ammonium acetate and acetonitrile)to give compound (029) (125 mg), which okharakterizovali LC/MS (LCRS (MH) m/z: 481,26); inhibition >70% of the proteasome CT-L at 20 mg/kg RO.

Example 4

Scheme 4: synthesis of example 035

Connection (031):

To a cooled to 0°C. a solution of Fmoc-L-4-thiazolidinone (030) (1.0 g, 2.5 mmol) in dichloromethane (4 ml) was added N-Mei (150 ml, 1.9 mmol), MSNT (755 mg, 2.55 mmol) and after the mixture became homogeneous, ribasso HMPB resin (800 mg, 0.51 mmol). The resulting reaction mixture was left to shake for two hours at room temperature. Then the resin was filtered off, washed with N,N-dimethylformamide (3×20 ml) and dichloromethane (3×20 ml) and left to air dry, receiving (031).

Connection (032):

The resin (031) (360 mg, 0.23 mmol) were placed in a 20%solution of piperidine in N,N-dimethylformamide (20 ml) and the resulting mixture was shaken for 1 hour at room temperature. Then the resin was filtered and washed with N,N-dimethylformamide (3×10 ml) and dichloromethane (3×10 ml).

To a cooled to 0°C. a solution of Fmoc-L-leucine (204 mg, of 0.58 mmol) in N,N-dimethylformamide (4 ml) was added NOWT (124 mg, of 0.92 mmol), HBTU (349 mg, of 0.92 mmol) and N,N-diisopropylethylamine (402 μl, 2.3 mmol). After the mixture became homogeneous, was added to the resin. The resulting mixture was left to shake at 5°C for 5 hours. Then the resin was filtered off, washed with N,N-dimethylformamide (3×20 ml) and dichloromethane (3×20 ml) and the resulting resin was allowed to dry in air, receiving (032).

Connection (033):

The resin (032) (0.23 mmol) were placed in a 20%solution of piperidine in N,N-dimethylformamide (20 ml) and the resulting mixture was shaken for 1 hour at room temperature. Then the resin was filtered and washed with N,N-dimethylformamide (3×10 ml) and dichloromethane (3×10 ml).

To a cooled to 0°C. dissolve the (022) (123 mg, of 0.58 mmol) in N,N-dimethylformamide (4 ml) was added NOWT (124 mg, of 0.92 mmol), HBTU (349 mg, of 0.92 mmol) and N,N-diisopropylethylamine (402 μl, 2.3 mmol). After the mixture became homogeneous resin was added and the resulting reaction mixture was left to shake at room temperature overnight. Then the resin was filtered off, washed with N,N-dimethylformamide (3×10 ml) and dichloromethane (3×10 ml) and the resulting resin was allowed to dry in air, receiving (033).

Connection (034):

To the resin (033) was added a 50%solution triperoxonane acid in dichloromethane (10 ml) and the resulting mixture was left to shake for 30 minutes. Then the resin was filtered and washed with dichloromethane (3×10 ml). Volatile (medium) was removed under reduced pressure, obtaining (34), as determined by LC/MS (LCRS (MH) m/z: 480,18), which was used without further purification.

Connection (035):

To a cooled to 0°C solution (034) and (006) (70 mg, 0.23 mmol), NOT (50 mg, and 0.37 mmol) and HBTU (140 mg, of 0.37 mmol) in tetrahydrofuran (50 ml) was added a solution of N,N-diisopropylethylamine (0.5 ml, 2.5 mmol) in tetrahydrofuran (5 ml). The mixture was stirred at room temperature for 5 hours and then was diluted with ethyl acetate (200 ml). Then washed it with saturated aqueous (solution) sodium bicarbonate (2×10 ml) and with brine (10 ml). The organic layers were dried over sulfate hydroxide is I, was filtered through celite-545 and the solvents were removed under reduced pressure. The obtained residue was purified HPLC (aqueous ammonium acetate and acetonitrile)to give compound (035) (15 mg), which okharakterizovali LC/MS (LCRS (MH) m/z: to 633.3); inhibition >90% of the proteasome CT-L at 40 mg/kg RO.

Example 5

Scheme 5: synthesis of example 039

Connection (036):

The resin (031) (800 mg, 0.23 mmol) were placed in a 20%solution of piperidine in N,N-dimethylformamide (20 ml) and the resulting mixture was shaken at room temperature for 1 hour. The resin was filtered and washed with N,N-dimethylformamide (3×10 ml) and dichloromethane (3×10 ml).

To a cooled to 0°C. a solution of Fmoc-L-Ser(OMe)-OH (435 mg, 1.3 mmol) in N,N-dimethylformamide (10 ml) was added NOWT (276 mg, 2.0 mmol), HBTU (710 mg, 2.0 mmol) and N,N-diisopropylethylamine (0.9 ml, 5.1 mmol). After the mixture became homogeneous, was added to the resin. The resulting mixture was left to shake at 5°C for 5 hours. Then the resin was filtered off, washed with N,N-dimethylformamide (3×20 ml) and dichloromethane (3×20 ml) and left to air dry, receiving (036).

Connection (037)

The resin (036) were placed in a 20%solution of piperidine in N,N-dimethylformamide (20 ml) and the resulting mixture was shaken at room temperature for 1 hour. The resin was filtered and washed with N,N-dimethylformamide (3) - Rev. 20 ml) and dichloromethane (3×20 ml).

To a cooled to 0°C solution (009) (162 mg, 1.3 mmol) in N,N-dimethylformamide (4 ml) was added NOWT (276 mg, 2.0 mmol), HBTU (710 mg, 2.0 mmol) and N,N-diisopropylethylamine (0.9 ml, 5.1 mmol). After the reaction mixture became homogeneous resin was added and the resulting reaction mixture was left to shake at room temperature overnight. Then the resin was filtered off, washed with N,N-dimethylformamide (3×10 ml) and dichloromethane (3×10 ml) and left to air dry, receiving (037).

Connection (038):

(037) was added a 50%solution triperoxonane acid in dichloromethane (10 ml) and the resulting mixture was left to shake for 30 minutes. Then the resin was filtered and washed with dichloromethane (3×10 ml). Volatile (medium) was removed under reduced pressure, obtaining (38), as determined by LC/MS (LCRS (MH) m/z: 383,09), which was used without further purification.

Connection (039):

To a cooled to 0°C solution (038) and (006) (156 mg, 0.51 mmol), NOT (111 mg, 0.82 mmol) and HBTU (311 mg, 0.82 mmol) in tetrahydrofuran (50 ml) was added a solution of N,N-diisopropylethylamine (0.5 ml, 2.5 mmol) in tetrahydrofuran (5 ml). The mixture was stirred at room temperature for 5 hours, and it became homogeneous. Then it was diluted with ethyl acetate (200 ml) and washed with saturated aqueous (solution) sodium bicarbonate (2×10 ml) and salt dissolve the Ohm (10 ml). The organic layers were dried over sodium sulfate and filtered through celite-545. The solvents were removed under reduced pressure and the residue was purified HPLC (aqueous ammonium acetate and acetonitrile), receiving (039) (22 mg), which okharakterizovali LC/MS (LCRS (MH) m/z: 536,21; inhibition >75% of the proteasome CT-L at 20 mg/kg RO.

Example 6

Scheme 6: synthesis of example 045 (method A)

Connection (041):

To a cooled to 0°C. a solution of Fmoc-L-alanine (040) (1.0 g, 3.2 mmol) in dichloromethane (30 ml) was added N-Mei (190 μl, 12.4 mmol), MSNT (950 mg, 3.2 mmol) and after the mixture became homogeneous, added HMPB resin (1 g, 0.64 mmol). The resulting reaction mixture was left to shake for two hours at room temperature. Then the resin was filtered off, washed with N,N-dimethylformamide (3×20 ml) and dichloromethane (3×20 ml), receiving (041).

Connection (042):

The resin (041) were placed in a 20%solution of piperidine in N,N-dimethylformamide (20 ml) and the resulting mixture was shaken at room temperature for 1 hour. The resin was filtered and washed with N,N-dimethylformamide (3×20 ml) and dichloromethane (3×20 ml).

To a cooled to 0°C. a solution of Fmoc-Ser(OMe)-OH (546 mg, 1.6 mmol) in N,N-dimethylformamide (10 ml) was added NOWT (346 mg, 2.6 mmol), HBTU (970 mg, 2.6 mmol) and N,N-diisopropylethylamine (1.1 ml, 6.4 mmol). After the reactions the fair mixture became homogeneous, was added to the resin. The resulting mixture was left to shake at 5°C for 5 hours. Then the resin was filtered off, washed with N,N-dimethylformamide (3×20 ml) and dichloromethane (3×20 ml) and left to air dry, receiving (042).

Connection (043):

The resin (042) (0.23 mmol) were placed in a 20%solution of piperidine in N,N-dimethylformamide (20 ml) and the resulting mixture was shaken at room temperature for 1 hour. The resin was filtered and washed with N,N-dimethylformamide (3×10 ml) and dichloromethane (3×10 ml).

To a cooled to 0°C solution (009) (203 mg, 1.6 mmol) in N,N-dimethylformamide (10 ml) was added NOWT (346 mg, 2.6 mmol), HBTU (970 mg, 2.6 mmol) and N,N-diisopropylethylamine (1.1 ml, 6.4 mmol). After the reaction mixture became homogeneous resin was added and the resulting reaction mixture was left to shake at room temperature overnight. Then the resin was filtered off, washed with N,N-dimethylformamide (3×20 ml) and dichloromethane (3×20 ml) and left to air dry, receiving (043).

Connection (044):

(043) was added a 50%solution triperoxonane acid in dichloromethane (10 ml) and the resulting mixture was left to shake for 30 minutes. Then the resin was filtered and washed with dichloromethane (3×10 ml). Volatile (medium) was removed under reduced pressure, obtaining (044), as determined by LC/MS (LCRS (MH) m/z: 300,11), which is the OE was used without further purification.

Connection (045):

To a cooled to 0°C. a solution of the above intermediate product (044) and (006) (195 mg, 0.64 mmol), NOT (137 mg, 1.0 mmol) and HBTU (357 mg, 1,0) in tetrahydrofuran (50 ml) was added a solution of N,N-diisopropylethylamine (0.5 ml, 2.5 mmol) in tetrahydrofuran (5 ml). The mixture was stirred at room temperature for 4 hours. Then it was diluted with ethyl acetate (200 ml) and washed with saturated aqueous (solution) sodium bicarbonate (2×10 ml) and with brine (10 ml). The organic layers were dried over sodium sulfate and filtered through celite-545. The solvents were removed under reduced pressure and the residue was purified HPLC (aqueous ammonium acetate and acetonitrile), receiving (045) (84 mg), which okharakterizovali LC/MS (LCRS (MH) m/z: 453,23; inhibition >80% proteasome CT-L at 20 mg/kg RO.

Example 7

Scheme 7: synthesis of example 045 (method)

Connection (047):

To a cooled to 0°C. a solution of methyl ester of N-Boc-serine (001) (6,57 g, 33 mmol), hydrochloride benzyl ester of L-alanine (046) (6,45 g, 30 mmol), NOT (of 5.05 g, 33 mmol) and HBTU (11.8 g, 33 mmol) in tetrahydrofuran (400 ml) for 10 minutes was added a solution of N,N-diisopropylethylamine (9.0 g, 70 mmol) in tetrahydrofuran (50 ml). The mixture became homogeneous and was stirred at room temperature for 5 hours. Then the greater part of the solvent is dalali under reduced pressure and the residue was diluted with ethyl acetate (500 ml). Was washed with saturated aqueous (solution) sodium bicarbonate (2×150 ml) and with brine (200 ml) and the organic layers were dried over sodium sulfate and filtered through celite-545. The solvents were removed under reduced pressure and the residue was purified flash chromatography (hexane and ethyl acetate), obtaining (047) (11.8 g), which okharakterizovali LC/MS (LCRS (MH) m/z: 381,19.

Connection (048):

To a cooled to 0°C solution (047) (11.8 g, was 31.0 mmol) in dichloromethane (100 ml) for 10 minutes was added triperoxonane acid (50 ml) and the resulting mixture was stirred and at the same temperature for another 3 hours. Then the solvents were removed under reduced pressure and the residue was placed overnight in a high vacuum, receiving TFA salt (048), which okharakterizovali LC/MS (LCRS (MH) m/z: 281,15), and used without further purification.

Connection (049):

To a cooled to 0°C solution (048), 5-methylisoxazol-3-carboxylic acid (009) (3,93 g, 31 mmol), NOT (4.7 g, 35 mmol) and HBTU (12.5 g, 35 mmol) in tetrahydrofuran (400 ml) for 10 minutes was added a solution of N,N-diisopropylethylamine (20 ml) in tetrahydrofuran (100 ml); the pH of the mixture was equal to ~8. The mixture was stirred at room temperature for 5 hours. Then the greater part of the solvent was removed under reduced pressure and diluted with ethyl acetate (1.0 l). It was further washed with saturated aqueous (solution) bicarbonate intothree the (2×100 ml) and with brine (100 ml) and the organic layers were dried over sodium sulfate and filtered through celite-545. The solvents were removed under reduced pressure and the residue was purified flash chromatography (hexane and ethyl acetate), obtaining (049) (10.8 g), which okharakterizovali LC/MS (LCRS (MH) m/z: 390,16.

Connection (044):

To a cooled to 0°C solution (049) (3.28 g, 8.4 mmol) in tetrahydrofuran (100 ml) was added 10%Pd/C (500 mg). The resulting mixture was stirred under the hydrogen pressure of 1 atmosphere for 4 hours. Then the mixture was filtered through celite-545 and cake on the filter was washed with tetrahydrofuran. The organic filtrate was concentrated under reduced pressure and placed in a high vacuum for 2 hours, obtaining (044), which okharakterizovali LC/MS (LCRS (MH) m/z: 281,15) and used without further purification.

Connection (045):

To a cooled to 0°C solution (044) and (006) (1.9 g, 8.5 mmol), NAWT (2.0 g, 13 mmol) and HBTU (5,4 g, 14 mmol) in tetrahydrofuran (200 ml) was added a solution of N,N-diisopropylethylamine (5,4 g, 42 mmol) in tetrahydrofuran (10 ml). The mixture was stirred at room temperature for another 5 hours. Then the greater part of the solvent was removed under reduced pressure and the obtained residue was diluted with ethyl acetate (400 ml). It was further washed with saturated aqueous (solution) sodium bicarbonate (2×50 ml) and with brine (50 ml) and the organic layers were dried over sodium sulfate and filtered through celite-545. The solvents were removed under reduced pressure and the residue was purified HPLC (aqueous ammonium acetate and acetonitrile), receiving (045) (1.35 g), which okharakterizovali LC/MS (LCRS (MH) m/z: 453,23).

Example 8

Scheme 8: synthesis of example 055

Connection (051)

To a cooled to 0°C. a solution of N-Boc-L-pyridylamine (050) (1.0 g, 3,76 mmol), hydrochloride benzyl ester of L-phenylalanine (002) (1.3 g, 3,76 mmol), NOT (0.68 g, 5.0 mmol) and HBTU (1.8 g, 5.0 mmol) in tetrahydrofuran (100 ml) was added a solution of N,N-diisopropylethylamine (1.6 ml) in tetrahydrofuran (10 ml). The mixture was stirred at room temperature for 3 hours and then was diluted with ethyl acetate (200 ml), washed with saturated aqueous (solution) sodium bicarbonate (2×50 ml) and with brine (100 ml); the organic layers were dried over sodium sulfate and filtered through celite-545. The solvent was removed under reduced pressure and the residue was purified flash chromatography (hexane and ethyl acetate), obtaining (051), which okharakterizovali LC/MS (LCRS (MH) m/z: 504,24).

Connection (052):

To a cooled to 0°C solution (051) in tetrahydrofuran (100 ml) was added 10%Pd/C (100 mg). The resulting mixture was stirred under the hydrogen pressure of 1 atmosphere for 4 hours. Then the mixture was filtered through celite-545 and cake on the filter was washed with tetrahydrofuran. The organic filtrate was then concentrated under reduced pressure and placed in a high vacuum, receiving (052), (data) LC/MS LCRS (MH) m/z: level of 414.2), which was used without further purification.

Connection (053):

To a cooled to 0°C solution (052) and (006) (0,85 g, 3.9 mmol), NOT (0,70 g, 5.3 mmol) and HBTU (1.70 g, 4.9 mmol) in tetrahydrofuran (100 ml) was added a solution of N,N-diisopropylethylamine (3 ml) in tetrahydrofuran (10 ml) and the mixture was stirred at room temperature overnight. Then it was diluted with ethyl acetate (200 ml), washed with saturated aqueous (solution) sodium bicarbonate (2×50 ml) and with brine (50 ml). The organic layers were dried over sodium sulfate, filtered through celite-545, the solvent was removed under reduced pressure and the residue was purified flash chromatography (hexane and ethyl acetate), obtaining (053) (1.51 g), which okharakterizovali LC/MS (LCRS (MH) m/z: 567,21).

Connection (054):

To a cooled to 0°C solution (053) (200 mg, 0,352 mmol) in dichloromethane (10 ml) was added triperoxonane acid (10 ml) and the resulting solution was stirred at the same temperature over the next hour. The solution was concentrated under reduced pressure and placed in a high vacuum, receiving (054), as confirmed by LC/MS (LCRS (MH) m/z: 467,26), which was used without further purification.

Connection (055):

To a cooled to 0°C solution (054), 5-methylisoxazol-3-carboxylic acid (009) (127 mg, 1.0 mmol), NOT (135 mg, 1.0 mmol) and HBTU (350 g, 1.0 mmol) in tetrahydrofuran (100 ml) was added races is theft N,N-diisopropylethylamine (0.5 ml) in tetrahydrofuran (2 ml). The mixture was stirred at room temperature for 5 hours. Then it was diluted with ethyl acetate (200 ml) and washed with saturated aqueous (solution) sodium bicarbonate (2×10 ml) and with brine (10 ml). The organic layers were dried over sodium sulfate and filtered through celite-545; solvents were removed under reduced pressure and the residue was purified HPLC (aqueous ammonium acetate and acetonitrile), receiving (055) (40 mg), which okharakterizovali LC/MS (LCRS (MH) m/z: 576,27; inhibition >80% proteasome CT-L at 20 mg/kg RO.

Example 9

Scheme 9: synthesis of example 061

Connection (057):

To a cooled to 0°C. a solution of N-Boc-L-n-valine (056) (1.0 g, 4.6 mmol), hydrochloride benzyl ester of L-phenylalanine (002) (1.4 g, 4.6 mmol), NAWT (1.0 g, 7.4 mmol) and HBTU (2.8 g, 7.4 mmol) in tetrahydrofuran (100 ml) was added a solution of N,N-diisopropylethylamine (3.2 ml, 18.4 mmol) in tetrahydrofuran (10 ml). The mixture was stirred at room temperature for 3 hours and then was diluted with ethyl acetate (200 ml), washed with saturated aqueous (solution) sodium bicarbonate (2×50 ml) and with brine (100 ml) and the organic layers were dried over sodium sulfate and filtered through celite-545. The solvent was removed under reduced pressure and the residue was purified flash chromatography (hexane and ethyl acetate), obtaining (057), which okharakterizovali LC/MS (LCRS (MH) m/z 455,25.

Connection (058):

To a cooled to 0°C solution (057) (1.30 grams, 2,875 mmol) in tetrahydrofuran (100 ml) was added 10%Pd/C (100 mg). The resulting mixture was stirred under the hydrogen pressure of 1 atmosphere for 4 hours. Then the mixture was filtered through celite-545 and cake on the filter was washed with tetrahydrofuran. Then the filtrate was concentrated under reduced pressure and placed in a high vacuum, receiving (058), as confirmed by LC/MS (LCRS (MH) m/z: 365,2), which was used without further purification.

Connection (059):

To a cooled to 0°C solution (058) and (006) (0,99 g, 4.6 mmol), NOT (of 0.62 g, 4.6 mmol) and HBTU (1.70 g, 4.9 mmol) in tetrahydrofuran (100 ml) was added a solution of N,N-diisopropylethylamine (2.4 g) in tetrahydrofuran (10 ml). The mixture was stirred at room temperature overnight and then was diluted with ethyl acetate (200 ml) and washed with saturated aqueous (solution) sodium bicarbonate (2×50 ml) and with brine (50 ml). The organic layers were dried over sodium sulfate and filtered through celite-545. The solvents were removed under reduced pressure and the residue was purified HPLC (aqueous ammonium acetate and acetonitrile), receiving (059) (1,21 g), which okharakterizovali LC/MS (LCRS (MH) m/z: 518,32).

Connection (060):

To a cooled to 0°C solution (059) (250 mg, 0.48 mmol) in dichloromethane (10 ml) was added triperoxonane acid (10 ml) and the resulting solution p is remedial at the same temperature over the next hour. The organic layers were concentrated under reduced pressure and placed in a high vacuum, receiving (060), as confirmed by LC/MS (LCRS (MH) m/z: 418,26), which was used without further purification.

Connection (061):

To a cooled to 0°C. a solution of (060) and (022) (122 mg, of 0.58 mmol), NOT (104 mg, 0.77 mmol) and HBTU (282 mg, to 0.72 mmol) in tetrahydrofuran (100 ml) was added a solution of N,N-diisopropylethylamine (0.35 ml) in tetrahydrofuran (2 ml) and the mixture was stirred at room temperature over the next 4 hours. Then it was diluted with ethyl acetate (200 ml) and washed with saturated aqueous (solution) sodium bicarbonate (2×10 ml) and with brine (10 ml). The organic layers were dried over sodium sulfate and filtered through celite-545. The solvents were removed under reduced pressure and the residue was purified HPLC (aqueous ammonium acetate and acetonitrile), receiving (061) (88,4 mg), which okharakterizovali LC/MS (LCRS (MH) m/z: 612,33); inhibition >80% proteasome CT-L at 40 mg/kg RO.

Example 10

Scheme 10: synthesis of example 068

Connection (063):

To a cooled to 0°C. a solution of N-Boc-HoSer(OMe)-OH (062) (1.0 g, 4.3 mmol), hydrochloride benzyl ester of L-phenylalanine (002) (1.3 g, 4.3 mmol), NOT (0.88 g, 6.5 mmol) and HBTU (2.3 g, 6.5 mmol) in tetrahydrofuran (100 ml) was added a solution of N,N-diisopropylethylamine (2.0 ml) in tetrahydrofuran (5 ml). The mixture was stirred is at room temperature over the next 3 hours then was diluted with ethyl acetate (200 ml) and washed with saturated aqueous (solution) sodium bicarbonate (2×50 ml) and with brine (100 ml). The organic layers were dried over sodium sulfate and filtered through celite-545. The solvents were removed under reduced pressure and the residue was purified flash chromatography (hexane and ethyl acetate), obtaining (063) (1,81 g), which okharakterizovali LC/MS (LCRS (MH) m/z: 471,24).

Connection (064):

To a cooled to 0°C solution (063) (1.35 g, 2,875 mmol) in tetrahydrofuran (100 ml) was added 10%Pd/C (100 mg). The resulting mixture was stirred under the hydrogen pressure of 1 atmosphere for 4 hours. Then the mixture was filtered through celite-545 and cake on the filter was washed with tetrahydrofuran. The organic filtrate was concentrated under reduced pressure and placed in a high vacuum, receiving (064), as confirmed by LC/MS (LCRS (MH) m/z: 381,19), which was used without further purification.

Connection (065):

To a cooled to 0°C solution (065) and (006) (0,99 g, 4.6 mmol), NOT (of 0.62 g, 4.6 mmol) and HBTU (1.70 g, 4.9 mmol) in tetrahydrofuran (100 ml) was added a solution of N,N-diisopropylethylamine (2.4 ml) in tetrahydrofuran (10 ml). The mixture was stirred at room temperature overnight and then was diluted with ethyl acetate (200 ml) and washed with saturated aqueous (solution) sodium bicarbonate (2×50 ml) and with brine (50 ml). The organic layers su is or over sodium sulfate and filtered through celite-545. The solvents were removed under reduced pressure and the residue was purified HPLC (aqueous ammonium acetate and acetonitrile), receiving (065) (1.11 mg), which okharakterizovali LC/MS (LCRS (MH) m/z: 534,31).

Connection (066):

To a cooled to 0°C solution (065) (230 mg, 0.43 mmol) in dichloromethane (20 ml) was added triperoxonane acid (10 ml) and the resulting solution was stirred at the same temperature over the next hour. Then the reaction mixture was concentrated under reduced pressure and placed in a high vacuum, receiving (066), as confirmed by LC/MS (LCRS (MH) m/z: 434,26), which was used without further purification.

Connection (068):

To a cooled to 0°C solution (066) and 5-isopropylthiazole-3-carboxylic acid (067) (81 mg, 0.52 mmol), NOT (93 mg, 0.69 mmol) and HBTU (262 mg, 0.69 mmol) in tetrahydrofuran (100 ml) was added a solution of N,N-diisopropylethylamine (0,30 ml) in tetrahydrofuran (2 ml) and the mixture was stirred at room temperature over the next 4 hours. Then it was diluted with ethyl acetate (200 ml) and washed with saturated aqueous (solution) sodium bicarbonate (2×10 ml) and with brine (10 ml). The organic layers were dried over sodium sulfate and filtered through celite-545. The solvents were removed under reduced pressure and the residue was purified HPLC (aqueous ammonium acetate and acetonitrile), receiving (068) (75,7 mg), which okharakterizovali LC/MS (LCRS (MH) m/z: 571,31) inhibition > 70% of the proteasome CT-L at 40 mg/kg RO.

Example 11

Scheme 11: synthesis of example 075 (method A)

Connection (070):

To a solution of methyl ester hydrochloride L-serine (069) (1.0 g, 6.4 mmol) in a mixture of water/dioxane (1:1, 80 ml) was added sodium hydroxide (768 mg, 19.2 mmol). After stirring the mixture at room temperature for 30 minutes, it was cooled to 0°C was added dropwise a solution of 9-fluorenylmethoxycarbonyl (1.65 g, 6.4 mmol) in dioxane (16 ml). The following 4 hours, the reaction mixture was allowed to mix at room temperature. Then the solvent was removed, the residue was diluted with water and 1 N. HCl brought the pH to ~1; the aqueous layer was extracted with ethyl acetate (4×100 ml). The organic layers were concentrated under reduced pressure and placed in a high vacuum, receiving (070) (1.8 g), which was confirmed by LC/MS (LCRS (MH) v/z: 342,13), which was used without further purification.

Connection (071):

Resin NMRV-VNA (500 mg, 0.32 mmol) was washed with dichloromethane. In a dry flask was dissolved in dichloromethane Fmoc-Ser(Me)-OH (070) (546 mg, 1.6 mmol), the solution was added 1-Mei (95 μl, 1.2 mmol) and then MSNT (474 mg, 1.6 mmol). After the reaction mixture became homogeneous (10 minutes), it was added to the resin NMRV-VNA in the form of a suspension in dichloromethane (5 ml). The resulting reaction mixture was left to shake what I'm in for the night. Then the resin was filtered and washed with DMF (3×20 ml), Meon (3×20 ml), DCM (3 x 20 ml) and left to air dry, receiving (071).

Connection (072):

The resin (071) (300 mg, 0,192 mmol) were placed in a 20%solution of piperidine in N,N-dimethylformamide (20 ml) and the resulting mixture was shaken at room temperature for 30 minutes. The resin was filtered and twice washed with N,N-dimethylformamide (3×20 ml) and dichloromethane (3×20 ml).

To a cooled to 0°C. a solution of Fmoc-Ser(Me)-OH (070) (0.48 mmol, 163 mg) in N,N-dimethylformamide (10 ml) was added NOWT (104 mg, 0.77 mmol), HBTU (291 mg, 1.92 mmol) and diisopropylethylamine (0,34 ml, 1.92 mmol). After the mixture became homogeneous, was added to the resin (0.13 mmol, 200 mg) and the resulting reaction mixture was left to shake things up during the night. Then the resin was filtered off, washed with DMF (10 ml), DCM (10 ml), Meon (10 ml), N2About (10 ml), DMF (10 ml), Meon (10 ml) and DCM (10 ml) and left to air dry, receiving (072).

Connection (073):

(072) (300 mg, 0,19 mmol) was added a 20%solution of piperidine in N,N-dimethylformamide (20 ml) and the resulting mixture was shaken at room temperature for 30 minutes. The resin was filtered and twice washed with N,N-dimethylformamide (3×20 ml) and dichloromethane (3×20 ml).

To a cooled to 0°C. the solution 009) (61 mg, 0.48 mmol) in N,N-dimethylformamide (2 ml) was added NOWT (104 mg, 0.77 mmol), HBTU (291 is g, 1.77 mmol) and N,N - diisopropylethylamine (0,34 ml, 1.92 mmol). After the mixture became homogeneous, was added to the resin (300 mg, 0,192 mmol) and the resulting reaction mixture was left to shake at room temperature overnight. Then the resin was filtered off, washed with DMF (10 ml), DCM (10 ml), Meon (10 ml), N2About (10 ml), DMF (10 ml), Meon (10 ml) and DCM (10 ml) and left to air dry, receiving (073).

Connection (074):

(073) was added a 50%solution triperoxonane acid in dichloromethane (10 ml) and the resulting mixture was left to shake for 30 minutes. Then the resin was filtered and washed with dichloromethane (3×10 ml). Volatile (medium) was removed under reduced pressure and the desired compound (074) okharakterizovali LC/MS (LCRS (MH) m/z: 330,12) and used without further purification.

Connection (075):

To a cooled to 0°C solution (074) and (006) (78 mg, 0.38 mmol), NOT (41 mg, 0.30 mmol) and HBTU (116 mg, 0.30 mmol) in acetonitrile (50 ml) was added a solution of N,N-diisopropylethylamine (0.1 ml, 0.6 mmol). The mixture was stirred at 0-4°C overnight and then was diluted with ethyl acetate (200 ml). It was further washed with saturated aqueous (solution) sodium bicarbonate (2×10 ml) and with brine (10 ml), organic layers were dried over sodium sulfate and filtered through celite-545. The solvents were removed under reduced pressure and the residue cleaned and HPLC (aqueous ammonium acetate and acetonitrile), receiving (075) (29 mg), which okharakterizovali LC/MS (LCRS (MH) m/z: 483,24); inhibition >80% proteasome CT-L at 20 mg/kg RO.

Example 12

Scheme 12: synthesis of example 075 (method)

Connection (076):

To a cooled to 0°C. a solution of methyl ester of N-Boc-serine-HE (43,8 g, 200 mmol), triethylamine (26.5 g, 260 mmol) and 4-(dimethylamino)pyridine in dichloromethane (1.2 l) for 30 minutes was added a solution of benzylbromide (41 g, 240 mmol) in dichloromethane (250 ml) and the resulting mixture was stirred at the same temperature over the next 3 hours. Then was added a saturated aqueous (solution) sodium bicarbonate (200 ml) and the organic layer was washed with saturated aqueous (solution) sodium bicarbonate (200 ml) and with brine (200 ml). The organic layers were dried over sodium sulfate and filtered through celite-545. The solvents were removed under reduced pressure and the residue was purified flash chromatography (hexane and ethyl acetate), obtaining (076) (54 g), which okharakterizovali LC/MS (LCRS (MH) m/z: 310,16).

Connection (077):

To a cooled to 0°C solution (076) (54 g, 174,6 mmol) in dichloromethane (200 ml) for 10 minutes was added triperoxonane acid (200 ml) and the resulting mixture was stirred at the same temperature for another 3 hours. Then the solvents were removed under reduced pressure and the residue was placed in a high vacuum, obtaining the Sol is TFA c (077), as confirmed by LC/MS (LCRS (MH) m/z: 210,11), which was used without further purification.

Connection (078):

To a cooled to 0°C solution (077) (43,8 g, 200 mmol), methyl ester of N-Boc-serine-OH (36,7 g, 167 mmol), NOT (27 g, 200 mmol) and HBTU (71,4 g, 200 mmol) in tetrahydrofuran (1.2 l) for 10 minutes was added a solution of N,N-diisopropylethylamine (75 g, 600 mmol) in tetrahydrofuran (250 ml); the pH of the mixture was equal to ~8. The mixture was stirred at room temperature for another 5 hours. Then the greater part of the solvent was removed under reduced pressure and the resulting material was diluted with ethyl acetate (1.0 l). It was further washed with saturated aqueous (solution) sodium bicarbonate (2×150 ml) and with brine (200 ml); the organic layers were dried over sodium sulfate and filtered through celite-545. The solvents were removed under reduced pressure and the residue was purified flash chromatography (hexane and ethyl acetate), obtaining (078) (65 g), which okharakterizovali LC/MS (LCRS (MH) m/z: 411,21).

Connection (079):

To a cooled to 0°C solution (079) (13,4 g, to 32.7 mmol)in tetrahydrofuran (300 ml) was added 10%Pd/C (2.7 g) and the resulting mixture was stirred for 4 hours under a hydrogen pressure of 1 atmosphere. The mixture was filtered through celite-545 and cake on the filter was washed with tetrahydrofuran. The organic layers were concentrated under reduced pressure and placed in high Vacu is m, receiving (079), as confirmed by LC/MS (LCRS (MH) m/z: 321,16), which was used without further purification.

Connection (080):

To a cooled to 0°C solution (079) and (006) (5.6 g, 26 mmol), NOT (6 g, up 41.4 mmol) and HBTU (14.8 g, up 41.4 mmol) in tetrahydrofuran (400 ml) was added a solution of N,N-diisopropylethylamine (23 ml) in tetrahydrofuran (40 ml) and the mixture was stirred at room temperature overnight. Then the greater part of the solvent was removed under reduced pressure, the residue was diluted with ethyl acetate (500 ml) and washed with saturated aqueous (solution) sodium bicarbonate (2×100 ml) and with brine (100 ml). The organic layers were dried over sodium sulfate and filtered through celite-545. The solvents were removed under reduced pressure and the residue was purified flash chromatography (hexane and ethyl acetate), obtaining (080) (9.2 grams), which okharakterizovali LC/MS (LCRS (MH) m/z: 474,27).

Connection (081):

To a cooled to 0°C solution (080) (200 mg, 0.43 mmol) in dichloromethane (10 ml) was added triperoxonane acid (10 ml) and the resulting solution was stirred at the same temperature for another hour. The organic layers were concentrated under reduced pressure and placed in a high vacuum, receiving (081), as confirmed by LC/MS (LCRS (MH) m/z: 374,22), which was used without further purification.

Connection (075):

To a cooled to 0°C solution (081) and 5-methylisoxazol-carboxylic acid (009) (65 mg, 0.5 mmol), NOT (65 mg, 0.5 mmol) and HBTU (175 mg, 0.5 mmol) in tetrahydrofuran (50 ml) was added a solution of N,N-diisopropylethylamine (0.5 ml) in tetrahydrofuran (2 ml) and the mixture was stirred at room temperature for another 5 hours. Then it was diluted with ethyl acetate (200 ml) and washed with saturated aqueous (solution) sodium bicarbonate (2×10 ml) and with brine (10 ml). The organic layers were dried over sodium sulfate and filtered through celite-545. The solvents were removed under reduced pressure and the residue was purified HPLC (aqueous ammonium acetate and acetonitrile), receiving (075) (85 mg), which okharakterizovali LC/MS (LCRS (MH) m/z: 483,24).

Example 13

Scheme 13: synthesis of example 083

Connection (083):

To a cooled to 0°C solution (081) (160 mg, 0.43 mmol) and isoxazol-3-carboxylic acid (082) (60 mg, 0.5 mmol), NOT (65 mg, 0.5 mmol) and HBTU (175 mg, 0.5 mmol) in tetrahydrofuran (50 ml) was added a solution of N,N-diisopropylethylamine (0.5 ml) in tetrahydrofuran (2 ml) and the mixture was stirred at room temperature for another 5 hours. Then it was diluted with ethyl acetate (200 ml) and washed with saturated aqueous (solution) sodium bicarbonate (2×10 ml) and with brine (10 ml). The organic layers were dried over sodium sulfate and filtered through celite-545. The solvents were removed under reduced pressure and the residue was purified HPLC (water AC the tat ammonium and acetonitrile), receiving (083) (74 mg), which okharakterizovali LC/MS (LCRS (MH) m/z: 469,22); inhibition >80% proteasome CT-L at 20 mg/kg RO.

Example 14

Scheme 14: synthesis of example 085

Connection (085):

To a cooled to 0°C solution (081) (160 mg, 0.43 mmol) and isoxazol-3-carboxylic acid (084) (65 mg, 0.5 mmol), NOT (65 mg, 0.5 mmol) and HBTU (175 mg, 0.5 mmol) in tetrahydrofuran (50 ml) was added a solution of N,N-diisopropylethylamine (0.5 ml) in tetrahydrofuran (2 ml) and the mixture was stirred at room temperature for another 5 hours. Then the reaction mixture was diluted with ethyl acetate (200 ml) and washed with saturated aqueous (solution) sodium bicarbonate (2×10 ml) and with brine (10 ml). The organic layers were dried over sodium sulfate and filtered through celite-545. The solvents were removed under reduced pressure and the residue was purified HPLC (aqueous ammonium acetate and acetonitrile), receiving (085) (71 mg), which okharakterizovali LC/MS (LCRS (MH) m/z: 483,24); inhibition >50% of the proteasome CT-L at 20 mg/kg RO.

Scheme 15: synthesis of example 088

Connection (087):

To a solution of (086) (obtained in the same way as (005), except that Cbz-phenylalanine was replaced with Cbz-leucine) (0,100 g, 0,295 mmol) in triperoxonane acid (10 ml) was added 10%Pd/C (20 mg). The resulting mixture was left to mix on the pressure of hydrogen at 1 atmosphere for 6 hours. Then the mixture was filtered through celite-545 and cake on the filter was washed with dichloromethane (50 ml). The filtrate was concentrated under reduced pressure and placed in a high vacuum over night, receiving (087), as confirmed by LC/MS (LCRS (MH) m/z: 206,1), which was used in the next transformation without further purification.

Connection (088):

To a cooled to 0°C solution (087) and (074) (166 mg, 0,354 mmol), NOT (54 mg, 0,354 mmol) and HBTU (134 mg, 0,354 mmol) in tetrahydrofuran (20 ml) was added a solution of N,N-diisopropylethylamine (0.2 ml, 1.18 mmol). The mixture was stirred at 0°C during the night, and it became homogeneous. Then it was diluted with ethyl acetate (20 ml) and washed with saturated aqueous (solution) sodium bicarbonate (2×10 ml) and with brine (10 ml). The organic layers were dried over sodium sulfate, filtered through celite-545 and concentrated under reduced pressure; the residue was purified HPLC (aqueous ammonium acetate and acetonitrile), receiving (088) (10 mg), which okharakterizovali LC/MS (LCRS (MH) m/z: 517,69); inhibition >80% proteasome CT-L at 20 mg/kg RO.

Scheme 16: synthesis of example 091

Connection (090):

To a solution of (089) (obtained in the same way as (005), except that Cbz-4-forfinally was replaced by Cbz-leucine) (0,100 g, 0.28 mmol) in triperoxonane acid (10 ml) was added 10%Pd/C (20 mg). The mixture of toulali mixed under pressure of hydrogen at 1 atmosphere for 6 hours. Then the mixture was filtered through celite-545 and cake on the filter was washed with dichloromethane (50 ml). The filtrate was concentrated under reduced pressure and placed in a high vacuum over night, receiving (090), as confirmed by LC/MS (LCRS (MH) m/z: 224,1), which was used in the next transformation without further purification.

Connection (091):

To a cooled to 0°C solution (090) and (074) (110 mg, 0,336 mmol), NOT (51 mg, 0,336 mmol) and HBTU (127 mg, 0,336 mmol) in tetrahydrofuran (20 ml) was added N,N-diisopropylethylamine (0.2 ml, 1.18 mmol). The mixture was stirred at 0°C during the night, and it became homogeneous. Then it was diluted with ethyl acetate (20 ml) and washed with saturated aqueous (solution) sodium bicarbonate (2×10 ml) and with brine (10 ml). The organic layers were dried over sodium sulfate, filtered through celite-545 and concentrated under reduced pressure. Then the residue was purified HPLC (aqueous ammonium acetate and acetonitrile), receiving (091) (60 mg), which okharakterizovali LC/MS (LCRS (MH) m/z: 535,69); inhibition >80% proteasome CT-L at 20 mg/kg RO.

Biological activity

Compounds were prepared in media for 10%PS80/Na-citrate (pH 3) and is administered orally (PO) mice (3 animals per group). One hour after dosing, animals were killed and selected the following tissues: blood, brain, adrenal, heart, and liver. Whole blood (~200 µl) was twice washed in PBS and whether Aravali hypotonic shock (hypotonic shock) (300 μl 50 mm Tris pH 8, 5 mm EDTA). The blood lysates were stored at -80°C until analysis. The blood lysates were cleared by centrifugation in microcentrifuge. The specific activity of the proteasome CT-L in each lysate was assessed by determining: a) the concentration of protein breakdown Bradford (Bradford) with bovine gamma globulin as standard and (b) the rate of cleavage of the substrate fluorogenic proteasome LLVY-AMC. The percentage of proteasome activity to similarly treated animals was calculated by dividing the average specific activity of each similarly dosed groups on the average specific activity of the group dosed media. The percentage inhibition of the proteasome was calculated by subtracting the percentage of proteasome activity of 100.

Equivalents

Specialists in this field are able to identify and establish, using no more than ordinary experimentation, many equivalents of the compounds described herein and methods of their use. Such equivalents are considered (included in the scope of the invention and covered by the following claims.

All the above-quoted references and publications cited here as a reference.

1. The compound having the structure of formula (I)or its pharmaceutically acceptable salt:

in which L is selected from C=O;
X is selected from O;
Z is absent;
each of R1and Rsup> 2independently from each other selected from hydrogen, C1-6-alkyl, C1-6-carbonylmethyl,1-6-alkoxyalkyl,1-6-hydroxyalkyl,1-6-cyanoalanine, C1-6-aralkyl,2-6-quinil, and C1-6-heteroalkyl where heteroaryl represents a 5 - or 6-membered ring containing one or two heteroatoms selected from N or S;
R3selected from C1-6-alkyl and C1-6-aralkyl, optionally substituted with halogen, C1-6-alkyl, C1-6-alkoxy or CF3;
R4represents hydrogen;
R5represents isoxazol replaced by amidon, C1-6-alkyl, C1-6-carbocyclization,1-6-alkoxy, C1-6-hydroxyalkyl, C1-6-alkoxyalkyl, C1-6-heterocyclization, where the heterocycle is a non-aromatic 6-membered ring containing N and O and optionally optionally substituted C1-6-alkyl, non-aromatic 4-membered ring containing one N,
or non-aromatic 5-membered ring containing one N, or C1-6-heteroalkyl where heteroaryl contains three atoms of N; furan or thiazole, optionally substituted C1-6-alkyl; and R6and R7represents hydrogen.

2. The connection of claim 1, wherein any of R1, R2and R3independently from each other represents Soboh the C 1-6-alkyl.

3. The compound according to claim 2, in which any one of R1, R2and R3independently from each other selected from methyl, ethyl, propyl, isopropyl, butyl, sec-butyl and isobutyl.

4. The connection of claim 1, wherein any of R1, R2and R3independently from each other represents propargyl.

5. The connection of claim 1, wherein any of R1and R2independently from each other represents C1-6-hydroxyalkyl.

6. The compound according to claim 5, in which any one of R1and R2independently from each other selected from hydroxymethyl and hydroxyethyl.

7. The connection of claim 1, wherein any of R' and R2independently from each other represents C1-6-alkoxyalkyl.

8. The connection according to claim 7, in which any one of R1and R2independently from each other selected from methoxymethyl and methoxyethyl.

9. The connection of claim 1, wherein any of R1and R2independently from each other represents C1-6-heteroalkyl.

10. The compound according to claim 1, in which at least one of R1and R2selected from C1-6-hydroxyalkyl and C1-6-alkoxyalkyl.

11. The connection of claim 10, in which at least one of R1and R2represents a C1-6-alkoxyalkyl.

12. Connection by claim 11, in which at least one of R1and R2selected from methoxime the silt and methoxyethyl.

13. The compound according to claim 1, in which R3selected from C1-6-alkyl and C1-6-aralkyl.

14. The connection 13, in which R3represents a C1-6-alkyl.

15. The connection 14, in which R3selected from methyl, ethyl, isopropyl, sec-butyl and isobutyl.

16. The connection indicated in paragraph 15, in which R3represents isobutyl.

17. The connection 13, in which R3represents a C1-6-aralkyl.

18. The connection 17 in which R represents phenylmethyl.

19. The compound according to claim 1, in which R5represents isoxazol-3-yl or isoxazol-5-yl.

20. The connection according to claim 19, in which R5represents isoxazol-3-yl, containing a substituent in the 5-position.

21. The connection according to claim 19, in which R5represents isoxazol-5-yl, containing the substituent in 3-position.

22. Connection claim 20 or 21, in which the Deputy is selected from methyl, ethyl, isopropyl and cyclopropylmethyl.

23. Connection claim 20 or 21, in which the Deputy is selected from C1-6-heteroalkyl and C1-6-geterotsiklicheskie.

24. Connection item 23, in which the Deputy is a 1,2,4-triazole-5-ylmethyl.

25. Connection item 23, in which the Deputy is azetidin-1-ylmethyl.

26. Connection item 23, in which the Deputy is selected from C1-6-alkoxygroup and C1-6 -alkoxyalkyl.

27. Connection p, in which the Deputy is selected from metoxygroup, ethoxypropan, methoxymethyl and methoxyethyl.

28. A compound selected from















29. Pharmaceutical composition having inhibitory activity against 20S proteasome and containing a therapeutically effective amount of a compound according to any one of claims 1 to 28 and a pharmaceutically acceptable diluent or carrier.

30. The pharmaceutical composition according to clause 29, which is orally bioavailable.

31. A method of treating cancer, introducing a therapeutically effective amount of the pharmaceutical composition according to item 30.

32. The compound of the formula

or its pharmaceutically acceptable salt.

33. Pharmaceutical composition having inhibitory activity against the 2OS Proteas who we are and containing a therapeutically effective amount of a compound according p and a pharmaceutically acceptable diluent or carrier.

34. A method of treating cancer, introducing a therapeutically effective amount of the pharmaceutical composition according p.



 

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FIELD: chemistry.

SUBSTANCE: invention relates to novel IAP inhibitors of general formula , where Q, X1, X2, Y, Z1, Z2, Z3, Z4, R1, R2, R3, R3', R4, R4', R5, R6, and R6', and n assume values given in the description.

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FIELD: medicine.

SUBSTANCE: invention refers to a compound having the structure of formula (I), or its pharmaceutically acceptable salt, wherein specified radicals are presented in the description, and also concerns a compound representing or its pharmaceutically acceptable salt. The present invention declares a pharmaceutical composition possessing inhibitory activity in the relation to 20S proteasome containing a pharmaceutically acceptable carrier or a diluent and a therapeutically effective amount of the compound, and also the invention refers to methods of treating the immune diseases, such as inflammatory intestinal disease, to treating cancer, to treating infection, to treating proliferative diseases, to treating neurodegenerative disease or asthma.

EFFECT: higher clinical effectiveness.

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FIELD: medicine.

SUBSTANCE: invention refers to a compound having the structure of formula (I), or its pharmaceutically acceptable salt, wherein specified radicals are presented in the description, and also concerns a compound representing or its pharmaceutically acceptable salt. The present invention declares a pharmaceutical composition possessing inhibitory activity in the relation to 20S proteasome containing a pharmaceutically acceptable carrier or a diluent and a therapeutically effective amount of the compound, and also the invention refers to methods of treating the immune diseases, such as inflammatory intestinal disease, to treating cancer, to treating infection, to treating proliferative diseases, to treating neurodegenerative disease or asthma.

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30 cl, 25 ex, 2 tbl

FIELD: medicine, pharmaceutics.

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EFFECT: obtaining pharmaceutical composition which possesses inhibiting activity with respect to NS3-4 HCV serinprotease, including formula I compound and pharmaceutically acceptable carrier.

30 cl, 25 ex, 2 tbl

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SUBSTANCE: in claim described are organic compounds of formula I where radicals are given in description, which are applicable for elimination, prevention or alleviation of one or more symptoms, associated with HCV disorders.

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23 cl, 11 dwg, 9 tbl, 15 ex

FIELD: chemistry.

SUBSTANCE: invention relates to novel azaindole derivatives, having JAK-2 and JAK-3 kinase inhibiting activity, or pharmaceutically acceptable salts thereof. In formula (I): R3 denotes H; X1 denotes N or CR4; R2 denotes H, COOH, COOR' or CONHR'; R4 denotes H, F, R, OH, OR', COR', COOH, COOR', CONH2 or CN; or R2 and R4, taken together, form a benzene ring optionally substituted with 1-2 R10; R' denotes C1-3-alkyl or C1-3-alkenyl, each optionally substituted 1-2 R5; each R5 is independently selected from CN, unsubstituted C1-2alkyl, or two groups R5 together with a carbon atom with which they are bonded form a cyclopropyl ring; each R10 is independently selected from halogen, OCH3 or OH; R1 denotes or , R is H or denotes C1-2alkyl, optionally substituted with 1-3 R11; R6 denotes C1-4alkyl, optionally substituted with 1-5 R12; values of radicals R7 -R9, ring A, R11 -R14. The invention also relates to a pharmaceutical composition containing said compounds and a method of treating or reducing severity of a pathological condition such as allergy, asthma, amyotrophic lateral sclerosis, multiocular sclerosis, graft rejection, rheumatoid arthritis, solid malignant tumour, haematologic malignant disease, leukaemia, lymphoma and myeloproliferative disorders.

EFFECT: high efficiency of using the compounds.

41 cl, 6 ex, 6 tbl

FIELD: chemistry.

SUBSTANCE: invention relates to 2,3-substituted pyrazine sulphonamides of formula (I), use thereof in treating allergic diseases, inflammatory dermatosis, immonological disorders and neurodegenerative disorders, as well as pharmaceutical compositions, having CRTH2 receptor inhibiting action and inhibiting chemoattractant receptor, homologous to the molecule expressed on T-helpers 2. in general formula .

A is selected from a group consisting of

, n denotes an integer independently selected from 0, 1, 2, 3 or 4; m equals 1 or 2; B is selected from a group consisting of phenyl or piperazinyl; R1 denotes hydrogen; R2 denotes phenyl, where R2 is optionally substituted with one or more substitutes selected from a group consisting of halogen, cyano, (C1-C6)alkyl; R3 is selected from a group consisting of (C1-C6)alkyl, aryl, heteroaryl, (C1-C6)alkylaryl, (C1-C6)alkylheteroaryl, (C3-C8)cycloalkyl and (C3-C8)heterocycloalkyl, where each of said (C1-C6)alkyl, aryl, heteroaryl, (C1-C6)alkylaryl, (C1-C6)alkylheteroaryl, (C3-C8)cycloalkyl and (C3-C8)heterocycloalkyl is optionally substituted with one or more substitutes selected from a group consisting of halogen, cyano, (C1-C6)alkyl, (C1-C6)alkoxy, heteroaryl, aryl, thioalkoxy and thioalkyl, or where said aryl, heteroaryl, (C1-C6)alkylaryl, (C1-C6)alkylheteroaryl, (C3-C8)cycloalkyl or (C3-C8)heterocycloalkyl can be condensed with one or more aryl, heteroaryl, (C3-C8)cycloalkyl or (C3-C8)heterocycloalkyl groups and can be substituted with one or more substitutes selected from a group consisting of (C1-C6)alkyl, alkoxy, aryl, heteroaryl, carboxyl, cyano, halogen, hydroxy, amino, aminocarbonyl, nitro, sulphoxy, sulphonyl, sulphonamide and trihaloalkyl; R7 is selected from a group consisting of hydrogen, (C1-C6)alkyl, (C2-C6)alkenyl, (C2-C6)alkynyl, aryl, heteroaryl, (C3-C8)cycloalkyl, (C3-C8)heterocycloalkyl, carboxyl, cyano, amino and hydroxy; aryl is selected from phenyl or naphthyl; and heteroaryl is selected from pyridyl, indolyl, 3H-indolyl, benzimidazolyl, quinolizinyl.

EFFECT: high efficiency of using the compounds.

4 cl, 10 dwg, 46 ex

FIELD: chemistry.

SUBSTANCE: invention relates to 2,3-substituted pyrazine sulphonamides of formula (I), use thereof in treating allergic diseases, inflammatory dermatosis, immonological disorders and neurodegenerative disorders, as well as pharmaceutical compositions, having CRTH2 receptor inhibiting action and inhibiting chemoattractant receptor, homologous to the molecule expressed on T-helpers 2. in general formula .

A is selected from a group consisting of

, n denotes an integer independently selected from 0, 1, 2, 3 or 4; m equals 1 or 2; B is selected from a group consisting of phenyl or piperazinyl; R1 denotes hydrogen; R2 denotes phenyl, where R2 is optionally substituted with one or more substitutes selected from a group consisting of halogen, cyano, (C1-C6)alkyl; R3 is selected from a group consisting of (C1-C6)alkyl, aryl, heteroaryl, (C1-C6)alkylaryl, (C1-C6)alkylheteroaryl, (C3-C8)cycloalkyl and (C3-C8)heterocycloalkyl, where each of said (C1-C6)alkyl, aryl, heteroaryl, (C1-C6)alkylaryl, (C1-C6)alkylheteroaryl, (C3-C8)cycloalkyl and (C3-C8)heterocycloalkyl is optionally substituted with one or more substitutes selected from a group consisting of halogen, cyano, (C1-C6)alkyl, (C1-C6)alkoxy, heteroaryl, aryl, thioalkoxy and thioalkyl, or where said aryl, heteroaryl, (C1-C6)alkylaryl, (C1-C6)alkylheteroaryl, (C3-C8)cycloalkyl or (C3-C8)heterocycloalkyl can be condensed with one or more aryl, heteroaryl, (C3-C8)cycloalkyl or (C3-C8)heterocycloalkyl groups and can be substituted with one or more substitutes selected from a group consisting of (C1-C6)alkyl, alkoxy, aryl, heteroaryl, carboxyl, cyano, halogen, hydroxy, amino, aminocarbonyl, nitro, sulphoxy, sulphonyl, sulphonamide and trihaloalkyl; R7 is selected from a group consisting of hydrogen, (C1-C6)alkyl, (C2-C6)alkenyl, (C2-C6)alkynyl, aryl, heteroaryl, (C3-C8)cycloalkyl, (C3-C8)heterocycloalkyl, carboxyl, cyano, amino and hydroxy; aryl is selected from phenyl or naphthyl; and heteroaryl is selected from pyridyl, indolyl, 3H-indolyl, benzimidazolyl, quinolizinyl.

EFFECT: high efficiency of using the compounds.

4 cl, 10 dwg, 46 ex

FIELD: chemistry.

SUBSTANCE: invention relates to an amine compound of formula (I), pharmaceutically acceptable addition salts, hydrates or solvates thereof, having immunodepressive effect , where R - H or P(=O)(OH)2; X - O or S; Y denotes -CH2CH2- or -CH=CH-; Z denotes C1-5-alkylene, C2-5-alkenylene or C2-5-alkynylene; R1 denotes CF3, R2 denotes C1-4-alkyl, substituted with OH or halogen; R3 and R4 independently denotes H < or C1-4-alkyl; A denotes optionally substituted C6-10-aryl, heteroaryl containing 5-10 ring atoms, where 1 or 2 atoms are selected from N, O and S, C3-7-cycloalkyl optionally condensed with optionally substituted benzene, or heterocycloalkyl containing 5-7 ring atoms, where 1 or 2 atoms are selected from N and O, where said substitutes are selected from C1-4-alkylthio, C1-4-alkylsulphanyl, C1-4-alkylsulphonyl, C2-5-alkylcarbonyl, halogen, cyano, nitro, C3-7-cycloalkyl, C6-10-aryl, C7-14-aralkyloxy, C6-10-aryloxy, optionally substituted with oxo or halogen, C2-3-alkyleneoxy, C3-4-alkylene or C1-2-alkylenedioxy, optionally substituted with halogen C1-4-alkyl or C1-4-alkoxy.

EFFECT: novel compound which is effective in reducing the level of lymphocytes in peripheral blood, suppresses tissue breakdown and exhibiting less side effects, such as bradycardia, is disclosed.

20 cl, 237 ex, 2 tbl

FIELD: chemistry.

SUBSTANCE: invention relates to an amine compound of formula (I), pharmaceutically acceptable addition salts, hydrates or solvates thereof, having immunodepressive effect , where R - H or P(=O)(OH)2; X - O or S; Y denotes -CH2CH2- or -CH=CH-; Z denotes C1-5-alkylene, C2-5-alkenylene or C2-5-alkynylene; R1 denotes CF3, R2 denotes C1-4-alkyl, substituted with OH or halogen; R3 and R4 independently denotes H < or C1-4-alkyl; A denotes optionally substituted C6-10-aryl, heteroaryl containing 5-10 ring atoms, where 1 or 2 atoms are selected from N, O and S, C3-7-cycloalkyl optionally condensed with optionally substituted benzene, or heterocycloalkyl containing 5-7 ring atoms, where 1 or 2 atoms are selected from N and O, where said substitutes are selected from C1-4-alkylthio, C1-4-alkylsulphanyl, C1-4-alkylsulphonyl, C2-5-alkylcarbonyl, halogen, cyano, nitro, C3-7-cycloalkyl, C6-10-aryl, C7-14-aralkyloxy, C6-10-aryloxy, optionally substituted with oxo or halogen, C2-3-alkyleneoxy, C3-4-alkylene or C1-2-alkylenedioxy, optionally substituted with halogen C1-4-alkyl or C1-4-alkoxy.

EFFECT: novel compound which is effective in reducing the level of lymphocytes in peripheral blood, suppresses tissue breakdown and exhibiting less side effects, such as bradycardia, is disclosed.

20 cl, 237 ex, 2 tbl

FIELD: medicine.

SUBSTANCE: invention refers to medicine, particularly the preparation fexofenadine for allergic diseases. The pharmaceutical composition contains an effective amount of fexofenadine as an active ingredient, and excipients - hydroxypropyl cellulose, a filler, desintegrant glidant, a solubiliser and stearic acid salt. The filler represents a combination of lactose and starch. As glidant, the composition contains colloidal silicon dioxide.

EFFECT: pharmaceutical composition is presented in the form of tablets and is characterised by a high degree of release of the active substance and storage stability.

7 cl, 1 tbl, 3 ex

FIELD: food industry.

SUBSTANCE: invention relates to production of a biologically active food additive and may be used for preventive alimentation. Milled horseradish root is infused in water during no less than 24 hours at a ratio of S: L = 1: (4.5 -5.0). The water extract is subjected to vacuum sublimation dehydration. Produced sublimated horseradish root powder in an amount of 25-35 g is mixed with 900-1100 g of natural floral honey. One introduces (while thoroughly stirring) magnolia vine oil into the mixture in an amount of 35-45 drops per 1000 g of the mixture.

EFFECT: simple mixing of the initial components, in a strictly specified ratio without modification of their chemical composition allows to enhance efficiency of the biologically active food additive impact on the immune system and to strengthen antiviral activity.

2 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: group of inventions relates to the field of medicine and deals with particles packed in an immunostimulating nucleic acid and intended for treatment of allergic hypersensitivity. The composition as per the invention concept contains a virus-like particle packed in immunostimulating nucleic acids with the virus-like particle representing an RNA bacteriofague virus-like particle packed in an immunostimulating nucleic acid containing a non-methylated CpG nucleotide. The compositions proposed by the invention are to be most appropriately applied for treatment of atopic eczema, asthma and mediate IgE-allergy(Type I allergy), primarily - pollen allergy and domestic dust allergy.

EFFECT: advantage of the inventions consists in creation of a medication for treatment of hypersensitivity (predominantly - of allergic nature).

24 cl, 39 ex, 32 tbl, 6 dwg

FIELD: medicine.

SUBSTANCE: invention refers to a compound having the structure of formula (I), or its pharmaceutically acceptable salt, wherein specified radicals are presented in the description, and also concerns a compound representing or its pharmaceutically acceptable salt. The present invention declares a pharmaceutical composition possessing inhibitory activity in the relation to 20S proteasome containing a pharmaceutically acceptable carrier or a diluent and a therapeutically effective amount of the compound, and also the invention refers to methods of treating the immune diseases, such as inflammatory intestinal disease, to treating cancer, to treating infection, to treating proliferative diseases, to treating neurodegenerative disease or asthma.

EFFECT: higher clinical effectiveness.

34 cl, 21 ex, 2 dwg

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to oncology. Method includes intramuscular introduction of immunomodulating medication reaferon. Optimal starting dose of reaferon is selected individually for each patient. For this purpose method of electroacupuncture diagnostics by R.Voll is used. Testing of information-wave markers of human body temperature 37.5 and 38.0 degrees Celcius and reaferon doses in 1, 2, 3 and more mln UNITS is carried out. Testing is performed separately on control points of measurement of meridians of lymphatic system, nerve degeneration, blood circulation, epithelial and parenchymatous degeneration, allergy, triple heater. After that selected is such dose of reaferon during combined, simultaneous testing of which with information-wave markers of human body temperature 37.5 and 38.0 degrees Celcius, indices of scale of apparatus for electroacupuncture diagnostics by R.Voll equal 50-60 conventional units of measurement.

EFFECT: method improves treatment efficiency due to reduction of side effects and complications.

1 ex

FIELD: chemistry.

SUBSTANCE: invention relates to novel derivatives of the anti-tumour antibiotic of the aureolic acid olivomycin A group, having anti-tumour activity and low toxicity, and synthesis method thereof. The invention also relates to a method of producing derivatives of the antibiotic of the aureolic acid olivomycin A group, involving selective oxidation of the side chain of olivomycin A aglycone through reaction with sodium periodate, followed by amidation of the obtained key intermediate 1'-des-(2,3-dihydroxy-n-butyroyl)-1'-carboxyolivomycin A with corresponding amines in the presence of a condensing agent.

EFFECT: disclosed compounds have marked anti-tumour activity and low toxicity compared with the original olivomycin A.

2 cl, 9 ex, 1 dwg, 6 tbl

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