RU2252935C2 - Serine protease inhibitors, pharmaceutical composition and therapeutical methods based on the same - Google Patents

Abstract

FIELD: pharmaceutical industry, medicine.

SUBSTANCE: invention relates to new compounds of formula

,

wherein A and B are independently CH or CR₃; X is C=O or (CR_4aR_4b)_m, (m = 1 or 2); Y is S(O)_n-R₂ (n = 1 or 2), S(O)_n-NR₂R₂, or S(O)_n-OR₂; N₁ and N₂ are nitrogen atoms; Q and R₁ are independently 1) optionally substituted C₁-C₁₀-alkyl; 2) optionally substituted aralkyl containing C₆-C₁₀-aryl, attached to C₁-C₁₀-alkyl; 3) optionally substituted aralkenyl containing C₅-C₁₀-aryl, attached to C₁-C₁₀-alkenyl; 4) optionally substituted C₆-C₁₀-aryl; 5) optionally substituted aryl, containing 5-10 ring atoms, selected from carbon and sulfur; each R₂ and R₃ are hydrogen; R_4a, R_4b, R₅, and R₆, are independently hydrogen; R₂ and R₃ are independently hydrogen or C₁-C₆-alkyl; as well as acid and base additive salts thereof. Also disclosed are method for production of claimed compounds, pharmaceutical composition inhibiting serine protease enzymes and therapeutic method based thereon.

EFFECT: new compounds and pharmaceutical composition for thrombosis preventing or abnormal thrombosis treatment.

11 cl, 7 tbl, 15 ex

Description

Discussion of the prior art

The technical field of the invention

One of the objects of the present invention are new compounds that are inhibitors of tissue factor (TF) / factor VIIa, factor VIIa, factor Xa, thrombin and / or kallikrein, as well as compositions containing these compounds. The compounds can be used to inhibit these factors and to treat mediated disorders. For example, the compounds can be used to prevent thrombosis or to treat abnormal thrombosis in mammals by inhibiting TF / factor VIIa, factor Xa, thrombin and / or kallikrein.

BACKGROUND OF THE INVENTION

Normal hemeostasis is the result of a complex balance between the processes of initiation of clot formation, clot formation and dissolution. The complex relationships between blood cells, specific plasma proteins, and the vascular surface support blood flow unless damage and blood loss occurs.

The onset of many serious diseases is associated with abnormal hemeostasis. For example, the formation of a local blood clot as a result of the destruction of an atherosclerotic plaque is the main cause of acute myocardial infarction and unstable angina. Treatment of an occlusive coronary thrombus with either thrombolytic therapy or percutaneous angioplasty may be accompanied by acute repeated thrombolytic closure of the damaged vessel. In addition, a large percentage of patients who underwent surgery, especially in the lower extremities, suffer from blood clots in the venous vascular system, which leads to a decrease in blood flow to the damaged area.

Therefore, there is a need for safe and effective therapeutic anticoagulants designed to limit or prevent blood clots.

Blood coagulation is a vital process for maintaining the body’s total water in case of tissue damage and is an important component of the host’s defense mechanisms. Coagulation or clot formation involves the sequential activation of a plurality of zymogens in a process leading to the formation of thrombin and the conversion of fibrinogen into an impermeable cross-linked fibrin clot. The formation of thrombin is the result of a blood coagulation cascade that has been well studied and described in detail (see, for example, Lawson J.H. et al., J. Biol. Chem. 269: 23357 (1994)). Reactions of this coagulation cascade include phases of initiation, amplification, and propagation. In addition, the cascade is divided into external and internal paths. The internal pathway includes factors XII, XI and IX and leads to the formation of a complex of factor IXa with its cofactor, i.e. factor VIIIa. This complex turns factor X into factor Xa. Factor Xa is an enzyme that forms a complex with its cofactor, i.e. factor Va, and quickly converts prothrombin to thrombin. Thrombin converts fibrinogen into fibrin monomers, which polymerize to form a clot. The external pathway includes factor VIIa and tissue factor, which form a complex (TF / factor VIIa), and convert factor X into factor Xa. As in the internal pathway, factor Xa converts prothrombin to thrombin.

Thrombin (factor IIa), as mentioned above, plays a major role in the coagulation cascade, converting fibrinogen to fibrin. As a result, chemical synthesis specialists have made numerous attempts to develop thrombin inhibitors (see, for example, US Pat. Nos. 5656600, 5656645, 5670479, 5646165, 5658939, 5658930 and WO 97/30073). In addition, compounds such as N-arylsulfonated phenylalanamides have been proposed as synthetic thrombin inhibitors.

Known factor Xa inhibitors include bisamidine compounds (Katakura S., Biochem. Biophys. Res. Commun., 197: 965 (1993)) and a compound based on the arginine skeleton (WO 93/15756; WO 94/13693). Phenyl and naphthyl sulfonamides (WO 96/100022; WO 96/16940; WO 96/40679) are also described as factor Xa inhibitors.

TF / factor VIIa is a serine protease complex that takes part in blood coagulation by activation of factor X and / or factor IX. Factor VIIa is formed from the precursor, i.e. factor VII, which is synthesized in the liver and secreted into the bloodstream, where it circulates as a single chain glycopeptide. The cDNA sequence of factor VII has been characterized (Hagen et al., Proc. Natl. Acad. Sci. U.S.A., 83: 2112-2416 (1986)).

Numerous natural and synthetic inhibitors of the TF / factor VIIa complex are known, and they have different activity and selectivity of action. Tissue factor pathway inhibitor (TFPI; Broze, Thromb. Haemostas., 74: 90 (1995)) and anticoagulant peptide c2 of nematodes (NAPc2; Stanssens et al., Proc. Natl. Acad. Sci. USA, 93: 2149 (1996) ) bind factor Xa before the formation of the quaternary complex of the inhibitor with the TF / factor VIIa complex. Small proteins that are direct inhibitors (Dennis et al., J. Biol. Chem. 35: 22317 (1994)) and inactive forms of TF / factor Vila are also known. (Kirchhofer et al., Arteriosclerosis, Thrombosis and Vascular Biol., 15: 1098 (1995); Jang et al., Circulation, 92: 3041 (1995)). In addition, synthetic peptides and soluble forms of mutant TFs have been obtained that retain binding affinity but have reduced cofactor activity (Roenning Throb. Res. 82: 73 (1996); Kelley et al. Blood, 89: 3219 (1997)). US Pat. No. 5,679,639 describes polypeptides and antibodies that inhibit the activity of serine proteases. US Pat. No. 5,580,560 describes a mutant factor Vila, which has a longer half-life. US Pat. Nos. 5,504,067 and 5,504,064 describe truncated TF for treating bleeding. It has also been found that fusion proteins including the Kunitz domain tissue factor are bifunctional anticoagulants (Lee et al., Biochemistry, 36: 5607-5611 (1997)). The TF / factor VIIa complex is considered to be a promising target for the development of inhibitors with a selective effect on bleeding associated with surgical intervention and the prevention of intravascular thrombosis (Harker et al., Thromb. Haemostas., 74: 464 (1995)).

Compounds that block or inhibit the enzymes of the coagulation cascade may find therapeutic use for treating or preventing thrombosis in mammals having a condition characterized by abnormal thrombosis. For example, for the arterial vasculature of an organ, the formation of an abnormal thrombus due to degradation of the formed atherosclerotic plaque is the main cause of acute myocardial infarction and unstable angina. Treatment of an occlusive coronary thrombus with thrombolytic therapy or using percutaneous intraluminal angioplasty (PTCA) may be accompanied by repeated closure of the vessel. As for the venous vascular network of the organ, many patients undergoing surgical intervention, especially in the abdominal and lower parts of the body, have known cases of thrombus formation, which reduces blood flow and can lead to pulmonary embolism. Disseminated intravascular coagulation syndrome in both venous and arterial systems usually occurs with septic shock, certain viral infections and cancer, and can lead to rapid and widespread formation of blood clots and impaired organ function (failure).

PTCA and recanalization are the most preferred procedures in the treatment of vascular occlusion. However, arterial thrombosis resulting from these procedures remains the main cause of organ dysfunction. For the currently most common heparin anticoagulant, it has been found that it is not fully effective in treating and preventing acute arterial thrombosis or retrombosis.

The synthesis and development of small molecules of inhibitors, which are based on the well-known three-dimensional structure of proteins, is the main modern way to create drugs. A number of thrombin inhibitors have been developed which have a hirudin type of structure (Stubbs and Bode, Current Opinion in Structural Biology, 4: 823-832 (1994)). New synthetic thrombin inhibitors as well as factor Xa and TF / factor VIIa inhibitors have been described (see, for example, Annual Reports in Medicinal Chemistry, Academic Press, San Diego, CA, 1995-1997).

US Pat. No. 5,589,173 discloses the use of a tissue factor antagonist and a thrombolytic agent for the treatment of myocardial infarction.

US Pat. No. 5,399,487 describes naphthalenesulfonamides that can be used to determine the activity of a proteolytic enzyme or as enzyme inhibitors.

Currently, there remains a need for the development of compounds that are effective inhibitors of coagulation cascade enzymes and which have increased inhibitory activity and / or selective action against certain cascade enzymes.

Summary of the invention

Thus, one of the objects of the present invention are new compounds that inhibit factors / enzymes of the coagulation cascade and which can be used to prevent or treat blood clot formation in arterial or venous vessels. These compounds can be used as inhibitors of coagulation factors and as anticoagulants in general.

In one embodiment, the subject of the invention is inhibitors that selectively inhibit factor VIIa, TF / factor VIIa compared to factor Xa, thrombin or kallikrein. According to this embodiment, the compounds preferably inhibit TF / factor VIIa about an order of magnitude (10X) better, more preferably about two orders of magnitude (100X) better, even more preferably about three orders of magnitude (1000X) better than factor Xa, thrombin and / or kallikrein.

In yet another embodiment, the invention provides compounds that specifically inhibit factor Xa compared to factor VIIa, TF / factor VIIa, thrombin or kallikrein. According to this embodiment, the compounds preferably inhibit Xa about an order of magnitude (10X) better, more preferably about two orders of magnitude (100X) better, even more preferably about three orders of magnitude (1000X) better than TF / factor VIIa, thrombin and / or kallikrein .

In yet another embodiment, an object of the invention is compounds that specifically inhibit thrombin compared to factor VIIa, TF / factor VIIa, Xa, or kallikrein. According to this embodiment, the compounds preferably inhibit thrombin about an order of magnitude (10X) better, more preferably about two orders of magnitude (100X) better, even more preferably about three orders of magnitude (1000X) better than TF / factor VIIa, factor Xa and / or kallikrein.

Another object of the invention is a method of inhibiting the activity of TF / factor VIIa, Xa or thrombin by bringing these enzymes into contact with an effective inhibitory amount of the new inhibitors of the present invention or a composition containing these compounds. Another object of the invention is a method of treating a disease mediated by TF / factor VIIa, Xa or thrombin, comprising administering to a mammal in need of such treatment an effective amount of one of the compounds of the invention or a composition containing the compound. And another object of the invention is a method of preventing thrombosis or treating abnormal thrombosis, comprising administering to a mammal in need of such treatment an effective amount of one of the compounds of the invention or a composition containing the compound and a carrier or excipient.

These and other objects will become apparent after studying the following description, which presents the compounds of the present invention having the following structure:

Where

A and B are independently CH, CR ₃ or N;

X is C = O or (CR _4a R _4b ) _m , where m is 1 or 2;

Y is S (O) _n -R ₁ , where n is 1 or 2, S (O) _n -NR ₂ R ₂ , where n is 1 or 2, S (O) _n -OR ₂ , where n is 1 or 2, C (O) R ₁ , C (S) R ₁ , C (O) -OR ₁ , C (O) -NR ₂ R ₂ ;

N ₁ and N ₂ are nitrogen atoms;

Q and R ₁ independently represent

(1) optionally substituted alkyl having from 1 to about 10 carbon atoms;

(2) an optionally substituted aralkyl containing an aryl moiety having from 6 to about 10 ring carbon atoms, which is bonded to an alkyl moiety comprising from 1 to about 10 carbon atoms;

(3) an optionally substituted heteroaralkyl containing a heteroaryl moiety having from 5 to about 10 ring atoms, which is bonded to an alkyl moiety comprising from 1 to about 10 carbon atoms;

(4) an optionally substituted carbocycloalkyl containing a carbocyclyl moiety having from 3 to about 10 ring carbon atoms, which is bonded to an alkyl moiety comprising from 1 to about 10 carbon atoms;

(5) an optionally substituted heterocycloalkyl containing a heterocyclyl moiety having from 3 to about 10 ring atoms, which is bonded to an alkyl moiety comprising from 1 to about 10 carbon atoms;

(6) optionally substituted alkenyl having from 2 to about 10 carbon atoms;

(7) an optionally substituted aralkenyl containing an aryl moiety having from 5 to about 10 ring atoms, which is bonded to an alkenyl moiety containing from 2 to about 10 carbon atoms;

(8) an optionally substituted heteroaralkenyl containing a heteroaryl moiety having from 5 to about 10 ring atoms, which is bonded to an alkenyl moiety comprising from 2 to about 10 carbon atoms;

(9) an optionally substituted carbocycloalkenyl containing a carbocyclyl moiety having from 3 to about 10 ring carbon atoms, which is bonded to an alkenyl moiety comprising from 2 to about 10 carbon atoms;

(10) an optionally substituted heterocycloalkenyl containing a heterocyclyl moiety having from 3 to about 10 ring atoms, which is linked to an alkenyl moiety containing from 2 to about 10 carbon atoms;

(11) optionally substituted aryl having from 6 to about 10 ring carbon atoms;

(12) an optionally substituted heteroaryl having from 5 to about 10 ring atoms, wherein the ring atoms are selected from carbon atoms and heteroatoms, the heteroatoms being nitrogen, oxygen or sulfur;

(13) an optionally substituted carbocyclyl having from 3 to about 10 carbon atoms;

(14) an optionally substituted heterocyclyl having from 3 to about 10 ring atoms, wherein the ring atoms are selected from carbon atoms and heteroatoms, the heteroatoms being nitrogen, oxygen or sulfur;

R ₂ each independently represents H, alkyl, substituted alkyl, C (O) R ₇ or C (NH) R ₇ or N ₂ R ₂ and N ₂ R ₂ together form a group N ₁ —CO — N ₂ ;

R ₃ is H, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, halogen or OH;

R _4a and R ₅ independently represent a group selected from the series consisting of H, unsubstituted or substituted alkyl, unsubstituted or unsubstituted alkoxyalkyl, unsubstituted or substituted haloalkyl, unsubstituted or substituted aryl, alkyl-OR ₇ , alkyl-NR ₇ R ₈ alkyl-OS (O) OR ₇ , alkyl-C (O) OR ₇ , alkyl-C (O) R ₇ , OC (O) R ₇ , C (O) OR ₇ , C (O) R ₇ , and groups in which alkyl, R ₇ or R _{8 is} substituted with 1-3 substituents selected from the group consisting of F, Cl, Br, I, OR ₇ , SR ₇ , NR ₇ R ₈ , OC (OR ₇ ), C (O ) OR ₇ , C (O) R ₇ , C (O) NR ₇ R ₈ , NHC (NH) NH ₂ , PO ₃ , unsubstituted or substituted indolyl, or unsubstituted or substituted imide angry;

R _4b is H, alkyl or substituted alkyl;

R ₆ each independently from each other is selected from the range including H, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl-OR ₇ , C ₁ -C ₆ -alkyl-NR ₇ R ₈ , C ₁ -C ₆ haloalkyl , halogen, cyano, OR ₇ , SR ₇ , NR ₇ R ₈ , C (O) OR ₇ , C (O) R ₇ and OC (O) R ₇ ;

R ₇ and R _{8 are} independently H or C ₁ -C ₆ alkyl; and their acid addition salts and base addition salts and prodrugs.

In addition, objects of the invention are compositions containing these compounds and the methods described below.

Detailed Description of Preferred Embodiments

Definitions

The term “disease mediated by factor VIIa, TF / factor VIIa, factor Xa, thrombin or kallikrein” refers to a disease or physiological condition, including blood coagulation, in which inhibition of one or more of these enzymes reduces or eliminates at least one of the physiological symptoms of the disease or condition.

The term "thrombosis" means the development or formation of a blood clot or thrombus in a blood vessel of a mammal or in a synthetic vessel, such as a plastic or glass tube or vessel. A thrombus, which is separated from its original place and found elsewhere, is called a thrombotic embolus.

The term "abnormal thrombosis" refers to thrombosis that occurs in a mammal other than a healthy mammal.

The term "alkyl" in the context of the present description individually or as a component of another concept means a branched or unbranched saturated aliphatic hydrocarbon group having the indicated number of carbon atoms, or, if the number of atoms is not specifically indicated, having up to 12 carbon atoms inclusive. Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 2-methylbutyl, 2,2-dimethylpropyl, n-hexyl, 2-methylpentyl, 2,2, dimethylbutyl, n-heptyl, 3-heptyl, 2-methylhexyl and the like. The terms "lower alkyl", "C ₁ -C ₆ alkyl" and "alkyl having 1-6 carbon atoms" are synonymous and are used interchangeably. Preferred “C ₁ -C ₆ alkyl” groups are methyl, ethyl, 1-propyl, isopropyl, 1-butyl or sec-butyl.

The terms "substituted alkyl" or "substituted C _n -C _m alkyl", where m and n are integers indicating the number of carbon atoms in the alkyl group, mean alkyl groups as defined above, which are substituted by 1, 2 or 3 atoms halogen (F, Cl, Br, I), trifluoromethyl, hydroxy, unsubstituted and substituted with C ₁ -C ₇ alkoxy, protected with hydroxy, amino (including alkyl and dialkylamino), protected with amino, unsubstituted and substituted with C ₁ -C ₇ acyloxy, unsubstituted and substituted C ₃ -C ₇ heterocyclyl, unsubstituted and substituted phenoxy, n tro-, carboxy, protected carboxy, protected and unprotected karboksialkoksigruppoy, unsubstituted and substituted acyl, carbamoyl, carbamoyloxy, cyano, methylsulfonylamino, unsubstituted and substituted benzyloxy, unsubstituted and substituted C ₃ -C ₆ carbocyclyl or C ₁ -C ₄ alkoxygroup. Substituted alkyl groups may be substituted with one (preferably), two or three identical or different substituents.

Examples of the above substituted alkyl groups include, but are not limited to: cyanmethyl, nitromethyl, hydroxymethyl, trityloxymethyl, propionyloxymethyl, aminomethyl, carboxymethyl, carboxyethyl, trifluoroethyl, trifluoropropyl, carboxypropyl, 2-aminopropyl, alkoxycarbonylmethylmethylmethyl, alkyloxymethylmethylmethylmethyl, carboxymethylmethylmethyl, alkyloxymethylmethylmethyl, alkyloxymethylmethylmethylmethylmethylmethylmethylmethylmethyl tert-butoxymethyl, acetoxymethyl, chloromethyl, bromomethyl, iodomethyl, trifluoromethyl, 6-hydroxyhexyl, 2,4-dichloro (n-butyl), 2-amino (isopropyl), 2-carbamoyloxyethyl, etc. The alkyl group may also be substituted with a carbocyclic group. Examples include cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl and cyclohexylmethyl, as well as the corresponding -ethyl, -propyl, -butyl, -pentyl, -hexyl groups, etc. Examples of preferred groups from this series include a substituted methyl group, for example a methyl group substituted with the same substituents as indicated for the substituted “C _n -C _m alkyl” group. Examples of substituted methyl groups include groups such as hydroxymethyl, protected hydroxymethyl (e.g. tetrahydropyranyloxymethyl), acetoxymethyl, carbamoyloxymethyl, trifluoromethyl, chloromethyl, carboxymethyl, bromomethyl and iodomethyl.

The term “alkoxy” refers to groups having the indicated number of carbon atoms, such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy groups and the like. The term “substituted alkoxy” means said alkoxy groups substituted with the same substituents as shown for the substituted “C _n -C _m alkyl” group, for example, 2,2,2-trifluoroethoxy, 2,2,2-trifluoropropoxy, etc.

The term "acyloxy" in the context of the present description refers to carboacyloxy groups having the indicated number of carbon atoms, such as formyloxy, acetoxy, propionyloxy, butyryloxy, pentanoyloxy, hexanoyloxy, heptanoyloxy and the like. The term “substituted acyloxy group” means the indicated acyloxy groups substituted with the same substituents as indicated for the substituted “C _n -C _m alkyl” group.

The terms “alkylcarbonyl,” “alkanoyl,” and “acyl” are used interchangeably herein to mean groups having the indicated number of carbon atoms, such as formyl, acetyl, propionyl, butyryl, pentanoyl, hexanoyl, heptanoyl, benzoyl, and the like.

The terms "carbocyclyl", "carbocyclic" and "carbocyclo" individually or as a fragment of a complex group, such as a carbocycloalkyl group, mean a mono-, bi-, or tricyclic aliphatic ring having 3-14 carbon atoms, preferably 3-7 carbon atoms . Preferred carbocyclic groups include cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl. The terms “substituted carbocyclyl” and “substituted carbocyclo group” mean that these groups are substituted with the same substituents as indicated for the substituted “C _n -C _m alkyl” group.

“Carbocycloalkyl group” means a carbocyclo group as defined above covalently bonded to an alkyl group as defined above.

The term "alkenyl" means a branched or unbranched hydrocarbon group having the indicated number of carbon atoms, which contains one or more carbon-carbon double bonds, each double bond independently of one another forms a cis, trans or non-geometric isomer. The term “substituted alkenyl” means these alkenyl groups substituted with the same substituents as indicated for the substituted “C _n -C _m alkyl” group.

The term "alkynyl" means a branched or unbranched hydrocarbon group having the indicated number of carbon atoms, which contains one or more carbon-carbon triple bonds. The term “substituted alkynyl” means these alkynyl groups substituted with the same substituents as indicated for the substituted “C _n -C _m alkyl” group.

The terms "alkylthio" and "substituted C ₁ -C ₁₂ alkylthio group" mean C ₁ -C ₁₂ alkyl and substituted C ₁ -C ₁₂ alkyl groups, respectively, which are attached to the sulfur atom, which in turn is the point of attachment of alkylthio or substituted alkylthio groups to the indicated group or substituent.

The term "aryl" in the context of the present description individually or as a component of another concept means a homocyclic aromatic group, non-condensed or condensed, which has the indicated number of carbon atoms, or, if the number of atoms is not specifically indicated, has up to 14 carbon atoms. Preferred aryl groups include phenyl, naphthyl, biphenyl, phenanthrenyl, naphthenyl, and the like. (see, for example, Lang’s Handbook of Chemistry (ed. Dean J.A), 13th ed., tables 7-2 [1985]).

The term “substituted phenyl” or “substituted aryl” means a phenyl group or an aryl group substituted with 1, 2, 3, 4 or 5, preferably 1-2, 1-3 or 1-4 substituents selected from the group consisting of halogen (F , Cl, Br, I), hydroxy, protected hydroxy, cyano, nitro, alkyl (preferably C ₁ -C ₆ alkyl), alkoxy (preferably C ₁ -C ₆ alkoxy), benzyloxy, carboxy, protected carboxy group, carboxymethyl, protected carboxymethyl, hydroxymethyl, protected hydroxymethyl, aminomethyl, protected aminomethyl, trifluoromethyl, alkylsulfonyl ino-, arylsulfonylamino, geterotsiklilsulfonilaminogruppu, heterocyclyl, aryl, or other groups specified. One of the methine (CH) and / or methylene (CH ₂ ) groups in these substituents, in turn, may be substituted with a group similar to those listed above. Examples of “substituted phenyl” include, but are not limited to, a mono- or di (halo) phenyl group such as 4-chlorophenyl, 2,6-dichlorophenyl, 2,5-dichlorophenyl, 3,4-dichlorophenyl, 3-chlorophenyl, 3-bromophenyl, 4-bromophenyl, 3,4-dibromophenyl, 3-chloro-4-fluorophenyl, 2-fluorophenyl and the like; a mono- or di (hydroxy) phenyl group such as 4-hydroxyphenyl, 3-hydroxyphenyl, 2,4-dihydroxyphenyl, its protected hydroxy derivative, and the like; a nitrophenyl group such as 3 or 4-nitrophenyl; a cyanphenyl group, for example 4-cyanophenyl; a mono- or di (lower alkyl) phenyl group such as 4-methylphenyl, 2,4-dimethylphenyl, 2-methylphenyl: 4- (isopropyl) phenyl, 4-ethylphenyl, 3- (n-propyl) phenyl and etc .; a mono- or di (alkoxy) phenyl group, for example, 3,4-dimethoxyphenyl, 3,4-diethoxyphenyl, 3-ethoxy-4-isopropoxyphenyl, 3-ethoxy-sec-butoxyphenyl, 3-methoxy-4-benzyloxyphenyl, 3- methoxy-4- (1-chloromethyl) benzyloxyphenyl, 3-ethoxyphenyl, 4- (isopropoxy) phenyl, 4- (tert-butoxy) phenyl, 3-ethoxy-4-methoxyphenyl and the like; 3 or 4-trifluoromethylphenyl; a mono- or dicarboxyphenyl or (protected carboxy) phenyl group such as 4-carboxyphenyl; mono- or di (hydroxymethyl) phenyl or (protected hydroxymethyl) phenyl, such as 3- (protected hydroxymethyl) phenyl or 3,4-di (hydroxymethyl) phenyl; mono- or di (aminomethyl) phenyl or (protected aminomethyl) phenyl, such as 2- (aminomethyl) phenyl or 2,4- (protected aminomethyl) phenyl; or mono- or di (N- (methylsulfonylamino) phenyl, such as 3- (N-methylsulfonylamino) phenyl. In addition, the term "substituted phenyl" refers to disubstituted phenyl groups in which the substituents are different from each other, for example 3-methyl -4-hydroxyphenyl, 3-chloro-4-hydroxyphenyl, 2-methoxy-4-bromophenyl, 4-ethyl-2-hydroxyphenyl, 3-hydroxy-4-nitrophenyl, 2-hydroxy-4-chlorophenyl and the like, as well as trisubstituted phenyl groups in which 1, 2 or 3 substituents are different, for example 3-methoxy-4-benzyloxy-6-methylsulfonylamino, 3-methoxy-4-benzyloxy-6-phenylsulf onylamino and tetrasubstituted phenyl groups in which the substituents are different, such as 3-methoxy-4-benzyloxy-5-methyl-6-phenylsulfonyl-amino. Preferred substituted phenyl groups include 3-methoxyphenyl, 3-ethoxyphenyl, 4-benzyloxyphenyl, 4-methoxyphenyl, 3-ethoxy-4-benzyloxyphenyl, 3,4-diethoxyphenyl, 3-methoxy-4-benzyloxyphenyl, 3-, methoxy-4- (1-chloromethyl) benzyloxyphenyl, 3-methoxy-4- (1-chloromethyl ) benzyloxy-6-methylsulfonylaminophenyl. In addition, the term "substituted phenyl" refers to phenyl groups including a fused aryl, phenyl or heteroaryl group. The fused ring may also be substituted with any number, preferably 1, 2 or 3, of the substituents indicated above for “substituted alkyl” groups.

The term “aralkyl” means 1, 2 or 3 aryl groups having the indicated number of carbon atoms attached to an alkyl group having the indicated number of carbon atoms, including but not limited to: benzyl, naphthylmethyl, phenethyl, benzhydryl (diphenylmethyl), trityl etc. A preferred aralkyl group is a benzyl group.

The term “substituted aralkyl” means an alkyl group, preferably a C ₁ -C ₈ alkyl group substituted on any of the carbon atoms with an aryl group, preferably a C ₆ -C ₁₀ aryl group, linked to the alkyl group at any position of the aryl ring, and substituted on the alkyl a fragment of 1, 2 or 3 groups selected from the group consisting of halogen (P, Cl, Br, I), hydroxy, protected hydroxy, amino, protected amino, C ₁ -C ₆ acyloxy, nitro, carboxy -, protected carboxy, carbamoyl, carbamoyloxy, cyano, C ₁ -C ₆ alkylthio, N- (methylene sulfonylamino) - or C ₁ -C ₄ alkoxy. Optionally, the aryl group may be substituted by 1, 2, 3, 4 or 5 groups selected from the group consisting of halogen, hydroxy, protected hydroxy, nitro, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, carboxy a protected carboxy group, carboxymethyl, protected carboxymethyl, hydroxymethyl, protected hydroxymethyl, aminomethyl, protected aminomethyl or N- (methylsulfonylamino) group. As indicated previously, if either a C ₁ -C ₈ alkyl moiety or an aryl moiety or both are disubstituted, then the substituents may be the same or different. This group may also be a substituted aralkyl moiety of a substituted aryloxy group.

Examples of “substituted aralkyl” and this group, when included, the composition of the “substituted aralkoxy group” include groups such as 2-phenyl-1-chloroethyl, 1-phenyl-1-chloromethyl, 1-phenyl-1-bromomethyl, 2 - (4-methoxyphenyl) ethyl, 2,6-dihydroxy-4-phenyl (n-hexyl), 5-cyan-3-methoxy-2-phenyl (n-pentyl), 3- (2,6-dimethylphenyl) - n-propyl, 4-chloro-3-aminobenzyl, 6- (4-methoxyphenyl) -3-carboxy (n-hexyl), 5- (4-aminomethylphenyl) -3- (aminomethyl) (n-pentyl), etc. P.

The term “carboxy-protecting group” as used herein means one of the ester derivatives of a carboxylic acid group, commonly used to block or protect a carboxylic acid group when reactions are carried out on other functional groups of the compound. Examples of such carboxy-protecting groups include fragments such as 4-nitrobenzyl, 4-methoxybenzyl, 3,4-dimethoxybenzyl, 2,4-dimethoxybenzyl, 2,4,6-trimethoxybenzyl, 2,4,6-trimethylbenzyl, pentamethylbenzyl, 3.4 -methylenedioxybenzyl, benzhydryl, 4,4'-dimethoxybenzhydryl, 2,2 ', 4,4'-tetramethoxybenzhydryl, alkyl such as methyl, ethyl, isopro-propyl, tert-butyl or tert-amyl, trityl, 4-methoxytrityl, 4, 4'-dimethoxytrityl, 4,4 ', 4' '- trimethoxytrityl, 2-phenylprop-2-yl, trimethylsilyl, tert-butyldimethylsilyl, phenacyl, 2,2,2-trichloroethyl, beta- (trimethylsilyl) ethyl, beta- ( di (n -butyl) methylsilyl) ethyl, para-toluenesulfonylethyl, 4-nitrobenzylsulfonylethyl, allyl, cinnamyl, 1- (trimethylsilylmethyl) prop-1-en-3-yl and the like. The types of carboxy-protecting groups used are not critical in the case where the derivatized carboxylic acid is stable under the conditions of the subsequent reaction (s) at other positions of the molecule, and they can be removed at the appropriate moment without disturbing the rest of the molecule. In particular, it is important not to bring the carboxy-protected molecule into contact with strong nucleophilic bases or to use reducing conditions in which highly active metal catalysts such as Raney nickel are used. (Such stringent removal conditions that also need to be avoided when removing amino protecting groups and carboxy protecting groups are described below). Preferred carboxy protecting groups are allyl and para-nitrobenzyl groups. Similar carboxy-protecting groups, which are used in the field of chemistry of cephalosporins, penicillins and peptides, can also be used to protect substituents of the carboxyl group. Additional examples of these groups can be found in E. Haslam, "Protective Groups in Organic Chemistry", eds. J.G.W. McOmie, Plenum Press, New York, N.Y., 1943, chapter 5 and T.W. Green, "Protective Groups in Organic Synthesis", John Wiley and Sons, New York, NY, 1981, chapter 5. The term "protected carboxy group" refers to a carboxy group substituted with one of the above carboxy protecting groups.

The term "amidoprotective group" in the context of the present description refers to any group commonly used in the field of peptide chemistry to protect peptide nitrogen atoms from undesirable side reactions. Such groups include para-methoxyphenyl, 3,4-dimethoxybenzyl, benzyl, O-nitrobenzyl, di (para-methoxyphenyl) methyl, triphenylmethyl, (para-methoxyphenyl) diphenylmethyl, diphenyl-4-pyridylmethyl, meta-2- (picolyl) - N'-oxide, 5-dibenzosuberyl, trimethylsilyl, tert-butyldimethylsilyl and the like. Further descriptions of these protecting groups can be found in "Protective Groups in Organic Synthesis", Theodora W. Greene, 1981, John Wiley and Sons, New York.

The terms “heterocyclic group”, “heterocyclic”, “heterocyclyl” or “heterocyclo” individually or as part of a complex group, such as a heterocycloalkyl group, are used interchangeably and refer to a mono-, bi- or tricyclic saturated or non-aromatic unsaturated ring having the indicated number of atoms, usually from 3 to about 10 ring atoms, where the ring atoms are carbon and 1, 2, 3 or 4 atoms of nitrogen, sulfur or oxygen. Typically, a 5-membered ring contains 0-2 double bonds, and a 6- or 7-membered ring has 0-3 double bonds, and heteroatoms representing nitrogen or sulfur can optionally be oxidized, and any heteroatom representing nitrogen may optionally be quaternized. Examples include pyrrolidinyl, oxiranyl, oxetanyl, tetrahydrofuranyl, 2,3-dihydrofuranyl, 2H-pyranyl, tetrahydropyranyl, thiranyl, thietanyl, tetrahydrothietanyl, aziridinyl, azetidinyl, 1-methyl-2-pyrrolyl, piperidinyl and 3,4,5,6- tetrahydropiperidinyl.

A “heterocycloalkyl” or “heterocycloalkenyl” group is a heterocyclo group as defined above covalently linked to an alkyl or alkenyl group as defined above.

Unless otherwise indicated, “heteroaryl” individually or as a fragment of a complex group, such as a heteroaralkyl group, means a mono-, bi- or tricyclic aromatic ring system having the indicated number of atoms, where at least one ring is 5-, 6 -or a 7-membered ring containing from 1 to 4 heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur, and preferably at least one heteroatom is nitrogen (see Lang's Handbook of Chemistry, above). The definition includes any bicyclic groups in which any of the above heteroaryl rings is fused to a benzene ring. Heteroaryls are preferred in which the heteroatom is nitrogen or oxygen.

Examples of heteroaryl (substituted or unsubstituted) groups falling within the term “heteroaryl” are the following ring systems: thienyl, furyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, triazolyl, thiadiazolyl, oxadiazolyl, tetrazolyl, tetrazolyl, tetrazolyl, tetrazolyl, tetrazolyl pyrimidyl, pyrazinyl, pyridazinyl, thiazinyl, oxazinyl, triazinyl, thiadiazinyl, oxadiazinyl, dithiazinyl, dioxazinyl, oxathiazinyl, tetrazinyl, thiatriazinyl, oxatriazinyl, dithiadiazinyl, imidazolidinyl, di irimidyl, tetrazolo [1,5-b] pyridazinyl and purinyl, as well as derivatives condensed with a benzene ring, for example, benzoxazolyl, benzofuryl, benzothiazolyl, benzothiadiazolyl, benzotriazolyl, benzoimidazolyl and indolyl.

5-membered ring systems containing a sulfur or oxygen atom and 1-3 nitrogen atoms can also be used according to the present invention. Examples of such preferred groups include thiazolyl, in particular thiazol-2-yl and thiazol-2-yl N-oxide, thiadiazolyl, in particular 1,3,4-thiadiazol-5-yl and 1,2,4-thiadiazol-5- sludge, oxazolyl, preferably oxazol-2-yl, and oxadiazolyl, such as 1,3,4-oxadiazol-5-yl and 1,2,4-oxadiazol-5-yl. Examples of systems included in another preferred group of 5-membered ring systems with 2-4 nitrogen atoms include imidazolyl, preferably imidazol-2-yl; triazolyl, preferably 1,3,4-triazol-5-yl; 1,2,3-triazol-5-yl, 1,2,4-triazol-5-yl, and tetrazolyl, preferably 1H-tetrazol-5-yl. Examples of systems included in a preferred group of benzene ring-condensed derivatives are benzoxazol-2-yl, benzothiazol-2-yl and benzimidazol-2-yl.

Also suitable specific examples of the above heterocyclic ring systems are 6-membered ring systems containing 1 to 3 nitrogen atoms. Examples thereof include pyridyl, such as pyrid-2-yl, pyrid-3-yl and pyrid-4-yl; pyrimidyl, preferably pyrimid-2-yl and pyrimid-4-yl; triazinyl, preferably 1,3,4-triazin-2-yl and 1,3,5-triazin-4-yl; pyridazinyl, in particular pyridazin-3-yl, and pyrazinyl. A preferred group are N-oxides of pyridine and N-oxides of pyridazine and pyridyl, pyrimid-2-yl, pyrimid-4-yl, pyridazinyl and 1,3,4-triazin-2-yl.

Substituents for optionally substituted heterocyclic ring systems and further examples of the 5- and 6-membered ring systems described above can be found in W. Druckheimer et al., US Pat. No. 4,278,793.

A particularly preferred “heteroaryl” group includes: 1,3-thiazol-2-yl, 4- (carboxymethyl) -5-methyl-1,3-thiazol-2-yl, 4- (carboxymethyl) -5-methyl- sodium salt 1,3-thiazol-2-yl, 1,2,4-thiadiazol-5-yl, 3-methyl-1,2,4-thiadiazol-5-yl, 1,3,4-triazol-5-yl, . 2-methyl-1,3,4-triazol-5-yl, 2-hydroxy-1,3,4-triazol-5-yl, 2-carboxy-4-methyl-1,3,4-triazol- sodium salt 5-yl, 2-carboxy-4-methyl-1,3,4-triazol-5-yl, 1,3-oxazol-2-yl, 1,3,4-oxadiazol-5-yl, 2-methyl 1,3,4-oxadiazol-5-yl, 2- (hydroxymethyl) -1,3,4-oxadiazol-5-yl, 1,2,4-oxadiazol-5-yl, 1,3,4-thiadiazol- 5-yl, 2-thiol-1,3,4-thiadiazol-5-yl, 2- (methylthio) -1,3,4-thiadiazol-5-yl, 2-amino-1,3,4-thiadiazol- 5-yl, 1H-tetrazol-5-yl, 1-methyl-1H-tetrazol-5-yl, 1- (1- (dimethylamino) et-2-yl) -1H-tetrazol-5-yl, 1- ( carboxymethyl) -1H-tetrazol-1-5-yl, sodium salt 1- (carboxymethyl) -1H-tetrazol-5-yl, 1- (methylsulfonic acid) -1H-tetrazol-5- sludge, 1- (methylsulphonic acid) -1H-tetrazol-5-yl sodium salt, 2-methyl-1H-tetrazol-5-yl, 1,2,3-triazol-5-yl, 1-methyl-1,2 3-triazol-5-yl, 2-methyl-1,2,3-triazol-5-yl, 4-methyl-1,2,3-triazol-5-yl, pyrid-2-yl N-oxide, 6-methoxy-2- (N-oxide) pyriaz-3-yl, 6-hydroxypyridaz-3-yl, 1-methylpyrid-2-yl, 1-methylpyrid-4-yl, 2-hydroxypyrimid-4-yl, 1 4,5,6-tetrahydro-5,6-dioxo-4-methylastriazin-3-yl, 1,4,5,6-tetrahydro-4- (formylmethyl) -5,6-dioxoastriazin-3-yl, 2 5-dihydro-5-oxo-6-hydroxyastriazin-3-yl, 2,5-dihydro-5-oxo-6-hydroxyastriazin-3-yl sodium salt, 2,5-dihydro-5-oxo-6 sodium salt hydroxy-2-methyl striazin-3-yl, 2,5-dihydro-5-oxo-6-hydroxy-2-methylastriazin-3-yl, 2,5-dihydro-5-oxo-6-methoxy-2-methylastriazin-3-yl, 2,5-dihydro-5-oxoastriazin-3-yl, 2,5-dihydro-5-oxo-2-methylastriazin-3-yl, 2,5-dihydro-5-oxo-2,6-dimethylastriazin-3- il, tetrazolo [1,5-b] pyridazin-6-yl and 8-aminotetrazolo [1,5-b] pyridazin-6-yl.

An alternative heteroaryl group includes: 4- (carboxymethyl) -5-methyl-1,3-thiazol-2-yl, 4- (carboxymethyl) -5-methyl-1,3-thiazol-2-yl sodium salt, 1 3,4-triazol-5-yl, 2-methyl-1,3,4-triazol-5-yl, 1H-tetrazol-5-yl, 1-methyl-1H-tetrazol-5-yl, 1- ( 1- (dimethylamino) et-2-yl) -1H-tetrazol-5-yl, 1- (carboxymethyl) -1H-tetrazol-5-yl, 1- (carboxymethyl) -1H-tetrazol-5-yl sodium salt, 1- (methylsulfonic acid) -1H-tetrazol-5-yl, sodium salt 1- (methylsulfonic acid) -1H-tetrazol-5-yl, 1,2,3-triazol-5-yl, 1,4,5, 6-tetrahydro-5,6-dioxo-4-methylastriazin-3-yl, 1,4,5,6-tetrahydro-4- (2-formylmethyl) -5,6-dioxoac triazin-3-yl, 2,5-dihydro-5-oxo-6-hydroxy-2-methylastriazin-3-yl sodium salt, 2,5-dihydro-5-oxo-6-hydroxy-2-methylastriazin-3- il, tetrazolo [1,5-b] pyridazin-6-yl and 8-aminotetrazolo [1,5-b] pyridazin-6-yl.

A “heteroaralkyl or“ heteroaralkenyl ”group refers to a heteroaryl group as defined above covalently linked to an alkyl group or alkenyl group as defined above.

"Pharmaceutically acceptable salts" include both acid addition salts and base addition salts. "Pharmaceutically acceptable acid addition salt" means salts that retain the biological effectiveness and properties of the free bases and are not undesirable from a biological or any other point of view, and are formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid , carbonic acid, phosphoric acid and the like, and organic acids selected from the range including classes of organic acids such as aliphatic, cycloaliphatic, aromatic f) araliphatic, heterocyclic, carboxylic and sulfonic acids, for example, formic acid, acetic acid, propionic acid, glycolic acid, gluconic acid, lactic acid, pyruvic acid, oxalic acid, malic acid, maleic acid, malonic acid, malonic acid, succinic acid acid, tartaric acid, citric acid, aspartic acid, ascorbic acid, glutamic acid, anthranilic acid, benzoic acid, cinnamic acid, mandelic acid, embonic acid, f niluksusnuyu acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, etc.

“Pharmaceutically acceptable base addition salts” include salts derived from inorganic bases, such as sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum and the like salts. Particularly preferred are salts of ammonium, potassium, sodium, calcium and magnesium. Salts derived from pharmaceutically acceptable organic non-toxic bases include salts of primary, secondary and tertiary amines, substituted amines, including naturally occurring substituted amines, cyclic amines and basic ion exchange resins such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine , 2-diethylaminoethanol, trimethamine, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, methylglucamine, theobromine, puri s, piperazine, piperidine, N-ethylpiperidine, and the resin polaminnye t.p.Osobenno preferred organic nontoxic bases are isopropylamine, diethylamine, etanoolamin, trimethamine, dicyclohexylamine "choline and caffeine.

The term "prodrug" in the context of the present description refers to a derivative of the original drug molecule, which enhances the required pharmaceutical characteristics or properties (for example, transport, bioavailability, pharmacodynamic characteristics, etc.) and which must be subjected to biotransformation, spontaneous or enzymatic within the body, to release the active parent drug.

Options for implementation

The invention generally relates to compounds having the following structure

In this structure, R ₂ , R ₅ , R ₆ ,, A, B, N ₁ , N ₂ , Q, X and Y have the above meanings. While alkyl is preferably unsubstituted or substituted C ₁ -C ₆ alkyl; alkenyl is preferably unsubstituted or substituted C ₂ -C ₆ alkenyl; alkynyl is preferably unsubstituted or substituted C ₂ -C ₆ alkynyl; aryl is preferably unsubstituted or substituted naphthyl or phenyl; more preferably phenyl; aralkyl is preferably unsubstituted or substituted benzyl. The variable m is preferably 1.

The group Y is preferably S (O) _n -R ₁ , where n is 1 or 2, or the group S (O) _n -NR ₂ R ₂ where n is 1 or 2, more preferably S (O) _n -R ₁ .

According to one preferred embodiment, R ₁ , for example, when Y is S (O) _n -R ₁ , is selected from the group consisting of C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, phenyl, naphthyl, benzyl and heteroaryl having 5-6 ring atoms selected from carbon atoms and 1-2 heteroatoms, where the heteroatoms are N, S or O, and R _{1 is} optionally substituted with 1-3 substituents selected from the group consisting of halogen, nitro, C ₁ -C ₆ alkyl, NR ₇ R ₈ , OR ₇ , SR ₇ , C ₁ -C ₆ alkyl-C (O) OR ₇ , C ₁ -C ₆ alkyl-C (O) R _7, C ₁ -C ₆ alkyl-C (O) R _7, C ₁ -C ₆ alkyl-OR _7, C ₁ -C ₆ halo keel, C ₁ -C ₆ alkyl-NR ₇ R _8, C (O) OR _7, OC (O) R _7, C (O) NR ₇ R _8, OC (O) NR ₇ R _8, NHC (O) R ₇ and NHC (O) NR ₇ R ₈ where R ₇ and R _{8 are} independently H or C ₁ -C ₆ alkyl. In this embodiment, all other variables R ₂ , R ₅ , R ₆ , A, B, Q, X, and Y may be independently selected from any groups satisfying the corresponding definitions above.

According to a second preferred embodiment, Q is phenyl optionally substituted with 1-5, preferably 2-4, more preferably 2-3, substituents selected from the group consisting of halogen, nitro, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, NR ₇ R ₈ , OR ₇ , SR ₇ , C ₁ -C ₆ alkyl-C (O) OR ₇ , OC ₁ -C ₆ alkyl-C (O) OR ₇ , C ₁ -C ₆ alkyl-OR ₇ , OC ₁ -C ₆ alkyl-OR ₇ , C ₁ -C ₆ alkyl-NR ₇ R ₈ , OC ₁ -C ₆ alkyl-NR ₇ R ₈ , C ₁ -C ₆ alkyl-C (O ) NR ₇ R ₈ , OC ₁ -C ₆ alkyl-C (O) NR ₇ R ₈ , C ₁ -C ₆ alkyl-C (O) R ₇ , OC ₁ -C ₆ alkyl-C (O) R ₇ , C ₁ -C ₆ haloalkyl, O-aralkyl (e.g. benzyloxy), C (O) OR ₇ , C (O) NR ₇ R ₈ , OC (O) NR ₇ R ₈ , NHC (O) R ₇ , NHC ( O) NR ₇ R ₈ , NR ₇ S (O) R ₁ , NR ₇ S (O) _n R ₇ , S (O) _n R ₇ , S (O) _n NR ₇ , where R ₇ and R _{8 are} independently H or C ₁ -C ₆ alkyl. In this embodiment, all other variables R ₂ , R ₅ , R ₆ , A, B, Q, X, and Y (and R ₁ ) can be independently selected from any groups satisfying the corresponding above definitions. Each alkyl, alkenyl and alkynyl moiety may also be substituted as described above.

According to a third preferred embodiment, Q has the structure

Where

R ₉ is H, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₁ -C ₆ alkoxy, hydroxy, NR ₇ R ₈ , SR ₇ or OR ₇ , where R ₇ and R ₈ independently of one another is H or substituted or unsubstituted C ₁ -C ₆ alkyl;

R ₁₀ , R ₁₁ and Z _{2 are} each independently selected from the range consisting of H, halogen, nitro, cyano, C ₁ -C ₆ alkyl, C ₆ -C ₁₀ aryl, NR ₇ R ₈ , OR ₇ , SR ₇ , C ₁ -C ₆ alkyl-C (O) R ₇ , C ₁ -C ₆ alkyl-C (O) NR ₇ R ₈ , C ₁ -C ₆ alkyl-C (O) OR ₇ , C ₁ -C ₆ alkyl-OS (O) R ₇ , C ₁ -C ₇ alkyl-OR ₇ , OS ₁ -C ₆ alkyl-C (O) R ₇ , OC ₁ -C ₆ alkyl-C (O) R ₇ , OC ₁ - C ₆ alkyl-OC (O) R ₇ , OC ₁ -C ₆ alkyl-OR ₇ , OC ₁ -C ₆ alkyl-C (O) NR ₇ R ₈ , C ₁ -C ₆ haloalkyl, OR ₁₂ , C ₁ - C ₆ alkyl-R ₁₂ , OC ₁ -C ₆ alkyl-R ₁₂ , C (O) OR ₇ , C (O) OR ₁₂ , C (O) NR ₇ R ₈ , OC (O) NR ₇ R ₈ , NR ₇ C (O) R ₇ , NR ₇ C (O) R ₁₂ , NR ₇ C (O) -NR ₇ R ₈ , NR ₇ C (O) R ₇ , NR ₇ C (O) OR ₁₂ , NR ₇ C (O) _n -R ₁ , NR ₇ S (O) _n -R ₇ and NR ₇ S (O) _n -R ₁₂ , where R ₇ and R _{8 are} independently H or unsubstituted substituted or substituted C ₁ -C ₆ alkyl, R ₁₂ is unsubstituted or substituted C ₆ -C ₁₀ aryl or heterocyclyl, as defined above, and n is 1 or 2;

Z ₁ is H, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, halogen or nitro. In this embodiment, all other variables R ₂ , R ₅ , R ₆ , A, B, X, and Y can be independently selected from any groups satisfying the above definitions. Each alkyl, alkenyl and alkynyl moiety may also be substituted as described above.

According to various aspects of the invention, Z ₁ and Z ₂ may be hydrogen; Z ₁ , Z ₂ and R ₁₁ may be hydrogen; or Z ₁ , R ₁₀ and R ₁₁ may be hydrogen; and the remaining ring substituents have the above meanings.

According to a further embodiment, the substituents at the 4 and 5 positions or at the 5 and 6 positions of the ring, when Q is substituted phenyl, can be bonded to each other to form an unsubstituted or substituted carbocyclic or heterocyclic ring. Examples of such compounds are shown below, where the symbol

preferably denotes a 5-membered or 6-membered carbocyclic or heterocyclic ring fused to the phenyl ring in the following positions.

или

or

Examples of suitable 5-membered or 6-membered carbocyclic or heterocyclic rings that may be fused to the phenyl ring include the ring systems below in which R ₆ is as defined above.

According to another preferred embodiment, Y is S (O) _n -R ₁ , where n is 1 or 2, preferably 2. In this embodiment, R ₁ may have the above meanings and each of the other variables independently of each other may have any of the above values.

Also preferred are compounds in which Q is substituted phenyl and R _{10 is} selected from the group consisting of C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ aminoalkyl, C ₁ -C ₆ haloalkyl, C ₁ - C ₆ hydroxyalkyl, phenyl, phenoxy, benzyl, benzyloxy, as well as a phenoxy and benzyloxy group substituted with a C ₁ -C ₆ alkyl, a C ₁ -C ₆ alkoxy group, a halogen, a C ₁ -C ₆ haloalkyl, a C ₁ -C ₆ hydroxyalkyl, With ₁ -C ₆ aminoalkyl, OS (O) -C ₁ -C ₆ alkyl, C (O) O-C ₁ -C ₆ alkyl, and C (O) OH, where each of the other variables can be independently selected from any of the above e values.

Also of interest are compounds in which R ₁₁ is NR ₇ C ₁ -C ₆ alkyl-C (O) NR ₇ R ₈ , NR ₇ S (O) _n R ₇ or NR ₇ S (O) _n R ₁₂ , n is 1 or 2 and / or in which Z ₁ = Z ₂ = H and / or in which R ₁₀ is OR ₇ , OR ₁₂ , OC ₇ -C ₁₀ aralkyl, OC ₁ -C ₆ alkyl-OR ₇ or OC ₁ -C ₆ alkyl-OR ₁₂ , where R ₇ and R ₁₂ are unsubstituted or substituted, as defined above. Suitable substituted R ₇ and R ₁₂ include those groups which are substituted as described above, for example, having 1 or 2 C ₁ -C ₆ alkoxy, C ₁ -C ₆ alkoxy-C ₁ -C ₆ alkoxy groups, halogen atoms, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ hydroxyalkyl, C ₁ -C ₆ aminoalkyl, OS (O) -C ₁ -C ₆ alkyl, C (O) O-C ₁ -C ₆ alkyl, C ₁ -C ₆ alkylC (O) OR ₇ -, C ₁ -C ₆ alkyl OC (O) OR ₇ -or C (O) OH groups. In these compounds, each of the other variables, independently of each other, can have any of the above values. Also of interest are those compounds in which Y is S (O) _n -R ₁ , where n is 1 or 2, i.e. representing disulfonamide compounds.

According to another embodiment, A and B are independently CH or CR ₃ , where R ₃ is H, C ₁ -C ₆ alkyl or OH, and where the other variables, independently of each other, can have any of the above values.

According to another embodiment, R ₆ is H or R ₃ is CH ₃ , where the other variables, independently of one another, can have any of the above values.

And according to another preferred embodiment, X denotes a carbonyl group (C = O), where the remaining variables, independently of each other, can have any of the above values. In this embodiment, preferably m is 1.

Table 1 below provides examples of some preferred groups at various positions of certain compounds of the invention. This table presents a group of specific compounds that are obtained by selecting all unique combinations of substituents taken one from each column for each variable, and combining these groups with the structure shown at the top of table 1.

Production Methods

Compounds of the present invention can be obtained using methods based on standard chemical techniques that are described and referenced in well-known manuals (e.g., March J. "Advanced Organic Chemistry" McGraw-Hill, New York, 1997; Collman JP, Hegedus LS, Norton JR, Finke RG "Principles and Applications of Organotransition Metall Chemistry", University of Science, Mill Valley, 1987; Larock RC, "Comprehensive Organic Transformations", Verlag, New York, 1989).

The main intermediate for the synthesis of compounds according to the invention has the following formula

In this formula, A, B, R ₂ , R _4a , R _4b , R ₅ , R ₆ , m and Q have the meanings, including the preferred meanings given above. This compound can be prepared using several alternative synthetic routes. After preparation, the cyano group can be converted to the amidino group (C (NH) NH ₂ ), for example, using known processes, such as the Pinner reaction. The cyano-containing compound having the above formula can be reacted with hydroxylamine, preferably in an alcohol solvent, followed by reduction in the presence of Raney Ni, preferably in an alcohol solvent, or can be first reacted with an ethanolic HCl solution and then with an alcoholic ammonia solution to obtain the corresponding amidino compounds. Alternatively, the desired amidino compound can be prepared using a modified Pinner reaction using pyridine / diethylamine (1/1) / hydrogen sulfide followed by the addition of methyl iodide / acetonitrile and then ammonium acetate / ethanol.

One way to synthesize compounds having the above formula is a condensation reaction using appropriately substituted precursors, as indicated in the scheme below.

This condensation reaction is carried out in the presence of a catalyst, preferably a catalyst such as Lewis acid, and an alkyl alcohol (ROH), preferably a lower alkyl alcohol such as methanol, ethanol, isopropanol, etc., followed by hydrolysis of the intermediate product, preferably with excess water, typically using up to about 10 equivalents of water. Suitable Lewis acids include ether, BF ₃ , AlCl ₃ , etc. W-NC is isonitrile in which W may be any suitable hydrocarbon group, typically an alkyl, carbocycloalkyl or aralkyl group, preferably having no more than about 12 carbon atoms. Particularly preferred isonitrile is benzyl isonitrile. The ester product can be purified by standard methods, including high pressure liquid chromatography (HPLC), column chromatography, recrystallization, etc.

The recovery of the resulting ester to produce an alcohol can be carried out using any known reducing agent ([H]), which preferably should reduce the ester earlier than nitrile. Acceptable reducing agents and methods are well known in the art (see, for example, Modern Synthetic Reactions. HO House, WA Benjami Inc., 2nd ed., 1977). A suitable reducing agent is lithium borohydride. Then the alcohol can be converted into an amine using known chemical reactions. Acceptable conditions include first the interaction of the alcohol with hydrogen azide, DEAD and triphenylphosphine (PPh ₃ ), and then with PPh ₃ and water, or first with phthalimide, DEAD and PPh ₃ , and then with hydrazine. These reactions are shown in the diagram below. Alternatively, the ester may be reacted with a reagent having a nucleophilic carbon atom to introduce suitable R _4a groups. Such reagents may include activated methylene carbon, such as, for example, methylene adjacent to one or more strong electron acceptor groups such as nitro (NO ₂ ), carboalkoxy (COOR _4a ), etc., Grignard reagents (R _4a MgHal, where Hal is halogen), etc., and then converted to alcohol or amine.

The conversion of the amine functional group to sulfonamide and the conversion of the nitrile functional group to amidine can be carried out in any desired order. A preferred reaction scheme is given below.

These transformations are carried out using known chemical reactions, purification and separation methods. An amine can be converted to sulfonamide by reaction with a suitably substituted sulfonyl chloride (ClSO ₂ R ₁ ) in the presence of a base. Nitrile can be reacted with hydroxylamine in an alcohol solvent, followed by reduction, for example, with Raney nickel and hydrogen, or by reacting with a mixture of HCl / alcohol and then ammonia / alcohol. An example of an acceptable reaction sequence is given below.

In this scheme, a - denotes BF ₃ OEt ₂ / EtOH, b - denotes LiВН ₄ / DME (dimethyl ether), c - denotes phthalimide, DIAD (diisopropylazodicarboxylate) / PPh ₃ / THF, d - denotes H ₂ NNH ₂ / EtOH, d is R ₁ SO ₂ Cl, e is H ₂ / Pt / C / EtOH and g is R ₇ SO ₂ Cl / NEt ₃ , NH ₂ OH-HCl / NEt ₃ , H ₂ / Ra-Ni / MeOH .

A similar synthesis scheme, which can be used to obtain the corresponding compounds in which X is carbonyl, is given below.

Compounds in which m is 2 can be obtained using the process shown in the scheme below, where the alcohol is a homologue of the alcohol indicated in the above scheme, and it can be converted to an amine (and other desired compounds) in a similar manner. In the scheme below, (a) denotes a base and (b) denotes a reducing agent, such as LiBH ₄ .

Compounds in which Y is C (O) —R ₁ ; C (O) -OR ₁ ; C (O) -NR ₁ R ₂ is prepared according to the process described above using the appropriate acyl halide (preferably acyl chloride), alkyl haloformate (preferably chloroformate) or isocyanate as shown in the diagram below:

An example of an acceptable reaction sequence is given below.

The esters resulting from the above condensation reactions can also function as intermediates for the synthesis of compounds in which X represents a carbonyl group.

The conversion of the ester to a carboxylic acid can be easily carried out by saponification with an alkali metal hydroxide such as lithium, sodium or potassium hydroxide. The combination of sulfonamide with an acid is first carried out by activating a carboxylate to combine it using, for example, carbonyldiimidazole or other conventional activating agents that are used to synthesize peptides. The second part of the coupling reaction is carried out by mixing an alkyl or arylsulfonamide with a strong base, for example, DBU (1,8-diazabicyclo [5.4.0] undec-7-ene) or sodium hydride, preferably in an anhydrous solvent such as a hydrocarbon or ether solvent for example tetrahydrofuran. Nitrile is converted to amidine using the previously described methods.

In this most preferred embodiment, Q is substituted phenyl, the substituents of which are the above Z ₁ , Z ₂ and R ₉ -R ₁₁ .

Another method for the preparation of intermediates that can be used to prepare the compounds of the invention is given below, and it involves the synthesis of imine derivatives from readily available aldehydes and ketones, followed by the addition of a nucleophilic reagent containing a nucleophilic carbon atom, designated as “Nu”. "Nu" may be a fragment such as CHR _4a NO ₂ , CHR _4a COOR, CH (NO ₂ ) (COOR), etc., which are obtained using well-known Grignard reactions, reactions in which the base is used to remove the proton from a carbon atom adjacent to an electron acceptor group (CO, COO, NO ₂ ), etc.

"Nu" can be converted into a group such as CHR _4a NH ₂ or CHR _4a CH ₂ OH or CHR _4a NH ₂ CH ₂ OH using the following known reduction reactions. In these intermediates, the amino group may be further sulfonated or acylated in any other way, as described above. The following is an example of an acceptable reaction sequence.

To obtain the above alcohol intermediate products, a different synthesis process may be used. As shown in the diagram below, the reaction of the primary styrene derivative with peracide, as a rule, allows to obtain a mixture of products containing non-hydrogen substituents R _4a and / or R ₅ , as shown below, which can be converted to alcohol by reaction with cyananiline without separation or appropriate cyanopyridine.

The alcohol can then be used to prepare the compounds of the invention according to the method described above.

If it is desired to obtain the corresponding compounds in which A and B are nitrogen, the aniline or substituted aniline used in the above reactions is replaced by the corresponding aminopyridines or substituted aminopyridines.

Compounds in which the sulfonamide nitrogen carries a substituent can be obtained by conventional alkylation of a nitrogen atom using known reactions, for example, by alkylation with dialkyl sulfate, alkyl halide, etc. according to known processes.

In a preferred embodiment, Q is a substituted aryl and more preferably a substituted phenyl group and has the structure below.

In this structural formula, Z ₁ , Z ₂ , R ₉ -R ₁₁ have the above general meanings and the meanings given in the preferred embodiments. The compounds of this embodiment are prepared according to the above Scheme I using an appropriately substituted benzaldehyde having a Q-CHO structure (R ₅ is H). These substituted benzaldehydes can be obtained from manufacturers or can be easily obtained from known benzaldehydes using well-known synthetic methods.

In one embodiment, Q is substituted with a nitro group. Preferably, the nitro group is located on the radical R ₁₁ (where Z ₁ , Z ₂ , R ₉ and R ₁₀ have the above general meanings and values given in the preferred embodiments), wherein the nitro group can be further reduced to give an amino group with an acceptable reducing agent. Typically, the cyanamine derivative or cyanosulfonamide derivative shown in Scheme 3 may be reacted with a reducing agent, which preferably should reduce the nitro group on R ₁₁ rather than the cyano group. Any reducing agents having these properties may be used, for example, hydrogen and a Pt / C catalyst. The aniline resulting from the reduction can then be reacted with a sulfonyl chloride (ClSO ₂ W, where W is as defined above) to produce a disulfonamide compound.

The synthesis of cyclic urea derivatives in which N ₁ -R ₂ and N ₂ -R ₂ together form a urea-specific bond, i.e. N ₁ -C (O) -N ₂ allows you to get additional compounds according to the invention, and the processes for their preparation are an additional variant of the method for producing enantiomerically pure compounds according to the invention. Derivatives of cyclic urea can be used, for example, to obtain dialkoxybissulfonamides and other compounds according to the invention, as shown in the diagram below.

Alternatively, the nitric acid in the scheme below can be replaced with sulfuric acid to give sulfonic acid derivatives, which can be further converted to sulfonamides and sulfones by known reactions.

Other compounds of the invention, including heterocyclic compounds, can be easily prepared using conventional starting materials that can be used according to the processes given in the above synthetic schemes. For example, using simple nitro- and hydroxy-substituted aldehydes as starting materials, after the condensation reaction described above, the corresponding esters are obtained which can be directly converted to cyclic derivatives of urethane or oxazole, which can then be subjected to the previously described treatment to give compounds of the invention. These reactions are schematically shown below for rings condensed in position 5 and position 6.

Compounds in which the ring is fused in the 4-position and 5-position with the phenyl ring are obtained by similar methods using the correspondingly substituted aldehyde as starting materials, as shown below.

Other fused heterocyclic compounds are prepared using conventional chemical synthesis reactions and appropriately substituted starting materials that are well known in the art of chemical synthesis, resulting in additional compounds of the invention. For example, fused furan ring systems can be prepared from the corresponding halo and hydroxy substituted aldehydes, as shown below.

Prodrugs of the compounds described above also fall within the scope of the invention. Suitable prodrugs include known amino protecting and carboxy protecting groups that cleave under physiological conditions, for example, by hydrolysis, to form the parent compound. A preferred class of prodrugs are compounds in which the nitrogen atom on the amino, amidino, aminoalkyleneamino, iminoalkyleneamino-guanidino group is substituted with a hydroxy group (OH), an alkylcarbonyl (-CO-W) group, an alkoxycarbonyl (-CO-OW) group, an acyloxyalkylalkoxycarbonyl (- CO-OWO-CO-W) group, where W denotes a monovalent or divalent group as defined above, or a group of the formula -C (O) -O-CP1P2-haloalkyl, where P1 and P2 have the same or different meanings and denote H, ( ness.) alkyl, ness. alkoxy, cyano, halo (ness.) alkyl or aryl. Preferably, the nitrogen atom is one of the nitrogen atoms of the amidino group of the compounds of the invention. These prodrug compounds are prepared by reacting the above-described compounds of the invention with an activated acyl derivative to bind the nitrogen atom of the compound of the invention with the carbonyl of the activated acyl derivative. Suitable activated carbonyl derivatives contain an easily cleavable leaving group bonded to carbonyl carbon and include acyl halides, acylamines, acyl pyridinium salts, acyl alkoxides, in particular acyl phenoxides such as para-nitrophenoxy acyl, dinitrophenoxy acyl, fluorophenoxy phenyl. The reactions are usually exothermic and are carried out in inert solvents at low temperatures, such as from -78 to about 50 ° C. The reactions are also usually carried out in the presence of an inorganic base, such as potassium carbonate or sodium bicarbonate, or an organic base, such as an amine, including pyridine, triethylamine, etc. One way to obtain prodrugs is described in WO 98/46576 published October 22, 1998.

Using the synthesis methods described above, the following representative compounds of the invention (m is 1) are shown in Table 2. In each of the radicals shown in the table, X may be carbonyl or (CR _4a R _4b ) _m , where m is 1 or 2; and the benzamidine ring may be substituted with halogen, hydroxy or alkyl.

The compounds of the invention contain one or more asymmetric carbon atoms. Therefore, the compounds may exist as diastereomers, enantiomers, or mixtures thereof. The synthetic routes described above can be used using racemates, diastereomers, or enantiomers as starting or intermediate products. Diastereomeric compounds can be separated by chromatography or crystallization. Similarly, enantiomeric mixtures can be resolved using the same or other methods known in the art. Each of the asymmetric carbon atoms may be in the R or S configuration, and both of these configurations fall within the scope of the invention.

Application

As indicated above, the compounds of the invention obtained and selected according to the above analyzes are inhibitors of serine proteases, for example, factor VIIa, TF / factor VIIa, factor Xa, kallikrein and / or thrombin. These compounds have the ability to inhibit the catalytic activity of these enzymes and therefore inhibit the coagulation cascade and prevent or limit the coagulation and / or formation of blood clots or emboli in the blood vessels and / or increase the coagulation time of blood. Thus, the compounds of the present invention inhibit the ability of TF / factor VIIa to convert factor X to factor Xa, inhibit the ability of factor Xa to convert prothrombin to thrombin (factor IIa); and / or the ability of thrombin to convert fibrinogen to fibrin monomers.

The selective effect of the compounds of the invention as inhibitors of these enzymes can be determined based on K _i values, as described in the examples below. Data on the selective activity of some representative compounds are shown below in tables 3-6.

Table 3

R ₁ X R ₉ R ₁₀ Z ₂ R ₁₁ K _i (TF / VIIa) Ph C = o Oet O-isoPr H H 0.003 Pr CH ₂ Oet OCH ₂ Ph H NHSO ₂ Me 0.004 Ph C = O Oet Oet H H 0.005 Ph C = O Oet H Oet H 0.007 Table 4 R ₁ X R ₉ R ₁₀ J ₂ R ₁₁ K _i (IIa) Ph CH ₂ Ome OCH (CH ₂ Cl) Ph H H 0.001 2-thiophene CH ₂ Ome OCH ₂ Ph H H 0.016 Ph C = o Oet O-isoPr H H 0.113 Ph CH ₂ Ome OCH (CH ₂ Cl) Ph H H 0.001 Table 5 R ₁ X R ₉ R ₁₀ Z ₂ R ₁₁ K _i (kallikrein) Pr CH ₂ Oet O-isoVu H NHSO ₂ Pr 0.001 Pr CH ₂ Oet O-isoPr H NHSO ₂ Me 0.001 Et C = o Oet O-isoPr H H 0.011 Ph C = O Oet Oet H H 0.002 Table 6 R ₁ X R ₉ R ₁₀ Z ₂ R ₁₁ K _i (Ha) Et C = O Oet O-isoPr N n 0.565 Bu C = O Oet O-isoPr n n 0.624 Ph C = O Oet O-isoPr N n 0.898 R ₁ X R ₉ R ₁₀ Z ₂ R ₁₁ K _i (Xa) Pr CH ₂ Ome OCH (CH ₂ Cl) Ph H H 0.140

The activity of the compounds of the invention as anticoagulants can be determined using the assays below. Analyzes to determine prothrombin time (PT) and activated partial thromboplastin time (APTT) coagulation can be performed using pooled normal plasma (human or various animal species) after adding increasing concentrations of inhibitors to the plasma. Clotting times are determined using an ACL 300 type automatic coagulation analyzer (Coulter Corp., Miami, Florida) and commercial reagents as follows.

PV analysis: Aqueous solutions of various concentrations of the inhibitor are added to the combined normal plasma in the ratio of 1 part of the inhibitor to 9 parts of the plasma. These mixtures are then added to the sample dishes. Innovin® (Dade International Inc., Miami, Florida), which is a mixture of human fat-free tissue factor and Ca ^{+ +} ions, is added to the reagents in dishes. The exact volumes of the sample and Innovin® are automatically transferred to the cells of the acrylic rotor, previously balanced at 37 ° C. After a 2-minute incubation period, coagulation is initiated by mixing the two components together using centrifugation. Coagulation is evaluated optically and clotting time is recorded in seconds. According to Janson et al. (Jansori TL et al., Haemostasis, 14: 440-444, 1984), fat-free human tissue factor is a strong initiator of coagulation in all species studied. In this system, the clotting time in control plasma samples (plasma plus diluent for an inhibitor) is typically 8-10 s. A plot of clotting time versus inhibitor concentration is constructed, and for each inhibitor, a concentration is determined at which the PV is 2 times higher than in the control plasma.

APTT analysis: The inhibitor and plasma are mixed and transferred to sample plates of an ACL 300 type device according to the process described above. Actin FS® and CaCl ₂ (Dade International Inc, Miami, Florida) are respectively added to the reagents in dishes 1 and 2. The exact volumes of the sample and activator (Actin FS®) are automatically transferred to the cells of the acrylic rotor, previously balanced at 37 ° C, and mixed by centrifugation. After a 2-minute activation period, coagulation is initiated by the addition of CaCl ₂ . Coagulation is monitored and data evaluated according to the method described for PV analysis. APTT for control plasma samples, as a rule, is 12-32 s, depending on what types of plasma are used for analysis.

Representative results of PV and APTT analyzes are shown in Table 7 below.

Table 7 Compound No. 2 × PV (μM) 2 × APTT (μM) 7 14 8 thirteen 16 57 33 5.5 eleven 72 thirty 60 589 22 40 596 8 140 628 125 90 672 34 78

The compounds of the invention can be used as in vitro diagnostic reagents for inhibiting coagulation in blood collection tubes. The use of stopper tubes with a vacuum inside as a blood collection device is well known (see Kasten BL, "Specimen Collection", Laboratory Test Handbook, 2nd ed., Lexi-Comp. Inc. Cleveland, pp. 16-17, eds. Jacobs et al., 1990). Such vacuum tubes may not contain coagulation inhibiting additives, in which case they are used to isolate serum from mammalian blood. They may also include coagulation-inhibiting additives such as heparin salts, citrate salts or oxalate salts, in which case they can be used to isolate serum from mammalian blood. The compounds of the invention can be introduced into blood collection tubes and function as TF / factor VIIa, factor Xa, thrombin and / or kallikrein inhibitors and inhibit coagulation of mammalian blood entering the tubes.

When used in blood collection tubes, the compounds of the present invention can be used individually, in the form of mixtures, or in combination with other coagulation inhibitor compounds that are known in the art. The amount of the compound of the invention should be an amount sufficient to prevent or inhibit clot formation when blood enters the tube. These compounds can be introduced into test tubes in the same way as known clot inhibitory compounds such as heparin salts are added. The liquid is typically lyophilized using known methods. Typically, the tubes should contain from about 2 to about 10 ml of mammalian blood and the compounds are added in an amount sufficient to prevent coagulation of this amount of blood. An acceptable concentration is 10-1000 nM.

These compounds also inhibit the formation of emboli and blood clots in the mammalian circulatory system and, therefore, can be used in vivo. It was found that thromboembolic disorders are directly related to the sensitivity of the mammal to the formation of emboli and blood clots. For example, the formation of a thrombus in a venous vessel leads to thrombophlebitis, which, as a rule, is treated with rest and the introduction of anticoagulants. Other conditions that can be treated using the anticoagulants of the invention include thrombolimphangiitis, thrombosinusitis, thrombosis, thromboangiitis and thrombarteritis.

Mammals that undergo medical procedures such as angioplasty and thrombolytic therapy are especially susceptible to blood clots. The compounds of the present invention can be used to inhibit blood clots after angioplasty. They can also be used in combination with antithrombolytic agents, such as tissue plasminogen activator and its derivatives (US Pat. Nos. 4,752,603; 4,766,075; 4,776,043; EP 199574; EP 238304; EP 228862; EP 297860; PCT WO 89/04368; PCT WO 89/00197 ), streptokinase and its derivatives or urokinase and its derivatives, for the prevention of arterial reocclusion after thrombolytic therapy. When administered in combination with the above thrombolytic agents, the compounds of the present invention can be administered before, simultaneously or sequentially with an antithrombolytic agent.

Mammals that undergo renal dialysis, blood oxygenation, cardiac catheterization and similar medical procedures, as well as mammals that are given certain prosthetic devices, are also sensitive to thromboembolic disorders. Physiological conditions, for known or unknown reasons, can also lead to thromboembolic disorders.

Thus, the compounds provided herein can be used to treat thromboembolic disorders in mammals. The compounds provided herein can also be used as adjuvants in anticoagulation therapy, for example, in combination with aspirin, heparin or warfarin and other anticoagulants. The various coagulation disorders described above are treated with the compounds of the invention in such a way as to prevent bleeding as a result of such a violation. The use of the compounds provided herein for such and similar disorders is apparent to those skilled in the art.

The compounds of the invention can also be used as intermediates in general or as precursors for inhibitors of coagulation of serine protease and, therefore, in addition to treating cardiovascular disease, these compounds can be used for metastatic disease or any disease in which coagulation inhibition is indicated.

In general, the inhibitors used according to the method of the invention are prepared by mixing at ambient temperature and at a suitable pH and with the desired degree of purity with physiologically acceptable carriers, i.e. carriers that are non-toxic to recipients in the applicable doses and concentrations. The pH of the composition mainly depends on the particular compound employed and its concentration, but is preferably in the range of from about 3 to about 8. An acceptable embodiment is the composition in acetate buffer, pH 5.

Inhibitor compounds for use in the present invention are preferably sterile. The compound should usually be stored as a solid composition, although lyophilized compositions or aqueous solutions are also suitable.

The composition according to the invention can be prepared, divided into doses and administered according to accepted medical practice. Factors that should be considered include the specific disorder to be treated, the specific mammal to be treated, the clinical condition of the particular patient, the cause of the disease, the delivery area of the agent, method of administration, schedule of use, and other factors known to practicing physicians. A “therapeutically effective amount” of a compound to be administered must meet these requirements and is the smallest amount necessary to prevent, alleviate symptoms or treat a disease caused by a coagulation factor. Such an amount is preferably lower than the amount toxic to the host or the amount that makes the host much more sensitive to bleeding.

In general, it has been proposed to use such an initial pharmaceutically effective amount of the inhibitor when administered parenterally so that the dose is about 0.01-100 mg / kg, preferably about 0.1-20 mg / kg of the patient’s body weight per day, with the usual initial dose range used the compound is from 0.3 to 15 mg / kg / day.

The compound of the invention is administered by any suitable route, including oral, topical, transdermal, parenteral, subcutaneous, intraperitoneal, intrapulmonary, and intrinasal and, if local immunosuppressive treatment is required, by introducing inside the lesion (including perfusion or other transplant contact with the inhibitor before transplantation). Parenteral perfusions include intramuscular, intravenous, intraarterial, intraperitoneal or subcutaneous administration.

The invention will become more apparent after studying the examples below. However, it should be obvious that they are not intended to limit the scope of the invention. All patent and literature sources are fully incorporated into this description by reference.

Examples

The compounds of the invention as a whole can be prepared using the reaction scheme below. Compounds other than the specific product shown are prepared according to the process described above using the appropriate starting materials. For example, additional compounds can be prepared using other starting styrenes, which can be easily synthesized from commercially available starting products and using standard reactions that are well known in the art.

Example 1

4-Benzyloxy-3-methoxystyrene (10 g, 42 mmol) was dissolved in dichloromethane (400 ml). Solid potassium bicarbonate (11 g, 110 mmol) was added and the reaction mixture was cooled to 0 ° C. Meta-chloroperbenzoic acid (12 g, approximately 42 mmol) was added and the reaction mixture was allowed to warm to room temperature and stirred for 16 hours. Reaction was monitored by thin layer chromatography. An additional amount of meta-chloroperbenzoic acid (4 g) was added and the reaction mixture was stirred for another 4 hours until complete absorption of the starting material. The reaction mixture was poured into a separatory funnel and washed first with water, then sodium bicarbonate and finally NaOH. The organic layer was separated and dried over anhydrous sodium sulfate. The solution was filtered and the solvent was evaporated in vacuo to give about 11 g of a crude product.

The crude product was then dissolved in acetonitrile (60 ml) and lithium perchlorate (8.5 g, 80 mmol) was added. The suspension was stirred for 5 minutes, during which time the reaction mixture became homogeneous. 4-aminobenzonitrile (9.5 g, 80 mmol) was added and the reaction mixture was heated to 60 ° C for 12 hours. Thin layer chromatography showed the presence of a new product with a lower Rf value. The solvent was removed in vacuo and the residue was dissolved in ethyl acetate, washed with water and dried over anhydrous sodium sulfate. The crude product was then subjected to flash chromatography (hexane: ethyl acetate, 1: 1) to obtain 6 g of 4- [1- (4-benzyloxy-3-methoxyphenyl) -2-hydroxyethylamino] benzonitrile A.

¹ H-NMR (CDCl ₃ ): 7.3-7.45 (m, 7H), 6.8 (m, 3H), 6.5 (d, 2H), 5.18 (s, 2H), 4 42 (m, 1H); 3.95 (dd, 1H); 3.85 (s, 3H); 3.8 (dd, 1H).

4- [1- (4-Benzyloxy-3-methoxyphenyl) -2-hydroxyethylamino] benzonitrile A- (470 mg, 1.25 mmol), phthalimide (1.47 g, 10 mmol) and triphenylphosphine (787 mg, 3 mmol ) was added to 40 ml of tetrahydrofuran. The mixture was stirred for 10 minutes and then cooled to 0 ° C. Then diisopropylazodicarboxylate (DIAD, 0.6 ml, 3 mmol) was slowly added. The reaction mixture was allowed to stir for 1 h. TLC showed the presence of a new product. The solvent was removed in vacuo and the residue was dissolved in 50 ml of ethyl acetate. The solution was washed three times with 2N. sodium hydroxide and twice with water. The organic layer was separated, dried over anhydrous sodium sulfate and filtered. The solvent was removed in vacuo and the residue was subjected to flash chromatography (hexane: ethyl acetate, 1: 1) to obtain 478 mg of 4- [1- (4-benzyloxy-3-methoxyphenyl) -2- (1,3-dioxo-1,3 -dihydroisoindol-2-yl) ethylamino] benzonitrile B (76% yield).

¹ H-NMR (CDCl ₃ ): 7.85 (m, 2H), 7.75 (m, 2H), 7.23-7.45 (m, 9H), 6.9 (m, 3H), 6 42 (d, 2H), 5.45 (d, 1H), 5.15 (s, 2H), 4.62 (m, 1H), 4.0 (m, 2H), 3.83 (s, 3H).

4- [1- (4-Benzyloxy-3-methoxyphenyl) -2- (1,3-dioxo-1,3-dihydroisoindol-2-yl) ethylamino] benzonitrile B was then dissolved in ethanol (60 ml) and hydrazine hydrate was added (2 g). The solution was heated to 60-70 ° C for 1.5 hours. TLC showed that the reaction was complete. The suspension was filtered to remove the by-product and ethanol was removed in vacuo. The residue was flash chromatographed on silica gel (ethyl acetate: 2N. NH ₃ in methanol, 9: 1) to obtain 372 mg of 4- [2-amino-1- (4-benzyloxy-3-methoxyphenyl) ethylamino] benzonitrile G (100% output).

¹ H-NMR (CDCl ₃ ): 7.3-7.45 (m, 7H), 6.32 (m, 3H), 6.5 (d, 2H), 5.52 (d, 1H), 4 3 (q, 1H); 3.83 (s, 3H); 3.08 (m, 2H); 1.95 (s, 2H).

4- [2-amino-1- (4-benzyloxy-3-methoxyphenyl) ethylamino] benzonitrile G (300 mg, 0.8 mmol) was dissolved in ethanol (3 ml) and hydroxylamine hydrochloride (350 mg, 5 mmol) was added and triethylamine (1 ml, 5.7 mmol). The reaction mixture was heated to 65-70 ° C for 2 hours. The residue was dissolved in ethyl acetate and water. The organic layer was separated, dried over anhydrous sodium sulfate and filtered. The solvent was removed in vacuo and replaced with 4 ml of methanol and 0.5 ml of acetic acid. Raney nickel (approximately 30 μl of a suspension in sodium hydroxide, Aldrich) was added and the reaction mixture was placed in a hydrogen atmosphere. The reaction mixture was stirred vigorously for 3 hours, the catalyst was filtered off and the solvent was removed in vacuo. The crude product was purified by flash chromatography on silica gel (ethyl acetate: acetone: methanol: ammonia, 2: 1: 1: 0.05) to give 160 mg of 4- [2-amino-1- (4-benzyloxy-3-methoxyphenyl) ethylamino ] benzamidine D. MS (M + H) = 391.

4- [2-amino-1- (4-benzyloxy-3-methoxyphenyl) ethylamino] benzamidine D (20 mg, 0.03 mmol) was dissolved in acetonitrile (2 ml) containing triethylamine (17 μl, 0.12 mmol) and water (0.3 ml). The desired sulfonyl chloride of the formula ClSO ₂ R (0.03 mmol) was added to this mixture and the reaction mixture was stirred for 4 hours. The solvent was removed in vacuo and the compounds were purified by preparative reverse phase HPLC (acetonitrile / water gradient with 0.1% trifluoroacetic acid added ), obtaining after lyophilization of the final product.

Examples 2a-2gg

Working similarly, other compounds of the invention were prepared, including:

a) 4- [benzenesulfonylamino-1- (4-benzyloxy-3-methoxyphenyl) ethylamino] benzamidine: MS (M + H) = 531,

b) N- [4- [2- (4-benzyloxy-3-methoxyphenyl) -2- (4-carbamimidoylphenylamino) ethylsulfamoyl] phenyl} acetamide: MS (M + H) = 588,

c) 4- [1- (4-benzyloxy-3-methoxyphenyl) -2- (4-nitrobenzenesulfonylamino) ethylamino] benzamidine: MS (M + H) = 576,

d) 4- [1- (4-benzyloxy-3-methoxyphenyl) -2- (4-fluorobenzenesulfonylamino) ethylamino] benzamidine: MS (M + H) = 549,

d) 4- [1- (4-benzyloxy-3-methoxyphenyl) -2- (4-fluorobenzenesulfonylamino) ethylamino] benzamidine: MS (M + H) = 565,

e) 4- [1- (4-benzyloxy-3-methoxyphenyl) -2- (3-nitrobenzenesulfonylamino) ethylamino] benzamidine: MS (M + H) = 576,

g) 4- [1- (4-benzyloxy-3-methoxyphenyl) -2- (2,5-dichlorobenzenesulfonylamino) ethylamino] benzamidine: MS (M + H) = 599,

h) 4- [1- (4-benzyloxy-3-methoxyphenyl) -2- (2-bromobenzenesulfonylamino) ethylamino] benzamidine: MS (M + H) = 609, 611,

i) 4- [1- (4-benzyloxy-3-methoxyphenyl) -2- (4-bromobenzenesulfonylamino) ethylamino] benzamidine: MS (M + H) = 609,

j) 4- [1- (4-benzyloxy-3-methoxyphenyl) -2- (4-isopropylbenzenesulfonylamino) ethylamino] benzamidine: MS (M + H) = 573,

k) 4- [1- (4-benzyloxy-3-methoxyphenyl) -2-phenylmethanesulfonylaminoethylamino] benzamidine: MS (M + H) = 545,

m) 4- [1- (4-benzyloxy-3-methoxyphenyl) -2- (4-carboxybenzenesulfonylamino) ethylamino] benzamidine: MS (M + H) = 575,

m) 4- [1- (4-benzyloxy-3-methoxyphenyl) -2- (3-carboxybenzenesulfonylamino) ethylamino] benzamidine: MS (M + H) = 575,

o) 4- [1- (4-benzyloxy-3-methoxyphenyl) -2- (2,4-dinitrobenzenesulfonylamino) ethylamino] benzamidine: MS (M + H) = 621,

p) 4- [1- (4-benzyloxy-3-methoxyphenyl) -2- (2,3,5,6-tetramethylbenzenesulfonylamino) ethylamino] benzamidine: MS (M + H) = 587,

p) 4- [1- (4-benzyloxy-3-methoxyphenyl) -2- (3,5-dichloro-2-hydroxybenzenesulfonylamino) ethylamino] benzamidine: MS (M + H) = 615,

c) 4- [1- (4-benzyloxy-3-methoxyphenyl) -2- (3,4-dimethoxybenzenesulfonylamino) ethylamino] benzamidine: MS (M + H) = 591,

t) 4- [1- (4-benzyloxy-3-methoxyphenyl) -2- (thiophen-2-sulfonylamino) ethylamino] benzamidine: MS (M + H) = 537,

y) N- {5- [2- (4-benzyloxy-3-methoxyphenyl) -2- (4-carbamimidoylphenylamino) ethylsulfamoyl] -4-methylthiazol-2-yl} acetamide: MS (M + H) = 595,

f) 4- [1- (4-benzyloxy-3-methoxyphenyl) -2- (naphthalene-2-sulfonylamino) ethylamino] benzamidine: MS (M + H) = 581,

x) 4- [1- (4-benzyloxy-3-methoxyphenyl) -2- (naphthalene-1-sulfonylamino) ethylamino] benzamidine: MS (M + H) = 581,

c) 4- [1- (4-benzyloxy-3-methoxyphenyl) -2- (2-phenylethanesulfonylamino) ethylamino] benzamidine: MS (M + H) = 557,

h) 4- [1- (4-benzyloxy-3-methoxyphenyl) -2- (3-trifluoromethylbenzenesulfonylamino) ethylamino] benzamidine: MS (M + H) = 599,

w) 4- [1- (4-benzyloxy-3-methoxyphenyl) -2- (2,3,4,5,6-pentafluorobenzenesulfonylamino) ethylamino] benzamidine: MS (M + H) = 521,

u) 4- [1- (4-benzyloxy-3-methoxyphenyl) -2-methanesulfonylaminoethylamino] benzamidine: MS (M + H) = 469,

e) 4- [1- (4-benzyloxy-3-methoxyphenyl) -2-ethanesulfonylaminoethylamino] benzamidine: MS (M + H) = 483,

aa) 4- [1- (4-benzyloxy-3-methoxyphenyl) -2-propanesulfonylaminoethylamino] benzamidine: MS (M + H) = 497,

bb) 4- [1- (4-benzyloxy-3-methoxyphenyl) -2-butanesulfonylaminoethylamino] benzamidine: MS (M + H) = 511,

cc) ethyl ester of [2- (4-benzyloxy-3-methoxyphenyl) -2- (4-carbamimidoylphenylamino) ethylsulfamoyl] acetic acid: MS (M + H) = 541,

g) [2- (4-benzyloxy-3-methoxyphenyl) -2- (4-carbamimidoylphenylamino) ethylsulfamoyl] acetic acid.

Example 3

4- [1- (4-Benzyloxy-3-methoxyphenyl) -2- (1,3-dioxo-1,3-dihydroisoindol-2-yl) ethylamino] benzonitrile B (1.8 g) was dissolved in a mixture containing ethanol (50 ml), acetic acid (3 ml), methanol (5 ml) and ethyl acetate (5 ml). This solution was added to a Parr flask containing 10% Pd / C (500 mg) and hydrogenated for 16 hours at a pressure of 35 psi. inch. The catalyst was removed by filtration through celite and the solvent was removed in vacuo to give 1 g of product, i.e. 4- [2- (1,3-dioxo-1,3-dihydroisoindol-2-yl) -1- (4-hydroxy-3-methoxyphenyl) ethylamino] benzonitrile (68%).

4- [2- (1,3-dioxo-1,3-dihydroisoindol-2-yl) -1- (4-hydroxy-3-methoxyphenyl) ethylamino] benzonitrile (1 g, 2.43 mmol) was dissolved in heterohydrofuran ( 30 ml) and triphenylphosphine (1.27 g, 4.84 mmol), (S) -2-chloro-1-phenylethanol (1.13 g, 7.26 mmol) were added. The reaction mixture was cooled to 0 ° C. and diethyl azodicarboxylate (0.842 g, 4.8 mmol) was added. The reaction mixture was allowed to warm to room temperature and the reaction mixture was stirred for 2.5 hours. The solvent was removed in vacuo and the residue was dissolved in ethyl acetate, washed several times with 0.5N. sodium hydroxide, washed once with brine and dried over anhydrous sodium sulfate. The crude product was purified by flash chromatography on silica gel (30% ethyl acetate in hexane) to give 1.1 g of 4- [1- [4- (2-chloro-1-phenylethoxy) -3-methoxyphenyl] -2- (1,3 -dioxo-1,3-dihydroisoindol-2-yl) ethylamino] benzonitrile (82%).

4- [1- [4- (2-Chloro-1-phenylethoxy) -3-methoxyphenyl] -2- (1,3-dioxo-1,3-dihydroisoindol-2-yl) ethylamino] benzonitrile (0.5 g ) was dissolved in ethanol (40 ml) and hydrazine hydrate (0.14 g) was added. The reaction mixture was kept for 2 hours at 65 ° C. The solvent was removed and replaced with ethyl acetate. The solution was washed twice with water and once with brine. The solution was dried over anhydrous sodium sulfate and the solvent was removed in vacuo to give 318 mg of the desired amine, i.e. 4- {2-amino-1- [4- (2-chloro-1-phenylethoxy) -3-methoxyphenyl] ethylamino} benzonitrile (83%).

4- [2-amino-1- [4- (2-chloro-1-phenylethoxy) -3-methoxyphenyl] ethylamino} benzonitrile (0.1 g, 0.237 mmol) was dissolved in dichloromethane (6 ml) and triethylamine (34 μl, 0.3 mmol). Phenylsulfonyl chloride (46 μl, 0.26 mmol) was added and the reaction mixture was stirred for 90 minutes. The reaction mixture was diluted with dichloromethane and washed once with saturated sodium bicarbonate solution and once with water. The solution was dried over sodium sulfate and the solvent was removed in vacuo. The product was purified on silica gel to give 100 mg of the desired product, i.e. N- [2- [4- (2-chloro-1-phenylethoxy) -3-methoxyphenyl] -2- (4-cyanophenylamino) ethyl] benzenesulfonamide.

N- [2- [4- (2-chloro-1-phenylethoxy) -3-methoxyphenyl] -2- (4-cyanophenylamino) ethyl] benzenesulfonamide (100 mg, 0.18 mmol) was dissolved in ethanol (3 ml) and hydroxylamine hydrochloride (62 mg, 0.89 mmol) was added. Triethylamine (90 mg, 0.89 mmol) was added to this mixture, followed by potassium carbonate (62 mg). The reaction mixture was heated to 80 ° C. for 48 hours. The reaction mixture was cooled and the solvent was removed in vacuo. The residue was dissolved in ethyl acetate and washed twice with water and once with brine. The solution was dried over sodium sulfate and the solvent was removed. The crude intermediate was dissolved in methanol (4 ml) and a few drops of acetic acid were added. Approximately 50-100 mg of Raney nickel suspended in sodium hydroxide (Aldrich) was added and the reaction mixture was placed in a hydrogen atmosphere. The suspension was stirred vigorously for 8 hours, the catalyst was filtered off and the solvent was removed in vacuo. The crude product was purified by reverse phase chromatography to give 4- [2-benzenesulfonylamino-1- [4- (2-chloro-1-phenylethoxy) -3-methoxyphenyl] ethylamino} benzamidine (32 mg). MS (M + H) = 579.

Example 4

4- {2-Propanesulfonylamino-1- [4- (2-chloro-1-phenylethoxy) -3-methoxyphenyl] ethylamino} benzamidine was obtained analogously to example 2 except that in the reaction with 4- {2-amino-1- [4- (2-Chloro-1-phenylethoxy) -3-methoxyphenyl] ethylamino} benzonitrile benzenesulfonyl chloride was replaced with propanesulfonyl chloride. MS (M + H) = 545.

Example 5

4-Benzyloxy-5-methoxy-2-nitrobenzaldehyde (12.2 g, 42 mmol) and 4-aminobenzonitrile (5 g, 42 mmol) were dissolved in methanol (165 ml) and stirred for 2 hours, and then heated to 60 ° C for 30 minutes The reaction mixture was allowed to cool to room temperature and benzylisonitrile (5 g, 42 mmol) was added. The reaction mixture was cooled to 0 ° C and boron trifluoroetherate (16 ml, 126 mmol) was added dropwise over 5 minutes. The reaction mixture was stirred for 20 minutes at 0 ° C and then allowed to warm to room temperature, after which it was stirred at ambient temperature for 2 hours. Water (4 ml) was added and the mixture was stirred at room temperature overnight. The next morning a yellow precipitate formed and the solid product was filtered off. The solid was washed with methanol and dried in air to give 8 g of the desired product. The solvent was removed from the filtrate in vacuo and replaced with ethyl acetate. The solution was washed with water and saturated sodium bicarbonate, dried over anhydrous magnesium sulfate, and the solvent was removed. The crude product was subjected to flash chromatography (hexane: ethyl acetate, 1: 1) to obtain an additional 7 g of the desired product, i.e. (4-Benzyloxy-5-methoxy-2-nitrophenyl) - (4-cyanophenylamino) acetic acid methyl ester.

¹ H-NMR (CDCl ₃ : 7.68 (s, 1H), 7.4 (m, 7H), 7.0 (s, 1H), 6.61 (d, 2H), 6.2 (s. 1H), 5.2 (s, 2H), 3.87 (s, 3H), 3.75 (s, 3H).

(4-Benzyloxy-5-methoxy-2-nitrophenyl) - (4-cyanophenylamino) acetic acid methyl ester (4.5 g, 10 mmol) was dissolved in dimethoxyethane and lithium borohydride (0.210 g, 10 mmol) was added. The reaction mixture was heated to reflux for 3 hours and cooled to room temperature. The reaction was stopped by adding acetic acid containing water and diluted with ethyl acetate. After transfer to a separatory funnel, the organic layer was washed several times with water.

The organic layer was dried over anhydrous sodium sulfate, filtered and the solvent was removed. The crude product was then subjected to flash chromatography to give 3.1 g of 4- [1- (4-benzyloxy-5-methoxy-2-nitrophenyl) -2-hydroxyethylamino] benzonitrile (74%).

¹ H-NMR (CDCl ₃ ): 7.77 (s, 1H), 7.3-7.5 (m, 7H), 7.15 (s, 1H), 6.42 (d, 2H), 5 4 (bs, 1H), 5.18 (dd, AB system, 2H), 4.15 (dd, 1H), 3.83 (s, 3H), 3.79-3.86 (dd, 1H )

4- [1- (4-Benzyloxy-5-methoxy-2-nitrophenyl) -2-hydroxyethylamino] benzonitrile (3.1 g, 7.4 mmol) was dissolved in tetrahydrofuran (120 ml) and triphenylphosphine (5.9 g , 22 mmol) and phthalimide (5.4 g, 37 mmol). The reaction mixture was cooled to 0 ° C. and diisopropylazadicarboxylate (DIAD, 4.6 g) was added dropwise. The reaction mixture was allowed to warm to room temperature and was stirred overnight. The solvent was removed in vacuo and replaced with ethyl acetate. The solution was washed several times with 1N. NaOH and dried over anhydrous sodium sulfate. After flash chromatography (hexane: ethyl acetate, 1: 1), the desired product was obtained, in which some DIAD was still present. The solid was washed several times with ethanol to give 3.2 g of the desired phthalimide, i.e. 4- [1- (4-benzyloxy-5-methoxy-2-nitrophenyl) -2- (1,3-dioxo-1,3-dihydroisoindol-2-yl) ethylamino] benzonitrile (3.2 g).

¹ H-NMR (CDCl ₃ : 7.83 (m, 2H), 7.75 (m, 2H), 7.37 (m, 7H), 6.91 (s, 1H), 6.41 (d, 2H), 6.17 (d, 1H), 5.65 (m, 1H), 5.18 (s, 2H), 4.27 (m, 2H), 3.61 (s, 3H).

4- [1- (4-Benzyloxy-5-methoxy-2-nitrophenyl) -2- (1,3-dioxo-1,3-dihydroisoindol-2-yl) ethylamino] benzonitrile (2.7 g, 5 mmol) dissolved in ethanol (100 ml) and hydrazine hydrate (0.65 ml, 20 mmol) was added. The reaction mixture was heated to 60 ° C. for 3 hours and then kept at room temperature for 48 hours. The precipitated solids were filtered off and the residue was subjected to flash chromatography to give 4- [2-amino-1- (4-benzyloxy-5- methoxy-2-nitrophenyl) ethylamino] benzonitrile (1.5 g).

¹ H-NMR (CDCl ₃ ): 7.75 (s, 1H), 7.3-7.5 (m, 7H), 7.05 (s, 1H), 6.45 (d, 2H), 5 80 (bs, 1H), 5.32 (m, 1H), 5.19 (s, 2H), 3.81 (s, 3H), 3.25 (dd, 1H), 3.0 (dd, 1H), 1.65 (bs, 2H).

4- [2-amino-1- (4-benzyloxy-5-methoxy-2-nitrophenyl) ethylamino] benzonitrile (0.227 g, 0.66 mmol) was dissolved in dichloromethane (4 ml) and triethylamine (0.14 ml, 1 mmol). The reaction mixture was cooled to 0 ° C. and 1-propanesulfonyl chloride (0.085 ml, 0.75 mmol) was added. The reaction mixture was stirred for 20 minutes and the product was purified by flash chromatography (hexane: ethyl acetate, 1: 1) to obtain 260 mg of the desired product, i.e. [2- (4-Benzyloxy-5-methoxy-2-nitrophenyl) -2- (4-cyanophenylamino) ethyl] propane-1-sulfonic acid amide.

¹ H-NMR (CDCl ₃ ): 7.77 (s, 1H), 7.3-7.5 (m, 7H), 7.12 (s, 1H), 6.41 (d, 2H), 6 , 0 (d, 1H), 5.3 (m, 1H), 5.17 (dd, AB, 2H), 4.75 (t, 1H), 3.85 (s, 3H), 3.65 ( m, 1H), 3.04 (m, 2H), 1.83 (m, 2H), 1.05 (t, 3H).

[2- (4-Benzyloxy-5-methoxy-2-nitrophenyl) -2- (4-cyanophenylamino) ethyl] propane-1-sulfonic acid amide (0.250 g) was dissolved in ethanol (10 ml) and added to Pt / C (5%). The reaction mixture was placed in a hydrogen atmosphere and stirred vigorously for 3 hours. The catalyst was filtered off and the product was chromatographed (hexane: ethyl acetate, 1: 2) to obtain 133 mg [2- (2-amino-4-benzyloxy-5-methoxyphenyl) -2 - (4-cyanophenylamino) ethyl] propane-1-sulfonic acid amide.

¹ H-NMR (CDCl ₃ ): 7.3-7.45 (m, 7H). 6.71 (s, 1H), 6.52 (d, 2H), 6.3 (s, 1H), 5.08 (s, 2H), 5.0 (t, 1H). 4.42 (q, 1H), 3.75 (s, 3H), 3.70 (bs, 2H), 3.45 (t, 2H), 2.97 (m. 2H), 1.80 (m, 2H) ), 1.03 (t, 3H).

[2- (2-Amino-4-benzyloxy-5-methoxyphenyl) -2- (4-cyanophenylamino) ethyl] propane-1-sulfonic acid amide (133 mg, 0.27 mmol) was dissolved in dichloromethane and triethylamine (0 05 ml, 0.35 mmol). The reaction mixture was cooled and methanesulfonyl chloride (0.023 ml, 0.3 mmol) was added dropwise. The reaction mixture was stirred for 2 hours and the product was purified by flash chromatography (hexane: ethyl acetate, 1: 1). The product was then dissolved in ethanol and hydroxylamine hydrochloride (35 mg, 0.5 mmol) was added. Sodium ethoxide (48 mg, 0.7 mmol) was added and the reaction mixture was heated for 48 hours. Ethanol was removed and water (4 ml) was added. The solid product was filtered and washed with water. The crude product was then dissolved in 4 ml of methanol containing 0.5 ml of acetic acid. Raney nickel (approximately 50 mg as a suspension in sodium hydroxide, Aldrich) was added and the reaction mixture was placed in a hydrogen atmosphere. The reaction mixture was stirred vigorously for 3 hours and the catalyst was filtered off. The crude product was subjected to reverse phase preparative chromatography to give the final product, i.e. 4- [1- (4-benzyloxy-2-methanesulfonylamino-5-methoxyphenyl) -2- (propan-1-sulfonylamino) ethylamino] benzamidine (12 mg): MS (M + H) = 590.

Examples 6a-6g

Using a process similar to that described in example 5, the following compounds were obtained:

a) 4- [2-benzenesulfonylamino-1- (2-benzenesulfonylamino-4-benzyloxy-5-methoxyphenyl) ethylamino] benzamidine. The process was consistent with the process described above, except phenylsulfonyl chloride was used instead of propanesulfonyl chloride and methanesulfonyl chloride. MS: (M + H) = 686.

b) 4- [2-Benzenesulfonylamino-1- (2-benzenesulfonylamino-4-benzyloxy-5-ethoxyphenyl) ethylamino] benzamidine. The process was consistent with the process described above, except that 3-ethoxy, 4-benzyloxy, 6-nitrobenzaldehyde was used as starting materials. MS: (M + H) = 700.

c) 4- [1- (4-Benzyloxy-5-ethoxy-2-methanesulfonylaminophenyl) -2- (propane-1-sulfonylamino) ethylamino] benzamidine. The process was consistent with the process described above, except that 3-ethoxy, 4-benzyloxy, 6-nitrobenzaldehyde was used as starting materials. MS: (M + H) = 604.

d) 4- [1- (4,5-Diethoxy-2-methanesulfonylaminophenyl) -2- (propan-1-sulfonylamino) ethylamino] benzamidine. The process was consistent with the process described above, except that 3,4-diethoxy, 6-nitrobenzaldehyde was used as starting materials. MS: (M + H) = 542.

e) [5-Benzyloxy-2- [1- (4-carbamimidoylphenylamino) -2- (propane-1-sulfonylamino) ethyl] -4-methoxyphenylsulfamoyl} acetic acid ethyl ester. The process was consistent with the process described above, except that chlorosulfonylacetic acid ethyl ester was used instead of methanesulfonyl chloride. MS: (M + H) = 676.

e) {5-Benzyloxy-2- [1- (4-carbamimidoylphenylamino) -2- (propan-1-sulfonylamino) ethyl] -4-methoxyphenylsulfamoyl} acetic acid. [5-Benzyloxy-2- [1- (4-carbamimidoylphenylamino) -2- (propane-1-sulfonylamino) ethyl] -4-methoxyphenylsulfamoyl} acetic acid ethyl ester (10 mg) was dissolved in water (2 ml) and tetrahydrofuran was added (2 ml) and LiOH (3 mg). The mixture was allowed to stir overnight. The product was purified using preparative reverse-phase HPLC. Received 3 mg of product. MS (M + H) = 648.

g) 4- [1- (3,4-Dimethoxy-2-methanesulfonylaminophenyl) -2- (propan-1-sulfonylamino) ethylamino] benzamidine. This compound was prepared according to a process similar to that described above, except that 2-bromo-3,4-dimethoxybenzaldehyde was used instead of 4-benzyloxy-5-methoxy-2-nitrobenzaldehyde. MS: (M + H) = 499.

Examples 7a-7g

Compounds 7a-7g are generally prepared as follows. Compound E (20 mg, 0.03 mmol) was dissolved in acetonitrile (2 ml) containing triethylamine (17 μl, 0.12 mmol) and water (0.3 ml). The corresponding acyl chloride, alkyl chloroformate or isocyanate (0.03 mmol) was added to this mixture and the reaction mixture was stirred for 4 hours. The solvent was removed in vacuo and the compounds were purified using preparative reverse phase HPLC (acetonitrile / water mixture containing 0.1% trifluoroacetic acid), obtaining the final product after lyophilization.

7a) N- [2- (4-Benzyloxy-3-methoxyphenyl) -2- (4-carbamimidoylphenylamino) ethyl] -2,2,2-trifluoroacetamide, MS (M + H) = 487.

7b) N- [2- (4-Benzyloxy-3-methoxyphenyl) -2- (4-carbamimidoylphenylamino) ethyl] acetamide, MS (M + H) = 433.

7c) N- [2- (4-Benzyloxy-3-methoxyphenyl) -2- (4-carbamimidoylphenylamino) ethyl] butyramide, MS (M + H) = 461.

7g) N- [2- (4-Benzyloxy-3-methoxyphenyl) -2- (4-carbamimidoylphenylamino) ethyl] -2-chloroacetamide, MS (M + H) = 467.

7e) [2- (4-Benzyloxy-3-methoxyphenyl) -2- (4-carbamimidoylphenylamino) ethyl] carboxylic acid methyl ester, MS (M + H) = 449.

7e) [2- (4-Benzyloxy-3-methoxyphenyl) -2- (4-carbamimidoylphenylamino) ethyl] carboxylic acid isobutyl ester, MS (M + H) = 491.

7g) [2- (4-Benzyloxy-3-methoxyphenyl) -2- (4-carbamimidoylphenylamino) ethyl] carboxylic acid 2,2,2-trichloroethyl ester, MS (M + H) = 565.

Example 8

Methyl ester of the above acid (920 mg, 2.85 mmol) was suspended in a mixture of THF / water (3/1) (40 ml) and cooled to 0 ° C. The solution was treated with 1 N. Ll OH (7.1 ml, 7.1 mmol) and stirred overnight. The reaction mixture was acidified with trifluoroacetic acid until a pH of 4.0 was reached. The solvent was removed in vacuo and the crude product was purified by flash chromatography (ethyl acetate containing 0.5% acetic acid) to give 1 g of a carboxylic acid.

Carbonyldiimidazole (131 mg, 0.8 mmol) was dissolved in anhydrous THF (1.6 ml), and the carboxylic acid obtained above (251 mg, 0.8 mmol) was added dropwise as a solution in THF (1.6 ml). The reaction mixture was stirred at room temperature for 30 minutes, kept at reflux for 30 minutes, and then cooled again to room temperature. N-propylsulfonamide (100 mg) was added and stirred for 10 minutes. DBU (123 mg) was added as a solution in THF (1.6 ml). The reaction mixture was acidified and extracted with ethyl acetate. The solvent was removed and the crude product was purified by flash chromatography (SiO ₂ , ethyl acetate) to give 195 mg of the above acylsulfonamide.

The nitrile obtained above (90 mg, 0.2 mmol) was dissolved in ethanol (2.5 ml). Diisopropylethylamine (202 mg, 1.56 mmol) was added, followed by hydroxylamine hydrochloride (83 mg, 1.2 mmol). The reaction mixture was heated to 70 ° C. for 21 hours. The reaction mixture was cooled and the solvent was removed in vacuo. The residue was dissolved in 30% acetonitrile / water (4 ml) and purified by reverse phase preparative chromatography (gradient of water / acetonitrile with the addition of 0.1% TFA) to obtain 14 mg of the hydroxyamidine above.

Hydroxyamidine was dissolved in ethanol (2 ml) and acetic acid (8 drops). Raney nickel (approximately 100 mg) was added and the reaction mixture was vigorously stirred under a hydrogen atmosphere for 2 hours and 45 minutes. The product was filtered through celite and the celite was washed first with a mixture of 30% acetonitrile / water containing 0.1% TFA, and then with acetonitrile. The solvent was removed in vacuo and the crude product was purified using reverse phase preparative chromatography (gradient of water / acetonitrile with the addition of 0.1% TFA) to give the desired product (6 mg). M + H = 435.

Example 9. Synthesis of enantiomerically pure 6-alkylsulfonamino-sulfonamides

To dimethylformamide (600 ml) was added 3-ethoxy-4-hydroxybenzaldehyde (40 g), followed by potassium carbonate (40 g, 1.2 eq.). Ethyl iodide (28.87 ml, 1.5 equiv.) Was added and the solution was heated to 60 ° C. for 6 hours. The solution was cooled to room temperature and the solvent was removed under reduced pressure. The solution was diluted with ethyl acetate (500 ml), washed with water, brine, dried over magnesium sulfate and evaporated to give the crude product 3,4-ethoxybenzaldehyde (49 g, 104%). 3,4-Ethoxybenzaldehyde (45 g) was dissolved in ethanol (300 ml) and the solution was cooled to 0 ° C. Potassium hydroxide (19.5 g, 1.5 equiv.) Was added to a separate flask containing ethanol (300 ml), followed by nitromethane (26 g, 1.5 equiv.) And the solution was stirred at room temperature for 10 min. and cooled to 0 ° C. The solution was added to 3,4-ethoxybenzaldehyde and stirred for 20 min and poured at 0 ° C into concentrated hydrochloric acid (200 ml). Ethanol was removed under reduced pressure, the solution was diluted with water (300 ml), and the reaction mixture was extracted with ethyl acetate. The organic layer was dried over magnesium sulfate and the solvent was removed to give a crude product. The crude product was purified by recrystallization from ethyl acetate (47 g, 87%). MS (M + H) = 238.

To tetrahydrofuran (350 ml) was added 1-nitro-2- (3,4-diethoxyphenyl) ethylene (16.64 g) and 4-aminobenznitrile (9.12 g, 1.1 eq.) And cooled to 0 ° C. Lithium diisopropylamide (47.8 ml, 1.02 equiv.) Was slowly added until a stable purple color was obtained. Zinc (50 g) was added in one portion and then acetic acid (35 ml). The solution was warmed to room temperature and stirred for 2 hours, after which acetic acid (35 ml) and zinc (10 g) were added. After another 2 h, acetic acid (25 ml) was added and the reaction mixture was stirred for 1 h. Concentrated hydrochloric acid (15 ml) was added and the solution was stirred for another 1 h. The solution was filtered through a pad of celite and water (250 ml) was added. . The solution was concentrated under reduced pressure to a volume of 300 ml and was added to citric acid (0.5 M, 500 ml) and ethyl acetate / hexane (500 ml). A layer of citric acid was collected and ammonium hydroxide was added until the solution became alkaline. This solution was extracted with ethyl acetate (3 × 300 ml) and the combined organic layers were dried over magnesium sulfate and evaporated under reduced pressure to obtain a crude product. The crude product was diluted with dichloromethane (350 ml) and cooled to 0 ° C. Phosgene (40.88 ml as a 20% solution in toluene, 1.1 equiv.) Was added, followed by a Hunig base (24.46 ml, 2 equiv.). The solution was stirred for 10 minutes and water (200 ml) was added. Dichloromethane was collected, dried over magnesium sulfate and purified by flash chromatography on silica gel (80% ethyl acetate / 20% hexane) to give the product as a white solid (9.76 g, 40%). MS (M + H) = 352.

To tetrahydrofuran (200 ml) was added 4- [5- (3,4-diethoxyphenyl) -2-oxoimidazolidin-1-yl-benzenitrile (2.10 g) and cooled to -78 ° C. N-Butyllithium (3.74 ml, 1 equiv.) Was added dropwise and the solution was stirred for 10 minutes. Solid (S) - (+) - 2- (6-methoxy-2-naphthyl) propionyl chloride (1.49 g, 1 equiv.) Was added in one portion and the reaction mixture was stirred for 1 h. The reaction mixture was warmed to room temperature temperature and evaporated under reduced pressure to a volume of 50 ml. The solution was diluted with ethyl acetate (300 ml) and washed with citric acid (0.5 M), water, brine and dried over magnesium sulfate. The solution was evaporated under reduced pressure and purified by flash chromatography on silica gel (50% ethyl acetate / 50% hexane) to give the product 4- {5- (3,4-diethoxyphenyl) -3- [2- (6-methoxynaphthalene- 2-yl) propionyl] -2-oxoimidazolidin-1-yl} benznitrile as a single diastereoisomer (1.20 g, 71%). This product was added to methanol (200 ml), after which lithium hydroxide (1 ml as a 10% aqueous solution) was added and stirred for 15 minutes. Acetic acid (10 drops) was added, methanol was removed under reduced pressure, and the product was purified by flash chromatography on silica gel (80% ethyl acetate / 20% hexane) to give the product (0.71 g, 95%) as a white solid colors. [α] _Na -55.0 (c 2.20, acetone). MS (M + H) = 352.

(R) - (-) - 4- [5- (3,4-diethoxyphenyl) -2-oxoimidazolidin-1-ylbenznitrile (0.710 g) was added to tetrahydrofuran (20 ml) and cooled to -78 ° C. N-Butyllithium (1.27 ml as a 1.6 M solution in hexane, 1 equiv.) Was added dropwise and the solution was stirred for 10 minutes. 1-Propylsulfonyl chloride (0.275 ml, 1.2 equiv.) Was added and the solution was stirred for 15 minutes and warmed to room temperature. Acetic acid (10 drops) was added, the solvent was removed under reduced pressure, and the product was purified by flash chromatography on silica gel (50% ethyl acetate / 50% hexane) to give the product (0.740 g, 80%). This product (0.300 g) was diluted with dichloroethane (10 ml) and cooled to 0 ° C. Nitric acid (0.137 ml, 5 equiv.) Was added dropwise and the solution was stirred for 1 h. The solution was diluted with dichloroethane (100 ml) and washed with water, saturated sodium bicarbonate, dried over magnesium sulfate and the solvent was evaporated under reduced pressure. This product was diluted with methanol (50 ml) and acetic acid (1 ml) and platinum (0.050 g, 5% carbon) were added. The solution was hydrogenated for 1 h, filtered through a pad of celite and the solvent was evaporated under reduced pressure. This product was dissolved in dichloromethane (5 ml) and Hünig base (0.177 ml, 1.5 equiv.) And chlorosulfonylacetic acid ethyl ester (0.189 g, 1.5 equiv.) Were added and the solution was stirred for 2 hours. The solution was purified using direct flash chromatography on silica gel (mixture of 50% ethyl acetate / 50% hexane) to give the product (0.160 g) ethyl ether (R) - [2- [3- (4-cyanophenyl) -2-oxo-1- (propane- 1-sulfonyl) imidazolidin-4-yl] -4,5-diethoxyphenylsulfamoyl} acetic acid. MS (M + H) = 624.

Ethyl ester (R) - {2- [3- (4-cyanophenyl) -2-oxo-1- (propan-1-sulfonyl) imidazolidin-4-yl] -4,5-diethoxyphenylsulfamoyl} acetic acid (0.160 g) diluted with ethanol (5 ml) and then lithium hydroxide (1 ml as a 10% aqueous solution, 10 equiv.) and the solution was stirred for 48 hours. The solution was purified by flash chromatography on silica gel (20% methanol mixture) / 80% dichloromethane) to give the product. This product was diluted with ethanol (1 ml) and hydroxylamine (0.035 g, 10 equiv.) And Hunig base (0.088 ml, 10 equiv.) Were added and the solution was heated to 60 ° C. for 6 hours. The reaction mixture was cooled to room temperature and stirred. within 12 hours. Ethanol (3 ml), acetic acid (0.5 ml) and Raney nickel (0.025 g) were added and the solution was hydrogenated for 1 hour. The reaction mixture was filtered through a pad of celite and the solvent was removed under reduced pressure. The product was purified by reverse phase preparative chromatography to give [2- [1- (4-carbamimidoylphenylamino) -2- (propan-1-sulfonylamino) ethyl] -4,5-diethoxyphenylsulfamoyl] acetic acid (52 mg). [α] _Na -42.1 (c 1.01, methanol). MS (M + H) = 587.

Example 10. 6-Alkoxy-substituted sulfonamides

Aminoacetonitrile (14.25 g, 92.5 mmol) was dissolved in 1,2-dichloroethane (150 ml) and the reaction mixture was placed under N ₂ . Triethylamine (32.76 g, 324 mmol) was added and the reaction mixture was cooled to 0 ° C. A solution of propane -1-sulfonyl chloride (13.19 g, 92.5 mmol) in 1,2-dichloroethane (20 ml) was added dropwise, the reaction mixture was allowed to warm to room temperature and stirred for 16 hours. Analysis by thin layer chromatography revealed the presence of a new product with a higher R _f . The solvent was removed in vacuo and the crude product was subjected to flash chromatography (methylene chloride: ethyl acetate, 9: 1) to give propan-1-sulfonic acid cyanmethylamide.

¹ H-NMR (CDCl ₃ ): 5.40 (s, 1H), 4.11 (s, 2H), 3.15 (m, 2H), 1.89 (q, 2H), 1.10 (t , 3H).

3-Ethoxy-4-hydroxybenzaldehyde (Aldrich, 100 g, 0.602 mol) was dissolved in N, N-dimethylformamide (1 L). The reaction mixture was placed in an atmosphere of N ₂ . Solid potassium carbonate (103 g, 0.662 mol) was added and the reaction mixture was stirred for 10 minutes. Iodoethane (175 g, 1.26 mol) was added and the reaction mixture was stirred for 16 hours at room temperature. The progress of the reaction was monitored by current chromatography, which revealed complete absorption of phenol and the formation of a new product. The reaction mixture was filtered to remove potassium carbonate and the solvent was removed in vacuo. The residue was dissolved in methylene chloride and filtered. Silica gel (200 g) was added and dichloromethane was removed in vacuo. The crude product adsorbed on silica gel was subjected to flash chromatography (hexane, 100%, 2 L, then ethyl acetate: hexane, 1: 3, 2 L, then ethyl acetate: hexane, 1: 1) to obtain 109.36 g of 3.4 diethoxybenzaldehyde.

¹ H-NMR (CDCl ₃ ): 9.83 (s, 1H), 7.41 (m, 2H), 6.96 (d, 1H), 4.17 (m, 4H), 1.50 (t , 3H); 1.47 (t, 3H).

3,4-Diethoxybenzaldehyde (31.45 g, 0.162 mol) was dissolved in methylene chloride (500 ml). 3-Chloroperbenzoic acid was added and the reaction mixture was heated to reflux for 4 hours. Analysis by thin-layer chromatography revealed aldehyde uptake. The reaction mixture was cooled to room temperature, diluted with methylene chloride and the reaction was stopped by adding a saturated aqueous solution of potassium carbonate. The layers were separated and the aqueous solution was extracted with methylene chloride. The methylene chloride extracts were combined, washed with water, brine and dried over anhydrous sodium sulfate, filtered and the solvent was removed in vacuo to give a crude oil.

The crude oil was dissolved in methanol (300 ml) and a 10% aqueous KOH solution (60 ml) was added. The reaction mixture was stirred at room temperature for 16 hours. Analysis by thin layer chromatography revealed absorption of the resulting intermediate. Methanol was removed in vacuo and the aqueous solution was acidified with 1.2N. Hcl. The solution was extracted with ethyl acetate. The ethyl acetate extracts were washed with water and brine, dried over anhydrous sodium sulfate, filtered and the solvent was removed in vacuo. The residue was subjected to flash chromatography (hexane: ethyl acetate, 3: 1) to obtain 22.06 g of 3,4-diethoxyphenol.

¹ H-NMR (CDCl ₃ ): 6.56 (d, 1H). 6.25 (d, 1H), 6.10 (dd, 1H), 3.82 (m, 4H), 1.22 (t, 3H), 1.20 (t, 3H).

Nitrobenzene (100 g) was cooled to 0 ° C and saturated with gaseous HCl. To this solution were added 3,4-diethoxyphenol (11.64 g, 64 mmol), propane-1-sulfonic acid cyanmethylamide (10.36 g, 64 mmol) and zinc chloride (17.45 g, 128 mmol). The reaction mixture was stirred for 1 h at 0 ° C and then allowed to warm to room temperature. Stirring was continued for 2 hours. Analysis by thin layer chromatography revealed an absorption of 3,4-diethoxyphenol. Water (100 ml) was carefully added and the reaction mixture was heated to 100 ° C for 1 h. The reaction mixture was cooled and extracted with methylene chloride. The extracts were washed with water, brine, dried over anhydrous sodium sulfate, filtered and the solvent was removed in vacuo. The residue was subjected to flash chromatography (2.5-5% ethyl acetate in methylene chloride). The purified product was triturated with ether, filtered and dried in air to give 1.73 g of [2- (4,5-diethoxy-2-hydroxyphenyl) -2-oxoethyl] propane-1-sulfonic acid amide.

¹ H-NMR (CDCl ₃ ): 11.86 (s, 1H), 6.93 (s, 1H), 6.46 (s, 1H), 5.26 (t, 1H) ', 4.55 ( d, 2H), 4.14 (q, 2H), 4.03 (q, 2H), 3.05 (m, 2H), 1.90 (m, 2H), 1.45 (m, 6H), 1.07 (t, 3H).

[2- (4,5-Diethoxy-2-hydroxyphenyl) -2-oxoethyl] propane-1-sulfonic acid amide (200 mg, 0.579 mmol) was dissolved in tetrahydrofuran (5 ml), placed under N _2, and cooled to 0 ° C. The borane complex: THF (1.74 ml, 1.0 M in THF, 1.74 mmol) was added dropwise. The reaction mixture was stirred for 15 minutes and allowed to warm to room temperature over 2 hours. Analysis by thin layer chromatography revealed uptake of the starting material and the formation of a new product with lower R _f . The reaction was stopped by adding 1.2n. aqueous Hcl (2.5 ml), the mixture was diluted with water and extracted with ethyl acetate. The extracts were washed with water and brine, dried over anhydrous sodium sulfate, filtered and the solvent was removed in vacuo to give 190 mg of a crude oil.

The crude oil was dissolved in acetonitrile (5 ml). 4-aminobenznitrile (194 mg, 1.64 mmol) and lithium perchlorate (233 mg, 2.19 mmol) were added. The reaction mixture was heated to 90 ° C. for 3.5 hours and then stirred for 18 hours at room temperature. Analysis by thin-layer chromatography revealed the presence of a new product with a higher R _f value compared to the product obtained by recovery. The reaction was stopped by adding water, and the mixture was extracted with ethyl acetate. The ethyl acetate extracts were washed with water and brine, dried over anhydrous sodium sulfate, filtered and the solvent was removed in vacuo. The residue was flash chromatographed (ethyl acetate: methylene chloride, 1: 9) to give 134 mg of [2- (4-cyanophenylamino) -2- (4,5-diethoxy-2-hydroxyphenyl) ethyl] amide propane-1-sulfonic acid.

¹ H-NMR (CDCl ₃ ): 7.34 (d, 2H), 6.71 (s, 1H), 6.61 (d, 2H), 6.41 (s, 1H), 5.58 (d , 1H), 4.79 (m, 1H), 4.57 (m, 1H), 3.99 (m, 4H), 3.48 (t, 2H), 3.00 (m, 2H), 1 80 (q, 2H), 1.41 (t, 3H), 1.34 (t, 3H), 1.03 (t, 3H).

[2- (4-Cyanophenylamino) -2- (4,5-diethoxy-2-hydroxyphenyl) ethyl] propane-1-sulfonic acid amide (100 mg, 0.233 mmol) was dissolved in dimethylformamide (1.5 ml) and treated with solid potassium bicarbonate (22 mg, 0.233 mmol) and then ethyl bromoacetate (0.37 ml, 0.233 mmol). The reaction mixture was stirred under N ₂ for 67 h. Analysis by thin layer chromatography revealed the presence of a new product with a higher R _f value. The reaction was stopped by adding water, and the mixture was extracted with ethyl acetate. The ethyl acetate extracts were washed with water and brine, dried over anhydrous sodium sulfate, filtered and the solvent was removed in vacuo. The residue was subjected to flash chromatography (ethyl acetate: methylene chloride, 1: 9) to obtain 78 mg of {2- [4-cyanophenylamino) -2- (propane-1-sulfonylamino) ethyl] -4,5-diethoxyphenoxy} ethyl acetate ethyl ester acids.

¹ H-NMR (CDCl ₃ ): 7.35 (d, 2H), 6.81 (s, 1H), 6.57 (d, 2H), 6.43 (s, 1H), 4.81 (t , 1H), 4.68 (ABq, 2H), 4.56 (t, 1H), 4.26 (q, 2H), 4.05 (q, 2H), 3.95 (m, 2H), 3 54 (t, 2H), 2.98 (m, 2H), 1.80 (m, 2H), 1.42 (t, 3H), 1.32 (t, 6H), 1.02 (t, 3H). MS (M + H) = 534.

[2- [4-Cyanophenylamino) -2- (propan-1-sulfonylamino) ethyl] -4,5-diethoxyphenoxy} acetic acid ethyl ester (76 mg, 0.142 mmol) was dissolved in tetrahydrofuran (3 ml). Water (1 ml) was added and the reaction mixture was cooled to 0 ° C. Aqueous LiOH (1.0 M, 0.42 ml, 0.42 mmol) was added to the reaction mixture. After stirring for 5 minutes, the reaction mixture was allowed to warm to room temperature and was stirred for 16 hours at room temperature. Ether absorption was detected by high pressure analytical liquid chromatography. An additional portion of LiOH (1.0 M, 0.14 ml) was added. The reaction mixture was stirred at room temperature for another 8 hours. The absorption of ether was incomplete, so freshly prepared LiOH (1.0 M, 0.14 ml) was added and the reaction mixture was stirred for 24 hours. Another portion of LiOH was added (1.0 M, 0.28 ml) and after 67 h, analysis using analytical HPLC-RP showed absorption of ether (total amount of 1.0 M LiOH was 0.98 ml, 6.9 eq.). The reaction mixture was acidified with acetic acid and THF was evaporated in a stream of N ₂ . The solution was made transparent by the addition of acetonitrile and then purified by reverse phase HPLC (acetonitrile / water gradient with 0.1% trifluoroacetic acid), to obtain, after lyophilization, 44 mg of acid trifluoroacetate {2- [4-cyanophenylamino) -2- (propane -1-sulfonylamino) ethyl] -4,5-diethoxyphenoxy} acetic acid.

¹ H-NMR (CD ₃ OD): 7.15 (d, 2H), 6.70 (s, 1H), 6.48 (d, 2H), 6.45 (s, 1H), 4.76 ( t, 1H), 4.60 (s, 2H), 3.85 (q, 2H), 3.73 (m, 2H), 3.43 (dd, 1H), 3.26 (dd, 1H partially closed peak solvent CH ₃ OH), 2.76 (m, 2H), 1.52 (m, 2H), 1.18 (t, 3H), 1.07 (t, 3H), 0.77 (t, 3H ) MS (M + H) = 506.

Acid trifluoroacetate {2- [4-cyanophenylamino) -2- (propane-1-sulfonylamino) ethyl] -4,5-diethoxyphenoxy] acetic acid (44 mg, 0.071 mmol) was dissolved in ethanol (1 ml) and treated with diisopropylethylamine (0 , 89 ml, 0.515 mmol) and solid hydroxylamine hydrochloride (25 mg, 0.355 mmol). The reaction mixture was placed in an atmosphere of N ₂ and heated for 3 hours to 60 ° C. After 3 hours, analysis by high pressure liquid chromatography revealed that the reaction was not yet complete. The reaction mixture was cooled to room temperature and stirred at room temperature for 16 hours, and the conversion process was monitored by HPLC. The reaction mixture was kept at 60 ° C for 8 hours, stirred at room temperature for 16 hours, kept at 70 ° C for 8 hours and stirred at room temperature for 16 hours. HPLC analysis revealed uptake of the starting material. The reaction mixture was acidified with acetic acid, Raney nickel was added and the reaction mixture was hydrogenated for 3 hours in a vessel of hydrogen at a pressure of about 1 atm. Analysis using analytical HPLC revealed absorption of an intermediate product, which is hydroxyamidine. The Raney nickel catalyst was filtered off and the solvent was removed in vacuo. The residue was purified by reverse phase preparative HPLC (acetonitrile / water gradient with 0.1% trifluoroacetic acid), yielding 5.6 mg of {2- [1- (carbamimidoylphenylamino) -2- (propan-1-sulfonylamino) after lyophilization ethyl] -4,5-diethoxyphenoxy} acetic acid in the form of bis trifluoroacetate. MS (M + H): 523.

Example 11. Synthesis of 6-alkylsulfonylalkylamino-substituted acylsulfonamide

4,5-Diethoxy-2-nitrobenzaldehyde (55.5 g, 206 mmol) and 4-aminobenznitrile (23 g, 195 mmol) were dissolved in methanol (700 ml) and stirred for 2 hours at 60 ° C. The reaction mixture was allowed to cool to 0 ° C. and tosylmethylisonitrile (45 g, 230 mmol) was added. Boron trifluoroethylate (78 ml, 620 mmol) was added dropwise over 10 minutes. The reaction mixture was stirred for 30 minutes at 0 ° C, allowed to cool to room temperature, and then stirred for 1.5 hours at ambient temperature. Water (18 ml) was added and the mixture was stirred at room temperature overnight. The next day, methanol was removed in vacuo and the residue was dissolved in ethyl acetate. The organic layer was washed with water and then dried over anhydrous sodium sulfate. Sodium sulfate was filtered off and ethyl acetate was removed in vacuo. The crude product was subjected to flash chromatography (hexane: ethyl acetate, 2: 1, then 1: 1) to obtain 46 g of the desired product, i.e. (4-ethoxy-5-ethoxy-2-nitrophenyl) - (4-cyanophenylamino) acetic acid methyl ester.

(4-Ethoxy-5-ethoxy-2-nitrophenyl) - (4-cyanophenylamino) acetic acid methyl ester (11 g, 27.5 mmol) was dissolved in ethyl acetate (300 ml) and was added to a flask containing 5% nitrogen atmosphere. th Pt / C (3 g). Nitrogen was removed and replaced with hydrogen (from a balloon) and the reaction mixture was stirred vigorously for 6 hours. The catalyst was filtered off and the solvent was removed in vacuo. The residue was dissolved in dichloromethane (approximately 300 ml) and pyridine (5.6 ml, 70 mmol) was added. The reaction mixture was cooled to 0 ° C. and methanesulfonyl chloride (2.5 ml, 33 mmol) was added dropwise. The reaction mixture was stirred overnight. The solution was washed with water and the solvent was removed in vacuo. The crude product was chromatographed on silica gel using flash chromatography (hexane: ethyl acetate, 1: 1) to give 5 g of the desired product, i.e. (4-cyanophenylamino) - [4,5-diethoxy-2- (methanesulfonylamino) phenyl] acetic acid methyl ester. (4-Cyanophenylamino) - [4,5-diethoxy-2- (methanesulfonylamino) phenyl] acetic acid methyl ester product (5 g, 10.7 mmol) was dissolved in anhydrous DMF (100 ml) and cesium carbonate was added. (7.25 g, 22 mmol) and iodomethane (1 ml, 16 mmol). The reaction mixture was stirred for 3 hours at room temperature and the solvent was removed in vacuo. The residue was dissolved in ethyl acetate, acidified with 1N. hydrochloric acid and the organic layer were washed once with water. The product was dried over anhydrous sodium sulfate and the solvent was removed in vacuo. The residue was subjected to flash chromatography (hexane: ethyl acetate, 1: 1) to obtain 3.6 g of the desired product, i.e. (4-cyanophenylamino) - [4,5-diethoxy-2- (methanesulfonylmethylamino) phenyl] acetic acid methyl ester.

The (4-cyanophenylamino) - [4,5-diethoxy-2- (methanesulfonylmethylamino) phenyl] acetic acid methyl ester obtained above (2.6 g, 5 mmol) was dissolved in methanol. 1N was added. LiOH (25 ml) and the reaction mixture was stirred at room temperature for 5 hours. Methanol was removed in vacuo and the reaction mixture was acidified with 1N. hydrochloric acid. The product was extracted with ethyl acetate and washed with water. Using flash chromatography (ethyl acetate with 5% acetic acid), 1.9 g of the desired acid were obtained, i.e. (4-cyanophenylamino) - [4,5-diethoxy-2- (methanesulfonylmethylamino) phenyl] acetic acid.

The above (4-cyanophenylamino) - [4,5-diethoxy-2- (methanesulfonylmethylamino) phenyl] acetic acid (350 mg, 0.75 mmol) was combined with carbonyldiimidazole (610 mg, 3.77 mmol) in anhydrous THF (6 ml). The reaction mixture was kept at 60 ° C for 1 h and then cooled to room temperature. To this solution phenylsulfonamide (650 mg, 4.14 mmol) and DBU (5 mmol) were added as a solution in 5 ml THF. The reaction mixture was stirred for 3 hours and THF was removed in vacuo. The residue was dissolved in ethyl acetate and acidified with 1N. hydrochloric acid. The organic layer was separated, washed with water and dried over anhydrous sodium sulfate. The crude product was purified by flash chromatography (hexane: ethyl acetate; 1: 1 and then ethyl acetate with 5% acetic acid) to give 302 mg of the desired product, i.e. N - {(4-cyanophenylamino) - [4,5-diethoxy-2- (methanesulfonylmethylamino) phenyl] acetyl} benzenesulfonamide.

The above N - {(4-cyanophenylamino) - [4,5-diethoxy-2- (methanesulfonylmethylamino) phenyl] acetyl} benzenesulfonamide (126 mg, 0.21 mmol) was dissolved in ethanol (1.8 ml) and heated to 60 ° C. Diisopropylethylamine (DIPEA; 260 μl, 1.5 mmol) was added, followed by hydroxylamine hydrochloride (74 mg, 1.04 mmol). The reaction mixture was stirred at 60 ° C. under nitrogen for 6 hours. The reaction mixture was then allowed to cool to room temperature. The solution was diluted with methanol (5 ml) and acetic acid (2 ml) and Raney Nickel 2800 (approximately 50 mg) was added as a suspension. Then the reaction mixture was intensively stirred in a hydrogen atmosphere for 1 h. The catalyst was filtered off and the solvent was removed. The crude products were purified using reverse phase preparative HPLC using a gradient of water-acetonitrile (0.1% TFA) to give 40 mg of the desired 4- {2-benzenesulfonylamino-1- [4,5-diethoxy-2- (methanesulfonylmethylamino) phenyl ] -2-oxoethylamino} benzamidine as a trifluoroacetic acid salt. MS: (M + H) = 604.

Example 12

4-Isopropoxy-5-ethoxybenzaldehyde (10.6 g, 50 mmol) and 4-aminobenzonitrile (5.9 g, 50 mmol) were dissolved in methanol (150 ml) and stirred at 60 ° C for 1.6 hours. the mixture was allowed to cool to 0 ° C and tosylmethylisonitrile (9.75 g, 50 mmol) was added. Boron trifluoro-etherate (19 ml, 150 mmol) was added dropwise over 10 minutes. The reaction mixture was stirred at 0 ° C for 30 minutes, allowed to warm to room temperature and then stirred at ambient temperature for 1.5 hours. Water (4.5 ml) was added and the mixture was stirred at room temperature for 2 days. Methanol was removed in. vacuum and the residue was dissolved in ethyl acetate. The organic layer was washed with water and then dried over anhydrous sodium sulfate. Sodium sulfate was filtered off and ethyl acetate was removed in vacuo. The crude product was subjected to flash chromatography (hexane: ethyl acetate, 1: 1) to obtain 12.5 g of the desired product, i.e. (4-Isopropoxy-5-ethoxyphenyl) - (4-cyanophenylamino) acetic acid methyl ester.

The above product, i.e. (4-Isopropoxy-5-ethoxyphenyl) - (4-cyanophenylamino) acetic acid methyl ester (6 g, 16.3 mmol) was treated with 1N. LiOH (approximately 50 ml) in THF (approximately 150 ml). The reaction mixture was stirred at room temperature for 6 hours and acidified with 1N. hydrochloric acid. THF was removed in vacuo and the product was extracted with ethyl acetate. The crude product was purified by reverse phase chromatography (ethyl acetate, 3% acetic acid) to obtain 4.85 g of the desired acid, i.e. (4-isopropoxy-5-ethoxyphenyl) - (4-cyanophenylamino) acetic acid.

The above (4-isopropoxy-5-ethoxyphenyl) - (4-cyanophenylamino) acetic acid (200 mg, 0.57 mmol) was combined with carbonyldiimidazole (200 mg, 1.2 mmol) in anhydrous THF (4 ml). The reaction mixture was allowed to stir at room temperature for 1 h. The corresponding alkyl or arylsulfonamide (2.2 mmol) and DBU (2.2 mmol) were added to this solution as a solution in 3 ml of THF. The reaction mixture was stirred overnight and THF was removed in vacuo. The residue was dissolved in ethyl acetate and acidified with acetic acid. The organic layer was separated, washed with water and dried over anhydrous sodium sulfate. The crude product was purified by flash chromatography (hexane: ethyl acetate, 1: 2) to give the desired product, i.e. N - {(4-cyanophenylamino) - [4-isopropoxy-5-ethoxyphenyl] acetyl] (alkyl or aryl) sulfonamide.

The N - {(4-cyanophenylamino) - [4-isopropoxy-5-ethoxyphenyl] acetyl} (alkyl or aryl) sulfonamide (approximately 0.24 mmol) obtained above was dissolved in ethanol (1-3 ml) and heated to 60 ° C. . Diisopropylethylamine (DIPEA; 260 μl, 1.5 mmol, 6 equiv.) And then hydroxylamine hydrochloride (84 mg, 1.25 mmol, 5 equiv.) Were added. The reaction mixture was stirred at 60 ° C. under nitrogen for about 6 hours. The reaction mixture was then allowed to cool to room temperature. The solution was diluted with methanol (5 ml) and acetic acid (2 ml) and Raney nickel (approximately 50 mg) was added as a suspension. Then the reaction mixture was intensively stirred in a hydrogen atmosphere for 1-6 hours. The catalyst was filtered off and the solvent was removed. The crude products were purified by reverse phase preparative HPLC using a gradient of water-acetonitrile (0.1% TFA) or by flash chromatography (ethyl acetate: acetone: water: acetic acid, 6: 2: 1: 1) to obtain the desired 4- {2- (alkyl or aryl) sulfonylamino-1- [4-isopropoxy, 5-ethoxyphenyl] -2-oxoethylamino] benzamidine as a salt of trifluoroacetic acid or acetic acid.

Using a similar process, the following compounds were prepared having different R ₁ groups:

R ₁ is ethyl: 4- {2-ethylsulfonylamino-1- [4-isopropoxy, 5-ethoxyphenyl] -2-oxoethylamino} benzamidine: MS (M + H) = 463.

R ₁ is propyl: 4- {2-propylsulfonylamino-1- [4-isopropoxy, 5-ethoxyphenyl] -2-oxoethylamino} benzamidine: MS (M + H) = 477.

R ₁ is n-butyl: 4- {2-butylsulfonylamino-1- [4-isopropoxy, 5-ethoxyphenyl] -2-oxoethylamino} benzamidine: MS (M + H) = 491.

R ₁ is CH ₂ CH ₂ CO ₂ Me: 3 - [(4-carbamimidoylphenylamino) - (3-ethoxy-4-isopropoxyphenyl) acetylsulfamoyl] proponic acid methyl ester: MS (M + H) = 521.

R ₁ is phenyl: 4- {2-benzenesulfonylamino-1- [4-isopropoxy, 5-ethoxyphenyl] -2-oxoethylamino} benzamidine: MS (M + H) = 511.

Example 13. Acyl sulfonamide substituted on the aminobenzamidine ring

2-Hydroxy-4-nitrobenzonitrile (11.2 g, 68 mmol) was dissolved in DMF (200 ml). Potassium carbonate (11 g, 80 mmol) and benzyl bromide (9 ml, 75 mmol) were added. The reaction mixture was stirred at room temperature overnight. DMF was removed in vacuo and the residue was dissolved in ethyl acetate and water. The organic layer was separated, washed with 1N. NaOH, then with water and dried over sodium sulfate. The crude product (5 g) was dissolved in ethyl acetate (75 ml) and added to a flask containing 5% Pt / C (500 mg). The reaction mixture was placed in a hydrogen atmosphere (balloon) and stirred vigorously for several hours until completion of the reaction (according to TLC). The catalyst was filtered off and the solvent was removed. The product was purified by flash chromatography to give 4.12 g of 4-amino-2-benzyloxybenzonitrile.

4,5-Diethoxybenzaldehyde (3.6 g, 17.8 mmol) and 4-amino-2-benzyloxybenzonitrile (3.7 g, 17.8 mmol) were dissolved in methanol (40 ml) and stirred for 2 hours Tosylmethylisonitrile (3.48 g, 17.8 mmol) was added. The reaction mixture was cooled to 0 ° C, and boron trifluoroethirate (6.7 ml, 54 mmol) was added dropwise. The reaction mixture was stirred at 0 ° C for 30 minutes, allowed to warm to room temperature and then stirred at ambient temperature for 3.5 hours. Water (1.6 ml) was added and the mixture was stirred at room temperature for 2 days. Methanol was removed in vacuo and the residue was dissolved in ethyl acetate. The organic layer was washed with water and then dried over anhydrous sodium sulfate. Sodium sulfate was filtered off and ethyl acetate was removed in vacuo. The crude product was subjected to flash chromatography (hexane: ethyl acetate, 1: 1) to obtain 4.2 g of the desired product, i.e. (3-Benzyloxy- (4-cyanophenylamino) -3,4-ethoxyphenyl) acetic acid methyl ester.

The product obtained above was treated with LiOH (1.96 g) in water (50 ml), methanol (100 ml) and THF (50 ml). The reaction mixture was stirred at room temperature for 3 hours and acidified with acetic acid. The solvent was removed in vacuo and the product was extracted with ethyl acetate. The crude product was purified by reverse phase chromatography (ethyl acetate, 3% acetic acid) to give 5 g of the desired acid, i.e. (3-benzyloxy-4-cyanophenylamino) -3,4-ethoxyphenyl) acetic acid.

(3-Benzyloxy-4-cyanophenylamino) -3,4-ethoxyphenyl) acetic acid (750 mg, 1.68 mmol) was dissolved in anhydrous THF (3 ml) and carbonyldiimidisole (CDI; 545 mg, 3.36 mmol) was added. The reaction mixture was heated to 40 ° C. for 1 h and then cooled to room temperature. To the mixture was added a solution of benzenesulfonamide (1.05 g, 6.7 mmol), DBU (1.02 g, 6.72 mmol) in THF (3 ml). The reaction mixture was stirred overnight and then 1N was added. hydrochloric acid. THF was removed in vacuo and the product was purified by flash chromatography (ethyl acetate, 2% acetic acid) to give the desired product. This product (215 mg) was dissolved in ethanol (5 ml) containing acetic acid (1 drop) and added to 5% Pd / C (100 mg). The reaction mixture was placed in a hydrogen atmosphere and vigorously stirred until completion of the reaction. The catalyst was filtered off and then the solvent was removed in vacuo. The crude product was purified by flash chromatography to give 150 mg of the desired product, i.e. N - [(3-hydroxy-4-cyanophenylamino) - (3,4-diethoxyphenylacetyl] benzenesulfonamide.

The N - [(3-hydroxy-4-cyanophenylamino) - (3,4-diethoxyphenylacetyl] benzenesulfonauide (75 mg, 0.15 mmol) obtained above was dissolved in ethanol (2 ml) and heated to 60 ° C. Diisopropylethylamine (DIPEA) was added. ; 185 μl, 1 mmol) followed by hydroxylamine hydrochloride (53 mg, 0.75 mmol). The reaction mixture was stirred at 60 ° C. under nitrogen for about 6 hours. The reaction mixture was then allowed to cool to room temperature. The solution was diluted with acetic acid (1 ml) and methanol (1 ml). Raney nickel 2800 (approximately 50 mg) was added as a suspension. Then the reaction mixture l was vigorously stirred in a hydrogen atmosphere for 0.5 h. The catalyst was filtered off and the solvent was removed. The crude products were purified using preparative reverse phase HPLC using a gradient of water-acetonitrile (0.1% TFA) to give the desired amidine, i.e. 4- [2-benzenesulfonylamino-1- (3,4-diethoxyphenyl) -2-oxoethylamino] -2-hydroxybenzamidine. MS (M + H) = 513.

Using a similar process, the corresponding halogen-containing compounds can be obtained from 2-chloro-4-nitrobenzonitrile and 2-bromo-4-nitrobenzonitrile.

Example 14. Analysis of antagonistic activity against tissue factor / factor VIIa

This assay can be used to determine the inhibition constant (K _i ) of a representative compound of the invention.

Materials:

Analysis buffer: 100 mM Hepes, pH 7.8, 140 mM NaCl, 0.1% PEG-8000, 0.02% Tween-80, 5 mM CaCl ₂ Coagulation Factor: recombinant human factor VIIa (NB No. 25942-16) Cofactor: soluble tissue factor (1-219) Substrate: Chromozym-tPA (Boehringer Mannheim, catalog number 1093 037). Reconstitute to 20 mM in H ₂ O. Dilute to 4 mM in assay buffer with CaCl ₂ before use. Samples: Dilute samples to reach a DMSO concentration of 3% in assay buffer (without CaCl ₂ )

Analysis Procedure:

1. Prepare a solution containing 2 μg / ml (90 nM) of tissue factor and 1.5 μg / ml (30 nM) of factor VIIa in an assay buffer containing CaCl ₂ .

2. Incubate for 15 min at room temperature.

3. Add 50 μl of sample to each well.

4. Add 50 µl of tissue factor / factor VIIa solution to each well.

5. Incubate for 15 min at room temperature, gently shaking.

6. Add 50 μl of substrate to each well.

7. Shake the tablet for 20-25 seconds.

8. Determine the absorbance at 405 nm every 10 s for a total of 5 minutes at room temperature.

9. Calculate the value of v _max at 10 points.

Example 15. Factor Xa, Thrombin, and Plasma Kallikrein Assays

These assays can be used to determine the inhibition constant (K _i ) of a representative compound of the invention.

Materials:

Analysis buffer: 100 mM Hepes, pH 7.8, 140 mM NaCl, 0.1% PEG-8000, 0.02% Tween-80 Coagulation Human factor XA, thrombin or plasma kallikrein (Hematologic Technologies). Factor: dilute to 0.45 μg / ml (9.8 nM) in assay buffer. Substrate: S-2222, S-2366 or S-2302 (see below, Chromogenix
Inc). Reconstitute to 5 mM in H ₂ O. Before use, dilute to 1.5 mM in assay buffer. Samples: Dilute samples until a DMSO concentration of 3% is reached in assay buffer.

Analysis Procedure:

1. Add 50 μl of sample to each well.

2. Add 50 μl of appropriately diluted coagulation factor to each well.

3. Incubate for 5 min at room temperature, gently shaking.

6. Add 50 μl of an appropriately diluted substrate to each well.

7. Shake the tablet for 20-25 seconds.

8. Determine the absorbance at 405 nm every 10 s for a total of 5 minutes at room temperature.

9. Calculate the value of V _max at 10 points.

Analysis - enzyme, substrate and final concentration

Analysis TF / F VIIa FHA Thrombin Plasma kallikrein The final concentration of coagulation factor 10 nM F VIIa 30 nM TF 3.3 nM 8.2 nM 1.5 nM Substrate Chromozym-tPA S-2222 S-2366 S-2302 The final concentration of the substrate 1.33 mm 0.5 mm 0.3 mm 0.3 mm

Claims (11)

1. The compound having the following structure:

where a and b independently from each other denote CH or CR ₃ , X is C = O or (CR _4a R _4b ) _m , where m is 1 or 2; Y is S (O) _n -R ₁ , where n is 1 or 2, S (O) _n -NR ₂ R ₂ , where n is 1 or 2, S (O) _n -OR ₂ , where n is 1 or 2; N ₁ and N ₂ are nitrogen atoms; Q and R ₁ independently represent (1) optionally substituted alkyl having from 1 to 10 carbon atoms; (2) an optionally substituted aralkyl containing an aryl moiety having from 6 to 10 ring carbon atoms, which is bonded to an alkyl moiety comprising from 1 to 10 carbon atoms; (3) an optionally substituted aralkenyl containing an aryl moiety having from 5 to 10 ring atoms, which is bonded to an alkenyl moiety comprising from 1 to 10 carbon atoms; (4) optionally substituted aryl having from 6 to 10 ring carbon atoms; (5) an optionally substituted heteroaryl having from 5 to 10 ring atoms, wherein the ring atoms are selected from the group consisting of carbon atoms and heteroatoms, the heteroatoms being sulfur; each R ₂ is H; R ₃ is H; R _4a , R _4b , R ₅ and R _{6 are} independently H; R ₇ and R _{8 are} independently H or C ₁ -C ₆ alkyl; and its acid addition salts and base addition salts. 2. The compound of claim 1, wherein Q is phenyl optionally substituted with one or more substituents selected from the group consisting of halogen, nitro, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl , NR ₇ R ₈ , OR ₇ , SR ₇ , C ₁ -C ₆ alkyl-C (O) OR ₇ , OC ₁ -C ₆ alkyl-C (O) OR ₇ , C ₁ -C ₆ alkyl-OR ₇ , OS ₁ -C ₆ alkyl-OR ₇ , C ₁ -C ₆ alkyl-NR ₇ R ₈ , OS ₁ -C ₆ alkyl-NR ₇ R ₈ , C ₁ -C ₆ alkyl-C (O) NR ₇ R ₈ , OS ₁ -C ₆ alkyl-C (O) NR ₇ R ₈ , C ₁ -C ₆ alkyl-C (O) R ₇ , OS ₁ -C ₆ alkyl-C (O) R ₇ , C ₁ -C ₆ haloalkyl , O-aralkyl, C (O) OR ₇ , C (O) NR ₇ R ₈ , OC (O) NR ₇ R ₈ , NHC (O) R ₇ , NHC (O) NR ₇ R ₈ , NR ₇ S ( O) ₂ R ₁ , NR ₇ S (O) ₂ R ₇ , NR ₇ S (O) ₂ C ₁ -C ₆ alkyl-C (O) OR ₇ , S (O) ₂ R ₇ and S (O) ₂ NR ₇ R ₈ . 3. The compound according to claim 1, in which Q has the structure

where R ₉ and R ₁₀ independently represent H, C ₁ -C ₆ alkoxy or O-C ₁ -C ₁₀ alkyl-C ₆ -C ₁₀ aryl; R ₁₁ is H, NR ₇ S (O) ₂ R ₇ or NR ₇ S (O) ₂ C ₁ -C ₆ alkyl-C (O) OR ₇ ; Z ₁ is H; Z ₂ is H or C ₁ -C ₆ alkoxy. 4. The compound according to claim 3, in which Z ₁ and Z ₂ denote hydrogen; Z ₁ , Z ₂ and R ₁₁ are hydrogen; or Z ₁ , R ₁₀ and R ₁₁ are hydrogen. 5. The compound according to claim 1, in which R ₉ and R ₁₀ independently represent OCH ₃ , OEt, OiPr, OiBu, OCH ₂ Ph or OCH (CH ₂ Cl) Ph. 6. The compound according to claim 1, in which X represents a carbonyl group, C = O. 7. The compound according to any one of claims 1 to 6, where Y is S (O) _n R ₁ , where n = 1 or 2. 8. A pharmaceutical composition for inhibiting serine protease enzymes, comprising a compound according to any one of claims 1 to 7 and a carrier or excipient. 9. A method of inhibiting the activity of TF / factor VIIa, factor Xa, thrombin or kallikrein, comprising bringing the TF / factor VIIa, factor Xa, thrombin or kallikrein into contact with an effective amount of the composition of claim 8. 10. A method of treating a disease mediated by TF / factor VIIa, factor Xa, thrombin or kallikrein, comprising administering to a mammal that needs it, an effective amount of a composition of claim 8. 11. A method of preventing thrombosis or treating abnormal thrombosis, comprising administering to a mammal that needs it, an effective amount of a composition of claim 8.