MXPA00011612A - Pyrazinone protease inhibitors - Google Patents

Abstract

Pyrazinone compounds are described, including compounds of Formula (I), wherein X is O, NR11 or CH=N, R3-R11, Ra, Rb, Rc, W, m and n are set forth in the specification, as well as hydrates, solvates or pharmaceutically acceptable salts thereof. The compounds of the invention are potent inhibitors of proteases, especially trypsin-like serine proteases, such as chymotrypsin, trypsin, thrombin, plasmin and factor Xa. Certain of the compounds exhibit antithrombotic activity via direct, selective inhibition of thrombin. Compositions for inhibiting loss of blood platelets, inhibiting formation of blood platelet aggregates, inhibiting formation of fibrin, inhibiting thrombus formation, and inhibiting embolus formation are described. Other uses of compounds of the invention are as anticoagulants either embedded in or physically linked to materials used in the manufacture of devices used in blood collection, blood circulation, and blood storage, such as catheters, blood dialysis machines, blood collection syringes and tubes, blood lines and stents. Additionally, the compounds can be detectably labeled and employed for in vivo imaging of thrombi.

Description

INHIBITORS OF PIETZINONE PROTEASE Field of Invention The present invention relates to novel compounds that function as inhibitors of the proteolytic enzyme, and particularly to a new class of thrombin inhibitors.

Background of the Invention Proteases are enzymes that split proteins into specific, simple peptide bonds. Proteases can be classified into four generic classes: serine, thiol or cysteine, acid or aspartyl, and metalloproteases (Cuypers et al, J. Biol. Chem. 25 7: 7086 (1982)). Proteases are essential for a variety of biological activities, such as digestion, formation and dissolution of blood clots, reproduction and immune reaction to external cells and organisms. Aberrant proteolysis is associated with a number of disease states in man and other animals. The human neutrophilic proteases, elastase and cathepsin G, are Ref. 124973 implicated as contributors to disease states marked by tissue destruction. These disease states include emphysema, rheumatoid arthritis, corneal ulcers and glomerular nephritis. (Barret, in En zyme Inhibi tors a s Drugs, Sanders, ed., University Park Press, Baltimore, (1980)). Additional proteases such as plasmid, esterase C-1, convertase C-3, urokinase, plasminogen activator, acrosin, and kallikrein, play major roles in the normal biological functions of mammals. In many cases, it is beneficial to break the function of one or more proteolytic enzymes in the course of a therapeutic treatment to a mammal.

Serine proteases include enzymes such as elastase (human leukocyte), cathepsin G, plasmid, esterase C-1, convertase C-3, urokinase, plasminogen activator, cytomotrypsin, trypsin, thrombin, factor Xa and kallikreins.

Human leukocyte elastase is released by polymorphonuclear leukocytes at sites of inflammation and thus is a contributing cause to a number of disease states. Cathepsin G is another human serine proteophyll protease. Compounds with the ability to inhibit the activity of these enzymes are expected to have a useful anti-inflammatory effect in the treatment of gout, rheumatoid arthritis and other inflammatory diseases, and in the treatment of emphysema. Cimothyrosine and trypsin are digestive enzymes. Inhibitors of these enzymes are useful in the treatment of pancreatitis. Inhibitors of urokinase and plasminogen activator are useful in the treatment of conditions of excess cell growth disease, such as hypertrophy of the benign prostate, carcinoma of the prostate and psoriasis.

Serine protease thrombin plays a central role in hemostasis and thrombosis, and as a multifactorial protein, it induces a number of effects on platelets, endothelial cells, smooth muscle cells, leukocytes, the heart, and neurons The activation of the coagulation cascade through either the intrinsic pathway (contact activation) or the extrinsic pathway (activation by exposure of the plasma to a non-endothelial surface, damage to vessel walls or tissue release factor) leads to a series of biochemical events that converge in thrombosis. The fibrinogen that starts the thrombin ends in a hemostatic plug (formation of clots), potentiates activated platelets through a unique proteolytic partition of the surface of the thrombin receptor cell (Coughlin, Semina rs in Hema tol ogy 31 (4): 270-277 (1994)), and automatically amplifies its own production through a feedback mechanism. In this way, inhibitors of thrombin function have a therapeutic potential in a housing of cardiovascular and non-cardiovascular diseases.

Factor Xa is another serine protease in the coagulation pathway. Factor Xa is associated with factor Va and calcium in a phospholipid membrane, thereby forming a prothrombinase complex. This prothrombinase complex then converts prothrombin to thrombin (Claeson, Bl ood Coa gula ti on and Fibrinolysi s 5: 411-436 (1994); Harker, Blood Coagulation and Fibrinolysis 5 (Suppl 1): S47-S58 (1994)). Factor Xa inhibitors are intended to offer an advantage over agents that directly inhibit thrombin since thrombin inhibitors still allow the generation of significant novel thrombin (Lefkovits and Topol, Circulation 90 (3): 1522-1536 (1994 ); Harker, Blood Coagulation and Fibrinolysis 5 (Suppl 1): S47-S58 (1994)).

In vivo diagnostic imaging methods for intravascular thrombi have been previously reported. These imaging methods use compounds that are detectably labeled with radioactive or paramagnetic atoms. For example, platelets labeled with the gamma emitter, In-111, can be used as an imaging agent to detect thrombi (Thakur, ML et al., Throm Res. 5: 345 (1976); Powers et al., Neurology 32: 938 (1982)). Thrombotic enzyme streptokinase labeled with Tc-99m has been proposed as an imaging agent (Wong, U.S. Patent No. 4,418,052 (1983)). The fibrin-binding domains of Staphylococcus to ureas derived from protein A labeled with gamma emitters, 1-125 and 1-131, have been proposed as imaging agents (Pang, U.S. Patent No. 5,011,686 (1991)). Monoclonal antibodies that are specific for fibrin (as contrasted with fibrinogen) and labeled with Tc-99m have been proposed as imaging agents (Berger et al., US Patent No. 5,024,829 (1991); Dean et al., US Pat. No. 4,980,148 (1990)). The use of paramagnetic contrast agents, gadolinium diethylenetriaminepentaacetic acid in magnetic resonance imaging of patients treated for thrombolysis for myocardial infarction has been reported (De Roos, A et al., Int. J Ca rd. Ima ging 7: 133 (1991 )). Radioactive labeling and paramagnetic labeling with alpha ketoamide derivatives have also been proposed as thrombus imaging agents (Abelman et al., U.S. Patent No. 5,656,600).

There is a continuing need for non-peptide compounds which are selective and potent protease inhibitors, and which possess greater biological availability and some side effects than currently available protease inhibitors. Accordingly, a new class of potent protease inhibitors, characterized by their potent inhibitory capacity and low mammalian toxicity, are potentially valuable therapeutic agents for a variety of conditions, including the treatment of a number of proteolytic disease states in mammals. .

Brief description of the invention.

The present invention is directed to novel aminoguanidine (hydrazinoamidine) and alkoxyguanidine (alkoxyaminoidine) pyrazinones having the Formula I (below). Processes for preparing the compounds of Formula I are also provided. The novel compounds of the present invention are potential inhibitors of proteases, especially trypsin-like serine proteases, such as chymotrypsin, trypsin, thrombin, plasmin and factor Xa. Some of the compounds exhibit antithrombotic activity by means of selective, direct inhibition of thrombin, or are useful intermediates for forming compounds having thrombotic activity. Also provided are methods for inhibiting or treating aberrant proteolysis in a mammal and methods for the treatment of thrombosis, ischemia, attack, restenosis or inflammation in a mammal by administering an effective amount of a compound of Formula I.

The invention includes a composition for inhibiting the loss of blood platelets, inhibiting the formation of aggregates of blood platelets, inhibiting fibrin formation, inhibiting the formation of thrombi, and inhibiting the formation of emboli in a mammal, comprising administering a compound of the invention in a pharmaceutically acceptable carrier. These compositions may optionally include anticoagulants, antiplatelet agents, and anti-itcholytic agents. The compositions may be added to the blood, blood products, or organs of mammals in order to effect the desired inhibitions.

Also provided are methods for inhibiting or treating aberrant proteolysis in a mammal, and methods for treating myocardial infarction; unstable angina; attack; restenosis; deep vein thrombosis; Disseminated intravascular coagulation caused by trauma, sepsis or tumor metastasis; hemodialysis; cardiopulmonary bypass surgery; respiratory distress syndrome in adults; endotoxic shock; rheumatoid arthritis; Ulcerative colitis; hardening; metastasis, hypercoagulability during chemotherapy; Alzheimer disease; Down's Syndrome; fiber formation in the eye; and wound healing. Other uses of the compounds of the invention are as anticoagulants either inserted or physically bound to materials used in the manufacture of devices that are used in blood collection, blood circulation, and blood storage, such as catheters, dialysis machines. of blood, syringes and blood collection tubes, blood lines and stenoses.

The invention also includes a method for reducing the thrombogenicity of a surface in a mammal by binding to the surface, either covalently or non-covalently, a compound of the invention.

In another aspect, the present invention includes compositions that are useful for in vi ve imaging for thrombi in mammals, comprising a compound of the present invention that is capable of being detected outside the body. Preferred are compositions comprising a compound of the present invention and a detectable label, such as a radioactive or paramagnetic atom.

In another aspect, the present invention provides diagnostic compositions that are useful for imaging thrombi in a mammal, comprising a pharmaceutically acceptable carrier and a diagnostically effective amount of a compound or composition of the present invention.

In another aspect, the present invention includes methods that are useful for imaging thrombi in a mammal.

Detailed description of the invention.

The compounds of the present invention include compounds of Formula I: or a solvent, or pharmaceutically acceptable salt thereof; where: W is hydrogen, R1, R ^ CO, RxCO, R1 (CH2) 5NHCO, or (R1) 2CH (CH2) 5NHCO, wherein s is 0-4; R1 is R2, R2 (CH2) iC (R12) 2, where t is 0-3, and each of R12 can be the same or different, (R2) (OR12) CH (CH2) P, where p is 1-4, (R2) (OR12) C (CH2) P, where p is 1-4, R2C (R12) 2 (CH2) t, where t is 0-3, and each of R12 may be the same or different, wherein (R12) 2 may also form a ring with C represented by C3.7 cycloalkyl, R2CF2C (R12) 2 (CH2) q, wherein q is 0-2, and each of R12 may be the same or different, wherein (R12) 2 may also form a ring with C represented by C3-7 cycloalkyl, R2CH2C (R12) 2 (CH2) qr where q is 0-2, and each of R12 may be the same or different, wherein (R12) 2 may also form a ring with C represented by C3-7 cycloalkyl, ( R2) 2CH (CH2) r, where r is 0-4 and each of R2 may be the same or different, and wherein (R2) 2 may also form a ring with C represented by C3-7 cycloalkyl, bicyclic alkyl C7_ 2 2, C? 0-66 tricyclic alkyl or a 5- to 7-membered mono- or bicyclic heterocyclic ring which may be saturated or unsaturated, and which contain from one to three heteroatoms selected from the group consisting of N, O and S, R20 (CH2) p, where p is 2-4, (R2) 2CF (CH2) r, where r is 0-4 and each of R12 may be the same or different, where (R12) ) 2 can also form a ring with C represented p or C3_7 cycloalkyl, C7-? 2-bicyclic alkyl, C? -6-6? tricyclic alkyl or a 5- to 7- membered mono- or bicyclic heterocyclic ring which may be saturated or unsaturated, and which contain from one to three heteroatoms selected from the group consisting of N, O and S, where s is 0 or 1, or 2 > phenyl, naphthyl, or biphenyl, each of which is substituted or unsubstituted with one or more of C? -4 alquiloalkyl, C? _4alkoxy, halogen, hydroxy, CF3, OCF3, COOH, CONH2, or S02NH2, a ring heterocyclic or non-heterocyclic mono- 5- to 7-membered or 9- to 10-membered bicyclic which can be saturated or unsaturated, wherein the heterocyclic ring contains from one to four heteroatoms selected from the group consisting of N, O and S , and wherein the heterocyclic or non-heterocyclic ring is substituted or unsubstituted by halogen or hydroxy, C-7 alkyl, substituted or unsubstituted by one or more of hydroxy, COOH, amino, aryl, C3.7 cycloalkyl, CF3, N (CH3) 2, alkylaryl C _. 3, heteroaryl, or heterocycloalkyl, CF3, C3_7 cycloalkyl, substituted or unsubstituted with aryl, C2_2 bicyclic alkyl, or Ccy-R3 tricyclic alkyl is hydrogen, alkyl, cycloalkyl, alkenyl, alkynyl, optionally substituted by aryl, optionally substituted by aralkyl, optionally substituted by heteroaryl, trifluoromethyl, halogen, hydroxyalkyl, cyano, nitro, carboxamido, C02Rx, -CH2ORx or -ORx, where Rx, in each instance, is independently one of hydrogen, alkyl or cycloalkyl wherein said alkyl or cycloalkyl groups may optionally have one or more unsaturations; R 'is hydrogen or halogen, R12 is hydrogen, phenyl, naphthyl, or biphenyl, each of which is unsubstituted or substituted by one or more of C? -4 alquiloalkyl, C _. alcoalkoxy, halogen, hydroxy, CF3, OCF3, COOH, or CONH2, a 5-7 membered monocyclic or bicyclic 9- to 10-membered heterocyclic ring which may be saturated or unsaturated, and which contain from one to four heteroatoms selected from the group consisting of N, O and S, C-alkyl ? -4, substituted or unsubstituted with one or more of hydroxy, COOH, amino, aryl, heteroaryl, or heterocycloalkyl, CF3, C? _4 cycloalkyl, C7_? 2 bicyclic alkyl, or Ccy-iß tricyclic alkyl? R5 is hydrogen, C4-4alkyl, or C2-4alkenyl; R6 is hydrogen, alkyl, alkenyl, aralkyl, aryl, hydroxyalkyl, aminoalkyl, (C2_10) alkyl, monoalkylamino, (C2-? O) dialkylamino, or carboxyalkyl; R7, R8, R9 and R10 are independently hydrogen, alkyl, aralkyl, aryl, hydroxyalkyl, aminoalkyl, monoalkylaminoalkyl, dialkylaminoalkyl or carboxyalkyl; or R7 and R8 are taken together to form - (CH2) U-, where u is 2 to 7, preferably 2 to 5, while R9 and R10 are as defined above; or R9 and R10 are taken together to form - (CH2) V-, where v is 2 to 7, preferably 2 to 5, while R7 and R8 are as defined above; or R7 and R9 are taken together to form - (CH2) and -, where y is 0 (one bond) or 1 to 7, preferably 0-4, while R8 and R10 are as defined above; is oxygen, NR, or CH = N (where N is attached to NR6) where R11 is hydrogen, alkyl, cycloalkyl or aryl, wherein said alkyl, cycloalkyl or aryl may be optionally substituted with amino, monoalkylamino, dialkylamino, alkoxy, hydroxy , carboxy, alkoxycarbonyl, aryloxycarbonyl, aralkoxycarbonyl, aryl, acylamino, cyano or trifluoromethyl; Rc R £ are independently hydrogen, alkyl, hydroxy, alkoxy, aryloxy, aralkoxy, alkoxycarbonyloxy, cyano or -C02R, where Rw is alkyl, cycloalkyl, phenyl, benzyl, where Rd and Re are independently hydrogen, C6-6alkyl, alkenyl or C2-6 phenyl, Rf is hydrogen, C2-C2alkenyl phenyl alkyl C2 2-6 / Rc- is hydrogen, C? _ßalkyl, alkenyl or C2_6 phenyl, and Rh is aralkyl or alkyl C? -6; n is from zero to 8; and m is from zero to 6.

In a class of compounds and pharmaceutically acceptable salts thereof, R3 is hydrogen, C4_4alkyl, C3_7cycloalkyl, or CF3; preferably C? -4 alkyl, and m and n are each from 0 to 4.

In a subclass of these classes of compounds and pharmaceutically acceptable salts thereof, R 4 is hydrogen or halogen.

In a group of these subclasses of pharmaceutically acceptable compounds and salts thereof, W is H or R1.

In a subset of these groups of compounds and pharmaceutically acceptable salts thereof, R1 is R2, R2 (CH2) tC (R12) 2, where t is 0-3, and each of R12 may be the same or different, R2C ( R12) 2 (CH2) t. wherein t is 0-3, and each R12 may be the same or different, wherein (R12) 2 may also form a ring with C represented by C3_7 cycloalkyl, R2CH2C (R12) 2 (CH2) q, wherein q is 0-2, and each of R12 may be the same or different, wherein (R12) 2 may also form a ring with C represented by cycloalkyl C3-7, (R2) 2CH (CH2) r, where r is 0-4 and each of R2 may be the same or different, and wherein (R2) 2 may also form a ring with CH represented by C3- cycloalkyl , C7-? 2 bicyclic alkyl, C? 0 -6r tricyclic alkyl or a 5- to 7- membered mono- or bicyclic heterocyclic ring which may be saturated or unsaturated, and which contains from one to three heteroatoms selected from the group consisting of N, O and S, R2CF2C (R12) 2 (CH2) q, where q is 0-2, and each of R12 may be the same or different, wherein (R12) 2 may also form a ring with C represented by cycloalkyl C3.7, or R20 (CH2) p, where p is 2-4; phenyl or naphthyl, each of which is unsubstituted or substituted by one or more of C? -4 alquilo ,alkyl, alco? alco4alkoxy, halogen, hydroxy, CF3, OCF3, or S02NH2, a monocyclic 5-membered heterocyclic ring; up to 7- member or 9- to 10-member bicyclic or a non-heterocyclic ring which may be saturated or unsaturated, wherein the heterocyclic rings contain from one to four heteroatoms selected from the group consisting of N, O and S , and wherein the heterocyclic or non-heterocyclic ring is substituted or unsubstituted with halogen or hydroxy, C? _7 alkyl, substituted or unsubstituted with one or more of hydroxy, COOH, C3-7 cycloalkyl, CF3, N (CH3) 2 , -C? -3alkaryl, heteroaryl, or heterocycloalkyl, CF3, or C3_7 cycloalkyl, substituted or unsubstituted with aryl; Y R12 is hydrogen, or Cx_4 alkyl, substituted or unsubstituted with one or more of hydroxy, COOH, amino, aryl, heteroaryl, or heterocycloalkyl.

In a family of these subgroups of compounds and pharmaceutically acceptable salts thereof, R3 is H, CH3, or CH2CH3; R4 is H or chloro; and W is PhCH2CH2, (CH3) 3C-, HOOCCH2, CF3CH2, (CH3) 2N (CH2) 2, PhCH20 (CH2) 2, PhCH (CH3), PhCH2CH (COOH), CH3 (CH2) 5, PhCH2, H, CH3 (CH2) 4, CH3CH2CH (CH3) CH2, (Ph) 2CHCH2, PhCH (CH3) CH2, (CH3) 2CH, PhCH (OH) CH2, PhC (CH3) 2CH2, (Ph) 2CHCH2, [> - CH2, > CH2CH2, , < ? (CH2) 2 < ? CF2CH2 A preferred group R5 is hydrogen Preferred compounds when X is NR11 are those wherein R11 is hydrogen or C6-6alkyl, optionally substituted by one, two or three, preferably one, amino, monoalkylamino, dialkylamino, alkoxy, hydroxy, alkoxycarbonyl, aryloxycarbonyl, aralkoxycarbonyl, carboalkoxy, phenyl, cyano, trifluoromethyl, acetylamino, pyridyl, thiophenyl, furyl, pyrrolyl or imidazolyl.

Suitable values of R 11 include hydrogen, methyl, ethyl, propyl, n-butyl, benzyl, phenethyl, 2-hydroxyethyl, 3-hydroxypropyl, 4-hydroxybutyl, carboxymethyl and carboxyethyl.

The most preferred compounds are those where X is oxygen.

Preferred compounds are those of Formula I, wherein R6 is hydrogen or C6_6 alkyl.

Preferred compounds are those of Formula I, wherein R7, R8, R9 and R10 are independently one of hydrogen, C6-6alkyl, ar (Ci-d) C6-? 0alkyl, C6-? Aryl or C2-10 hydroxyalkyl or C2.7 carboxyalkyl. Useful values of R7, R8, R9 and R10 include hydrogen, methyl, ethyl, propyl, n-butyl, benzyl, phenylethyl, 2-hisroxyethyl, 3-hydroxypropyl, 4-hydroxypropyl, 4-hydroxybutyl, 2-carboxymethyl, 3- carboxyethyl, and 4-carboxypropyl.

Also preferred are compounds wherein any of R7 and R8, R9 and R10 combine as an alkylene bond, - (CH2) U- and - (CH2) V- respectively, where u and v are each 2-5, more preferably 2 or 3 , Preferred values of Ra, Rb and Rc in Formula I are hydrogen, hydroxy, C 1-6 alkyl, C 1-6 alkoxy, cyano or -C02R, where Rw, in each instance, is preferably one of C? -4 alkyl, C- or benzyloxycarbonyl cycloalkyl. Suitable values of Ra, R and Rc include hydrogen, methyl, ethyl, propyl, n-butyl, hydroxy, methoxy, ethoxy, cyano, -C02CH3, -C02CH2CH3 and -C02CH2CH2CH3. In the most preferred embodiments, R £ Rc s on each one i drogeno.

Also preferred in Ra, Rb and Rc e s the C02Rw group, where Rw e s one of where Rd-R are as defined above. When Ra, Rb and Rc are the group -C02Rw, where Rw is one of these portions, the resulting compounds are pro-drugs having the desirable formulation and biological availability characteristics. A preferred value for each of Rd, Rc and Rg is hydrogen, Rf is methyl, and preferred values for Rh include benzyl and tert-butyl.

Preferred values of n in Formula I include from zero to 6, more preferably from zero to 4 and even more preferably zero, 1 or 2.

The preferred values of m are from zero to 4, more preferably zero, 1 or 2.Especially preferred compounds are represented by formulas II, III and IV: or a solvate, hydrate or pharmaceutically acceptable salt thereof; where it is as defined, and has the preferred values, as per Formula I, above; X is -0-, -NH- or -CH = N- (an amidinohydrazone group, where NH is adhered to the NH group of the parent formula). X is preferably bonded to the ring in the 3- or 4- position of the ring, more preferably in the 4- position.

R3 is hydrogen, C? -3 alkyl, halogen or C? _2 alkoxy; R4 is hydrogen or halogen; n is 0, 1, 2 or 3, preferably 0, 1 or 2; m is 0, 1, 2 or 3, preferably 0 or 1; n 'is 0, 1, 2 or 3, preferably 0, 1 or 2; and m 'is 0, 1, 2 or 3, preferably 0 or 1.

Specific compounds within the scope of the invention include the following: 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (phenethylamino) -pyrazinone, 1- . { N- [2- (amidinoaminoo) ethyl] amino} carboni lmeti 1-6-methyl-3- (2, 2-diphenylethylamino) -pyrazinone, 1-. { N- [2- (amidinoaminooxy) ethyl] amino Jcarbonylmethyl-6-methyl-3- (4-methylphenethylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino Jcarbonylmethyl-6-methyl-3- (4-methoxyphenethylamino) -pyrazinone, 1- . { - [2- (amidinoaminooxy) ethyl] amino} carboni lmet il-6-methyl-3- (l-phenylcyclobutyl) methylamino-pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmet il-6-met i 1-3- [2- (1-naphthalene) ethyl] amino-pyrazinone, 1-. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2-phenyl-l-butylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmet-6-met-il-3- (2- [3,4-methylenedioxyphenyl] ethylamino) -pyrazone, 1-. { N- [2- (amidinoaminooxy) ethyl] amino Jcarbonylmethyl-6-met il-3- (2- [2-pyridyl] ethylamino) -pyrazinone, 1- . { - [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-met il-3- (2- [2-methyl-phenyl] -ethylamino-pyrazine, 1- {N- [2- (amidinoaminooxy) ethyl] amino.}. carbonylmethyl-6-methyl-3- (2- [3-methylphenyl] ethylamino) -pyrazinone, 1-. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2- [2-trifluoromethylphenyl] ethylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino Jcarbonylmethyl-6-methyl-3- (2- [3-trifluoromethylphenyl] ethylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methy1-3- (2- [4-trifluoromethylphenyl] ethylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxi) et il] amino} carbonylmethyl-6-methyl-3- (2- [3,5-dimethylphenyl] ethylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2-indanylamino) -pyrazinone, 1-. { N- [2- (amidinoaminooxy) ethyl] amino Jcarbonylmethyl-6-methyl-3- (2- [3, 4-difluorophenyl] ethylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino Jcarbonylmethyl-6-met il-3- (2- [5-indanyl] ethylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonyl lmethyl-6-methyl-3- (2-fluorophenethylamino) -pyrazinone, 1-. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (3,4-dimethoxyphenethylamine) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmeti1-6-methyl-3- (4-fluorofenet-ylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (4-ethylphenethylamino) -pyrazinone, 1-. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2-phenylpropylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (3,4-dimethylphenethylamino) -pyrazinone, 1-. { N- [2- (amidinoaminooxy) ethyl] amino} car onylmethyl-6-methyl-3- (2-naphthalene-ethylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2, 2-diphenylpropylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2- (3-indolyl) -ethylamino-pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2- [1- (4-methylnaphthalene)] ethylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino Jcarbonylmethyl-6-methyl-3- (2- [2, 4-difluorophenyl] ethylamino) -pyrazinone, 1-. { N- [2- (amidino-N '-methylaminooxy) ethyl] amino} car onylmethyl-6-met il-3- (2- [4-methylphenyl] ethylamino) -pyrazinone, 1- . { - [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-5-chloro-6-methyl-3- (phenethylamino) -pyrazinone, 1-. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-5-chloro-6-methyl-3- (2, 2-diphenylethylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (phenethyl) -pyrazinone, 1-. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (1-methyl-2-phenylethyl) -pyrazinone, 1- . { N- [2 - (amidinohydrazino) ethyl] amino} car onylmethyl-6-ethyl-3- (phenethyl) -pyrazinone, l-. { N- [2- (amidinohydrazino) ethyl] amino} carbonylmethyl-6-methyl-3- (4-methylphenylethyl) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (l-methyl-2- (4-methylamino-3-pyridi1) ethyl-pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (l-methyl-2- (3-pyridyl) ethyl-pyrazinone, 1- { N- [2- (amidinoaminooxy) ethyl] amino.}. carbonylmethyl-6-methyl-3 - (2-cyclopropyl-2- (3, 4-dimethoxyphenyl) ethyl) -pyrazinone, 1-. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmet-6-methyl-3- (2-cyclobutylethyl) -pyrazinone, 1- . { - [2- (amidinoaminooxy) ethyl] amino} carboni lmet i 1-6-methyl-3- (2-cyclobutyl-2,2-difluoroethyl) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmet i 1-6-methyl-3- (2- (3-fluorophenyl) ethyl) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2, 2-difluoro-2-phenylethyl) -pyrazinone, 1- . { N- [2- (amidinoaminooxi) et il] amino} carbonylmethyl-6-methyl-3- (2-phenylcyclopropyl) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-met i 1-3- (2- (4-chlorophenyl) -2-cyclopropylethyl) -pyrazinone, 1-. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methy1-3- (2-cyclopropyl-2- (3-pyridyl) ethyl) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2-benzylcyclopropyl) -pyrazinone, 1-. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2-cyclopropyl-2- (3-fluorophenyl) ethyl) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2-cyclopropyl-2- (3,4-difluorophenyl) ethyl) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2, 2-difluoro-2-phenylethylene) -pyrazinone, as well as the pharmaceutically acceptable salts thereof, for example the hydrochloride, acetate and trifluoroacetate and salts thereof.

It will also be understood that the present invention contemplates including stereoisomers as well as optical isomers, for example, mixtures of enantiomers as well as individual enantiomers and diastereomers, which result as a consequence of a structural asymmetry in selected compounds of the present series. The compounds of the present invention can also have polymorphic crystalline forms, with all the polymorphic crystalline forms being included in the present invention.

The compounds of Formula I can also be solvates, especially hydrated. Hydration may occur during the manufacture of the compounds or compositions comprising the compounds, or hydration may occur over time due to the hygroscopic nature of the compounds.

Certain compounds within the scope of Formula I are derivatives referred to as prodrugs. The term "pro-drug" denotes a derivative of a known direct acting drug, which derivative has increased release characteristics and a therapeutic value compared to the drug, and is transformed into the active drug by a chemical or enzymatic process. Useful pro-drugs are those where Ra, Rb and / or Rc are -C02Rw, where Rw is as defined above. See U.S. Patent No. 5,466,811 and Saulnier et al., Bi org. Med. Ch em. Le t t. 4: 1985-1990 (1994).

When any variable occurs more than once in any constituent or in Formula I, its definition in each occurrence is independent of its definition every other occurrence. Also, combinations of substituents and / or variables are permissible only if such combinations result in stable compounds.

In another aspect, the present invention includes compositions that are useful for in vi ve image of thrombi in a mammal, comprising a compound of the present invention that is capable of being detected outside the body. Preferred are compositions comprising a compound of the present invention and a detectable label, such as a radioactive or paramagnetic atom.

In another aspect, the present invention provides diagnostic compositions that are useful for in vitro thrombus imaging in a mammal, comprising a pharmaceutically acceptable carrier and a diagnostically effective amount of a compound or composition of the present invention.

In another aspect, the present invention includes methods that are useful for viii image of thrombi in a mammal.

According to a preferred aspect, useful compounds are those wherein the substituent R1 is replaced with a detectable label, such as a radioactive iodine atom, such as 1-125, 1-135 or 1-123. In this regard, R1 is preferably phenyl, which has a substitution 1-123 for, 1-125 for or 1-131 for, or benzyl, having a substitution of 1-123 meta, 1-125 meta or 1-131 meta.

The detectable label can also be a radioactive or paramagnetic chelate in which an appropriate ligand (L) is linked to a Rl substituent, either directly or via a divalent linking group A. "Alternatively, the group -A" - L replaces the groups W in Formula I. By an appropriate ligand it means an organic portion that is capable of chelating a radioactive or paramagnetic metal ion.

In these compounds, the divalent linking groups A "which are capable of covalently linking to an amino-free group and the chelating medium, eg, A" can be -C (= S) -, -C (= 0) - , -C (= NH) - (CH2) 6-C (= NH) -, -C (= 0) - (CH2) 6-C (= 0) -, and similar Also, in the compounds represented by Formula I, the chelating ligand, L, includes groups capable of covalently linking or non-covalently linking a radioactive or paramagnetic atom. Chelating media include those that are regularly used to make complexes of radioactive or paramagnetic atoms. These include chelating media containing 3 up. 12, preferably 3 to 8, methylene phosphonic acid groups, methylene carbohydroxamic acid groups, carboxyethyleneidene groups, or especially carboxymethyl groups, which are bonded to a nitrogen atom. If only one or two of the acid groups are bonded to the nitrogen atom, then the nitrogen is bonded to another nitrogen atom having such groups by optionally substituted ethylene groups or by up to four separate ethylene units, which are separated by one atom of nitrogen, oxygen or sulfur. Preferred as a means of completing the diethylenetrimine-N, N, N ', N ", N" -pentaacetic acid (DTPA). DTPA is well known in the art as a chelating medium for the radioactive atom of indium-111 (In-111), technetium-99m (Tc-99m) and the paramagnetic gadolinium atom (Gd). Khaw, et al., Sci in 209: 295 (1980); Paik C.H. and collaborators, U.S. Pat. No. 4,652,440 (1987); Gries, H. and collaborators, U.S. Pat. No. 4,957,939 (1990). A preferred chelating ligand, L, is 1- (p-aminobenzyl) -diethylenetriaminepentaacetic acid. Also included as chelating media are the compounds containing sulfhydryl or amine moieties, the total of which in any combination is at least four. These sulfhydryl or amine moieties are separated one from each other by at least two atoms which may be either carbon, nitrogen, oxygen or sulfur. Especially preferred for chelating media, L, is the metallothionein which is well known in the art as a chelating medium for Tc-99M.

The term "alkyl" as used herein by itself or as part of another group, refers to both straight or branched chain radicals of up to 12 carbons, such as methyl, ethyl, propyl, isopropyl, butyl, t- butyl, isobutyl, pentyl, hexyl, isohexyl, heptyl, 4, -dimetilpentilo, octyl, 2,2,4-ilpentilo TriMet, nonyl, decyl, undecyl, dodecyl.

The term "alkenyl" as used herein means a straight or branched chain radical of 2-20 carbon atoms, unless the length of the chain is limited thereto, including, but not limited to, ethenyl, 1- propenyl, 2-propenyl, 2-methyl-1-propenyl, 1-butenyl, 2-butenyl, and the like. Preferably, the alkenyl chain is from 2 to 10 carbon atoms in length, more preferably, 2 to 8 carbon atoms in length, even more preferably from 2 to 4 carbon atoms in length.

The term "alkynyl" is used herein to mean a straight or branched chain radical of 2-20 carbon atoms, unless the length of the chain is limited thereto, where it is at least one triple bond between two of the carbon atoms in the chain, including, but not limited to, acetylene, 1-propylene, 2-propylene, and the like. Preferably, the alkynyl chain is 2 to 10 carbon atoms in length, more preferably 2 to 8 carbon atoms in length, even more preferably 2 to 4 carbon atoms in length.

In all cases of the present where it is an alkenyl or alkynyl portion as a substituent group, the unsaturated bond, that is, the vinylene or acetylene bond preferably does not directly bind to a nitrogen, oxygen or sulfur moiety.

The term "alkoxy" is used herein to mean a straight or branched chain radical of 1 to 20 carbon atoms, unless the length of the chain is limited thereto, bound to an oxygen atom, including, but not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, and the like. Preferably the alkoxy chain is from 1 to 10 carbon atoms in length, more preferably from 1 to 8 carbon atoms in length.

The term "aryl" as used herein, by itself or as part of another group, refers to monocyclic or bicyclic aromatic groups, containing from 6 to 12 carbons in the ring portion, preferably 6-10 carbons in the ring portion, such as phenyl, naphthyl or tetrahydronaphthyl.

The term "heteroaryl" as used herein refers to groups having from 5 to 14 ring atoms; 6, 10 or 14 p electrons sharing in a cyclic formation; and containing carbon atoms and 1, 2 or 3 oxygen, nitrogen or sulfur (where examples of heteroaryl groups are: thienyl, benzo [b] thienyl, naphtho] 2, 3-b] thienyl, thianthrenyl, furyl, pyranyl, isobenzofuranyl, benzoxazolyl, chromenyl, xanthenyl, phenoxatinyl, 2H-pyrrolyl, pyrrolyl, imidazolyl, pyrazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolizinyl, isoindolyl, 3H-indolyl, indolyl, indazolyl, purinyl, 4H-quinolizinyl, isoquinolyl, quinolyl, phthalazinyl, naphthyridinyl, quinazolinyl, cinolonilo, pteridinyl, 4aH-carbazolyl, carbazolyl, ß-carbolinyl, phenanthridinyl, acridinyl, perimidinyl, phenanthrolinyl, phenazinyl, isothiazolyl, phenothiazinyl, isoxazolyl, furazanyl and phenoxazinyl groups).

The term "aralkyl" or "arylalkyl" as employed herein by itself or as part of another group, alkyl Cl-6 as discussed above having an aryl substituent, such as benzyl, phenylethyl or 2 refers -naphthylmethyl.

The term "cycloalkyl" as used herein, by itself or as part of another group, refers to cycloalkyl groups containing from 3 to 9 carbon atoms, preferably from 3 to 7 carbon atoms. Typical examples are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and cyclononyl.

The term "Bicyclic alkyl C -? 2" is intended to include bicyclo [2.2.1] heptyl (norbornyl), bicyclo [2.2.2] octyl, 1,1,3-trimethyl-ilicyclo [2.2.1] heptyl (bornyl), and the like.

The term "Cι-ie tricyclic alkyl" is intended to include tricyclo [5, 2, 1, O 2 '6] decyl, adamantyl, and the like.

The term "halogen" or "halo" as used herein, by itself or as part of another group, refers to chlorine, bromine, fluorine or iodine with chlorine being preferred.

The term "monoalkylamine" as used herein by itself or as part of another group, refers to a group that is substituted with an alkyl group having from 1 to 6 carbon atoms.

The term "dialkylamine" as used herein, by itself or as part of another group, refers to an amino group which is substituted with two alkyl groups, each having from 1 to 6 carbon atoms.

The term "hydroxyalkyl" as used herein refers to any of the above alkyl groups substituted by one or more hydroxyl portions.

The term "carboxyalkyl" as used herein refers to any of the above alkyl groups substituted by one or more carboxylic acid moieties.

The term "heterocycle" or "heterocyclic ring", as used herein, except where noted, represents a stable heterocyclic ring system of 5 to 7 mono or bicyclic members or a stable heterocyclic ring system of 7 to 10 bicyclic members any ring may be saturated or unsaturated, and which consists of carbon atoms and from one to three heteroatoms selected from the group consisting of N, O and S, and wherein the nitrogen and sulfur heteroatoms may optionally be oxidized, and the The nitrogen heteroatom can optionally be quaternized, and includes any bicyclic group in which any of the heterocyclic rings defined above is fused to the benzene ring, especially rings containing one to three nitrogen atoms, one of oxygen or sulfur, or one of oxygen or sulfur combined with one or two nitrogen atoms, the heterocyclic ring can be linked to any teroatomo or carbon atom that results in the creation of a stable structure. Examples of such heterocyclic groups include piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolodinyl, 2-oxoazepinyl, azepinyl, pyrrolyl, 4-piperidonyl, pyrrolidinyl, pyrazolyl, pyrazolidinyl, imidazolyl, imidazolinyl, imidazolidinyl, pyridyl, pyrazinyl. , pyrimidinyl, pyridazinyl, oxazolyl, oxazolidinyl, isoxazolyl, isoxazolidinyl, morpholinyl, thiazolyl, thiazolidinyl, isothiazolyl, quinuclidinyl, isothiazolidinyl, indolyl, quinoolinilo, isoquinolinyl, benzimidazolyl, thiadiazoyl, benzopyranyl, benzothiazolyl, benzoxazolyl, furyl, tetrahydrofuryl, tetrahydropyranyl, thienyl, benzothienyl , tiamorgolinyl, thiamorpholinyl sulfoxide, thiamorpholinyl sulfone, and oxadiazolyl. Morpholino is the same as morpholinyl.

The term "heteroatom" as used herein means an oxygen atom ("O"), a sulfur atom ("S"), or a nitrogen atom ("N"). It will be recognized that when the heteroatom is nitrogen, it can form a NRaRb portion, where Ra and Rb are, independently of one another, hydrogen or Ci to C8 alkyl, or together with the nitrogen to which they are bound, form a saturated or not saturated of 5, 6, 6, 7 members.

Schemes 1 and 2 summarize the synthetic steps for producing compounds of Formula I.

Scheme 2 1. 5 (from scheme 1) 2. optional removal R », R °, Rc In Schemes 1 and 2, W, R3, R7, R8, R9, R10, Ra, R, Rc, n and m are as defined above; R4 = Cl or Br; P is an amino protecting group, such as benzyloxycarbonyl (Cbz), and P 'is an ester protecting group, such as benzyl.

In Scheme 1, protected aminoalcohol 1 is coupled to N-hydroxyphthalimide 2 using a Mitsunobu coupling procedure (Mitsunobu, 0. Synthesis 1 (1981)) to provide compound 3. Preferred coupling conditions include the use of trialkylphosphine or triarylphosphine, such as triphenylphosphine, in a suitable solvent, such as tetrahydrofuran or methylene chloride, and a dialkyl azodicarboxylate. , such as diethyl azodicarboxylate. The unveiling of the phthalamide protecting group is carried out using standard conditions well known in the art (Greene, TW, Wuts, PGM, Protective Groups in Organic Synthesis, 2nd edition, John Wiley and Sons, Inc., New York (1991)), such such as methylamine or hydrazine, in a suitable solvent, such as ethanol or isopropanol. The guanidylation of the resulting alkoxyamine for 4 using substituted guanidylating reagents such as N, N'-bis (tert-butoxycarbonyl) -S-methylisothiourea (Bergeron, RJ And McManis, JS, J. Org. Chem. 52: 1100 (1987) ) or N-Ra, N-Rb, N-Rc-lH-pyrazole-1-carboxamide (Beratowicz, MS et al., Tetrahedron Letter 34: 3389 (1993)). The deprotection of the amino group to give the intermediate 5 is carried out using a standard procedure well known in the art (Greene, TW, Wuts, PGM, Protective Goups in Organi c Synthesis, 2nd edition, John Wiley and Sons, Inc. New York (1991)), such as palladium on carbon, in a suitable solvent, such as methanol or ethanol. In some cases, it is advantageous to add an acid, such as hydrochloric acid.

In Scheme 2, an ester-protected glycine, such as P '= benzyl, or ethyl, is concentrated with an aldehyde, such as acetaldehyde, and a cyanide, such as cyanotrimethylsilane (TMSCN), in an appropriate solvent, such as chloride of methylene to provide the aminonitrile 6. the aminonitrile is reacted with oxalyl chloride or oxalyl bromide in a suitable solvent, such as 1,2-dichlorobenzene to give the pyrazinone 7. The 3-chloro or 3-bromo pyrazinone 7 it is displaced by an appropriate amine, such as phenethylamine, 2,2-diphenylethylamine or 4-methoxyphenethylamine, in an appropriate solvent, such as ethyl acetate, to give compound 8. ester 8 is converted to acid 9 p or standard procedures known in the art (Greene, TW, Wuts, PGM, Protective Groups in Organi c Syn thes is, 2nd edition, John Wiley and Sons, Inc. New York (1991)), such as hydrolysis using a base, such as LiOH or NaOH, in an appropriate solvent, such as tetrahyd rofuran / methanol / water. The remaining chloride or bromide is optionally removed by hydrogenolysis using a catalyst, such as carbon palladium or Raney nickel, in an appropriate solvent, such as water to provide 9. Alternatively, in the case of P '= benzyl, the deprotection of the benzyl ester and the removal of the remaining chloride or bromide can be carried out simultaneously using palladium on carbon in a suitable solvent, such as tetrahydrofuran / ethanol to give the 9. the acid is then coupled to the amine 5 using typical amino acid coupling procedures, such as BOP or PyBOP, in a suitable solvent, such as N, N-dimethylformamide, and a base, such as di-isopropylethylamine (DIEA), followed by optional removal of Ra, R, Rc, in the case of Ra and Rb = tert-butyloxycarbonyl (Boc) and Rc = hydrogen. The Boc groups are removed by treatment with an acid, such as trifluoroacetic acid or hydrochloric acid, in an appropriate solvent, such as methylene chloride or dioxane to provide the objective compound 10.

Compounds where X is -CH = NR1X- or NR11 can be formed using the steps shown in Scheme 3.

Scheme 3 Acid 9 (as synthesized in Scheme 2) is then coupled to an amino alcohol 11 using standard amino acid coupling procedures, such as BOP or PyBOP in a solvent such as N, N-dimethylformamide, and a base, such as DIEA , to form compound 12. The corresponding aldehyde 13 is synthesized using routine procedures for the oxidation of alcohols (see for example Carey, FA, and Sunderberg, RJ, Advanced Organic Chemistry, Part B: Reactions and Synthesis, 3rd edition, Plenum Press, New York (1990)) such as Swern oxidation (Mancuso, AJ et al., Journal of Organic Chemistry 3329 (1976)) pyridinium chlorochromate (Corey, EJ and Suggs, JW Tetrahedron Letters 2647 (1975) pyridinium dichromate (Corey , EJ and Schmidt, G. Tetrahedron Letters 399 (1979)), or pyridine sulfur trioxide / dimethylsulfoxide complex (Tetrahedron Letters 28: 1603 (1987)).

The aldehyde 13 is then converted to amidinohydrazine 14 using standard conditions, for example, treatment with an aminoguanidine or a substituted aminoguanidine, such as aminoguanidine or 2-hydrazinoimidazoline, optionally in the presence of an acid such as nitric acid, hydrogen chloride, or hydrogen bromide, in a suitable solvent, for example, ethanol or methanol, which, in addition, may contain other solvents such as dichloromethane or tetrahydrofuran.

The conversion of amidinohydrazone 14 to aminoguanidine 15 is carried out under reduced conditions well known in the art, for example, lithium borohydride in an appropriate solvent such as tetrahydrofuran or methanol at various temperatures up to reflux temperature. As an alternative method, catalytic hydrogenation with palladium on carbon catalyst can be employed.

Ra, Rb, and Rc in compounds 14 and 15 can optionally be removed using conditions well known in the art. In the case of Ra and Rb = tert-butyloxycarbonyl (Boc) and Rc = hydrogen, the Boc groups are removed by treatment with acid, such as trifluoroacetic acid or hydrochloric acid, in an appropriate solvent, such as methylene chloride or dioxane to provide compounds 16 and 17 respectively.

The pharmaceutically acceptable salts of the compounds of Formula I (in the form of water soluble or dispersible products in water or oil) include the conventional non-toxic salts or the quaternary ammonium salts which are formed, for example, from inorganic or organic acids or bases. . Examples of such acid addition salts include acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camforate, camphorsulfate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, fumarate, glucoheptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate. , hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, laureate, ethanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oxalate, pamoate, pectinate, persulfate, 3-phenylpropionate, picrate, pivalate, propionate, succinate, sulfate, tartrate, thiocyanate , tosylate, and undecanoate, the base salts include ammonium salts, alkali metal salts such as sodium and potassium salts, earth metal alkano metal salts such as clathium or magnesium salts, salts with organic bases such as dicyclohexylamine salts, N -methyl-glucamine, and salts with amino acids such as arginine, lysine, and the like. Also, basic groups containing nitrogen can be quaternized with agents such as lower alkyl halides, such as methyl, ethyl, propyl and butyl chloride, bromide and iodide; dialkyl sulfates such as dimethyl, diethyl, dibutyl; and diamyl sulfates, long chain halides such as decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides, aralkyl halides such as benzyl and phenethyl bromides and others. Preferred acids for the formation of acid addition salts include HCl and acetic acid.

The compounds of the present invention represent a novel class of potent inhibitors of metallo, acid, thiol and serine proteases. Examples of the serine proteases inhibited by the compounds within the alkaline of the invention include neutrophil lecocyte elastase, a proteolytic enzyme involved in the pathogenesis of emphysema; chymotrypsin and trypsin, digestive enzymes; pancreatic elastase, and cathepsin G, a chymotrypsin-like potease also associated with leukocytes; thrombin and factor Xa, proteolytic enzymes in the blood coagulation pathway. The inhibition of thermolysin, a metalloprotease, and pepsin, an acid protease, also contemplates the use of the compounds of the present invention. The compounds of the present invention are preferably used to inhibit trypsin-like proteases.

For their end-use applications, the potency and other biochemical parameters of the inhibitory characteristics of the enzyme of the compounds of the present invention are already met by standard biochemical techniques well known in the art. For example, an end-use application of the compounds that inhibit cimothyrosine and trypsin is in the trent of pancreatitis. The current dose ranges for their end-use applications may, of course, depend on the nature and severity of the disease status of the patient or animal to be treated, as determined by the attending specialist. A range of useful doses is expected to be around 0.01 to 10 mg per kg per day for an effective therapeutic effect.

The compounds of the present invention that are distinguished by their ability to inhibit thrombin can be employed for a number of therapeutic purposes. As thrombin inhibitors, the compounds of the present invention inhibit thrombin production. Therefore, these compounds are useful for the trent or prophylaxis of those characterized by abnormal veins or arterial thrombosis that involves both the production and the action of thrombin. These conditions include, but are not limited to, deep vein thrombosis, disseminated intravascular coagulation that occurs during septic shock, viral infections, and cancer.; myocardial infarction; attack; deviation of the coronary artery; fiber formation in the eye; hip replacement; and thrombus formation resulting from both thrombotic therapy or percutaneous transluminal coronary angioplasty (PCTA). Other uses include the use of said thrombin inhibitors as anticoagulants either inserted into or physically bound to the materials used in the manufacture of devices that are used in the collection of blood, blood circulation, and blood storage, such as catheters, blood dialysis machines, blood collection syringes and tubes, and blood lines. The compounds of the present invention can also be used as an anticoagulant in extracorporeal blood circuits.

Grafted metals have been shown to reduce restenosis, but are thrombogenic. One strategy to reduce the thrombigenicity of these grafts is to coat, insert, absorb or covalently bind to an agent that inhibits thrombin to the graft surface. The compounds of the present invention can be used for this purpose. The compounds of the invention can be linked to, or inserted into, soluble and / or biodegradable polymers and subsequently coated on graft materials. Such polymers can include polyvinylpyrrolidone, polyhydroxy-propylmethacrylamide-phenol, polyhydroxyethyl-aspartamide-phenol, or polyethylene-oxide-polylysine substituted with palmitoyl residues, polylactic acid, polyglycolic acid, polylactic and polyglycolic acid copolymers, polyepsilone caprolactone, polyhydroxy butyric acid, polyorthoesters , polyacetals, polydihydropyrans, polycyanoacrylates and amphipathic block copolymers or crosslinked hydrogels. See European Application 761 251, European Application 604,022, Canadian Patent 2,164,684 and PCT Published Applications WO 96/11668, WO 96/32143 and WO 96/38136.

By virtue of the effects of thrombin on host-type cells, such as smooth muscle cells, endothelial cells and neutrophils, the compounds of the present invention find further use in the treatment or prophylaxis of adult respiratory distress syndrome; inflammatory answer; healing of wounds; reperfusion damage; arterosclerosis; and restenosis after an injury such as spherical angioplasty, atherectomy, and arterial graft location.

The compounds of the present invention may be useful in the treatment of neoplasty and metastasis as well as in neurodegenerative diseases, such as Alzheimer's and Parkinson's disease.

When used as thrombin inhibitors, the compounds of the present invention can be administered in an effective amount within the dose range of from about 0.1 to about 500 mg / kg, preferably from 0.1 to 10 mg / kg of body weight, in a simple regimen or divided into 2-4 daily doses.

When used as thrombin inhibitors, the compounds of the present invention can be used in combination with thrombolytic agents such as tissue plasminogen activator, streptokinase, and urokinase. Additionally, the compounds of the present invention may be used in combination with other antithrombotic or anticoagulant drugs such as, but not limited to, thromboxane receptor antagonist fibrinogen antagonists.

Thrombin inhibitors can be coupled with soluble polymers as carriers of target drugs. Such polymer may include polyvinyl pyrrolidone, pyran copolymer, polyhydroxy-propyl methacrylamide-phenol, polyhydroxyethyl-aspartamide-phenol, or polyethylene oxide-polylysine substituted with palmitoyl residues. Additionally, thrombin inhibitors can be coupled to a class of biodegradable polymers useful for performing a controlled release of a drug, for example, polylactic acid, polyglycolic acid, polylactic and polyglycolic acid copolymers, polyepsilone caprolactone, polyhydroxy butyric acid, polyorthoesters , polyacetals, polydihydropyrans, polycyanoacrylates and cross-linked or amphiphmatic block copolymers of hydrogels.

Human leukocyte elastase is released by polymorphonuclear leukocytes at sites of inflammation and thus is a contributing cause to a number of disease states. The compounds of the present invention are expected to have an anti-inflammatory effect useful in the treatment of gout, rheumatoid arthritis and other inflammatory diseases, and in the treatment of emphysema. The inhibitory properties of leukocyte elastase of the compounds of the present invention are determined by the method described below, cathepsin G has also been implicated in disease states of arthritis, gout and emphysema, and, in addition, glomerulonephritis and infestations lung infections caused by lung infections. In their end-use applications, the inhibitory properties of the enzyme of the compounds of Formula I are already successful by standard biochemical techniques that are well known in the art.

The inhibitory properties for cathepsin G of the compounds within the scope of the present invention are determined by the following method. A partially purified human cathepsin G preparation is obtained by the procedure of Baugh et al., Bi or chesmi try 15: 836 (1979). The leukocyte granules are a major source for the preparation of leukocyte elastase and cathepsin G (activity resembling chymotrypsin). The leukocytes are lysed and the granules are isolated. The leukocyte granules are extracted with 0.20 M sodium acetate, pH 4.0, and the extracted ones are again dialyzed with 0.05 M Tris buffer, pH 8.0 containing 0.05 M NaCl overnight at 4 ° C. A fraction of the protein is precipitated during dialysis and isolated by centrifugation. This fraction contains more of the cimotrypsin-like activity of leukocyte granules. The specific substrates are prepared for each enzyme, namely N-Sus-Ala-Ala-Pro-Val-p-nitroanilide and Su-Ala-Ala-Pro-Phe-p-nitroanilide. The latter is not hydrolyzed by leukocyte elastase. Enzyme preparations are assayed in 2.00 mL of Hepes 0.10 M buffer solution, pH 7.5, containing 0.50 M NaCl, 10% dimethylsulfoxide and Su-Ala-Ala-Pro-Phe-p-nitroanilide 0.0020 M as a substrate. Hydrolysis of the p-nitroanilide substrate is observed at 405 nm and at 25 ° C.

The range of useful doses for the application of compounds of the present invention as inhibitors of neutrophil elastase and as inhibitors of cathepsin G depends on the nature and severity of the disease state, as determined by the attending specialist, with a range from 0.01 to 10 mg / kg of body weight, per day, being useful for the aforementioned disease states.

The compounds of the present invention that inhibit urokinase or plasminogen activator are potentially useful in the treatment of excess cell growth disease states. Such compounds of the present invention may also be useful in the treatment of benign prostatic hypertrophy and prostatic carcinoma, the treatment of psoriasis, and as abortifacients. For their end-use applications, the potency and other biochemical parameters of the inhibitory characteristics for the enzyme of the compounds of the present invention are already successful by standard biochemical techniques well known in the art. The current dose ranges for this application depend on the nature and severity of the disease status of the patient or animal to be treated as determined by the attending specialist. It is expected that a general dose range is around 0.01 to 10 mg per kg per day for an effective therapeutic effect.

Additional uses for the compounds of the present invention include the analysis of commercial reactive enzymes for concentration in active sites. For example, chymotrypsin is supplied as a standard reagent for use in the clinical quantification of cyphotrypsin activity in pancreatic juices and faeces. Such analyzes are diagnostic for gastrointestinal and pancreatic disorders. Pancreatic elastase is also commercially available as a reagent for the quantification of al-antitrypsin in plasma. Al-antitrypsin plasma increases its concentration during the course of several inflammatory diseases and deficiency in al-antitrypsin is associated with the increased incidence of lung disease. The compounds of the present invention can be used to increase the acuity and reproducibility of these assays by titrammetric standardization of the commercial elastase supplied as a reagent. See, U.S. Patent No. 4,499,082.

The activity of the protease in certain proteins extracted during the purification of particular proteins is a recurrent problem that can be complicated and compromises the results of the protein isolation procedures. Certain proteases present in such extracts can be inhibited during the purification steps by the compounds of the present invention, which hermetically bind to various ptoteolytic enzymes.

The pharmaceutical compositions of the invention can be administered to any animal that can experience the beneficial effects of the compounds of the invention. Among the first of such animals are humans, although the invention is not intended to be limiting.

The pharmaceutical compositions of the present invention can be administered by any means that accomplishes its intended purpose. For example, administration can be by parenteral, subcutaneous, intravenous, intramuscular, intraperitonal, transdermal, buccal, or ocular routes. Alternatively, or concurrently, the administration may be by the oral route. The dose administered will depend on the age, health, and weight of the recipient, type of concurrent treatment, if any, frequency of treatment, and nature of the desired effect.

In addition to the pharmacologically active compounds, novel pharmaceutical preparations may contain pharmaceutically acceptable carriers comprising excipients and auxiliaries that facilitate processing of the active compounds into preparations that can be used pharmaceutically.

The pharmaceutical preparations of the present invention are made so that they are, per se, known, for example, by means of conventional blends, granulation, pill making, dissolving, or lyophilization process. In this way, pharmaceutical preparations for oral use can be obtained by combining the active compounds with solid excipients, optionally grinding the resulting mixture and processing the mixture of granules, then adding appropriate auxiliaries, if desired or necessary, to obtain tablets or dragees of core .

Suitable excipients are, in particular, fillers such as saccharides, for example, lactose or sucrose, mannitol or sorbitol, cellulose and / or calcium phosphate preparations, for example, tricalcium phosphate or calcium hydrogen phosphate, as well as binders. , such as, starch paste, using, for example, corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth, methyl cellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, and / or polyvinyl pyrrolidone. If desired, disintegrating agents may be added, such as the above-mentioned starches and also carboxymethyl starch, cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof, such as sodium alginate. The auxiliaries are, all of the above, flow regulating agents and lubricants, for example, silica, talc, stearic acid or salts thereof, such as, magnesium stearate or calcium stearate, and / or polyethylene glycol. Core pellets are provided with appropriate coatings that, if desired, are resistant to gastric juices. For this purpose, solutions may be used, which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, polyethylene glycol, and / or titanium dioxide, lacquered solutions and appropriate organic solvents or solvent mixtures. In order to produce coatings resistant to gastric juices, solutions of appropriate cellulose preparations, such as acetylcellulose phthalate or hydroxypropylmethyl cellulose phthalate, are used. Dry fillers or pigments may be added to the tablets or dragee coatings, for example, for identification in order to characterize the combinations of the doses of active compounds.

Other pharmaceutical preparations that can be used orally include soft-setting capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. Soft-fit capsules can contain the active compounds in the form of granules which can be mixed with fillers such as lactose, binders such as starches, and / or lubricants such as talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the active compounds are preferably dissolved or suspended in suitable liquids, such as fatty oils or liquid paraffin. In addition, stabilizers can be added.

Formulations suitable for parenteral administration include aqueous solutions of the active compounds in a water soluble form, for example, water soluble salts, alkaline solutions and complexes including cyclodextrin. Especially preferred alkali salts are the ammonium salts prepared, for example, with Tris, choline hydroxide, Bis-Tris propane, N-methylglucamine, or arginine. One or more modified or unmodified cyclodextrins may be employed to stabilize and increase the water solubility of the compounds of the present invention. Cyclodextrins useful for this purpose are described in U.S. Pat. Nos. 4,727,064, 4, 764, 604 and 5,024, 998.

In addition, suspensions of the active compounds may be administered as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils, for example, sesame oil, or synthetic fatty acid esters, for example ethyl oleate or triglycerides or polyethylene glycol-400 (the compounds are soluble in PEG-400). Aqueous injection suspensions may contain substances that increase the viscosity of the suspension, for example, sodium carboxymethyl cellulose, sorbitol, and / or dextran. Optionally, the suspension may also contain stabilizers.

The compounds of Formula I can be labeled with radioactive iodine using an exchange reaction. The exchange of all hot by cold iodine is well known in the art. Alternatively, a compound labeled with radioactive iodine can be prepared from the corresponding bromine compound by means of a tributyltin intermediate. See, U.S. Patent No. 5,122,361, incorporated herein by reference.

The present invention also includes compositions that are useful for in vitro imaging of thrombi in a mammal, wherein the compositions comprise a compound of Formula I in a complex with a radioactive atom.

For compounds of Formula I, appropriate radioactive atoms include Co-57, Cu-67, Ga-67, Ga-68, Ru-97, Tc-99m, In-lU In-113m, Hg-197, Au- 198, and Pb-203. Some radioactive atoms have superior properties for use in radiochemical imaging techniques. In particular, technetium-99m (Tc-99m) is an ideal radioactive atom for imaging due to its nuclear properties. This is a gamma emitter and has a simple proton energy of 140 ke V, a half-life of about 6 hours, and is now available from a Mo-99 / Tc-99 generator. Rhenium-186 and -188 also have gamma emission that allows an image to be observed. Preferred compositions contain the radioactive atom Tc-99m.

The compositions of the present invention are conveniently prepared by making a complex of a compound of Formula I with radioisotopes that are suitable for external detection. The emitters gamma, indium-lllm and tecnio-99m, are preferred as radioactive atoms because they are detectable with a gamma camera and have a favorable half-life in vi vo.

The compounds of Formula I can be labeled by any of the many techniques known in the art to provide a composition of the present invention. For example, these compounds can be labeled through a chelating agent such as diethylenetriaminepentaacetic acid (DTPA) or metallothionein, both of which can be covalently linked to the compound of Formula I.

In general, the compositions of the present invention contain technetium-99m are prepared by forming an aqueous mixture of tecnetium-99m and a reducing agent and a water-soluble ligand, and then contacting the mixture with a compound of the present invention represented by Formula I. For example, the imaging compounds of this invention are made by reacting technetium-99m (in an oxidized state) with the compounds of the present invention having chelating means in the presence of a reducing agent to form a stable complex between tecnetium-99m in a reduced state (IV or V valence state).

One embodiment of the composition of the present invention is prepared by labeling a compound of Formula I having a chelating medium DTPA with tecnet io-99m. This can be performed by combining a predetermined amount (such as 5 μg to 0.5 mg) of the compound of the present invention with an aqueous solution containing citrate buffer solution and tin reducing agent, then adding freshly eluted sodium pertechnetate containing a predetermined level of radioactivity (like 15 mCi). After allowing an incubation of the mixture at room temperature, the reaction mixture is filled in a protected syringe through a sterile filter (0.2-0.22 microns), then dispensed in a 0.9% saline for injection, if desired .

Another embodiment of the compositions of the present invention is prepared by labeling a compound of Formula I having metallothionein chelating means with technetium-99m. This can be accomplished by combining pertechnetate-99m aqueous sodium with aqueous tin glycoheptonate to form a soluble complex of technetium-99m (in reduced state) with two glucoheptonate molecules, then combining this solution with a compound of Formula I having linked metallothionein to this. After the mixture is incubated for a period of time and under conditions that allow an exchange of the technetium-99m of the glucoheptonate complex for the metallothionein of the compound of Formula I, the technetium-labeled composition of the present invention is formed.

The source of tecnetium-99m is preferably soluble in water. The preferred sources are alkaline and alkaline earth metal pertechnetate (Tc04-). Technetium-99m is most preferably obtained in the form of fresh sodium pertechnetate from a sterile technetium-99m generator (as of a conventional Mo-99 / Tc-99m generator). However, any other physiologically acceptable tectherium-99m source can be used.

The reducing agents for use in the method are physiologically acceptable to reduce the technetium-99m from its oxidized state to the valence state IV and V or to reduce the rhenium from its oxidized state. Reductive agents that can be used with tin chloride, tin fluoride, tin glucoheptonate, tin tartrate, and sodium dithionite. Preferred agents are tin reductive agents, especially tin chloride or tin glucoheptonate. The amount of reducing agent is an amount necessary to reduce tecnetium-99m to provide binding to the chelating means of a compound of Formula I in its reduced radioisotope state. For example, tin chloride (SnCl2) is the reductive agent and can be used in the range of 1-1,000 μg / mL. Especially preferred concentrations are around 30-500 pg / mL.

Complexes of citric acid with tecnetium-99m rapidly form a complex of technetium-99m-citrate. During contact with a compound of Formula I, the substantially quantitative transfer of tecnetium-99m from its citrate complex to the chelating media of the compound of Formula I is carried out rapidly and under mild conditions. The amount of citric acid (such as sodium citrate) can be in the range from about 0.5 mg / ml to the maximum amount soluble in the medium. Preferred amounts of citric acid are in the range of 15 to 30 μg / ml.

The amount of compound of Formula I having a chelating medium may be in the range from 0.001 to about 3 mg / mL, preferably about 0.017 to about 0.15 mg / mL. Finally, technetium-99m in the form of pertechnetate can be used in amounts of preferably about 1-50 mCi. The amount of mCi per gram of the compound of the present invention is preferably about 30-150.

The reaction between the compound of Formula I and the metal ion-transfer ligand complex is preferably carried out in an aqueous solution at a pH at which the compound of Formula I is stable. By "stable" it means that the compound remains soluble and retains its inhibitory activity against a-thrombin. Usually, the pH for the reaction will be from around 5 to 9, the preferred pH being above 6-8. The technetium-99m-citrate complex and a compound of Formula I are incubated, preferably at a temperature of from about 20 ° C to about 60 ° C, more preferably from about 20 ° C to about 37 ° C, by a sufficient amount of time to allow the transfer of the metal ion of the citrate complex to the chelating means of the compound of Formula I. Generally, less than one hour is sufficient to complete the reaction transfer under these conditions.

Alternative compositions of the present invention include a labeled In-111 compound of the present invention.

The present invention also includes compositions of the compounds of the present invention that are useful for thrombus in vitro imaging of a mammal, comprising a compound represented by Formula I in complex to a paramagnetic atom.

Preferred paramagnetic atoms are divalent or trivalent ions of elements with an atomic number of 21 to 29, 42, 44 and 58 to 70. Suitable ions include chromium (III), manganese (II), iron (III), iron (II) ), cobalt (II), nickel (II), copper (II), praseodymium (II I), neodymium (III), samarium (III) and ytterbium (III). Due to their very strong magnetic moments, gadolinium (111), terbium (III), disoprosium (III), holmium (III), and erbium (III) are preferred. Galdolinium (III) is especially preferred for the paramagnetic atom.

The compositions of the present invention can be prepared by combining a compound of Formula I with a paramagnetic atom. For example, the metal oxide or metal salt (eg, nitrate, chloride or sulfate) of an appropriate paramagnetic atom is dissolved or suspended in a medium comprising water and an alcohol, such as methyl, ethyl or isopropanol alcohol. . This mixture is added to a solution of an equimolar amount of the compound of Formula I in a similar aqueous medium and stirred. The reaction mixture is heated moderately until the reaction is complete. The insoluble compositions formed can be isolated by filtration, although the soluble compositions can be isolated by evaporation of the solvent. If the acid groups in the chelating media are still present in the composition of the present invention, the inorganic or organic bases, and even the amino acids, can be added to convert the acid complex to a neutral complex to facilitate isolation or purification of the composition homogeneous Organic bases or basic amino acids can be used as neutralizing agents, as well as inorganic bases such as sodium, potassium or lithium hydroxides, carbonates or bicarbonates.

The present invention also includes diagnostic compositions that are useful for in vitro imaging of thrombi in a mammal, comprising a pharmaceutically acceptable carrier and a diagnostically effective amount of compositions derived from the compounds of Formula I.

The "diagnostically effective amount" of the composition required as a dose will depend on the route of administration, the type of mammal to be treated, and the physical characteristics of the specific mammal under consideration. These factors and their relationship to determine their dose are well known by specialists skilled in medical diagnostic techniques. Also, the diagnostically effective amount and method of administration can be adapted to perform effective optimization, but they depend on such factors as weight, diet, current medication and other factors that those skilled in medical techniques recognize. At any point, the dose for the image should be sufficient to detect the presence of the imaging agent at the site of a thrombus in question. Typically, the radiological image requires that the dose provided by the position of the pharmaceutical composition of the present invention be about 5 to 20 μCi, preferably about 10 μCi. The magnetic resonance image requires that the dose provided be around 0.001 to 5 mmol / kg, preferably about 0.005 to 0.5 mmol / kg of a compound of Formula I in complex with a paramagnetic atom. In both cases, it is known in the art that the current dose depends on the thrombus site.

"Pharmaceutically acceptable carriers" for in vi vo use are well known in the pharmaceutical art, and are described, for example, in Rem in gt on 's Pha rma ce u t i ca l Sci in ces, Mark Publishing Co. (A.R. Gennaro editores, 1985). The pharmaceutical compositions of the present invention can be formulated with a pharmaceutically acceptable carrier to provide sterile solutions or suspensions for administration by injection. In particular, the injections can be prepared in convenient forms, either in liquid solutions or suspensions, solid forms suitable for solutions or suspensions in a liquid before injection, or as emulsions. Suitable excipients are, for example, water, saline, dextrose, mannitol, lactose, lecithin, albumin, sodium glutamate, cysteine hydrochloride, or the like. In addition, if desired, the injectable pharmaceutical compositions may contain minor amounts of non-toxic auxiliary substances, such as wetting agents, pH regulating agents, and the like. If desired, preparations that increase absorption (e.g., liposomes) can be used.

The present invention also encompasses diagnostic compositions prepared for storage or administration. These can, additionally, contain preservatives, stabilizers and dyes. For example, sodium benzoate, sorbic acid and p-hydroxybenzoic acid esters can be added as preservatives. Id, at 1449. In addition, antioxidants and suspending agents can be used.

The imaging methods of the present invention also offer several advantages over prior imaging techniques for the detection or observation of the presence, size, regression or increase of a thrombus. In particular, the present invention provides compounds, compositions and diagnostic compositions that have been designed to bind extremely tightly to thrombin associated with a thrombus and thereby reduce the "background" due to radioactivity or circulating paramagnetism as a consequence of the imaging agent. not linked Additionally, the in vivo imaging by coronary injection of the compounds, compositions or diagnostic compositions of the present invention is expected to be at least instantaneous, since these imaging agents saturate the thrombin bound to the thrombus immediately.

Accordingly, the present invention also includes methods for in vi ve image of a thrombus in a mammal, comprising the steps of: (1) administering to the mammal a diagnostically acceptable amount of a compound, composition or diagnostic composition of the present invention and (2) detect a thrombus in the blood flow.

The term "image in vi vo" as used herein, refers to methods of detecting a thrombus in a mammal, as well as the observation of the size, location and number of thrombi in a mammal, as well as their dissolution or growth. of the thrombus.

By using the compounds, compositions or diagnostic compositions in vi by this method, "administration" is performed parenterally, either in a systemic manner or to the local objective. Systemic administration is carried out by injecting the compounds, compositions or diagnostic compositions of the present invention into a convenient and accessible vein or artery. This includes, but is not limited to, administration through the vein of the cubutal ankle. Administration to the local target is performed by injecting the compounds, compositions or diagnostic compositions of the present invention in a manner close to the flow of a vein or artery suspected to contain thrombi distant from the site of injection. This includes, but is not limited to, directing the injection into the vasculature of the coronary artery for the image of the coronary thrombus, in the carotid artery for a thrombus image in the cerebral vasculature, or in a pedal vein for an image of thrombosis in the deep vein of the leg.

Also, the manner of delivery of a composition of the present invention to the thrombus site is considered within the scope of the term "administration". For example, a compound represented by Formula I having chelating agents linked to it can be injected into the mammal, followed some time later by the radioactive atom thereby forming in vi ve in the thrombus syrin the composition comprising the compound of the invention. formula in complex with the radioactive atom. Alternatively, a composition comprising the compound of the formula in a complex with the radioactive atom can be injected into the mammal.

The "diagnostically effective amount" of the compounds, compositions or diagnostic compositions used in the methods of the present invention, as previously mentioned, depend on the route of administration, the type of mammal to be treated, and the physical characteristics of the specific mammal under treatment. These factors and their relationship to determine this dose are well known to specialists skilled in medical diagnostic techniques. In any case, the dose for visual image should be sufficient to detect the presence of the imaging agent at the site of a thrombus in question. Typically, the radiological image may require that the dose provided by the diagnostic composition of the present invention be about 5 to 20 μCi, preferably about 10 μCi. The magnetic resonance image may require that the dose provided by the diagnostic composition be about 0.001 to 5 mmol / kg, preferably about 0.005 to 0.5 mmol / kg of a compound of Formula I in complex with a paramagnetic atom. In both cases, it is known in the art that the current dose depends on the thrombus site.

The detection of the thrombus by the image is made possible by the presence of radioactivity or paramagnetic atoms located in such thrombi.

The radioactive atoms associated with the compositions and diagnostic compositions of the present invention are preferably imaged using radiation detection means capable of detecting gamma radiation, such as a gamma camera or the like. Typically, radiation imaging cameras employ a conversion medium (where the high-energy gamma ray is absorbed, displacing an electron that emits a photon during its return to the orbital state), photoelectric detectors placed in a space detection chamber (to determine the position of the emitted photons), and a circuit to analyze the photons detected in the camera and produce an image.

The paramagnetic atoms associated with the compositions and diagnostic compositions of the present invention are detected in magnetic resonance imaging (MRI) systems. In such systems, a strong magnetic field is used to linearize the nuclear spin vectors of the atoms in the patient's body. The field is disturbed by the presence of paramagnetic atoms located in a thrombus and an image of the patient in the nucleus is read back to its equilibrium alignments.

The following examples are illustrative, but not limiting, of the methods and compositions of the present invention. Other appropriate modifications and adaptations of the variety of conditions and parameters normally found and obvious to those skilled in the art are within the spirit and scope of the invention.

Example 1 L- Trifluoroacetate. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (phenethylamino) -pyrazinone to . N- [2-benzyloxycarbonylamino) atoxy] phthalimide To a solution of benzyl N- (2-hydroxyethyl) carbamate (5.9 g, 30 mmol), N-hydroxy phthalimide (4.9 g, 30 mmol), and triphenylphosphine (7.9 g, 30 mmol) in tetrahydrofuran (100 mL) were added. he added diethyl azodicarboxylate (5.2 g, 30 mmol). The reaction mixture was stirred at room temperature overnight. Ethyl acetate (200 L) was added, washed with saturated NaHCO 3 (2x100 mL) and brine (100 mL), dried over Na 2 SO 4, and filtered. After evaporating the filtrate, the residue was purified by flash column chromatography (methylene chloride for 4% ethyl acetate in methylene chloride) to give the title compound as a white solid (9.3g, 91%).

X H-NMR (300 MHz, CDCl 3) d 7.84 (m, 2 H), 7.37 (m, 5 H), 5.97 (brs, 1 H), 5.14 (s, 2 H), 4.27 (t, J = 4.9 Hz, 2 H), 3.51 (q, J = 5.2 Hz, 2H). b. 2- (benzyloxycarbonylamino) ethoxyamine To a solution of N- [2- (benzyloxycarbonylamino) ethoxy] phthalimide (1.36g), 4.0 mmol), as prepared in the previous step, in ethanol (20 mL) and tetrahydrofuran (20 mL) was added 40% methylamine (2.0 mL, 25 mmol) and stirred at room temperature for 1 hour. After evaporating the solvent, the residue was passed through silica gel (3: 1 ethyl acetate: hexane for ethyl acetate) to give the title compound as a white solid (800 mg, 95%). X H-NMR (300 MHz, CDCl 3) d 7.36 (m, 5 H), 5.47 (br s, 2 H), 5.21 (br s, 1 H), 5.10 (s, 2 H) 3.72 (t, J = 5.0 Hz, 2 H) , 3.44 (q, J = 5.0 Hz, 2H). c. [N, N '-Di (tert-butoxycarbonyl)] 2- (benzyloxycarbonylamino) ethoxyguanidine Boc To a solution of 2- (benzyloxycarbonylamino) ethoxyamine (780 mg, 3.7 mmol), as prepared in the previous step, in N, N-dimethylformamide (20 mL) was added [N, N '-di (tert-butoxycarbonyl )] amidinopyrazole (1.25 g, 4.0 mmol). The mixture was stirred at room temperature overnight, and the solvent was evaporated under high vacuum. The residue was purified by flash column chromatography (0-5% ethyl acetate in methylene chloride) to give the title compound as a colorless oil (1.55g, 93%). XH-NMR (300 MHz, CDC13) d 9.08 (s, 1H), 7.67 (s, lH), 7.33 (m, 5H), 6.21 (br s, 1H), 5.21 (br s, 1H), 5.11 (s) , 2H), 4.12 (t, J = 4.8 Hz, 2H), 3.54 (q, J = 4.9 Hz, 2H), 1.49 (s, 9H), 1.46 (s, 9H). d. [N, N '-Di (tert-butoxycarbonyl)] 2 -aminoethoxyguanidine A mixture of [N, N '-di (tert-butoxycarbonyl)] 2-benzyloxycarbonylamino) ethoxyguanidine (730 mg, 1.5 mmol), as prepared in the previous step, 10% palladium on carbon (70 mg) in ethanol ( 20 mL) and tetrahydrofuran (20, mL) was hydrogenated under hydrogen (sphere) for 30 minutes. The catalyst was removed by filtration through Celite and the filtrate was concentrated in vacuo. The residue was purified on a Waters Sep-Pak (10g, 95: 5 methylene chloride: methanol saturated with ammonia) to give the title compound as a colorless oil (290mg, 61%) H-NMR (300 MHz, CDC13) d 9.08 (br s, 1H), 4.08 (t, J = 5.2 Hz, 2H), 2.99 (q, J = 5.1 Hz, 2H), 1.50 (s, 9H), 1.48 (s, 9H). and. benzyl-N- (1-cyanoethyl) glycine hydrochloride Trimethylsilyl cyanide (TMSCN) was added (4.0 mL, 30 mmol) carefully to a stirred solution of free base benzyl glycine (5.0 g, 30mmol) and acetaldehyde (1.7 mL, 30mmol) in methylene chloride (15 mL) under an argon atmosphere. After 15 hours, the volatile components were removed in vacuo, and the residue was dissolved in ethyl acetate (200 L), washed with brine, (100 mL), dried (Na 2 SO 4) and evaporated to an oil. The oil was redissolved in ether (30 mL), and IMM HCl in ether (33 mL) was added dropwise to give the title compound (6.60 g, 100%) as an off white crystalline precipitate. mp: 137-138 ° C; ^ -N ROOO MHz, CD3OD) d 7.31-7.48 (m, 5H), 5.32 (s, 2H), 4.68 (q, J = 7.0 Hz, 1H), 4.22 (s, 2H), 1.73 (d, J = 7.1 Hz, 3H); Cl MS m / z = 192 (M + H). Analysis calculated for C_.2H14N202. HCl: C. 56.49; H, 5.95; N, 11.00 Found: C, 56.32; H, 5.88, N, 10.89. l-benzyloxycarbonylmethyl-3,5-dichloro-6-methylpyrazinone A stirred mixture of oxalyl chloride (5.3 mL, 60 mmol) and benzyl-N- (1-cyanoethyl) glycine hydrochloride (3.82 g, 15 mmol), as prepared in the preceding step, in 1,2-dichlorobenzene (30 ml) was heated to 100 ° C overnight. The solvent was evaporated in vacuo and the residue was purified by flash column chromatography (20-30% ethyl acetate in hexane) to give a solid. 10% ethyl acetate in hexane (100 mL) was added and the solid was collected to give the title compound as an orange crystalline solid (2.7g, 55%). ^ -NMROOO MHz, CDC13) d 7.38 (m, 5H), 5.24 (s, 2H), 4.89 (s, 2H), 2.34 (S, 3H). g. 3- (2-phenethylamino) -5-chloro-6-methyl-1- (benzyloxycarbonylmethyl) -pyrazinone Phenethylamine (1.5 mL, 12 mmol) was added to a stirred solution of 1-benzyloxycarbonylmethyl-3,5-dichloro-6-methypypyrazinone (1.31 g, 4.0 mmol), as prepared in the previous step, in ethyl acetate ( 10 mL) and the resulting mixture was heated at reflux temperature under nitrogen. After 2 hours, the reaction mixture was cooled, diluted with methylene chloride (100mL), washed with 5% citric acid (2x50mL), brine (50mL) dried (Na2SO4) and filtered. After evaporating the filtrate in vacuo, the solid was collected and washed with 20% ethyl acetate in hexane to give the title compound as a crystalline solid (1.5 g, 91%). ^ -NMROOO MHz, CDC13) d 7.22-7.38 (m, 10H), 6.11 (t, J = 5.8 Hz 1H), 5.21 (s, 2H), 4.79 (s, 2H), 3.68 (q, J = 7.0 Hz, 2H), 2.93 (t, J = 7.0 Hz, 2H), 2.22 (s, 3H). h. Mixture of 3- (2-phenethylamino) -6-methyl-1-carboxymethylpyrazine and 3- (2-phenethylamino) -5-chloro-6-meth i 1- 1-carboxymethylpyrazinone A mixture of 3- (2-phenethylamino) -5-chloro-6-methyl-1- (benzyloxycarbonylmethyl) -pyrazinone (412 mg, 1.0 mmol), as prepared in the preceding step, and palladium on carbon (10%, 100mg) in tetrahydrofuran (10 mL) and ethanol (40 mL) was stirred under hydrogen (sphere) for two days. The reaction was filtered through celite, the alkaline flux was washed with ethanol, and the filtrate was evaporated in vacuo to give a mixture of 3- (2-phenethylamino) -6-methyl-1-carboxymethylpyrazinone and 3- (2 -phenethylamino) -5-chloro-6-met il-1 -carboxy-ethylpyrazinone as a slightly red solid (210 mg) which was used directly in the next step without further purification. i. l-. { N- [2- (N ', N "-Bis (tert-butoxycarbonyl) amidinoaminooxy) ethyl] amino.} Carbonyl methyl-6-methyl-3- (enedylamino) -pyrazinone and 1- { N- [2 - (N ', N "-Bis (tert-butoxycarbonyl) amidinoaminooxy) ethyl] amino} carbonyl methyl-5-chloro-6-methyl-3- (phenethylamino) -pyrazinone To a solution of the mixture of 3- (2-phenethylamino) -6-methyl-1-carboxymethylpyrazinone and 3- (2-phenethylamino) -5-chloro-6-methyl-1-carboximetylpyrazinone (200 mg) , as prepared in the preceding step, [N, N '-di (tert-butoxycarbonyl)] 2-aminoethoxyguanidine (225 mg, 0.7 mmol), as prepared in step d of Example 1, di-isopropylethylamine (180 μL , 1.0 mmol) in N, N-dimethylformamide (10 mL) was added a Castro reagent (BOP) (442 mg, 1.0 mmol). The mixture was stirred at room temperature overnight, the solvent was removed under high vacuum, and the residue was dissolved in methylene chloride (100mL), washed with 10% citric acid (2 x 50 L) and brine (50%). mL), and dried over Na2SO4. After evaporating the solvent in vacuo, the residue was purified by column chromatography (2: 1 to 3: 1 ethyl acetate: hexane) to give l-. { N- [2- (N ', N "-bis (tert-butoxycarbonyl) amidinoaminooxy) ethyl] amino}. Carbonyl met il-5-chloro-6-met il-3- (phenet-lamino) -pyrazinone as a colorless foam (100 mg, 23%). 1 H-NMR (300 MHz, CDCl 3 / CD 3 OD) d 8.42 (t, J = 4.8 Hz, 1 H), 7.26 (m, 5 H), 6.68 (s, 1 H), 4.75 ( s, 2H), 4.11 (t, J = 4.6Hz, 2H), 3.60 (, 4H), 2.91 (m, 2H), 2.15 (s, 3H), 1.52 (s, 9H), 1.49 (s, 9H) 1- {N- [2- (NT, N "-bis (tert-butoxycarbonyl) amidinoaminooxy) ethyl] aminotcarbonyl methyl-6-methyl-3- (phenethylamino) -pyrazinone was also isolated as a colorless foam (130). mg, 32%). ^ -NMROOO MHz, CDCl3 / CD3OD) d 8.47 (t, J = 4.9 Hz, 1H), 7.44-7.87 (m, 3H), 7.25 (m, 5H), 4.80 (s, 2H), 4.11 (m, 2H ), 3.65 (t, J = 7.0 Hz, 2H), 3.59 (m, 2H), 2.93 (t, J = 7.1 Hz, 2H), 2.28 (s, 3H), 1.52 (s, 9H), 1.48 (s) , 9H). j. l- trifluoroacetate. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (phenethylamino) -pyrazinone A mixture of 1-. { N- [2- (N ', N "-bis (tert-butoxycarbonyl) amidinoaminooxy) ethyl] amino.}. Carbonyl met il-6-met il-3- (phenethylamino) -pyrazinone (120 mg, 0.2 mmol) , as prepared in the preceding step, and trifluoroacetic acid (2 mL) in methylene chloride (4 mL) was stirred at room temperature for 1 hour.After evaporating the solvent in vacuo, the residue was purified in a Waters Sep. Pak (5g, 10% methylene chloride in methanol) to give the title compound as a white solid (90 mg, 89%). ^ - MROOO MHz, DMSO-d6) d 10.95 (s, 1H), 8.44 (t, J = 5.6 Hz, 1H), 7.73 (br s, 4H), 7. 20-7.30 (m, 6H), 6.67 (s, 1H), 4.61 (s, 2H), 3. 81 (t, J = 5.4 Hz, 2H), 3.50 (q, J = 6.7 Hz, 2H), 3. 39 (q, J = 5.4 Hz, 2H), 2.86 (t, J = 6.9Hz, "H), 2.07 (s, 3H). Mass spectrum (MALDI-TOF, a-cyano-4-hydroxycinnamic acid matrix) Calculated for C? 8 H 25 N 703: 388.2 (M + H), 410.2 (M + Na); Found: 388. 3, 410.4.

Example 2 L- Trifluoroacetate. { N- [2-amidinoaminooxy) ethyl] amino} carbonylmethyl-5-chloro-6-methyl-3- (phenethylamino) -pyrazinone The title compound was prepared from 1-. { N- [2- (N ', N "-bis (tert-butoxycarbonyl) amidinoaminooxy) ethylamino} carbonylmethyl-5-chloro-6-methyl-3- (phenethyl amino) -pyrazinone, as prepared in step i of Example 1, using the procedure of Example 1, step J. 1 H-NMR (300 MHz, DMSO-d6) d 10.96 (s, 1H), 8.46 (br s, 1H), 7.73 (br s, 4H) , 7.42 (t, J = 5.3 Hz, 1H), 7.18-7.33 (m, 5H), 4.66 (s, 2H), 3.81 (br s, 2H), 3.49 (m, 2H), 3.36 (m, 2H) , 2.86 (t, J = 7.2 Hz, "H), 2.18 (s, 3H). Mass spectrum (MALDI-TOF, a-cyano-4-hydroxycinnamic acid matrix) Calculated for C? 8H2 ClN703: 422.2 (M + H), 444.2 (M + Na); Found: 422-1, 444.0.

Example 3 L- Trifluoroacetate. { N- [2-amid noaminoo i) ethyl] amino} carbonylmethyl-6-methyl-3- (2, 2-diphenylethylamino) -pyrazinone The title compound was prepared from l-benzyloxycarbonylmethyl-3,5-dichloro-6-methypypyrazinone, as prepared in step f of Example 1, and 2, 2-diphenylethylamine using the procedure of Example 1, steps gj, 1H-NMR (300 MHz, DMS0-d6) d 10.89 (br s, 1H), 8.42 (t, J = 5.5 Hz, 1H), 7.70 (br s, 4H). 7.17-7.31 (m, 10H), 6.74 (br s, 1H), 6.69 (s, 1H), 4.57 (s, 2H), 4.48 (t, J = 7.8 Hz, 2H), 3.91 (t, j = 6.6 Hz, 2H), 3.79 (t, J = 5.4 Hz, 2H), 3.36 (t, J = 5.4 Hz, 2H), 2.06 (s, 3H). MALDI-TOF mass spectrum, α-cyano-4-hydroxycinnamic acid matrix) Calculated for C 24 H 29 N 703: 464.3 (M + H), 486.2 (M + Na): Found: 464.3, 486.3.

Example 4 L- Trifluoroacetate. { N- [2- (amidinoaminooxy) ethyl] a int)} carbonylmethyl-5-chloro-6-methyl-3- (2, 2-diphenylenylamino) -pyrazinone The title compound was prepared from l-benzyloxycarbonylmethyl-3,5-dichloro-6-methypypyrazinone, as prepared in step f of Example 1, and 2, 2-diphenylethylamine using the procedure of Example 1, steps gj. 1 H-NMR (300 MHz, DMS0-d 6) d 10.90 (br s, 1H), 8.41 (br s, 1H). 7.71 (br s, 4H), 7.18-7.32 (m, 10H), 4.63 (s, 2H), 4.52 (t, J = 7.7 Hz, 2H), 3.89 (t, J = 6.3 Hz, 2H), 3.79 ( t, J = 5.3 Hz, 2H), 3.37 (t, J = 5.3 Hz, 2H), 2.17 (s, 3H). Mass spectrum (MALDI-TOF; α-cyano-4-hydroxycinnamic acid matrix) Calculated for C 24 H 28 C 1 N 703: 498.2 (M + H), 520.2 (M + Na); Found: 498.2, 520.2. Example 5 L- Trifluoroacetate. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (4-methylphenethylamino) -pyrazinone The title compound was prepared from l-benzyloxycarbonylmethyl-3,5-dichloro-6-methypyroxine inone, as prepared in step f of Example 1, and -met i 1 phenethylamine using the procedure of Example 1, steps gj, XH-NMR (300 MHz, DMSO-d6) d 10.92 (s, 1H), 8.44 (t, J = 5.6 Hz, 1H), 7.71 (br s, 4H), 7.11 (s, 4H), 6.67 (s, 1H), 4.61 (s, 1H), 3.81 (t, J = 5.4 Hz, 2H), 3.50 (m, 2H), 3.38 (m, 2H), 2.81 (t, J = 6.9 Hz, 2H) , 2.27 (s, 3H), 2.07 (s, 3H), 2.07 (s, 3H). Mass spectrum (MALDI-TOF, a-cyano-4-hydroxycinnamic acid matrix) Calculated for C? 9H27N703: 402.2 (M + H), 424.2.

Example 6 L- Trifluoroacetate. { N- [2-amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (4-methoxyphenethylamino) -pyrazinone The title compound was prepared from l-benzyloxycarbonylmethyl-3,5-dichloro-6-methypypyzinone, as prepared in step f of Example 1, and -methoxy phenethylamine using the procedure of Example 1, steps gj . 1 H-NMR (300 MHz, DMSO-d 6) d 10.88 (s, 1 H), 8.42 (t, J = 5.5 Hz, 1 H), 7.68 (br s, 4 H), 7.15 (d, J = 8.6 Hz, 2 H) , 6.85 (d, J = 8.7 Hz, 2H), 6.67 (s, 1H), 4.61 (s, 2H), 3.81 (t, J = 5.3 Hz, 2H), 3.72 (s, 3H), 3.50 (m, 2H), 3.38 (m, 2H), 2.79 (t, J = 6.9 Hz, 2H), 2.07 (s, 3H), Mass spectrum (MALDI-TOF, a-cyano-4-hydroxycinnamic acid matrix) calculated for C? 9 H 27 N 704: 418.2 (M + H), 440.2 (M + Na); Found: 418.3, 440.4. Example 7 L- Trifluoroacetate. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (1-phenylcyclobutyl) methylamino-pyrazinone (1-phenylcyclobutyl) ethylamine To a solution of 1-phenylcyclobutanecarbonitrile (0.96g, 6.09 mmol) in anhydrous tetrahydrofuran (40 mL) was added a 1 N solution of lithium aluminum hydride (LAH) in tetrahydrofuran (12 mL, 12 mmol). After stirring for 2 hours at room temperature, the excess LAH was quenched slowly with water (10 mL) and diluted with additional tetrahydrofuran (20 mL). This was then reacted with 0.25 N aqueous NaOH (5 mL) at room temperature for 20 hours and filtered, the filtrate was concentrated in vacuo, and the residue was purified by flash column chromatography (10% methanol in dichloromethane, saturated with gas of ammonia) to give the title compound as a yellow oil (0.52g, 53%). XH NMR (300 MHz, CDCl 3) d 7.32 (m, 2H), 7.19 (m, 1H), 7.10 (m, 2H), 2.93 (s, 2H), 2.38-2.26 (m, 2H), 2.18-2.01 ( m, 3H), 1.92-1.82 (m, 1H), 1.14 (bs, 2H). b. 3- (1-Phenylcyclobutyl) methylamino-5-chloro-6-methyl-1- (benzyloxycarbonylmethyl) pyrazinone A solution of the product from the preceding step (0.52 g, 3.21 mmol) and 1-benzyloxycarbonyl Imet i 1-3, 5-dichloro-6-methypypyrazinone (0.36 g, 1.08 mmol), as prepared in step f of Example 1 , in ethyl acetate (15 mL) was refluxed for 3 hours and the solvent was removed in vacuo. The residue was dissolved in dichloromethane, washed with 10% aqueous citric acid and brine, dried over Na 2 SO 4, and filtered. The evaporated filtrate was then purified by flash column chromatography (20% ethyl acetate in hexanes) to give the title compound as a yellow oil (0.35 g, 71%). H NMR (300 MHz, CDCl 3) d 7.32 (m, 7H), 7.17 (m, 3H), 5.86 (t, J = 5.5 Hz), 5.17 (s, 2H), 4.73 (s, 2H), 3.72 ( d, 2H, J = 5.7 Hz), 2.35 (m, 2H), 2.22 (M, 2H), 2.17 (s, 3H), 1.89 (m, 1H). c. 3- (1-Phenylcyclobutyl) methylamino-6-methyl-1-carboxymethyl-pyrazinone The product from the preceding step (0.33g, 0.74 mmol), palladium (O) in 10% carbon (0.13g), and solid potassium hydroxide (0.2g, 3.6 mmol) were dissolved in a 1: 1: 1 mixture of methanol, water, and tetrahydrofuran (60 mL), the gas was removed by bubbling with nitrogen and placed under a pressure aspirator, stirred under a hydrogen sphere at room temperature. After 24 hours the reaction was filtered over Celite, the filtrate was evaporated, and the residue was partially purified by flash column chromatography (20% methanol in dichloromethane) to give the title compound (0.16g, 65%) as a solid. which was used without further purification. d. 1-. { N- [2- (N ', N "-bis { Tert-butoxycarbonyl}. Amidinoaminooxy) ethyl] amino.}. Carbonyl methyl-6-methyl-3- (1-phenyl-cyclobutyl) methylamino-pyrazinone The product of the preceding step (0.16g, 0.48 mmol), Castro's reagent (0.24g, 0.54 mmol), and [N, N '-di (tert-butoxycarbonyl)] -2-aminoethoxyguanidine (0.16 g, 0.52 mmol), as prepared in step d of Example 1, they were dissolved in N, N-dimethylformamide (10 mL) and reacted with triethylamine (0.25 mL, 1.80 mmol) at room temperature. After 24 hours the solvent was removed in vacuo, the residue was dissolved in dichloromethane, and the solution was washed with 10% aqueous citric acid, saturated NaHCO3, and brine, dried over Na2SO4, and filtered. The filtrate was evaporated, purified by flash column chromatography (33% ethyl acetate in dichloromethane) to give the title compound as a clear solid (O.llg, 37%) 1ti NMR (300 MHz, CDCl 3) d 9.15 ( s, 1H), 8.36 (t, J = 5 Hz), 7.57 (s, 1H), 7.32 (m, 2H), 7.18 (m, 3H), 6.66 (s, 1H), 5.77 (m, 1H), 4.72 (s, 2H), 4.11 (m, 2H9, 3.71 (d, 2H, J = 5.7 Hz), 3.60 (dd, 2H, J = 8.8 Hz, 5.0 Hz), 2.36 (m, 2H), 2.26 (, 3H), 2.12 (s, 3H), 1.90 (m, 1H), 1.51 (s, 9H), 1.46 (s, 9H) Mass Spectrum (MALDI-TOF, a-cyano-4-hydroxycinnamic acid matrix) calculated for C3_.H45N707: 428.2 (M-2Boc + H) Found: 428.9. and. l- trifluoroacetate. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (l-phenylcyclobutyl) methylami o-pyrazinone The product from the preceding step (0.10 g, 0.17 mmol) was dissolved in dichloromethane (5 mL) and reacted with trifluoroacetic acid (2 mL) at room temperature. After 6 hours, the volatiles were removed in vacuo and the residue was purified on a silica Sep-Pak Waters (elution gradient: 10-20% methanol in dichloromethane) to give the title compound as a light yellow hygroscopic solid (0.10. g, 100%). XH NMR (300 MHz, CDC13 / CD30D) d 7.33 (m, 2H), 7.19 (m, 3H), 6.56 (d, 1H, J = 1.0Hz), 4.65 (s, 2H), 3.94 (t, 2H, J = 5 Hz), 3.75 (s, 2H), 3.48 (t, 2H, J = 5 Hz), 2.43 (m, 2H), 2.25 (m, 3H), 2.12 (s, 3H), 1.93 (m, 1H), Mass spectrum (MALDI-TOF, gentisic acid matrix) calculated for C2? H29N703: 428.2 (M + H), 450.2 (M + Na). Found: 428.8, 450.7.

Example 8 L- Hydrochloride. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- [2- (l-naphthalene) ethyl] amino-pyrazinone to. 2- (l-naphthalenoethyl) phthalimide A solution of l-naphthalene-ethanol (2.51 g, 14.6 mmol), triphenylphosphine (4.57 g, 17.4 mmol), and phthalimide (2.37 g, 16.1 mmol) in tetrahydrofuran (120 mL) was reacted with diethylazodicarboxylate (2.80 mL, 17.8 mmol). mmol) at room temperature for 2 hours. The evaporated product was washed with diethyl ether and dried in a vacuum desiccator overnight giving the title compound (4.34g, 99%) as an ivory solid. 1 H NMR (300 MHz, CDCl 3) d 8.31 (d, 1 H, J = 8.5 Hz), 7.86 (, 3 H), 7.74 (m, 3 H), 7.60 (dd, 1 H, J = 8.4 Hz, 1.5 Hz), 7.50 (ddd, 1H, J = 8.0 Hz, 6.9 Hz, 1.2 Hz), 7.41 (m, 2H), 4.04 (m, 2H), 3.44 (m, 2H). b. 2- (l-naphthalene) ethylamine The product from the preceding step (4.30 g, 14.3 mmol) was dissolved in methanol (30 mL) and reacted with 40% aqueous methylamino (20 mL) at 70 ° C for 20 hours. The reaction was concentrated in vacuo, the residue was dissolved in dichloromethane, and the resulting solution was extracted with 10% aqueous HCl and water. The aqueous layer was made basic with solid KOH and the resulting solution was extracted with dichloromethane. The last organic layer was washed with brine, dried over Na 2 SO, filtered, and the filtrate was evaporated to give the title compound as a brown oil (0.83 g, 34%). X H NMR (300 MHz, CDCl 3) d 8.06 (m, 1 H), 7.87 (m, 1 H), 7.74 (d, 1 H, J = 8.0 Hz), 7.50 (m, 2 H), 7.37 (m, 2 H), 3 , 23 (t, 2H, J = 6.6 Hz), 3.11 (t, 2H, J = 6.6 Hz). c. 3- [2- (l-naphthalene) ethyl] amino-5-chloro-6-methyl-1- (benzyloxycarbonylmethyl) pyrazinone A solution of the product from the preceding step (0.82 g, 4.76 mmol), triethylamine (2.0 mL, 14 mmol), and 1-benzyloxycarbonylmethyl-3,5-dichloro-6-methylpyrazinone (1.05 g, 3.21 mmol). As prepared in step f of Example 1, in ethyl acetate (80 mL) it was refluxed for 20 hours. After evaporation, the residue was dissolved in dichloromethane and washed with 10% aqueous citric acid, saturated NaHCO3, and brine, dried over NaSO4, and filtered. The filtrate was then evaporated in vacuo to give the title compound as a brown solid. (1.49g, 68%). 1 R NMR (300 MHz, CDCl 3) d 8.33 (d, 1 H, J = 8.6 Hz), 7.86 (dd, 1 H, J = 8 1 Hz), 7.75 (d, 1 H, J = 7.8 Hz), 7.58 (ddd, 1H, J = 8.4 Hz, 6.9 Hz, 1.5 Hz), 7.50 (ddd, 1H, J = 8.0 Hz, 6.9 Hz, 1.2Hz), 7.37 (m, 7H), 6.26 (t, 1H, J = 5.9 Hz) , 5.22 (s, 2H), 4.80 (s, 2H), 3.78 (m, 2H), 3.40 (m, 2H), 2.24 (s, 3H). d. 3- [2- (l-naphthalene) ethyl] amino-6-methyl-1-carboxymethyl-pyrazinone A mixture of the product from the previous step (1.48g, 3.20 mmol), palladium (O) in 10% carbon (500 mg), and solid KOH (2.05 g, 36.5 mmol) in 2: 1: 1 tet rahydrofuran / methanol / water, was bubbled with nitrogen gas, aspirated, and stirred vigorously under hydrogen (sphere) at room temperature. ambient. After 16 hours the reaction was filtered over Celite, the alkaline flux was washed with methanol / water, and the filtrate was evaporated. The residue was treated with 10% aqueous HCl, cooled, and filtered, the precipitate was washed with water and ether and dried in vacuo overnight to give the title compound (0.84 g, 78%) as a golden solid. which appeared to be an inseparable mixture-TLC of 5-chloro and 5-hydro products by NMR. X H NMR (400 MHz, CDCl 3) d 8.54 (d ', 0.5 H, J = 8.3 Hz). 8.25 (d, 1H, J = 8.2 Hz), 7.93 t, 2H, J = 7.1 Hz), 7.82 (t, 2H, J = 8.6 Hz). 7.51 (m, 7H), 6.75 (s, 1H), 4.77 (s, 1H), 4.74 (s, 2H), 3.79 (m, 2H), 3.56 (m, 1H), 3.40 (m, 2H), 3.31 (m, 1H), 2.25 (s, 1.5H), 2.17 (s, 3H). and. 1- . { N- [2- (N ', N "-bis { Tert-b-toxicarbonyl}. Amidinoaminooxy) ethyl] amio.}. Carbonyl-methyl-6-methyl-3- [2- (l-naphthalene) ethyl. ] amino-pyrazinone The product of the preceding step (0.84 g, 2.49 mmol), Castro's reagent (1.15 g, 2.60 mmol), and [N, N '-di (tert-butoxycarbonyl)] -2-aminoethoxy-guanidine (0.91 g, 2.55 mmol), as prepared in step d of Example 1, it was dissolved in N, N-dimethylformamide (50 mL) and treated with triethylamine (1.0 mL, 7.2 mmol) at room temperature. After stirring the reaction overnight it was evaporated in vacuo and the residue was purified by flash column chromatography (ethyl acetate) to give the title compound (0.79g)., 50%) as a slightly orange solid. 1 H NMR (400 MHz, CDCl 3) d 9.17 (s, 1 H), 8.41 (m, 1 H), 8.13 (d, 1 H, J = 8.4 Hz). 7.84 (m, 1H), 7.73 (dd, 1H, J = 7.5 Hz, 1.6 Hz), 7.58 (m, 1H), 7.50 (m, 2H), 7.37 (, 3H), 6.72 (d, 1H, J = 0.9 Hz), 6.15 (m, 1H), 4.77 (s, 2H), 4.12 (m, 2H), 3.76 (m, 2H), 3.61 (dd, 2H, J = 8.7 Hz, 5.1 Hz), 3.39 (t , 2H, J = 7.3 Hz), 2.14 (d, 3H, J = 0.7 Hz), 1.51 (s, 9H), 1.47 (s, 9H).

F. l- hydrochloride. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- [2- (l-naphthalene) ethyl] amino-pyrazinone The product of the preceding step (0.79 g, 1.25 mmol) was dissolved in dichloromethane (10 mL) and reacted with trifluoroacetic acid (5 mL) at room temperature overnight. The reaction was concentrated in vacuo, the residue was purified by flash column chromatography (15% methanol in dichloromethane, saturated with ammonia gas), and the evaporated column fractions were treated with 4N HCl in ethanol and concentrated under high vacuum. giving the title compound (0.43g, 73%) as a pale yellow solid. 1 H NMR (300 MHz, CDCl 3) d 11.1 (s, 1 H), 8.71 (t, 1 H, J = 5.3 Hz), 8.26 (d, 1 H, J = 8.2 Hz), 7.94 (d, 1 H, J = 8.1 Hz ), 7.83 (d, 1H, J = 8.2 Hz), 7.78 (bs, 4H), 7.56 (m, 3H), 7.46 (t, 1H, J = 7.5 Hz), 6.73 (s, 1H), 4.69 (s) , 2H), 3.80 (m, 4H), 3.42 (m, 4H), 2.14 (s, 3H). Mass spectrum (LCMS, ESI pos.) Calculated for C22H27N703: 438.2 (M + H). Found: 438.2.

Example 9 L- Trifluoroacetate. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl -6-methyl-3- (2-phenyl-1-butylamino) -pyrazinone The title compound was prepared as a pale yellow solid from 1-phenyl-cyclopropane carbonitrile, in a manner analogous to that of Example 7, except that the cyclopropane ring was opened during the catalytic hydrogenation. XH NMR (300 MHz, CDC13 / CD30D) d 7.24-707 (m, 5H), 6.47 (s, 1H), 3.81 (t, 2H, J = 4.7 Hz), 3.62 (dd, 1H, J = 13.5 Hz, 6.3 Hz), 3.42 (dd, 1H, J = 13.6 Hz, 9.0 Hz), 3.35 (t, 2H, J = 4.6 Hz), 2.73 (m, 1H), 2.02 (s, 3H), 1.72-1.44 (m, 2H), 0.69 (t, 3H, J = 7.3 Hz). Mass spectrum (MALDI-TOF, gentisic acid matrix) calculated for C2oH29N703: 416.2 (M + H), 438.2 (M + Na). Found: 416.4, 438.4.

Example 10 L- Trifluoroacetate. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2- [3,4-methylenedioxyphenyl] ethylamino) -pyrazinone The title compound was prepared as a pale yellow oil from 3-methoxodioxifenet ylamine hydrochloride, analogously to Example 1. X H NMR (300 MHz, CDCl 3 / CD 3 OD) d 6.70 (m, 3 H ), 6.63 (s, 1H), 5.91 (s, 2H), 4.67 (s, 2H), 3.95 (t, 2H, J = 4.8 Hz), 3.61 (t, 2H, J = 7.3 Hz), 3.49 (t , 2H, J = 4.7 Hz), 2.86 (t, 2H, J = 7.3 Hz), 2.17 (s, 3H), Mass spectrum (MALDI-TOF, a-cyano-4-hydroxycinnamic acid matrix) calculated for C_ .9H25N703: 432.2 (M + H), 454.2 (M + Na). Found: 432.6, 454.8.

Example 11 L- Trifluoroacetate. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2- [2-pyridyl] ethylamino) -pyrazinone The title compound was prepared as an orange oil from 2- (2-aminoethyl) pyridine, in a manner analogous to that of Example 1, XH NMR (300 MHz, CDC13 / CD30D) d 8.46 (d, 1H, J = 4.2 Hz), 7.74 (t, 1H, J = 7 Hz), 7.34 (d, 1H, J = 7.7 Hz), 7.25 (dd, 1H, J = 7.1 Hz, 4.9 Hz), 6.71 (s, 1H) 3.99 (t, 2H, J = 4.5 Hz), 3.72 (t, 2H, J = 6.7 Hz) 3.54 (m, 2H), 3.11 (t, 2H, J = 6.7 Hz), 2.18 (s, 3H). Mass spectrum (MALDI-TOF, gentisic acid matrix) calculated for C? 7H24N803: 389.2 (M + H), 411.2 (M + Na). Found: 389.8, 411.6.

Example 12 l- Trifluoroacetate. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2- [2-methylphenylethylamino) -pyrazinone The title compound was prepared as a pale orange solid from 2-methybenzyl cyanide, in a manner analogous to Example 7. XH NMR (300 MHz, DMS0-d6) d 11.13 (s, 1H), 8.48 ( t, 1H, J = 5.5 Hz), 7.83 (bs, 4H), 7.12 (m, 4H), 6.68 (d, 1H, J = 1.0 Hz), 4.63 (s, 2H), 3.82 (t, 2H, J = 5.4 Hz), 3.47 (, 2H), 3.39 (q, 2H, J = 5 Hz), 2.85 (m, 2H), 2.31 (s, 3H), 2.08 (s, 3H). Mass spectrum (LCMS, ESI pos.) Calculated for C? 9H2 N703: 402.2 (M + H). Found: 402.2.

Example 13 L- Hydrochloride. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2- [3-methylphenyl] ethylamino) -pyrazinone The title compound was prepared as a pale yellow solid from 3-methylbenzylcyanidin, in a manner analogous to Example 7. XH NMR (400 MHz, DMSO-d6) dl 1.16 (s, 1H), 8.74 (t, 1H, J = 5.5 Hz), 7.80 (bs, 4H), 7.19 (t, 1H, J = 7.5 Hz), 7.16 (s, iH), 7.12 (d, 1H, J = 7.5 Hz), 7.03 (d, 1H, J = 7.4 Hz), 6.70 (s, 1H), 4.67 (s, 2H), 3.82 (t, 2H, J = 5.2 Hz), 3.69 (m, 2H), 3.39 (q, 2H, J = 5.3 Hz), 2.89 (t, 2H, J = 7.6 Hz), 2.28 (s, 3H), 2.14 (s, 3H). Mass spectrum (LCMS, ESI pos.) Calculated for C? 9H2 N703: 402.2 (M + H). Found: 402.2.

Example 14 L- Hydrochloride. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2- [2-trifluoromethylphenyl] ethylamino) -pyrazinone The title compound was prepared as a pale yellow solid from 2- (trifluoromethyl) phenethyl alcohol, in a manner analogous to Example 8. XH NMR (400 MHz, DMS0-d6) d 11.09 (s, 1H) , 8.66 (t, 1H, J = 5.5 Hz), 7.76 (bs, 4H), 7.70 (m, 2H), 7.64 (t, 1H, J = 7.5 Hz), 7.46 (t, 1H, J = 7.6 Hz) , 6.72 (s, 1H), 4.67 (s, 2H), 3.82 (t, 2H, J = 5.3 Hz), 3.71 (m, 2H), 3.39 (q, 2H, J = 5.5 Hz), 3.11 (t, 2H, J = 7.3 Hz), 2.13 (s, 3H). Mass spectrum (LCMS, ESI pos.) Calculated for C? 9H24N703F3: 456.2 (M + H). Found: 456.1.

Example 15 L- Trifluoroacetate. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2- [3-trifluoromethylphenyl] ethylamino) -pyrazinone The title compound was prepared as a white solid from 3- (trifluoromethyl) phenethyl alcohol, in a manner analogous to Example 8.? H NMR (400 MHz, DMSO-d6) d 11.01 (s, 1H ), 8.47 (t, 1H, J = 5.6 Hz), 7.75 (bs, 4H), 7.57 (m, 4H), 6.68 (s, 1H), 4.62 (s, 2H), 3.81 (t, 2H, J = 5.4 Hz), 3.57 (q, 2H, J = 6.6 Hz), 3.37 (m, 2H), 2.98 (t, 2H, J = 7.3 Hz), 2.10 (s, 3H). Mass spectrum (LCMS, ESI pos.) -calculated for C? 9H24N703F3: 456.2 (M + H). Found: 456.2.

Example 16 L- Trifluoroacetate. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2- [4-trifluoromethylphenyl] ethylamino) -pyrazinone The title compound was prepared as a brown solid from 4-trif luoromethyl) phenylacetonitrile, in a manner analogous to that of Example 7. 1 H NMR (400 MHz, DMSO-d6) d 10.98 (s, 1H), 8.45 (t, 1H, J = 5.5 Hz), 7.74 (bs, 4H), 7.66 (D, 2H, J = 8.1 Hz), 7.47 (d, 2H, J = 8.0 Hz), 6.68 (s, 1H), 4.62 (s, 2H), 3.81 (t, 2H, J = 5.4 Hz), 3.38 (q, 2H, J = 5.4 Hz), 2.97 (t, 2H, J = 7 Hz), 2.08 (s, 3H). Mass spectrum (LCMS, ESI pos.) Calculated for d9H24N703F3: 456.2 (M + H). Found: 456.2.

Example 17 L- Trifluoroacetate. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2- [3,5-dimethylphenyl] ethylamino) -pyrazinone The title compound was prepared as a pale yellow solid from the example 3,5-dimethylphenylacetonityl, in a manner analogous to Example 7. XH NMR (400 MHz, DMSO-d6) d 10.88 (s, 1H), 8.44 (t, 1H, J = 5.7 Hz), 7.68 (bs, 4H), 6.84 (s, 3H), 6.68 (s, 1H), 4.61 (s, 2H), 3.81 (t, 2H, J = 5.4 Hz ), 2.77 (t, 2H, J = 7.6 Hz), 2.24 (s, 6H), 2.08 (s, 3H). Mass spectrum (LCMS, ESI pos.) Calculated for C2oH29N703: 416.2 (M + H). Found: 416.2.

Example 18 L- Hydrochloride. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2-indanylamino) -pyrazinone The title compound was prepared as a white solid from 2-aminoindane hydrochloride, in a manner analogous to that of Example 1. XH NMR (400 MHz, DMSO-d6) d 10.95 (s, 1H), 8.54 (m, 1H), 7.69 (bs, 4H), 6.71 (s, 1H), 4.64 (s, 2H), 3.81 (t, 2H, J = 5.3 Hz), 3.27 (dd, 2H, J = 16 Hz, 7.6 Hz), 3.05 (dd, 2H, J = 16Hz, 6.7 Hz), 2.11 (s, 3H). Mass spectrum (LCMS, ESI pos.) Calculated for C? 9H25N703: 400.2 (M + H). Found: 400.3.

Example 19 L- Hydrochloride. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2- [3, 4-difluorophenyl] ethylamino) -pyrazinone The title compound was prepared as a white solid from 3,4-di fluorophenylacetonitrile, in a manner analogous to that of Example 7. XH NMR (400 MHz, DMS0-d6) d 11.05 (s, 1H) 8.63 ( t, 1H, J = 5.5 Hz), 7.74 (bs, 4H), 7.46 (t, 1H, J = 10 Hz), 7.36 (dt, 1H, J = 10.9 Hz, 8.5 Hz), 6.70 (s, 1H) , 4.66 (s, 2H), 3.82 (t, 2H, J = 5.4 Hz), 3.66 (m, 2H), 3.38 (m, 2H), 2.91 (t, 2H, J = 7.4 Hz), 2.13 (s, 3H). Mass spectrum (LCMS, ESI pos.) Calculated for C? 8H23N703F2: 424.2 (M + H). Found: 424.2.

Example 20 L- Hydrochloride. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2- [5-indanyl] ethylamino) -pyrazine The title compound was prepared as a pale yellow solid from 5-indanyl-acetonitrile (F. Lauria and W. Logemann, US Patent No. 3,452,085) in a manner analogous to that of Example 7. XH NMR (400 MHz, DMSO-d6) d 11.13 (s, 1H), 8.71 (t, 1H, J = 5.5 Hz), 7.78 (bs, 4H), 7.19 (s, 1H), 7.14 (d, 1H, J = 7.7 Hz), 7.07 (m, 1H), 6.70 (s, 1H), 4.67 (s, 2H), 3.82 (t, 2H, J = 5.3 Hz), 3.65 (m, 2H), 2.84 (m, 6H), 2.13 (s) , 3H), 1.99 (pentet, 2H, J = 7.4 Hz). Mass spectrum (LCMS, ESI pos.) Calculated for C2? H29N703: 428.2 (M + H). Found: 428.3.

Example 21 Salt of the trifluoroacetic acid of l-. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2-fluorophenethylamino) -pyrazinone The title compound was prepared from l-benzyloxycarbonylmethyl-3,5-dichloro-6-methypypyrazinone, as prepared in step f of Example 1, and 2-fluorophenethylamino using the procedure of Example 1, steps g-j. 1 H NMR (300 MHz, DMS0-d 6) d 10.91 (s, 1 H), 8.43 (t, J = 5.6 Hz, 1 H), 7.70 (brs, 4 H), 7.30 (m, 2 H), 7.15 (m, 3 H) , 6.66 (s, 1H), 4.61 (s, 2H), 3.81 (t, J = 5.4 Hz, 2H), 3.37 (m, 4H), 2.90 (t, J = 7.2 Hz, 2H), 2.06 (s, 3H). Mass spectrum (MALDI-TOF, cyano-4-hydroxycinnamic acid matrix) calculated for C18H24N703: 406.2 (M + H), 428.2 (M + Na): Found: 406.3, 428.3.

Example 22 Salt of trifluoroacetic acid of l-. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (3,4-dimethoxyphenethylamino) -pyrazinone The title compound was prepared from 1-benzyloxycarbonylmethyl-3,5-dichloro-6-methypypyrazinone, as prepared in step f of Example 1, and 3-dimethoxyphenethylamino using the procedure of Example 1, steps gj- 1 H NMR (300 MHz, DMSO-d 6) d 10.95 (s, 1 H), 8.44 (t, J = 5.6 Hz, 1 H), 7.72 (br s, 4 H), 6.85 (t, J = 8.1 Hz, 2 H), 6.73 (d, J = 8.1 Hz, 2H), 6.67 (s, 1H), 4.61 (s, 2H), 3.81 (t, J = 5.3 Hz, 2H), 3.73 (s, 3H), 3.71 (s, 3H) ), 3.39 (m, 4H), 2.79 (t, 3 = 1. 2 Hz, 2H), 2.07 (s, 3H). Mass spectrum (MALDI-TOF, cyano-4-hydroxycinnamic acid matrix) calculated for C2oH29N705: 448.2 (M + H), 470.2 (M + Na): Found: 448.6, 470.4. E j a lo 23 Salt of trifluoroacetic acid of l-. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (4-fluorophenethylamino) -pyrazinone The title compound was prepared from l-benzyloxycarbonylmethyl-3,5-dichloro-6-methylpyrazinone, as prepared in step f of Example 1, steps gj, 1U NMR (300 MHz, DMSO-d6) d 10.97 ( s, 1H), 8.45 (t, J = 5.6 Hz, 1H), 7.74 (br s, 4H), 7.26 (m, 3H), 7.15 (t, J = 8.8 Hz, 2H), 6.68 (s, 1H) , 4.61 (s, 2H), 3.81 (t, J = 5.4 Hz, 2H), 3.39 (m, 4H), 2.85 (t, J = 7.3 Hz, 2H), 2.07 (s 3H). Mass spectrum (MALDI-TOF, cyano-4-hydroxycinnamic acid matrix) calculated for C? 8H24N703: 406.2 (M + H), 428.2 (M + Na): Found: 406.6, 428.5.

Example 24 Salt of trifluoroacetic acid of l-. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (4-ethylphenethylamino) -pyrazinone The title compound was prepared from l-benzyloxycarbonylmethyl-3,5-dichloro-6-methypypyrazinone, as prepared in step f of Example 1, and 4-ethephenimethylamine using the procedure of Example 7, steps b. . 1 H NMR (300 MHz, DMSO-d 6) d 10.97 (s, 1 H), 8.45 (t, J = 5.6 Hz, 1 H), 7.73 (br s, 4 H), 7.18 (m, 1 H), 7.14 (s, 4 H) ), 6.67 (s, 1H), 4.61 (s, 2H), 3.81 (t, J = 5.2 Hz, 2H), 3.39 (m, 4H), 2.82 (t, J = 7.5 Hz, 2H), 2.56 (q , J = 7.6 Hz, 2H), 2.07 (s, 3H), 1.16 (t, J = 7.5 Hz, 3H). Mass spectrum (MALDI-TOF, cyano-4-hydroxycinnamic acid matrix) calculated for C20H29N7O3: 416.2 (M + H), 438.2 (M + Na): Found: 416.2, 438.2.

Example 25 Salt of trifluoroacetic acid of l-. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2-phenylpropylamino) -pyrazinone The title compound was prepared from l-benzyloxycarbonylmethyl-3,5-dichloro-6-ethopyrazinone, as prepared in step f of Example 1, and (-methephenylethylamide using the procedure of Example 7, steps be • H NMR (300 MHz, DMSO-d6) d 10.98 (s, 1H), 8.43 (t, J = 5.6 Hz, 1H), 7.74 (br s, 4H), 7.28- (m, 5H), 7.20 (m, 1H) , 6.66 (s, 1H), 4.59 (s, 2H), 3.80 (t, J = 5.4 Hz, 2H), 3.44 (t, J = 6.5 Hz, 2H), 3.38 (t, J = 5.4 Hz, 2H) , 3.13 (q, J = 7.1 Hz, 2H), 2.06 (s, 3H), 1.19 (d, J = 7.0 Hz, 3H), Mass spectrum (MALDI-TOF, calculated cyano-4-hydroxycinnamic acid matrix) for C? 9H27N703: 402.2 (M + H), 424.2 (M + Na): Found: 402.4, 424.5.

Example 26 Salt of trifluoroacetic acid of l-. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (3,4-dimethylphenethylamino) -pyrazinone The title compound was prepared from 3,4-dimethylbenzylcyanide using the procedure of Example 7, steps a-e. lti NMR (300 MHz, DMSO-d6) d 11.10 (s, 1H), 8.47 (t, J = 5.5 Hz, 1H), 7.81 (br s, 4H), 7.55 (m, 1H), 7.03 (t, J = 7.9 Hz, 2H), 6.94 (d, J = 7.8 Hz, 1H), 6.68 (s, 1H), 4.62 (s, 2H), 3.82 (t, J = 5.3 Hz, 2H), 3.48 (m, 2H) ), 3.38 (m, 2H), 2.78 (t, J = 7.5 Hz, 2H), 2.19 (s, 3H), 2.17 (s, 3H), 2.08 (s, 3H). Mass spectrum (MALDI-TOF, cyano-4-hydroxycinnamic acid matrix) calculated for C2oH29N703: 416.2 (M + H), 438.2 (M + Na): Found: 416.0, 437.9.

Example 27 Salt of trifluoroacetic acid of l-. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2-naphthalene-ethylamino) -pyrazinone The title compound was prepared from 2-naphthalene ethanol using the procedure of Example 8, steps a-f. H NMR (400 MHz, DMSO-d6) d 10.90 (s, 1H), 8.43 (t, J = 5.5 Hz, 1H), 7.87 (m, 3H), 7.85 (s, 1H), 7.69 (br s, 4H ), 7.47 (m, 3H), 6.69 (s, 1H), 4.61 (s, 2H), 3.80 (t, J = 5.4 Hz, 2H), 3.61 (m, 4H), 3.04 (t, J = 7.2 Hz , 2H), 2.07 (s, 3H). Mass spectrum (LCMS, ESI) calculated for C22H27N703: 438.2 (M + H). Found: 438.2.

Example 28 Salt of trifluoroacetic acid of l-. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2, 2-diphenylpropylamino) -pyrazinone The title compound was prepared from 1-benzyl-1-loxi carbonyl-1-methyl-3,5-dichloro-6-methypypyrazinone, as prepared in step f of Example 1, and 2, 2-diphenylpropylamide hydrochloride using the procedure of Example 1, steps gj. X H NMR (400 MHz, DMSO-d 6) d 10.89 (s, 1 H), 8.41 (t, J = 5.4 Hz, 1 H), 7.70 (br s, 4 H), 7.32 (m, 4 H), 7.26 (m, 6 H) ), 6.68 (s, 1H), 5.75 (m, 1H), 4.56 (s, 2H), 4.00 (d, J = 5.0 Hz, 2H), 3.79 (t, J = 5.4 Hz, 2H), 3.36 (m , 2H), 2.05 (s, 3H), 1.67 (s, 3H). Mass spectrum (LCMS, ESI) calculated for C25H3? N703: 478.2 (M + H). Found: 478.2.

Example 29 Salt of trifluoroacetic acid of l-. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2- (3-indolyl) -ethylamino) -pyrazinone The title compound was prepared from l-benzyloxycarbonylmethyl-3,5-dichloro-6-met i lpira zinone, as prepared in step f of Example 1, and tryptamine using the procedure of Example 1, steps g-j. 1ti NMR (400 MHz, DMSO-d6) d 10.98 (s, 1H), 8.44 (t, J = 5.6 Hz, 1H), 7.74 (br s, 4H), 7.00-7.60 (m, 6H), 6.69 (s) , 1H), 4.62 (s, 2H), 3.81 (t, J = 5.3 Hz, 2H), 3.39 (m, 4H), 2.97 (t, J = 7.4 Hz, 2H), 2.08 (s, 3H). Mass spectrum (LCMS, ESI) calculated for C24H26N803: 427.2 (M + H). Found: 427.2.

Example 30 Salt of trifluoroacetic acid of l-. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2- [1- (4-methylnaphthalene) ethylamino) -pyrazinone The title compound was prepared as a white solid from l- (4-meth ilnaphthalene) acetohitrii, in a manner analogous to that of Example 7.: H NMR (400 MHz, DMSO-d6) d 10.94 (s, 1H), 8.45 (t, J = 5.6 Hz, 1H), 8.33 (d, J = 7.2 Hz, 1H), 8.04 (d, J = 7.1 Hz, 1H), 7.72 (br s, 4H), 7.58 (m , 2H), 7.28 (s, 2H), 6.73 (s, 1H), 4.63 (s, 2H), 3.82 (t, J = 5.4 Hz, 2H), 3.57 (m, 2H), 3.40 (t, J = 6.4 Hz, 2H), 3.29 (t, J = 7.3 Hz, 2H), 2.62 (s, 3H), 2.09 (s, 3H). Mass spectrum (MALDI-TOF, cyano-4-hydroxycinnamic acid matrix) calculated for C23H29N703: 452.2 (M + H), 474.2 (M + Na): Found: 452.2, 474.3.

Example 31 l- Trifluoroacetate. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2- [2, 4-difluorophenyl] ethylamino) -pyrazinone The title compound was prepared as a white solid from 2,4-difluorophenylethanol, in an analogous manner for Example 8. 1 H NMR (400 MHz, DMSO-d 6) d 11.03 (s, 1H), 8.46 (t, J = 5.6 Hz, 1H), 7.77 (br s, 4H), 7.34 (q, J = 7.8 Hz, 1H), 7.18 (t, J = 9.5 Hz, 1H), 7.03 (t, J = 8.0 Hz, 1H) , 6.67 (s, 1H), 4.62 (s, 2H), 3.81 (t, J = 5.4 Hz, 2H), 3.51 (m 2H), 3.39 (t, J = 5.5 Hz, 2H), 2.89 (t, J = 7.1 Hz, 2H), 2.08 (s, 3H). Mass spectrum (MALDI-TOF, cyano-4-hydroxycinnamic acid matrix) calculated for C_.9H23F2N703: 424.2 (M + H), 446.2 (M + Na): Found: 424.3, 446.5.

Example 32 1- Hydrochloride. { N- [2- (amidino-N 'methylaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2- [4-methylphenyl] ethylamino) -pyrazinone To a mixture of the product of Example 5 (0.09 g, 0.19 mmol) and sodium bicarbonate (0.53 g, 6.30 mmol) in N, -dimethylformamide (10 mL) was added with 2 mL of 4 N HCl in ethanol, dissolved in methanol / dichloromethane, and filtered again. The filtrate was then concentrated in vacuo to give the title compound as a yellow solid (0.08g, 86%). XH NMR (400 MHz, DMSO-d6) d 8.62 (t, 1H, J = 5.3 Hz), 7.90 (bs, 3H), 7.18 (d, 2H, J = 7.9 Hz), 7.11 (d, 2H, J = 7.9 Hz), 6.70 (s, 1H), 4.64 (s, 2H), 3.91 (t, 2H, J = 5.2 Hz), 3.39 (q, 2H, J = 5.2 Hz), 3.25 (s, 3H), 2.86 (t, 2H, J = 7.5 Hz), 2.27 (s, 3H), 2.12 (s, 3H). Mass spectrum (LCMS, ESI) calculated for C20H29N7O3: 416.2 (M + H). Found: 416.2; MS / MS Found 374.1 (M-C (= NH) NH2).

Example 33 Salt of the trifluoroacetic acid of l-. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2, 2-difluoro-2-phenylethylamino) -pyrazinone The title compound was prepared from l-benzyloxycarbonylmethyl-3,5-dichloro-6-methypypyrazinone, as prepared in step f of Example 1, and 2, 2-difluoro-2-phenylethylamine using the procedure of Example 1, steps gj. 1 H NMR (400 MHz, DMSO-d 6) d 10.89 (s, 1 H), 8.46 (t, J = 5.5 Hz, 1 H), 7.69 (br s, 4 H), 7.53 (m, 2 H), 7.50 (m, 3 H) ), 6.84 (t, J = 6.5 Hz, 1H), 6.64 (s, 1H), 4.62 (s, 2H), 4.06 (dt, J = 6.5, 14.4 Hz, 2H), 3.81 (t, J = 5.4 Hz , 2H), 3.38 (m, 2H), 2.06 (s, 3H). Mass spectrum (LCMS, ESI) calculated for C? 8H23F2N703: 424.3 (M + H). Found: 424.4.

Example 34 Preparation of tablets Tablets containing 25.0, 50.0 were prepared. and 100.0 mg, respectively, of the following active compounds as illustrated below: to. 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-met il-3- (4-methoxyphenethylamino) pyrazinone; Y b. 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2, 2-diphenylethylamino) pyrazinone.

DOSE FOR TABLET CONTAINING FROM 25-100 MG OF ACTIVE COMPOUND Amount-mg Active compound 25.0 50.0 100.00 Microcrystalline cellulose 37.25 100.0 200.0 Edible corn starch 37.25 4.25 8.5 Modified Magnesium stearate 0.50 0.75 1.5 All the active compounds, cellulose, and a portion of the corn starch were mixed and granulated to 10% of the corn starch paste. The resulting granulation was sieved, dried and mixed with the corn starch residue and the magnesium stearate. The resulting granulation is then compressed into tablets containing 25.0, 50.0 and 100.0 mg, respectively, of active ingredient per tablet.

Example 35 Preparation of the intravenous solution An intravenous dosage form from the above-mentioned active compounds was prepared as follows.

Active compound 0.5-10.0 mg Sodium citrate 5-50 mg Citric acid 1-15 mg Sodium chloride 1-8 mg Water per injection (USP) q.s. for 1 ml Using the above amounts, the active compound was dissolved at room temperature in a previously prepared solution of sodium chloride, citric acid and sodium citrate in water by injection (USP, see page 1636 of United States Pharmacopeia / National Formulary for 1995, published by the United States Pharmacopeial Convention, Inc., Rockville, Maryland (1994).

Example 36 In Vitro Inhibition of Purified Enzymes Reagents: All buffer salts are obtained from Sigma Chemical Company (San Luis, MO), and where the highest purity is available. The substrates of enzyme, N-benzoyl-Phe-Val-Arg-p-nitroanilide (Sigma B7632), N-benzoyl-Ile-Glu-Gly-Arg-p-nitroanilide hydrochloride (Sigma B2291), Np-Tosil- Gly-Pro-Lys-p-nitroanilide (Sigma T6140), N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide (Sigma S7388) and N-CBZ-Val-Gly-Arg-p-nitroanilide (Sigma C7271 ) are obtained from Sigma. The N-succinyl-Ala-Ala-Pro-Arg-p-nitroanilide (BACHEM L-1720) and N-succinyl-Ala-Ala-Pro-Val-p-nitroanilide (BACHEM-1770) are obtained from BACHEM (King of Prussia, PA).

Human thrombin, human factor Xa, and human plasmid are obtained from Enzyme Research Laboratories (South Bend, Indiana), bovine a-cimotripsima (Sigma C4129), bovine trypsin (Sigma T8642), and human kidney cell urokinase (Sigma U5004) are obtained from Sigma. Human leukocyte elastase is obtained from Elastin Products (Pacific, MO).

Ki determination: All analyzes are based on the ability of the test compound to inhibit the catalytic hydrolysis of the enzyme from a substrate p-nitroanilidine peptide. In a typical Ki determination, the substrate is prepared in DMSO, and diluted in a test buffer consisting of 50 mM HEPES, 200 mM NaCl, pH 7.5. The final concentrations for each of the substrates are listed below. In general, substrate concentrations are lower than the experimentally determined value for Km. The test compounds are prepared as a 1.0 mg / ml solution in DMSO. The dilutions are prepared in DMSO producing 8 final concentrations that span 200 times the concentration range. The enzyme solutions are prepared at the concentrations listed below in assay buffer.

In a typical K_. Determination, in each well of a 96-well tray, 280 mL of a substrate solution, 10 mL of a test compound solution is dripped, and the tray is allowed to equilibrate thermally at 37 ° C in a tray-reading molecular device by > 15 minutes. The reactions are initiated by the addition of an aliquot of 10 mL of enzyme and the increased absorbance at 405 nm is recorded for 15 minutes.

The corresponding data for less than 10% of the hydrolysis of the total substrate are used in the calculations. The ratio of velocity (ratio of change in absorbency as a function of time) for a sample that does not contain test compound is divided by the velocity of a sample containing the test compound, and plotted as a function of the concentration of the test compound. The data is adapted for a linear regression, and the value of the inclination of the line is calculated. The reverse side of the slope is the experimentally determined Ki value.

Thrombin: Thrombin activity is evaluated as the ability to hydrolyze the substrate N-succinyl-Ala-Ala-Pro-Arg-p-nit roanilide. The substrate solutions are prepared at a concentration of 32 mM (32 mM << Km = 180 mM) in a test buffer. The final concentration of DMSO was 4.3%. The purified human a-thrombin was diluted in assay buffer at a concentration of 15 nM. The final reagent concentrations were: [thrombin] = 0.5 nM, [substrate N-succinyl-Ala-Ala-Pro-Arg-p-nit roanilide] = 32 nM.

Factor Xa: The Fxa activity was evaluated as the ability to hydrolyze the hydrochloride substrate of N-benzoyl-Ile-Glu-Gly-Arg-p-nitroanilide. Substrate solutions were prepared at a concentration of 51 nM (51 <Km = 1.3 mM) in a test buffer. The final DMSO concentration was 4.3%. The purified activated human factor X was diluted in assay buffer at a concentration of 300 nM. The final reagent concentrations were: [FXa] = 10 nM, [N-benzoyl-Ile-Glu-Gly-Arg-p-nitroanilide hydrochloride] = 51 nM.

Plasmin: The activity of plasmin was evaluated as the ability to hydrolyze N-p-Tosyl-Gly-Pro-Lys-p-nit roanilide. The substrate solutions were prepared at a concentration of 37 mM (37 mM <Km = 243 mM) in assay buffer. The final concentration of DMSO was 4.3%. The purified human plasmid was diluted in assay buffer at a concentration of 240 nM. The final reagent concentrations were: [Plasmin] = 8 nM, [N-p-Tosyl-Gly-Pro-Lys-p-nitroanilide] = 37 mM.

Chymotrypsin: The activity of chymotrypsin was evaluated as the ability to hydrolyze N-succinyl-Ala-Ala-Pro-Phe-p-nit roanilide. The substrate solutions were prepared at a concentration of 14 mM (14 mM << Km = 62 mM) in a test buffer. The final concentration of DMSO was 4.3%. The purified chymotrypsin of bovine was diluted in assay buffer at a concentration of 81 nM, the final reagent concentrations were: [Chymotrypsin] = 2.7 nM, [N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide [ = 14 m.

Trypsin: The activity of trypsin was evaluated as the ability to hydrolyze N-benzoyl-Phe-Val-Arg-p-nitroanilide. The substrate solutions were prepared at a concentration of 13 mM (13 mM <Km = 291 M) in assay buffer. The final concentration of DMSO was 4.3%. purified bovine trypsin was diluted in assay buffer to a concentration of 120 nM. The final reagent concentrations were: [Trypsin] = 4 nM, [N-benzoyl-Phe-Val-Arg-p-nitroanilide] = 13 mM.

Elastase: The activity of elastase was evaluated as the ability to hydrolyze N-succinyl-Ala-Ala-pro-Val-p-nitroanilide. The substrate solutions were prepared at a concentration of 19 mM (19 mM <Km = 89 mM) in assay buffer. The final concentration of DMSO was 4.3%. Purified human leukocyte elastase was diluted in assay buffer at a concentration of 750 nM. The final reagent concentrations were: [Elastase] = 25 nM, [N-succinyl-Ala-Ala-Pro-Val-p-nitroanilidine] = 19 mM.

Urokinase: The activity of urokinase was evaluated as the ability to hydrolyze N-CBZ-Val-Gly-Arg-p-nit roanilide. The substrate solutions were prepared at a concentration of 100 mM (100 mM <Km = 1.2 mM) in assay buffer. The final concentration of DMSO was 4.3%. Purified human kidney urokinase was diluted in assay buffer at a concentration of 1.2 mM. The final reagent concentrations were: Urokinase] = 40 nM, and [N-CBZ-Val-Gly-Arg-p-nit roanilide] = 100 mM.

The results of the compound of Examples 1 to 6 are shown in the following table.

Table 1 Assay, Ki (μM) (Thrombin) Compound (Ex No.) Ki (trorabine) 0.046 0.330 0.006 0.085 0.020 0.013 The results indicate that the compounds of the present invention are highly potent and selective inhibitors of thrombin.

Having now fully described the invention, it will be understood by those of ordinary skill in the art that it may be executed within a broad and equivalent range of conditions, formulations, and other parameters without affecting the scope of the invention or any of its forms of realization of it. All patents and publications cited herein are fully incorporated herein by reference in their entirety.

It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is that which is clear from the present description of the invention.

Having described the invention as above, the content of the following is claimed as property.

Claims (3)

Claims: A compound of the formula a pharmaceutically acceptable solvent, hydrate or salt thereof; characterized because: W is hydrogen, R1, R ^ CO, R ^ O, R1 (CH2) 5NHCO, or (R1) 2CH (CH2) 5NHCO, wherein s is 0-4; is R2, R2 (CH2) iC (R12) 2, where t is 0-3, and each of R12 may be the same or different, (R2) (OR12) CH (CH2) p, where p is 1-4 , (R2) (OR12) C (CH2) P, where p is 1-4, R2C (R12) 2 (CH2) t, where t is 0-3, and each of R12 is the same or different, and wherein (R12) 2 together with C in which it is adhered, can also form a cycloalkyl ring of 3- to 7-membered, R2CH2C (R12) 2 (CH2) q, where q is 0-2, and each one of R12 is the same or different, where (R12) 2 together with C in which it is adhered, can also form a cycloalkyl ring of 3- up 7- members, R2CF2C (R12) 2 (CH2) q, where q is 0-2, and each of R12 is the same or different, where (R12) 2 together with C in which it is adhered, can also form a cycloalkyl ring of 3- up 7- members, (R2) 2CH (CH2) r, where r is 0-4 and each of R2 can be the same or different, and where (R2) 2 together with C in which it is adhered, can also form a C3_7 cycloalkyl ring, C7-? 2 bicyclic alkyl, tricyclic alkyl C? -i? or a saturated or unsaturated 5- to 7-membered mono- or bicyclic heterocyclic ring, and which contain from one to three heteroatoms selected from N, O and S, R20 (CH2) p, where p is 2-4 (R2) 2CF (CH2) r, where r is 0-4 and each of R12 may be the same or different, wherein (R12) 2 may also form a ring with C represented by C3_7 cycloalkyl, C7-? 2 bicyclic alkyl, C? or? 6f tricyclic alkyl or a 5- to 7- membered mono- or bicyclic heterocyclic ring which may be saturated or unsaturated, and which contain from one to three heteroatoms selected from the group consisting of N, O and S, where s is 0 or 1, R2CF2C (R12) 2 i is phenyl, naphthyl, or biphenyl, each of which is unsubstituted or substituted by one or more of C? -4 alquiloalkyl, C? _4alkoxy, halogen, hydroxy, CF3, OCF3, COOH, CONH2, or S02NH2, a heterocyclic or non-heterocyclic 5- to 7-membered or bicyclic 9- to 10-membered heterocyclic ring which may be saturated or unsaturated, wherein the heterocyclic ring contains from one to four heteroatoms selected from the group consists of N, O, and S, and is optionally substituted with halogen or hydroxy, C-7 alkyl, optionally substituted with one or more of hydroxy, COOH, amino, aryl, C3-7 cycloalkyl, CF3, N (CH3) 2 , C3_3 alkylaryl, heteroaryl, or heterocycloalkyl, CF3, C3_7 cycloalkyl, optionally substituted with aryl, C7-? 2-bicyclic alkyl, or C? -16 / tricyclic alkyl / hydrogen, alkyl, cycloalkyl, alkenyl, alkynyl, optionally substituted by aryl, optionally substituted by aralkyl, optionally substituted by heteroaryl, trifluoromethyl, halogen, hydroxyalkyl, cyano, nitro, carboxamido, C02Rx, -CH2ORx or -0RX, where Rx, in each instance, is independently one of hydrogen, alkyl or optionally unsaturated alkyl or optionally unsaturated cycloalkyl; R 'is hydrogen or halogen, R12 is hydrogen, phenyl, naphthyl, or biphenyl, each of which is unsubstituted or substituted by one or more of C4-4alkyl, C4-4alkoxy, halogen, hydroxy, CF3, 0CF3, COOH, or CONH2, a 5- to 7-membered monocyclic or bicyclic 9- to 10-membered heterocyclic ring which may be saturated or unsaturated, and which contain from one to four heteroatoms selected from the group consisting of N, O, and S, alkyl C_ . 4, substituted or unsubstituted with one or more of hydroxy, COOH, amino, aryl, heteroaryl, or heterocycloalkyl, CF 3, C 4 cycloalkyl, C 7 ι 2 bicyclic alkyl, or Cι-ie tricyclic alkyl; R- is hydrogen, C? _4 alkyl, or C2- alkenyl R is hydrogen, alkyl, alkenyl, aralkyl, aryl, hydroxyalkyl, aminoalkyl, (C2-? 0) alkyl, monoalkylamino, (C2-? O) dialkylamino, or carboxyalkyl; R7, R8, R9 and R10 are independently hydrogen, alkyl, aralkyl, aryl, hydroxyalkyl, aminoalkyl, monoalkylaminoalkyl, dialkylaminoalkyl or carboxyalkyl; or R7 and R8 are taken together to form - (CH2) U-, where u is 2 to 7, preferably 2 to 5, while R9 and R10 are as defined above; or R9 and R10 are taken together to form (CH2) V-, where v is up to 7, preferably 2 to 5, while R7 and R8 are as defined above; or R7 and R9 are taken together to form - (CH2) and -, where y is 0 (one bond) or 1 to 7, preferably 0-4, while R8 and R10 are as defined above; X is oxygen, NR11, or CH = N (where N is linked to NR6) where R11 is hydrogen, alkyl, cycloalkyl or aryl, wherein the alkyl, cycloalkyl or aryl may be optionally substituted with amino, monoalkylamino, dialkylamino, alkoxy, hydroxy, carboxy, alkoxycarbonyl, aryloxycarbonyl, aralkoxycarbonyl, aryl, acylamino, cyano or trifluoromethyl; Ra, R and Rc are independently hydrogen, alkyl, hydroxy, alkoxy, aryloxy, aralkoxy, alkoxycarbonyloxy, cyano or -C02Rw, where Rw is alkyl, cycloalkyl, phenyl, benzyl, where Rd and Re are independently hydrogen, C-. 6 alkyl, alkenyl or C2-6 phenyl / Rf is hydrogen, alkyl C1-6, alkenyl phenyl • 2 - ts r R? is hydrogen, C6-6 alkyl, alkenyl or C2-6 phenyl, and aralkyl or alkyl C? -6; n is from zero to 8; and m is from zero to 6. 2. The compound according to claim 1, characterized in that R3 is hydrogen, C4_4alkyl, C3_7 cycloalkyl or CF3; m is from zero to 4; and n is from zero to 4. 3. The compound according to claim 2, characterized in that R 3 is C 4 alkyl. 4. The compound according to claim 2, characterized in that R 4 is hydrogen or halogen. 5. The compound according to claim 4, characterized in that W is H or R1 6. The compound according to claim 5, characterized in that Rx is R 'R (CH2) tC (R, 12) 2, where t is 0-3, and each of R12 may be the same or different, R2C (R12) 2 (CH2) t, where t is 0-3, and each of R12 may be the same or different, and wherein (R12) 2 may also form a heterocyclic ring from 3 to 7 members with the C to which they are linked, R2CH2C (R12) 2 (CH2) q, where q is 0-2, and each R12 can be the same or different, where (R12) 2 together with the C to which they are linked, can also form a ring with 3 to 7 cycloalkyl members, (R2) 2CH (CH2) r, where r is 0-4 and each of R2 can be the same or different, and where (R2) 2 can also form, together with the C to which it is linked, a C3-7 cycloalkyl, C7_2 bicycloalkyl, triocycloalkyl, or 5- or 7- membered mono- or bicyclic heterocyclic which can be be saturated or unsaturated, and which contains from one to three heteroatoms selected from the group consisting of N, O and S, R2CF2C (R12) 2 (CH2) q, where q is 0-2, and each of R12 may be the same or different, where (R12) 2 together with the C to which it is linked, can also form a C3_7 cycloalkyl ring, or R20 (CH2) p, wherein p is 2-4; phenyl or naphthyl, each of which is unsubstituted or substituted by one or more of C ?4 alkyl, C ?4 alkoxy, halogen, hydroxy, CF3, OCF3, or S02NH2, a 5- to 7- membered monocyclic ring or a 9- to 10-membered bicyclic saturated or unsaturated, having from zero to 4 heteroatoms selected from N, O and S, and wherein the ring is substituted or unsubstituted with halogen or hydroxy, C? _7 alkyl, substituted or unsubstituted with one or more of hydroxy, COOH, C3-7 cycloalkyl, CF3, N (CH3) 2, -C? _3alkaryl, heteroaryl, or heterocycloalkyl, CF3, or C3_7 cycloalkyl, substituted or unsubstituted with aryl; Y , 12 is hydrogen, or C? _4 alkyl, substituted or unsubstituted with one or more of hydroxy, COOH, amino, aryl, heteroaryl, or heterocycloalkyl. 7. The compound according to claim 6, characterized in that R3 is hydrogen, CH3, or CH2CH3; R4 is hydrogen or chlorine; and is PhCH2CH2, (CH3) 3C-, HOOCCH2, CF3CH2, (CH3) 2N (CH2) 2, PhCH20 (CH2) 2. PhCH (CH3), PhCH2CH (COOH), CH3 (CH2) 5, PhCH2, H, CH3 (CH2) 4, CH3CH2CH (CH3) CH2, (Ph) 2CHCH2, PhCH (CH3) CH2, (CH3) 2CH, PhCH (OH) CH2, PhC (CH3) 2CH2, (Ph) 2CHCH2, or W is CH 2, CH 2 CH 2, on * (CHafe, (CH 2) 2 f CF2CH2 8. The compound according to any of claims 1-7, characterized in that R5 is hydrogen. 9. The compound according to any of claims 1-7, characterized in that X is R 11 or -CH = N- wherein R 11 is hydrogen or C 1-6 alkyl, optionally substituted by one to three amino, monoalkylamino, dialkylamino, alkoxy, hydroxy, alkoxycarbonyl, aryloxycarbonyl, aralkoxycarbonyl, carboalkoxy, phenyl, cyano, trifluoromethyl, acetylamino, pyridyl, thiophenyl, furyl, pyrrolyl or imidazolyl. 10. The compound according to any of claims 1-7, characterized in that X is oxygen. 11. The compound according to any of claims 1-7, characterized in that it is hydrogen or alkyl Cl-6 • 12. The compound according to any of claims 1-7, characterized in that R: R 10 are independently one of hydrogen, C alquilo-6 alkyl, aryl-Cß-io-alkyl (Ci-β) aryl Cβ-io. C2-? o hydroxyalkyl or C2-7 carboxyalkyl. 13. The compound according to claim 12, characterized in that R7, R8, R9 and R10 are independently one of hydrogen, methyl, ethyl, propyl, n-butyl, benzyl, phenylethyl, 2-hydroxyethyl, 3-hydroxypropyl, 4-hydroxybutyl , 2-carboxymethyl, 3-carboxyethyl or 4-carboxy-boroyl, or wherein either R7 and R8 or R9 and R10 form a 3 to 7-membered carbocyclic ring with C to which they are linked. 14. The compound according to any of claims 1-7, characterized in that Ra, Rb and Rc are independently one of hydrogen, C? -6 alkyl, C? -io-cyano alkoxy, or -C02Rw where Rw, in each instance, is alkyl C? _4, C4.7 cycloalkyl or benzyloxycarbonyl. 15. The compound according to claim 14, characterized in that Ra, Rb and Rc are independently one of hydrogen, methyl, ethyl, propyl, n-butyl, hydroxy, methoxy, ethoxy, cyano, -C02CH3, -C02CH2CH3 or -C02CH2CH2CH3. 16. The compound according to claim 15, characterized in that Ra, Rb and Rc are each hydrogen. 17. The compound according to claim 14, characterized in that Ra, Rb and Rc are independently one of where Rd-Rh are as defined in claim 1. 18. The compound according to claim 17, characterized in that Ra, Rb and Rc are each hydrogen Rf is methyl; and Rh is benzyl or tert-butyl. 19. The compound according to claim 1, characterized in that it has the Formulas II, III or IV: IV or a solvate, hydrate or pharmaceutically acceptable salt thereof; wherein n 'and m' are each independently 0-3, and R3, R4, W, X, n and m are as defined in claim 1. 20. The compound according to claim 19, characterized in that W is PhCH2CH2, (CH3) 3C-, HOOCCH2, CF3CH2, (CH3) 2N (CH2) 2, PhCH20 (CH2) 2, PhCH (CH3), PhCH2CH (COOH), CH3 (CH2) 5, PhCH2, H, CH3 (CH2) 4, CH3CH2CH (CH3) CH2, (Ph) 2CHCH2, PhCH (CH3) CH2, (CH3) 2CH, PhCH (0H) CH2, PhC (CH3) 2CH2, (Ph) 2CHCH2, or W is CH 2, CH 2 CH 2 O N- (CH 2.) 2, (CH2) 2 - (CH2) 2, or 21. The compound according to claim 1, characterized in that it is one of 1 -. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-met i 1-3- (fenet i lamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-met il-3- (2,2-diphenylethylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxi) et il] amino} carbonylmethyl-6-methyl-3- (4-methylphenethylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carboni lmet il-6-met il-3- (4-methoxy phenethylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxi) et il] amino} carbonylmethyl-6-met il-3- (1-phenylcyclobutyl) methylamino-pyrazinone, 1- . { N- [2- (amidinoaminooxi) et il] amino} carbonylmethyl-6-methyl-3- [2- (1-naphthalene) ethyl] amino-pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-met i 1-3- (2-phenyl-1-butylamino) -pyrazinone, 1-. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-met il-3- (2- [3,4-methylenedioxyphenyl] ethylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-met il-3- (2- [2-pyridyl] ethylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxi) et il] amino} carbonylmethyl-6-methyl-3- (2- [2-methylphenyl] ethylamino-pyrazinone, 1-. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2- [3-methylphenyl] ethylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carboni lmet il-6-methyl-3- (2- [2-trifluoromethylphenyl] ethylamino) -pyrene zinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2- [3-trifluoromethylphenyl] ethylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2- [4-trifluoromethylphenyl] ethylamino) -pylin zinone, 1-. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2- [3,5-dimethylphenyl] ethylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-met il-3- (2-indanylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2- [3, 4-difluorophenyl] ethylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl 1-6-methyl-3- (2- [5-indanyl] ethylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carboni lmet il-6-met il-3- (2-fluoro-phenethylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxi) et il] amino} carbonylmethyl-6-methyl-3- (3,4-dimethoxyphenethylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxi) et il] amino} carbonylmethyl-6-met il-3- (4-fluoro-phenethylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-met il-3- (4-et-ilphene-ylamino) -pyrazinone, 1-. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-met il-3- (2-phenylpropylamino) -pyrazinone, 1- . { - [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (3,4-dimethyl-phenethylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxi) et il] amino} carbonylmethyl-6-methyl-3- (2-naphthalene-ethylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxi) et il] amino} carbonylmethyl-6-methyl-3- (2, 2-diphenylpropylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-met il-3- (2- (3-indolyl) -et i lamino-pira zinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2- [1- (4-methylnaphthalene)] ethylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2- [2, 4-difluorophenyl] ethylamino) -pyrazinone, 1-. { N- [2- (amidino-N '-methylaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2- [4-methylphenyl] ethylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxi) et il] amino} carbonylmethyl-5-chloro-6-methyl-3- (phenethylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-5-chloro-6-met il-3- (2, 2-diphenylethylamino) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-met il-3- (phenethyl) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-met il-3- (1-met il-2-phenylethyl) -pyrazinone, 1- . { N- [2- (amidinohydrazino) ethyl] amino} carbonylmethyl-6-ethyl-3- (phenethyl) -pyrazinone, l-. { N- [2- (amidinohydrazino) ethyl] amino} carbonylmethyl-6-methyl-3- (4-met-il-phenylethyl) -pyrazinone, 1-. { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (l-methyl-2- (4-methylamino-3-pyridyl) et ilpyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (l-methyl-2- (3-pyridyl) ethyl-pyrazine, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2-cyclopropyl-2- (3,4-dimethoxyphenyl) ethyl) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-met il-3- (2-cyclobutylethyl) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl 1-6-methyl-3- (2-cyclobutyl-2, 2-difluoroethyl) -pyrene zinone, 1- . { - [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2- (3-fluorophenyl) ethyl) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2, 2-difluoro-2-phenylethyl) -pyrazinone, 1-. { N- [2- (amidinoaminooxy) ethyl] amino Jcarbonylmethyl-6-met il-3- (2-phenylcyclopropyl) -pyrazinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2- (4-chlorophenyl) -2-cyclopropylethyl) -pyran zinone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} charcoal lmet il-6-methyl-3- (2-cyclopropyl-2- (3-pyridyl) ethyl) -pyrazinone, 1- . { N- [2- (amine-inoamin- or i) ethyl] amino} carbonylmethyl-6-methyl-3- (2-benzylcyclopropyl) -pyrazinone, 1- . { - [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-met il-3- (2-cyclopropyl-2- (3-fluorophenyl) ethyl) -pira z inone, 1- . { N- [2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-3- (2-cyclopropyl-2- (3,4-difluorophenyl) ethyl) -pyrazinone,

1- . { - [

2- (amidinoaminooxy) ethyl] amino} carbonylmethyl-6-methyl-

3- (2,2-difluoro-2-phenylethylene) -pyran zinone, pharmaceutically acceptable salts thereof 22. A compound according to any of claims 1-7, characterized in that R1 comprises a radioactive atom. 23. The compound according to claim 22, characterized in that the radioactive atom is a radioactive iodine atom selected from 1-125, 1-131 and 1-123. 24. A pharmaceutical composition, characterized in that they comprise a compound of claim 1 and a pharmaceutically acceptable carrier. 25. A pharmaceutical composition, characterized in that they comprise a compound of claim 9 and a pharmaceutically acceptable carrier. 26. A pharmaceutical composition, characterized in that they comprise a compound of claim 21 and a pharmaceutically acceptable carrier. 27. A method for inhibiting the protease, characterized in that they comprise contacting the protease with a compound of claim 1. 28. The method according to claim 27, characterized in that the protease is leukocyte neutrophil elastase, chymotrypsin, trypsin, pancreatic elastase, cathepsin G, thrombin, factor Xa, thermolysin, or pepsin. 29. The method according to claim 27, characterized in that the protease is thrombin. 30. A method for reducing the t-rhombhogenicity of a surface, characterized in that it comprises covalently coating, inserting, absorbing or binding a compound of claim 1 to the surface. 31. A method for the treatment of aberrant proteolysis in a mammal, characterized in that it comprises administering a compound of claim 1 to the mammal. 32. A method for the treatment of thrombosis associated with ischemia, viral infections, stroke, cancer, restenosis, myocardial infarction, disseminated intramuscular coagulopathy that occurs during septic shock, unstable angina, disseminated intramuscular coagulation caused by trauma, coronary artery bypass, replacement hip, thrombotic therapy, sepsis, hemodialysis, adult respiratory distress syndrome, rheumatoid arthritis, ulcerative colitis, ulcerative colitis; hardening, metastasis, hypercoagulability during chemotherapy, Alzheimer's disease, Down syndrome; fibrin formation in the eye, wound healing, or inflammation in a mammal, characterized in that they comprise administering a compound of claim 1 to the mammal. 33. A method for reducing blood coagulation in a mammal, characterized in that it comprises administering a compound of claim 1 to the mammal. 34. A method for detecting thrombi in a mammal, characterized in that it comprises: (a) administering a compound of claim 22 to the mammal; and (b) detecting the compound bound to the thrombus. 35. The method according to claim 30, characterized in that the surface is a graft. 36. A pharmaceutical dosage form, characterized in that it comprises from about 0.1 to about 500 milligrams of a compound of claim 1. 37. The pharmaceutical dosage form according to claim 36, characterized in that it is adapted for parenteral or oral administration. Summary of the Invention Pyrazine compounds are disclosed, including compound of Formula (I) wherein X is 0, NR, 11 or CH = N, R -R, 11, R% R, R, W, m and n are placed in the specification, as well as hydrates, solvates or pharmaceutically acceptable salts thereof. The compounds of the invention are potent inhibitors of proteases, especially of trypsin-like serine proteases, such as chymotrypsin, trypsin, thrombin, plasmin and factor Xa. Some of the compounds exhibit antithrombotic activity by means of the selective, direct inhibition of thrombin. Compositions are described for inhibiting the loss of blood platelets, inhibiting the formation of blood platelet aggregates, inhibiting fibrin formation, inhibiting the formation of thrombi, and inhibiting the formation of emboli. Other uses of the compounds of the invention as anticoagulants both inserted or physically bound to materials used in the manufacture of devices that are used in blood collection, blood circulation, and blood storage, such as catheters, dialysis machines, Blood, syringes and blood collection tubes, blood lines and keys. Additionally, the compounds can be labeled for detection and used for thrombus imaging.